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Article(id=1240375118623461910, tenantId=1146029695717560320, journalId=1227665162245664772, issueId=1240375105386238038, articleNumber=null, orderNo=null, doi=10.20043/j.cnki.MPM.202307011, pmid=null, cstr=null, oa=null, hot=null, price=null, onlineType=0, articleFormat=0, articleType=null, articleTypeStr=null, receivedDate=1688400000000, receivedDateStr=2023-07-04, revisedDate=null, revisedDateStr=null, acceptedDate=null, acceptedDateStr=null, onlineDate=1773658073334, onlineDateStr=2026-03-16, pubDate=1711296000000, pubDateStr=2024-03-25, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1773658073334, onlineIssueDateStr=2026-03-16, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1773658073334, creator=13701087609, updateTime=1773658073334, updator=13701087609, issue=Issue{id=1240375105386238038, tenantId=1146029695717560320, journalId=1227665162245664772, year='2024', volume='51', issue='6', pageStart='961', pageEnd='1152', issueExtLink='null', onlineDate='null', pubDate='null', beforeIssueId=null, nextIssueId=null, price=null, status=1, issueComplete=1, articleOrder=1, issueType=-1, specialIssue=null, createTime=1773658070179, creator=13701087609, updateTime=1773658539618, updator=13701087609, preIssue=null, nextIssue=null, ext={EN=IssueExt(id=1240377074414833974, tenantId=1146029695717560320, journalId=1227665162245664772, issueId=1240375105386238038, language=EN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=), CN=IssueExt(id=1240377074414833975, tenantId=1146029695717560320, journalId=1227665162245664772, issueId=1240375105386238038, language=CN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=)}, issueFiles=null}, startPage=1083, endPage=1087, ext={EN=ArticleExt(id=1240375120833860265, articleId=1240375118623461910, tenantId=1146029695717560320, journalId=1227665162245664772, language=EN, title=Effects of TTR Gly83Arg mutation on the change of retinol and TTR protein, columnId=1228016572065837304, journalTitle=Modern Preventive Medicine, columnName=Experimental Technology and Applications, runingTitle=null, highlight=null, articleAbstract=
Objective

To explore the relationship between retinol and TTR protein, and to provide ideas for preventing Hereditary vitreous amyloidosis (HVA).

Methods

Twenty Gly83Arg mutant mice were constructed and bred (reference construction [2]) as the mutant group. After the irregular turbidity in the vitreous of the mutant mice eyeball was observed under slit lamp, the lens camera was used for photography, and B-ultrasound examination was conducted, and white clump-like turbidity was found in the posterior ocular region. After the turbidity in the vitreous cavity of the mice was confirmed, the mice were killed by cervical vertebrae dislocation, and DNA in liver cells was extracted and sequenced to confirm the successful construction of mutant mice. At the same time, 20 wild type C57BL/6 mice were fed as the control group, 4 mice in the mutant group and 4 mice in the control group were randomly selected from the number table, and the retinol concentration was determined by high performance liquid chromatography combined with mass spectrometry. Three mice in the mutant group and three mice in the control group were selected from a random number table. After grinding the eyeball tissues, RNA and protein were extracted. TTR gene mRNA expression, SDS-PAGE protein gel color development was detected by fluorescent quantitative PCR, and TTR gene protein expression was detected by Western Blot. The difference at P < 0.05 was statistically significant.

Results

The mutation site was consistent with the site successfully modeled by our research group in the previous stage. Heterozygous mutation occurred at the G base at the 107th position of exon 3 (C.107G →C), and the codon of the amino acid at the 83rd position was mutated from GGC to CGC (Gly83Arg). The concentration of retinol in eye of mutant mice was lower than that of normal mice in control group, the content of retinol in liver tissue of mutant mice was higher than that in control group, and the content of retinol in blood of mutant mice was lower than that in control group (eyeball: t=6.600, P=0.022; liver: t=9.045, P<0.001; blood: t=4.343, P=0.012). Fluorescence quantitative PCR results showed that mRNA expression in eyeball tissue of mutant group was lower than that of control group (t=5.764，P=0.004). SDS-PAGE protein gel color showed that protein bands appeared in the mutant group near 11kD. Western Blot showed that TTR gene protein expression in mutant group was lower than that in control group (t=37.48，P<0.001).

Conclusion

The deposition of vitreous amyloid protein in the eyeball tissue of Gly83Arg mutant mice may be related to the increase of mutant TTR protein and the decrease of retinol, which has important significance for future prevention and treatment.

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目的

探索视黄醇与TTR蛋白之间的联系，为预防遗传性玻璃体淀粉样变性疾病（Hereditary vitreous amyloidosis，HVA）提供思路。

方法

构造并饲养Gly83Arg突变模型小鼠20只（参照文献构建）作为突变组，裂隙灯下观察到突变小鼠眼球玻璃体中出现不规则混浊现象后，再用眼前段照相机进行摄影，并行B超检查，发现眼部后段出现白色团样混浊，在确认小鼠玻璃体腔出现混浊后，选择颈椎脱臼法处死小鼠，并提取肝细胞中的DNA后进行测序，确证突变组小鼠构建成功。并同时饲养野生型C57BL/6小鼠20只作为对照组，随机数字表选取突变组4只，对照组4只小鼠眼球，肝脏组织以及血液，在高效液相色谱联合质谱测定视黄醇浓度；随机数字表选取突变组3只，对照组3只小鼠，取眼球组织研磨后提取RNA及蛋白质，采用荧光定量PCR检测TTR基因mRNA表达、SDS-PAGE蛋白凝胶显色，Western Blot检测TTR基因蛋白质表达，结果均通过独立样本t检验。

结果

突变位点与课题组前期成功造模小鼠位点一致，第3外显子的第107位置的G碱基处产生了杂合突变（c.107G→C），第83位置的氨基酸的密码子由GGC突变成CGC，即Gly83Arg。高效液相色谱联合质谱测定突变组小鼠眼球视黄醇浓度低于对照组正常小鼠，突变小鼠肝脏组织视黄醇含量要大于对照组小鼠肝脏组织视黄醇含量，并且突变组小鼠血液中视黄醇含量低于照组小鼠（眼球：t=6.600，P=0.022；肝脏：t=9.045，P<0.001；血液：t=4.343，P=0.012）；荧光定量PCR结果显示突变组小鼠眼球组织的mRNA表达低于对照组（t=5.764，P=0.004）；SDS-PAGE蛋白凝胶显色表示在接近11kD处，突变组出现蛋白条带；Western Blot显示突变组小鼠眼球组织TTR基因蛋白表达低于对照组（t=37.48，P<0.001）。

结论

Gly83Arg突变型模型小鼠眼球组织中玻璃体淀粉样蛋白的沉积，可能与变异的TTR蛋白增多以及视黄醇的减少有关，这对以后的防治有重要的意义。

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蔡善君，E-mail：
, copyrightStatement=本刊刊出的所有文章不代表中华预防医学会和本刊编委会的观点，除非特别声明。, copyrightOwner=中华预防医学会和四川大学华西公共卫生学院, extLink=null, articleAbsUrl=null, sourceXml=9euKRE/RpNGq6R9jBme8ww==, magXml=6RVeL8gIsapo74GH86Pzsg==, pdfUrl=null, pdf=v9lYLzMB8oI+wGbFxeD3mQ==, pdfFileSize=705809, pdfExtLink=null, richHtmlUrl=null, mobilePdfUrl=null, reviewReport=null, pdfFirstPage=null, abstractGraph=yFZolsVUWTp1QBYR22tVKA==, abstractGraphContent=null, abstractVideo=null, citation=null, cebUrl=null, magXmlContent=ZKUpPAJUU5MSmLcLi8Ut/g==, mapNumber=null, authorCompany=null, fund=null, authors=

辛诚（1995—），男，硕士，医师，研究方向：眼底病的研究

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辛诚（1995—），男，硕士，医师，研究方向：眼底病的研究

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A novel mechano-enzymatic cleavage mechanism underlies transthyretin amyloidogenesis[J]. EMBO Molecular Medicine, 2015, 7(10): 1337-1349., articleTitle=A novel mechano-enzymatic cleavage mechanism underlies transthyretin amyloidogenesis, refAbstract=null), Reference(id=1240746311226544475, tenantId=1146029695717560320, journalId=1227665162245664772, articleId=1240375118623461910, doi=null, pmid=null, pmcid=null, year=2020, volume=9, issue=2, pageStart=395, pageEnd=402, url=null, language=null, rfNumber=[2], rfOrder=1, authorNames=Liz MA, Celho T, Bellotti V, journalName=Neurology and Therapy, refType=null, unstructuredReference=Liz MA, Celho T, Bellotti V, et al. A narrative review of the role of transthyretin in health and disease[J]. Neurology and Therapy, 2020, 9(2): 395-402, articleTitle=A narrative review of the role of transthyretin in health and disease, refAbstract=null), Reference(id=1240746311549505892, tenantId=1146029695717560320, journalId=1227665162245664772, articleId=1240375118623461910, doi=null, pmid=null, pmcid=null, year=2012, volume=25, issue=6, pageStart=646, pageEnd=651, url=null, language=null, rfNumber=[3], rfOrder=2, authorNames=Beirão M, Matos E, Beirão I, journalName=Transplant International, refType=null, unstructuredReference=Beirão M, Matos E, Beirão I, et al. No ocular involvement in familial amyloidotic polyneuropathy ATTR V30M domino liver recipients[J]. Transplant International, 2012, 25(6): 646-651., articleTitle=No ocular involvement in familial amyloidotic polyneuropathy ATTR V30M domino liver recipients, refAbstract=null), Reference(id=1240746311629197671, tenantId=1146029695717560320, journalId=1227665162245664772, articleId=1240375118623461910, doi=null, pmid=null, pmcid=null, year=1990, volume=189, issue=null, pageStart=394, pageEnd=402, url=null, language=null, rfNumber=[4], rfOrder=3, authorNames=Fex G, Johannesson G, journalName=Methods in Enzymology, refType=null, unstructuredReference=Fex G, Johannesson G. Transfer of retinol from retinol-binding protein complex to liposomes and across liposomal membranes[J]. Methods in Enzymology, 1990, 189: 394-402., articleTitle=Transfer of retinol from retinol-binding protein complex to liposomes and across liposomal membranes, refAbstract=null), Reference(id=1240746311700500843, tenantId=1146029695717560320, journalId=1227665162245664772, articleId=1240375118623461910, doi=null, pmid=null, pmcid=null, year=1990, volume=29, issue=16, pageStart=3878, pageEnd=3883, url=null, language=null, rfNumber=[5], rfOrder=4, authorNames=Noy N, Xu ZJ, journalName=Biochemistry, refType=null, unstructuredReference=Noy N, Xu ZJ. Interactions of retinol with binding proteins: implications for the mechanism of uptake by cells[J]. Biochemistry, 1990, 29(16): 3878-3883., articleTitle=Interactions of retinol with binding proteins: implications for the mechanism of uptake by cells, refAbstract=null), Reference(id=1240746311801164145, tenantId=1146029695717560320, journalId=1227665162245664772, articleId=1240375118623461910, doi=null, pmid=null, pmcid=null, year=1991, volume=30, issue=26, pageStart=6380, pageEnd=6386, url=null, language=null, rfNumber=[6], rfOrder=5, authorNames=Noy N, Blaner WS, journalName=Biochemistry, refType=null, unstructuredReference=Noy N, Blaner WS. Interactions of retinol with binding proteins: studies with rat cellular retinol-binding protein and with rat retinol-binding protein[J]. Biochemistry, 1991, 30(26): 6380-6386., articleTitle=Interactions of retinol with binding proteins: studies with rat cellular retinol-binding protein and with rat retinol-binding protein, refAbstract=null), Reference(id=1240746311897633144, tenantId=1146029695717560320, journalId=1227665162245664772, articleId=1240375118623461910, doi=null, pmid=null, pmcid=null, year=1990, volume=29, issue=16, pageStart=3883, pageEnd=3888, url=null, language=null, rfNumber=[7], rfOrder=6, authorNames=Noy N, Xu ZJ, journalName=Biochemistry, refType=null, unstructuredReference=Noy N, Xu ZJ. Kinetic parameters of the interactions of retinol with lipid bilayers[J]. Biochemistry, 1990, 29(16): 3883-3888., articleTitle=Kinetic parameters of the interactions of retinol with lipid bilayers, refAbstract=null), Reference(id=1240746311994102141, tenantId=1146029695717560320, journalId=1227665162245664772, articleId=1240375118623461910, doi=null, pmid=null, pmcid=null, year=2018, volume=169, issue=null, pageStart=13, pageEnd=19, url=null, language=null, rfNumber=[8], rfOrder=7, authorNames=Ran LX, Zheng ZY, Xie B, journalName=Experimental Eye Research, refType=null, unstructuredReference=Ran LX, Zheng ZY, Xie B, et al. A mouse model of a novel missense mutation (Gly83Arg) in a Chinese kindred manifesting vitreous amyloidosis only[J]. Experimental Eye Research, 2018, 169: 13-19., articleTitle=A mouse model of a novel missense mutation (Gly83Arg) in a Chinese kindred manifesting vitreous amyloidosis only, refAbstract=null), Reference(id=1240746312090571137, tenantId=1146029695717560320, journalId=1227665162245664772, articleId=1240375118623461910, doi=null, pmid=null, pmcid=null, year=1976, volume=590, issue=null, pageStart=1, pageEnd=64, url=null, language=null, rfNumber=[9], rfOrder=8, authorNames=Andersson R, journalName=Acta Medica Scandinavica. Supplementum, refType=null, unstructuredReference=Andersson R. Familial amyloidosis with polyneuropathy. A clinical study based on patients living in northern Sweden[J]. Acta Medica Scandinavica. Supplementum, 1976, 590: 1-64., articleTitle=Familial amyloidosis with polyneuropathy. A clinical study based on patients living in northern Sweden, refAbstract=null), Reference(id=1240746312220594564, tenantId=1146029695717560320, journalId=1227665162245664772, articleId=1240375118623461910, doi=null, pmid=null, pmcid=null, year=1952, volume=75, issue=3, pageStart=408, pageEnd=427, url=null, language=null, rfNumber=[10], rfOrder=9, authorNames=Andrade C, journalName=Brain : a Journal of Neurology, refType=null, unstructuredReference=Andrade C. A peculiar form of peripheral neuropathy; familiar atypical generalized amyloidosis with special involvement of the peripheral nerves[J]. Brain : a Journal of Neurology, 1952, 75(3): 408-427., articleTitle=A peculiar form of peripheral neuropathy; familiar atypical generalized amyloidosis with special involvement of the peripheral nerves, refAbstract=null), Reference(id=1240746312346423688, tenantId=1146029695717560320, journalId=1227665162245664772, articleId=1240375118623461910, doi=null, pmid=null, pmcid=null, year=2011, volume=26, issue=4, pageStart=230, pageEnd=238, url=null, language=null, rfNumber=[11], rfOrder=10, authorNames=Shi Y, Li J, Hu J, journalName=eye Science, refType=null, unstructuredReference=Shi Y, Li J, Hu J, et al. A new Arg54Gly transthyretin gene mutation associated with vitreous amyloidosis in Chinese[J]. eye Science, 2011, 26(4): 230-238., articleTitle=A new Arg54Gly transthyretin gene mutation associated with vitreous amyloidosis in Chinese, refAbstract=null), Reference(id=1240746312442892688, tenantId=1146029695717560320, journalId=1227665162245664772, articleId=1240375118623461910, doi=null, pmid=null, pmcid=null, year=2014, volume=21, issue=2, pageStart=140, pageEnd=142, url=null, language=null, rfNumber=[12], rfOrder=11, authorNames=Yin J, Xia XY, Shi Y, journalName=Amyloid : the International Journal of Experimental and Clinical Investigation: the Official Journal of the International Society of Amyloidosis, refType=null, unstructuredReference=Yin J, Xia XY, Shi Y, et al. Chinese familial transthyretin amyloidosis with vitreous involvement is associated with the transthyretin mutation Gly83Arg: a case report and literature review[J]. Amyloid : the International Journal of Experimental and Clinical Investigation: the Official Journal of the International Society of Amyloidosis, 2014, 21(2): 140-142., articleTitle=Chinese familial transthyretin amyloidosis with vitreous involvement is associated with the transthyretin mutation Gly83Arg: a case report and literature review, refAbstract=null), Reference(id=1240746312522584466, tenantId=1146029695717560320, journalId=1227665162245664772, articleId=1240375118623461910, doi=null, pmid=null, pmcid=null, year=2006, volume=13, issue=3, pageStart=170, pageEnd=177, url=null, language=null, rfNumber=[13], rfOrder=12, authorNames=Liepnieks JJ, Wilson DL, Benson MD, journalName=Amyloid: the International Journal of Experimental and Clinical Investigation : the Official Journal of the International Society of Amyloidosis, refType=null, unstructuredReference=Liepnieks JJ, Wilson DL, Benson MD. Biochemical characterization of vitreous and cardiac amyloid in Ile84Ser transthyretin amyloidosis[J]. Amyloid: the International Journal of Experimental and Clinical Investigation : the Official Journal of the International Society of Amyloidosis, 2006, 13(3): 170-177., articleTitle=Biochemical characterization of vitreous and cardiac amyloid in Ile84Ser transthyretin amyloidosis, refAbstract=null), Reference(id=1240746312589693334, tenantId=1146029695717560320, journalId=1227665162245664772, articleId=1240375118623461910, doi=null, pmid=null, pmcid=null, year=2010, volume=128, issue=2, pageStart=206, pageEnd=210, url=null, language=null, rfNumber=[14], rfOrder=13, authorNames=Hara R, Kawaji T, Ando E, journalName=Archives of Ophthalmology, refType=null, unstructuredReference=Hara R, Kawaji T, Ando E, et al. Impact of liver transplantation on transthyretin-related ocular amyloidosis in Japanese patients[J]. Archives of Ophthalmology, 2010, 128(2): 206-210., articleTitle=Impact of liver transplantation on transthyretin-related ocular amyloidosis in Japanese patients, refAbstract=null), Reference(id=1240746312656802203, tenantId=1146029695717560320, journalId=1227665162245664772, articleId=1240375118623461910, doi=null, pmid=null, pmcid=null, year=2003, volume=4, issue=1, pageStart=1, pageEnd=10, url=null, language=null, rfNumber=[15], rfOrder=14, authorNames=Marill J, Idres N, Capron CC, journalName=Current Drug Metabolism, refType=null, unstructuredReference=Marill J, Idres N, Capron CC, et al. Retinoic acid metabolism and mechanism of action: a review[J]. Current Drug Metabolism, 2003, 4(1): 1-10., articleTitle=Retinoic acid metabolism and mechanism of action: a review, refAbstract=null), Reference(id=1240746312719716768, tenantId=1146029695717560320, journalId=1227665162245664772, articleId=1240375118623461910, doi=null, pmid=null, pmcid=null, year=2011, volume=3, issue=1, pageStart=63, pageEnd=103, url=null, language=null, rfNumber=[16], rfOrder=15, authorNames=D’Ambrosio DN, Clugston RD, Blaner WS, journalName=Nutrients, refType=null, unstructuredReference=D’Ambrosio DN, Clugston RD, Blaner WS. Vitamin A metabolism: an update[J]. Nutrients, 2011, 3(1): 63-103., articleTitle=Vitamin A metabolism: an update, refAbstract=null), Reference(id=1240746312828768673, tenantId=1146029695717560320, journalId=1227665162245664772, articleId=1240375118623461910, doi=null, pmid=null, pmcid=null, year=2016, volume=75, issue=2, pageStart=212, pageEnd=215, url=null, language=null, rfNumber=[17], rfOrder=16, authorNames=Noy N, journalName=Proceedings of the Nutrition Society, refType=null, unstructuredReference=Noy N. Vitamin a in regulation of insulin responsiveness: mini review[J]. Proceedings of the Nutrition Society, 2016, 75(2): 212-215., articleTitle=Vitamin a in regulation of insulin responsiveness: mini review, refAbstract=null)], funds=[Fund(id=1240746310886805831, tenantId=1146029695717560320, journalId=1227665162245664772, articleId=1240375118623461910, awardId=31871261, language=CN, fundingSource=国家自然科学基金(31871261), fundOrder=null, country=null)], companyList=[AuthorCompany(id=1240746307703328889, tenantId=1146029695717560320, journalId=1227665162245664772, articleId=1240375118623461910, xref=null, ext=[AuthorCompanyExt(id=1240746307707523194, tenantId=1146029695717560320, journalId=1227665162245664772, articleId=1240375118623461910, companyId=1240746307703328889, language=EN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=Ophthalmology Department of Zunyi Medical University, Zunyi, Guizhou 563000, China), AuthorCompanyExt(id=1240746307715911803, tenantId=1146029695717560320, journalId=1227665162245664772, articleId=1240375118623461910, companyId=1240746307703328889, language=CN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=遵义医科大学眼科，贵州 遵义 563000)])], figs=[ArticleFig(id=1240746309171335401, tenantId=1146029695717560320, journalId=1227665162245664772, articleId=1240375118623461910, language=EN, label=Fig.1, caption=Comparison of retinol content in eye tissues of TTR Gly83Arg mutant mice and control mice, figureFileSmall=wzVldUfcwSS7um0yhZs3hw==, figureFileBig=VkpLtzYHKSwQKpMlTsuBIQ==, tableContent=null), ArticleFig(id=1240746309301358832, tenantId=1146029695717560320, journalId=1227665162245664772, articleId=1240375118623461910, language=CN, label=图1, caption=TTR Gly83Arg突变小鼠与对照组小鼠各个组织视黄醇含量比较

注：★：P<0.05，差异具有统计学意义；眼部：t=6.600，P=0.022；肝脏：t=9.045，P<0.001；血液：t=4.343，P=0.012。

, figureFileSmall=wzVldUfcwSS7um0yhZs3hw==, figureFileBig=VkpLtzYHKSwQKpMlTsuBIQ==, tableContent=null), ArticleFig(id=1240746309519462653, tenantId=1146029695717560320, journalId=1227665162245664772, articleId=1240375118623461910, language=EN, label=Fig.2, caption=Comparison of TTR mRNA expression in eye tissues of TTR Gly83Arg mutant mice and control mice, figureFileSmall=oIQhF0FFCcyFShQirwfc6g==, figureFileBig=TSwz2mR+wsMghXxMj4SeJg==, tableContent=null), ArticleFig(id=1240746309628514561, tenantId=1146029695717560320, journalId=1227665162245664772, articleId=1240375118623461910, language=CN, label=图2, caption=TTR Gly83Arg突变小鼠眼球组织与对照组小鼠眼球组织中的TTR mRNA相对表达水平比较

注：★★：P<0.05，差异具有统计学意义（t=5.764，P=0.004）。

, figureFileSmall=oIQhF0FFCcyFShQirwfc6g==, figureFileBig=TSwz2mR+wsMghXxMj4SeJg==, tableContent=null), ArticleFig(id=1240746309724983562, tenantId=1146029695717560320, journalId=1227665162245664772, articleId=1240375118623461910, language=EN, label=Fig.3, caption=Protein SDS-PAGE analysis of eyeball tissues of TTR Gly83Arg mutant mice and control mice, figureFileSmall=7nb1OtLJ36jXJ0w3ETrhtQ==, figureFileBig=mBesNeegd8Id5q3Rj2ULww==, tableContent=null), ArticleFig(id=1240746309804675342, tenantId=1146029695717560320, journalId=1227665162245664772, articleId=1240375118623461910, language=CN, label=图3, caption=TTR Gly83Arg突变组小鼠与对照组小鼠眼球组织的蛋白质SDS-PAGE分析 在接近11 kD处，突变组可见一染色条带，染色较对照组深, figureFileSmall=7nb1OtLJ36jXJ0w3ETrhtQ==, figureFileBig=mBesNeegd8Id5q3Rj2ULww==, tableContent=null), ArticleFig(id=1240746309909532949, tenantId=1146029695717560320, journalId=1227665162245664772, articleId=1240375118623461910, language=EN, label=Fig.4, caption=Protein Western blot analysis of eyeball tissues of TTR Gly83Arg mutant mice and control mice, figureFileSmall=bKBHuEEMvzIOUUjsSRQIlg==, figureFileBig=XdkQqaO+Twexvu+yFMm98g==, tableContent=null), ArticleFig(id=1240746310018584859, tenantId=1146029695717560320, journalId=1227665162245664772, articleId=1240375118623461910, language=CN, label=图4, caption=TTR Gly83Arg突变组小鼠与对照组小鼠眼球组织的TTR蛋白相对表达水平比较

注：★★：P<0.05，差异具有统计学意义（t=37.48，P<0.001）。

, figureFileSmall=bKBHuEEMvzIOUUjsSRQIlg==, figureFileBig=XdkQqaO+Twexvu+yFMm98g==, tableContent=null), ArticleFig(id=1240746310136025375, tenantId=1146029695717560320, journalId=1227665162245664772, articleId=1240375118623461910, language=EN, label=Table 1, caption=

Analysis of retinol concentration in various tissues of mutant and control groups（）

, figureFileSmall=null, figureFileBig=null, tableContent=
组别组织样本数视黄醇浓度（ng/ml）
突变组眼部3232.498 3±3.910 7a
肝脏35 414.280 0±853.370 0a
血清348.840 0±41.200 0a
对照组眼部3281.799 8±5.510 0
肝脏3931.950 0±92.290 0
血清3270.380 0±78.160 0
), ArticleFig(id=1240746310266048812, tenantId=1146029695717560320, journalId=1227665162245664772, articleId=1240375118623461910, language=CN, label=表1, caption=

突变组与对照组各组织中视黄醇浓度分析（）

, figureFileSmall=null, figureFileBig=null, tableContent=
组别组织样本数视黄醇浓度（ng/ml）
突变组眼部3232.498 3±3.910 7a
肝脏35 414.280 0±853.370 0a
血清348.840 0±41.200 0a
对照组眼部3281.799 8±5.510 0
肝脏3931.950 0±92.290 0
血清3270.380 0±78.160 0
), ArticleFig(id=1240746310379295026, tenantId=1146029695717560320, journalId=1227665162245664772, articleId=1240375118623461910, language=EN, label=Table 2, caption=

Analysis of relative expression of TTR mRNA (TTR/GAPDH) in eyeball tissues of mutation group and control group ()

, figureFileSmall=null, figureFileBig=null, tableContent=
组别样本数眼球TTR mRNA相对表达量
（TTR/GAPDH）
对照组31.001 2±0.059 7
突变组30.619 8±0.097 9a
P0.004 0
), ArticleFig(id=1240746310479958327, tenantId=1146029695717560320, journalId=1227665162245664772, articleId=1240375118623461910, language=CN, label=表2, caption=

突变组与对照组眼球组织TTR mRNA相对表达量（TTR/GAPDH）分析（）

, figureFileSmall=null, figureFileBig=null, tableContent=
组别样本数眼球TTR mRNA相对表达量
（TTR/GAPDH）
对照组31.001 2±0.059 7
突变组30.619 8±0.097 9a
P0.004 0
), ArticleFig(id=1240746310593204540, tenantId=1146029695717560320, journalId=1227665162245664772, articleId=1240375118623461910, language=EN, label=Table 3, caption=

Analysis of TTR protein relative expression (TTR/GAPDH) between mutation group and control group（）

, figureFileSmall=null, figureFileBig=null, tableContent=
组别样本数眼球TTR蛋白相对表达量
（TTR/GAPDH）
对照组30.999 9±0.013 9
突变组30.594 9±0.012 5a
P0.000 003 024 17
), ArticleFig(id=1240746310693867840, tenantId=1146029695717560320, journalId=1227665162245664772, articleId=1240375118623461910, language=CN, label=表3, caption=

突变组与对照组TTR蛋白相对表达量（TTR/GAPDH）分析（）

, figureFileSmall=null, figureFileBig=null, tableContent=
组别样本数眼球TTR蛋白相对表达量
（TTR/GAPDH）
对照组30.999 9±0.013 9
突变组30.594 9±0.012 5a
P0.000 003 024 17
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TTR蛋白Gly83Arg突变对视黄醇以及TTR蛋白变化规律的影响
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辛诚 , 蔡善君
现代预防医学 | 实验技术及其应用 2024,51(6): 1083-1087
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现代预防医学 | 实验技术及其应用 2024, 51(6): 1083-1087
TTR蛋白Gly83Arg突变对视黄醇以及TTR蛋白变化规律的影响
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辛诚, 蔡善君
作者信息
  • 遵义医科大学眼科，贵州 遵义 563000

    • 辛诚（1995—），男，硕士，医师，研究方向：眼底病的研究

通讯作者:

蔡善君，E-mail：
Effects of TTR Gly83Arg mutation on the change of retinol and TTR protein
Cheng XIN, Shan-jun CAI
Affiliations
  • Ophthalmology Department of Zunyi Medical University, Zunyi, Guizhou 563000, China
出版时间: 2024-03-25 doi: 10.20043/j.cnki.MPM.202307011
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摘要
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目的

探索视黄醇与TTR蛋白之间的联系，为预防遗传性玻璃体淀粉样变性疾病（Hereditary vitreous amyloidosis，HVA）提供思路。

方法

构造并饲养Gly83Arg突变模型小鼠20只（参照文献构建）作为突变组，裂隙灯下观察到突变小鼠眼球玻璃体中出现不规则混浊现象后，再用眼前段照相机进行摄影，并行B超检查，发现眼部后段出现白色团样混浊，在确认小鼠玻璃体腔出现混浊后，选择颈椎脱臼法处死小鼠，并提取肝细胞中的DNA后进行测序，确证突变组小鼠构建成功。并同时饲养野生型C57BL/6小鼠20只作为对照组，随机数字表选取突变组4只，对照组4只小鼠眼球，肝脏组织以及血液，在高效液相色谱联合质谱测定视黄醇浓度；随机数字表选取突变组3只，对照组3只小鼠，取眼球组织研磨后提取RNA及蛋白质，采用荧光定量PCR检测TTR基因mRNA表达、SDS-PAGE蛋白凝胶显色，Western Blot检测TTR基因蛋白质表达，结果均通过独立样本t检验。

结果

突变位点与课题组前期成功造模小鼠位点一致，第3外显子的第107位置的G碱基处产生了杂合突变（c.107G→C），第83位置的氨基酸的密码子由GGC突变成CGC，即Gly83Arg。高效液相色谱联合质谱测定突变组小鼠眼球视黄醇浓度低于对照组正常小鼠，突变小鼠肝脏组织视黄醇含量要大于对照组小鼠肝脏组织视黄醇含量，并且突变组小鼠血液中视黄醇含量低于照组小鼠（眼球：t=6.600，P=0.022；肝脏：t=9.045，P<0.001；血液：t=4.343，P=0.012）；荧光定量PCR结果显示突变组小鼠眼球组织的mRNA表达低于对照组（t=5.764，P=0.004）；SDS-PAGE蛋白凝胶显色表示在接近11kD处，突变组出现蛋白条带；Western Blot显示突变组小鼠眼球组织TTR基因蛋白表达低于对照组（t=37.48，P<0.001）。

结论

Gly83Arg突变型模型小鼠眼球组织中玻璃体淀粉样蛋白的沉积，可能与变异的TTR蛋白增多以及视黄醇的减少有关，这对以后的防治有重要的意义。

视黄醇  /  转甲状腺素蛋白（TTR）  /  突变
Objective

To explore the relationship between retinol and TTR protein, and to provide ideas for preventing Hereditary vitreous amyloidosis (HVA).

Methods

Twenty Gly83Arg mutant mice were constructed and bred (reference construction [2]) as the mutant group. After the irregular turbidity in the vitreous of the mutant mice eyeball was observed under slit lamp, the lens camera was used for photography, and B-ultrasound examination was conducted, and white clump-like turbidity was found in the posterior ocular region. After the turbidity in the vitreous cavity of the mice was confirmed, the mice were killed by cervical vertebrae dislocation, and DNA in liver cells was extracted and sequenced to confirm the successful construction of mutant mice. At the same time, 20 wild type C57BL/6 mice were fed as the control group, 4 mice in the mutant group and 4 mice in the control group were randomly selected from the number table, and the retinol concentration was determined by high performance liquid chromatography combined with mass spectrometry. Three mice in the mutant group and three mice in the control group were selected from a random number table. After grinding the eyeball tissues, RNA and protein were extracted. TTR gene mRNA expression, SDS-PAGE protein gel color development was detected by fluorescent quantitative PCR, and TTR gene protein expression was detected by Western Blot. The difference at P < 0.05 was statistically significant.

Results

The mutation site was consistent with the site successfully modeled by our research group in the previous stage. Heterozygous mutation occurred at the G base at the 107th position of exon 3 (C.107G →C), and the codon of the amino acid at the 83rd position was mutated from GGC to CGC (Gly83Arg). The concentration of retinol in eye of mutant mice was lower than that of normal mice in control group, the content of retinol in liver tissue of mutant mice was higher than that in control group, and the content of retinol in blood of mutant mice was lower than that in control group (eyeball: t=6.600, P=0.022; liver: t=9.045, P<0.001; blood: t=4.343, P=0.012). Fluorescence quantitative PCR results showed that mRNA expression in eyeball tissue of mutant group was lower than that of control group (t=5.764，P=0.004). SDS-PAGE protein gel color showed that protein bands appeared in the mutant group near 11kD. Western Blot showed that TTR gene protein expression in mutant group was lower than that in control group (t=37.48，P<0.001).

Conclusion

The deposition of vitreous amyloid protein in the eyeball tissue of Gly83Arg mutant mice may be related to the increase of mutant TTR protein and the decrease of retinol, which has important significance for future prevention and treatment.

Retinol  /  Transthyretin (TTR)  /  Mutation
辛诚, 蔡善君. TTR蛋白Gly83Arg突变对视黄醇以及TTR蛋白变化规律的影响. 现代预防医学, 2024 , 51 (6) : 1083 -1087 . DOI: 10.20043/j.cnki.MPM.202307011
Cheng XIN, Shan-jun CAI. Effects of TTR Gly83Arg mutation on the change of retinol and TTR protein[J]. Modern Preventive Medicine, 2024 , 51 (6) : 1083 -1087 . DOI: 10.20043/j.cnki.MPM.202307011
正文
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遗传性玻璃体淀粉样变性（Hereditary vitreous amyloidosis，HVA）是遗传性淀粉样变性中一种罕见的常染色体显性遗传性疾病，特点是仅有眼部有淀粉样物质异常的聚集，主要与TTR蛋白的基因突变有关[1]，但目前关于其形成机制善不明确。
TTR蛋白绝大部分在肝细胞中合成，其余则从眼球中视网膜色素上皮细胞（Retinal pigment epithelium，RPE）以及脉络层中合成[3]，并且由于存在的血-视网膜屏障，阻止了血清中的TTR蛋白进入视网膜内[4]，但在人类的眼球中，TTR蛋白的表达却可能与视黄醇的浓度直接相关。视黄醇是重要的营养物质，主要由外界补充（包括饮食等其他方式），并且大部分储存于肝细胞中，视黄醇从肝细胞到血液中需要通过和视黄醇结合蛋白4（Retinol binding protein 4，RBP4）以及TTR蛋白结合形成TTR-RBP4-视黄醇三元复合物来进行运输[5-8]。前期研究证实，突变模型小鼠由于TTR蛋白的基因突变，导致TTR蛋白在肝脏及血液中的表达量下降[2]，理论上肝脏中转运至眼部的视黄醇含量以及在眼部中TTR蛋白也会发生相应的变化。因此，本研究拟探索突变模型鼠眼部，肝脏组织以及血液中视黄醇表达量，以及眼部组织中TTR蛋白的表达量，明确视黄醇与TTR蛋白之间的关系，深入认识该突变所致的HVA的形成机制，并为HVA的预防提供思路。
1 动物与材料
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1.1 实验动物
课题组前期已成功构建模型[2]，饲养Gly83Arg模型小鼠20只（NEO-；FLP-；TTR+/-），对照组小鼠（C57BL/6）20只，所有小鼠饲养在相同的控制条件下，包括光照（12h光／12h暗循环）、湿度（50％～60％）和温度（22～24℃），并且可以获得标准食物和水，本次实验已通过伦理审查，审查编号：ZMU21-2303-135。
1.2 主要设备
高分辨度动物超声摄像系统，低温冷冻离心机 （Sigma 3K15），Real-time检验仪 （ABI-7500），液质联用仪（AB SCIEX 4000 Q TRAP），电泳仪BIO-RAD （公司型号 mini protean 3 cell），电转仪PS-9 （大连竞迈科技股份集团公司），酶标仪（Thermo 型号MK3），移液枪（Thermo），水浴锅 (莱卡公司HI1210），一体式化学发光成像仪（ChemiScope 5300 Pro）
1.3 主要试剂
BCA蛋白定量试剂盒（凯基KGDBCA），丽春红S （上海国药 3761-53-3），Tween-20（Amresco BYL40713），ECL发光液 （北京鼎国 ECL-0011），甲醇/乙腈/甲酸 （HPLC级），正己烷（HPLC级），维生素 A( 50 mg /L) 标准品（成都瑞芬思），维生素 A 乙酸酯（VA-D6）内标标准品（成都瑞芬思），Trizol试剂盒（invitrogen），氯仿/异丙醇/无水乙醇 （上海国药），SYBRGreen PCR试剂盒 （Thermo F-415XL），逆转录试剂盒（Thermo K1622），GAPDH （Proteintech 60004-1-Ig），Anti-Prealbumin（abcam ab215202），辣根酶标记后的山羊抗兔IgG（中杉金桥ZB2301），辣根酶标记后的山羊抗小鼠IgG（中杉金桥ZB2305），M5 HiPer“染立显”蛋白染胶液（聚合美MF767-01）
2 研究方法
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2.1 模型小鼠的构建与验证
前期通过与上海南方生物模式研究中心合作，在Ensembl数据库选出TTR基因序列, ET克隆法构建TTR基因点突变载体。用pBR322-MK-TTR（KI）打靶载体对C57BL/6N小鼠ES细胞进行基因打靶。获得F0代甲状腺素蛋白基因Gly83Arg点突变的C57BL/6N嵌合体小鼠。将F0代和Flp转基因小鼠、C57BL/6J野生型小鼠交配得到F2代无Flp的TTR基因Gly83Arg点突变的杂合子小鼠。F2代小鼠和纯系C57BL/6小鼠交配获得F3代纯合子及杂合子，建立了TTR基因Gly83Arg点突变小鼠模型。将构建的突变小鼠通过裂隙灯下眼前段照相确认发现不规则混浊，小鼠眼部B超：模型小鼠乙醚吸入麻醉后，在小鼠B超机下进行拍摄，证实突变组小鼠眼部出现混浊。基因测序：发现小鼠眼球玻璃体中出现混浊后处死小鼠，取出肝，脑，肾等组织。选择1只杂合子小鼠的肝脏进行基因测序，结果应显示第3外显子的107位置的碱基处出现杂合突变，第83位置的氨基酸的密码子由GGC突变成CGC，测序服务由华大基因提供。
2.2 高效液相色谱联合质谱检测视黄醇含量
实验条件：色谱条件：色谱柱Agilent ZOBAX SB-C18 (4.6×150 mm 5 μm)。流动相由 A、B两相组成，A 为甲醇，B为0.2‰甲酸水溶液，比例：93（A）:7（B）流速1 ml/min。柱温：40℃。进样量为10 μl质谱条件：离子源：电喷雾离子源，扫描方式：多反应监测（MRM）正离子扫描模式，m/z（269.1/93.3），CUR 40psi；IS 5 500 V；TEM 550 ℃；EP 10 V;GS1：55psi;GS2 55psi;CAD 中等;DP 50 V;CE 26V;CXP 10 V。配置标准品并绘制标准曲线后对样品进行测定。实验方法：（1）在暗室里处死小鼠后将两只眼球放入含0.5 ml PBS的1.5 ml离心管中。（2）将离心管置于低温条件后通过细胞破碎仪下将其内容物均质化（功率220，时间2分钟，温度60.0℃）。（3）将200 μl匀浆液转移到5 ml离心管中。（4）加入20 μl内标乙酸视黄酯（10 μg/ml）和100 μl 乙醇。（5）短暂涡旋离心管。（6）加入1 ml 己烷并涡旋30 s两次。（7）在台式低速离心机中以 4 000 rpm 的速度离心5分钟。（8）离心后将上层相转移到新的5 ml离心管中，剩余部分重复（6）（7）步骤两次，取出的上层相均加入至同一个5 ml离心管中，并用锡箔纸包裹离心管。（9）在N2温和气流下干燥上清液（时间不超过30分钟）。（10）将样品溶解在100 μl 流动相（甲醇）中，涡旋振荡后在台式低速离心机中以 4 000 rpm 的速度离心3分钟，转移到进样小瓶。注：整个过程避光进行。
2.3 RT-qPCR法检测眼球组织TTR基因的表达
Real-time PCR所使用的的引物由上海生工公司Primer Premier 5.0软件设计、合成，以管家基因GAPDH为内参。TTR：上游：5’-CACAATCGCCCTCAAACC-3；下游：5’-ATGCCAAGTGTCTTCCAGTA-3’；GAPDH：上游：5’-AGCGTCAAAGGTGGAGGGAGTGG-3’；下游：5’-TCAAGGGCATCCTGGGCTACAC-3’。实验方法：组织总RNA抽提利用Trizol试剂盒；并利用该试剂盒逆转录为cDNA，逆转录体系：采用20 μl逆转录反应体系，将PCR反应条件设置为：37℃持续15 min，85 ℃持续5秒。Real time-PCR反应体系：采用20 μl体系，10.0 μl GoTaq Green Master Mix ，1.0 μl上游引物溶液（F），1.0 μl下游引物溶液（R）2.0 μl cDNA,6.0 μl dH2O。PCR扩增条件：95℃ 10 min，(95℃ 10 s, 60℃ 15 s, 72℃ 20 s) 并进行45个循环。将GAPDH设置为内参，并采用 2△△CT法对目的基因的相对表达量进行计算。
2.4 SDS-PAGE蛋白凝胶显色以及Western blot检测眼球组织TTR蛋白表达
眼球样本中提取总蛋白，并采用BCA法测量蛋白质浓度。SDS-PAGE蛋白凝胶显色：制备蛋白样品后变性并完成配胶，电泳，染色以及脱色。Western blot检测：制备蛋白样品后并使其变性，进行上样，配胶，电泳，切胶，转膜，封闭，孵育一抗：根据试剂盒说明书确定一抗稀释浓度比例，用封闭液稀释抗体至合适浓度，4℃过夜孵育。二抗孵育：待一抗孵育结束后，TBST冲洗3次后，将PVDF膜放入以1：5 000稀释的HRP标记的兔二抗或者鼠二抗溶液中，37℃孵育1 h。最后进行曝光及显色。SDS-PAGE蛋白凝胶显色以及Western blot检测均采用ImageJ软件分析蛋白质条带的灰度值并进行量化。
2.5 统计学方法
运用graphpad prism 8.4.3数据进行统计学处理。样本资料为正态分布，计量资料以表示，不同小鼠眼部，肝脏组织以及血液中视黄醇表达，TTR基因mRNA、蛋白表达均使用独立样本t检验，检验水准：α=0.05。
3 结果
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3.1 模型小鼠前期验证工作
观察到小鼠眼球中玻璃体腔出现不规则混浊后，提取肝组织后进行测序，突变组小鼠肝组织基因测序显示第3外显子的107位置的碱基处出现杂合突变，第83位置的氨基酸的密码子由GGC突变成CGC，即Gly83Arg，与课题组前期构建的模型小鼠突变位点一致。
3.2 两组小鼠不同组织中视黄醇含量
检测结果TTR Gly83Arg突变组小鼠眼球组织中的视黄醇含量较对照组小鼠眼球组织含量低（t=6.600，P=0.022）；肝脏组织视黄醇含量要大于对照组（t=9.045，P<0.001）；血液中视黄醇含量要低于照组（t=4.343，P=0.012）（表1，图1）。
3.3 RT-PCR检测说明
突变组小鼠眼球组织中的TTR蛋白mRNA表达水平较对照组小鼠眼球组织含量低，两组差异有统计学意义（t=5.764，P=0.004）（表2，图2）。
3.4 SDS-PAGE分析
提示突变组小鼠眼球组织中TTR蛋白变性后的残基蛋白表达水平提高，突变组在11kd处出现蛋白条带（图3），Western Blot检测提示，突变组小鼠眼球组织中TTR蛋白表达低于对照组（表3，图4），两组差异有统计学意义（t=37.48，P<0.001）。
4 讨论
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转甲状腺素淀粉样变主要与TTR蛋白出现突变有关，TTR蛋白四聚体稳定性降低解聚为单体，这些具有细胞毒性的变性蛋白沉积在各种不同组织中造成不同程度的病变[1]，是一种多系统疾病，眼部是其中可能累及的一个器官位置，目前有32个TTR基因突变涉及到眼部，并且仅累及眼部的造成玻璃体出现淀粉样沉积的有Arg54Gly[9]、Lys35Thr+Leu55Arg[10]、Gly83Arg[11]、Trp41Leu[12]，这与个基因遗传学分子特征有关。在本次研究中，课题组之前已成功构建了Gly83Arg模型小鼠[3]。本次研究首先验证了所采用模型鼠构建成功，基因测序的突变位点与前期所构建的模型鼠的突变位点相同，第3外显子的107位碱基处出现杂合突变G→C（c.107G→C）。
在对Gly83Arg突变小鼠眼部组织研究中发现，SDS-PAGE结果显示突变组的小鼠在接近11kD处出现条带，这与JURIS J等人对在TTRIle84Ser淀粉样变性中结果相似[13]，我们推断该条带的形成可能是由于变异的TTR蛋白增多并所形成的残基聚集形成的，在TTR蛋白相关的淀粉样变性所导致的玻璃体混浊中，玻璃体中的淀粉样蛋白主要是由于眼部组织自身合成变异TTR蛋白增多所导致的[14]。同样我们采用了Western Blot检测与RT-PCR检测了眼部组织中TTR蛋白的表达量，在Gly83Arg小鼠中，其TTR蛋白及mRNA表达水平降低。
同样本次实验对Gly83Arg突变小鼠的眼部，肝脏组织以及血液中视黄醇的含量进行了检测，发现突变型小鼠的眼球组织以及血液中的视黄醇含量减少，但在肝脏组织中增多。视黄醇-RBP4-TTR三元复合物将视黄醇从肝脏转运至血液中[15-16]，而由于突变组小鼠肝脏中的TTR蛋白减少[2]，导致视黄醇积聚于肝脏组织中，从而肝脏进入血液循坏中的视黄醇含量也减少。而在眼部中主要介导视黄醇转运功能的是STRA6受体，并且该受体受TTR蛋白的影响[17]，首先需要视黄醇对其进行激活。在Gly83Arg小鼠中，血液循环中的低视黄醇水平影响了STRA6转运视黄醇的功能，并且增多的变异TTR蛋白抑制了RBP4与STRA6的结合，从而导致了眼部组织中的视黄醇含量减低。
在Gly83Arg突变小鼠的眼球组织中，由于眼球组织中突变TTR蛋白的增多以及血液中转运的视黄醇含量减少，导致了眼球组织中视黄醇含量的减少，这些或许能为认识Gly83Arg所致的淀粉样病变中淀粉样蛋白仅存在于眼部组织的形成机制提供新的思路，也提示该病的治疗不仅可以采取玻璃体切割，或许也可以通过视黄醇对Gly83Arg突变所引起的玻璃体淀粉样变进行干预，为该疾病以后的治疗甚至预防提供了新的路线。
基金
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  • 国家自然科学基金(31871261)
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[1]
Marcoux J, Mangione PP, Porcari R, et al. A novel mechano-enzymatic cleavage mechanism underlies transthyretin amyloidogenesis[J]. EMBO Molecular Medicine, 2015, 7(10): 1337-1349.
[2]
Liz MA, Celho T, Bellotti V, et al. A narrative review of the role of transthyretin in health and disease[J]. Neurology and Therapy, 2020, 9(2): 395-402
[3]
Beirão M, Matos E, Beirão I, et al. No ocular involvement in familial amyloidotic polyneuropathy ATTR V30M domino liver recipients[J]. Transplant International, 2012, 25(6): 646-651.
[4]
Fex G, Johannesson G. Transfer of retinol from retinol-binding protein complex to liposomes and across liposomal membranes[J]. Methods in Enzymology, 1990, 189: 394-402.
[5]
Noy N, Xu ZJ. Interactions of retinol with binding proteins: implications for the mechanism of uptake by cells[J]. Biochemistry, 1990, 29(16): 3878-3883.
[6]
Noy N, Blaner WS. Interactions of retinol with binding proteins: studies with rat cellular retinol-binding protein and with rat retinol-binding protein[J]. Biochemistry, 1991, 30(26): 6380-6386.
[7]
Noy N, Xu ZJ. Kinetic parameters of the interactions of retinol with lipid bilayers[J]. Biochemistry, 1990, 29(16): 3883-3888.
[8]
Ran LX, Zheng ZY, Xie B, et al. A mouse model of a novel missense mutation (Gly83Arg) in a Chinese kindred manifesting vitreous amyloidosis only[J]. Experimental Eye Research, 2018, 169: 13-19.
[9]
Andersson R. Familial amyloidosis with polyneuropathy. A clinical study based on patients living in northern Sweden[J]. Acta Medica Scandinavica. Supplementum, 1976, 590: 1-64.
[10]
Andrade C. A peculiar form of peripheral neuropathy; familiar atypical generalized amyloidosis with special involvement of the peripheral nerves[J]. Brain : a Journal of Neurology, 1952, 75(3): 408-427.
[11]
Shi Y, Li J, Hu J, et al. A new Arg54Gly transthyretin gene mutation associated with vitreous amyloidosis in Chinese[J]. eye Science, 2011, 26(4): 230-238.
[12]
Yin J, Xia XY, Shi Y, et al. Chinese familial transthyretin amyloidosis with vitreous involvement is associated with the transthyretin mutation Gly83Arg: a case report and literature review[J]. Amyloid : the International Journal of Experimental and Clinical Investigation: the Official Journal of the International Society of Amyloidosis, 2014, 21(2): 140-142.
[13]
Liepnieks JJ, Wilson DL, Benson MD. Biochemical characterization of vitreous and cardiac amyloid in Ile84Ser transthyretin amyloidosis[J]. Amyloid: the International Journal of Experimental and Clinical Investigation : the Official Journal of the International Society of Amyloidosis, 2006, 13(3): 170-177.
[14]
Hara R, Kawaji T, Ando E, et al. Impact of liver transplantation on transthyretin-related ocular amyloidosis in Japanese patients[J]. Archives of Ophthalmology, 2010, 128(2): 206-210.
[15]
Marill J, Idres N, Capron CC, et al. Retinoic acid metabolism and mechanism of action: a review[J]. Current Drug Metabolism, 2003, 4(1): 1-10.
[16]
D’Ambrosio DN, Clugston RD, Blaner WS. Vitamin A metabolism: an update[J]. Nutrients, 2011, 3(1): 63-103.
[17]
Noy N. Vitamin a in regulation of insulin responsiveness: mini review[J]. Proceedings of the Nutrition Society, 2016, 75(2): 212-215.
2024年第51卷第6期
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doi: 10.20043/j.cnki.MPM.202307011
  • 接收时间：2023-07-04
  • 首发时间：2026-03-16
  • 出版时间：2024-03-25
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  • 收稿日期：2023-07-04
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国家自然科学基金(31871261)
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    遵义医科大学眼科，贵州 遵义 563000

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蔡善君，E-mail：
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