Article(id=1322923066203976627, tenantId=1146029695717560320, journalId=1255847919539208197, issueId=1322923011518628427, articleNumber=null, orderNo=null, doi=10.13193/j.issn.1673-7717.2025.03.015, pmid=null, cstr=null, oa=null, hot=null, price=null, onlineType=0, articleFormat=0, articleType=null, articleTypeStr=null, receivedDate=null, receivedDateStr=null, revisedDate=null, revisedDateStr=null, acceptedDate=null, acceptedDateStr=null, onlineDate=1793339037705, onlineDateStr=2026-10-30, pubDate=1741536000000, pubDateStr=2025-03-10, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1793339037705, onlineIssueDateStr=2026-10-30, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1793339037705, creator=13701087609, updateTime=1793339037705, updator=13701087609, issue=Issue{id=1322923011518628427, tenantId=1146029695717560320, journalId=1255847919539208197, year='2025', volume='43', issue='3', pageStart='1', pageEnd='258', issueExtLink='null', onlineDate='null', pubDate='1741536000000', pubDateStr='2025-03-10', beforeIssueId=null, nextIssueId=null, price=null, status=1, issueComplete=1, articleOrder=1, issueType=-1, specialIssue=0, createTime=1793339024666, creator='13701087609', updateTime=1793340602374, updator='13701087609', preIssue=null, nextIssue=null, articleTotal=null, ext={EN=IssueExt(id=1322929628976906432, tenantId=1146029695717560320, journalId=1255847919539208197, issueId=1322923011518628427, language=EN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=), CN=IssueExt(id=1322929628981100737, tenantId=1146029695717560320, journalId=1255847919539208197, issueId=1322923011518628427, language=CN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=)}, issueFiles=null, downloadFileDto=null}, startPage=92, endPage=95, ext={EN=ArticleExt(id=1322923066422080436, articleId=1322923066203976627, tenantId=1146029695717560320, journalId=1255847919539208197, language=EN, title=Study on Anti-Aging Effect of Cistanche deserticola Polysaccharide by Improving Homeostasis of Intestinal Flora, columnId=1256183361521967297, journalTitle=Chinese Archives of Traditional Chinese Medicine, columnName=Target on National Project, runingTitle=null, highlight=null, articleAbstract=
Objective

By studying the anti-aging effect of Cistanche deserticola polysaccharides(CDPS)in vivo,it aimed to explore the possible mechanisms underlying this anti-aging effect.

Methods

Seventy-five Kunming species(KM)mice aged 12 months were randomly divided into a blank control group(WT),a low-dose group of Cistanche deserticola polysaccharides(CDPS. L),a medium dose group of Cistanche deserticola polysaccharides(CDPS. M),a high-dose group of Cistanche deserticola polysaccharides(CDPS. H),and a positive drug group(P). The mice were given corresponding drugs or the same volume of distilled water by gavage until obvious signs of death appeared. The other mice that did not show obvious signs of death were uniformly sampled 180 days after administration. The levels of glutathione peroxidase(GSH-Px),total superoxide dismutase(TSOD)and malondialdehyde(MDA)in serum were detected to observe the antioxidant effect of Cistanche deserticola polysaccharides.16SrRNA sequencing was used to analyze the structure of gut microbiota. The effects on cognitive ability and intestinal function of mice were observed through HE staining of pathological sections of the hippocampus and intestines.

Results

Compared with that o the WT group,the survival rate of the mice treated with CDPSwas improved,and the effect of CDPS. M and CDPS. H groups was better than that of Pgroup,but it was not significant. Compared with those of the WT group mice,the GSH-Px activity and T-SOD activity of the CDPSgroups were significantly increased,and the MDA levels in each treatment group were significantly reduced compared to that in the WT group,and the GSH-Px activity of CDPS. H group was significantly improved compared to that in Pgroup. CDPScan improve the intestinal microbiota homeostasis in mice,increase the levels of Akkermansia muciniphila,Alloprevotella and Prevotellaceae probiotics,and improve intestinal function. Compared with those of the other groups,the intestinal villus length and wall thickness of mice were longer. CDPSalso can reduce hippocampal neuronal death caused by aging.

Conclusion

CDPShas anti-aging effects,which may be due to its antioxidant properties and its ability to improve intestinal function,thereby affecting the homeostasis of gut microbiota,resulting in an increase in the levels of Akkermansia muciniphila,Alloprevotella and Prevotellaceae probiotics. Additionally,the effect of CDPSon reducing hippocampal neuronal death in mice may also be one of the anti-aging mechanisms of CDPS.

, authors=Chunsheng ZHANG1, 3, Bing LI1, luyu WANG1, Zhihua ZHOU1, Ting ZHANG1, Li ZHU1, Shuang LIU4, Xin WANG5, Gang LI1, 2, authorsList=Chunsheng ZHANG, Bing LI, luyu WANG, Zhihua ZHOU, Ting ZHANG, Li ZHU, Shuang LIU, Xin WANG, Gang LI, authorCompany=null, correspAuthors=Gang LI, authorNote=null, correspAuthorsNote=null, copyrightStatement=null, copyrightOwner=null, extLink=null, articleAbsUrl=null, sourceXml=null, magXml=null, pdfUrl=null, pdf=null, pdfFileSize=null, pdfExtLink=null, richHtmlUrl=null, mobilePdfUrl=null, reviewReport=null, pdfFirstPage=null, abstractGraph=null, abstractGraphContent=null, abstractVideo=null, citation=null, cebUrl=null, magXmlContent=null, mapNumber=null, fund=null), CN=ArticleExt(id=1322923069572002753, articleId=1322923066203976627, tenantId=1146029695717560320, journalId=1255847919539208197, language=CN, title=肉苁蓉多糖通过改善肠道菌群稳态产生抗衰老作用的研究, columnId=1256183362172142382, journalTitle=中华中医药学刊, columnName=国家项目点击, runingTitle=null, highlight=null, articleAbstract=
目的

通过体内研究肉苁蓉多糖(Polysaccharides of Cistanche deserticola,CDPS)的抗衰老作用,探究产生这种抗衰老作用可能发生的机制。

方法

75只12月龄的昆明种(Kunming species,KM)小鼠按照完全随机的方法分为空白对照组(Wild Type group,WT)、肉苁蓉多糖低剂量组(CDPS.L)、肉苁蓉多糖中剂量组(CDPS.M)、肉苁蓉多糖高剂量组(CDPS.H),双歧杆菌四联活菌片组(Positive drug group,P),分别给予相应的药物或相同体积的蒸馏水灌胃直至小鼠出现明显死亡证候,其余未出现明显死亡证候的小鼠在给药后180 d统一取材。进行血清中的谷胱甘肽过氧化物酶(Glutathione peroxidase,GSH-Px),总超氧化物歧化酶(Total Superoxide Dismutase,T-SOD)和丙二醛(Malonaldehyde,MDA)的检测来观察肉苁蓉多糖的抗氧化作用;采用16SrRNA测序分析肠道菌群结构;通过小鼠海马和肠的病理切片HE染色观察对小鼠认知能力和对肠道功能的影响。

结果

与WT组小鼠相比,CDPS给药组小鼠的生存率均提高,且CDPS.M及CDPS.H组的效果优于P组,但是并不具有显著性;与WT组小鼠相比,CDPS给药组小鼠的GSH-Px活力、T-SOD活力均有明显提高,各给药组MDA水平较WT组小鼠明显下降,且CDPS.H组的GSH-PX活力较P组有明显改善;CDPS给药可以改善小鼠的肠道菌群稳态,使Akkermansia muciniphila、Alloprevotella和Prevotellaceae这三种益生菌的水平升高,且可以改善小鼠的肠道功能,与其他组相比小鼠肠道的肠绒毛长度较长,肠壁厚度也较厚。CDPS给药同时可以减少衰老引起的海马神经元死亡。

结论

CDPS具有抗衰老作用,这种作用可能是因为其具有抗氧化作用,及其可以通过改善肠道功能,进而影响肠道菌群稳态,使Akkermansia muciniphila、Alloprevotella和Prevotellaceae这三种益生菌的水平升高而实现的,并且CDPS减少小鼠海马神经元死亡的作用也可能是CDPS抗衰老作用机制之一。

, authors=张春生1, 3, 李冰1, 王鲁豫1, 周志华1, 张婷1, 朱莉1, 梁彦1, 刘爽4, 王鑫5, 李刚1, 2, authorsList=张春生, 李冰, 王鲁豫, 周志华, 张婷, 朱莉, 梁彦, 刘爽, 王鑫, 李刚, authorCompany=null, correspAuthors=李刚, authorNote=

张春生(1999-),男,内蒙古赤峰人,硕士在读,研究方向:神经药理学,中蒙药药理学。

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李刚(1979-),男,内蒙古呼和浩特人,教授,硕士研究生导师,博士,研究方向:神经药理学,中蒙药药理学。E-mail:。
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张春生(1999-),男,内蒙古赤峰人,硕士在读,研究方向:神经药理学,中蒙药药理学。

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张春生(1999-),男,内蒙古赤峰人,硕士在读,研究方向:神经药理学,中蒙药药理学。

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注:A:各组小鼠的生存曲线。B:截至给药后180天,WT、CDPS.L、CDPS.M、CDPS.H和P组的生存率。C:各组小鼠血清GSH-PX活性。D:各组小鼠血清中T-SOD活性。E:各组小鼠血清MDA含量。Mean±SD. n=15.*P<0.05,**P<0.01,***P<0.001,****P<0.000 1。

, figureFileSmall=q/8iLUDedbUYFiDK3Gv3Iw==, figureFileBig=xiPlOtDEg0vfNz1ZcpG5ew==, tableContent=null), ArticleFig(id=1322923075137843211, tenantId=1146029695717560320, journalId=1255847919539208197, articleId=1322923066203976627, language=EN, label=null, caption=null, figureFileSmall=KQpB6gdvFuLbt1nb76PVFQ==, figureFileBig=0ZCk6OZJRd8WNdgX8STJug==, tableContent=null), ArticleFig(id=1322923075242700812, tenantId=1146029695717560320, journalId=1255847919539208197, articleId=1322923066203976627, language=CN, label=图2, caption=肉苁蓉多糖对衰老小鼠肠道菌群的影响

注:A:基于OTU的维恩图。B:门级别的物种丰度直方图。C:物种丰度聚类在门水平上的热图。

, figureFileSmall=KQpB6gdvFuLbt1nb76PVFQ==, figureFileBig=0ZCk6OZJRd8WNdgX8STJug==, tableContent=null), ArticleFig(id=1322923075318198285, tenantId=1146029695717560320, journalId=1255847919539208197, articleId=1322923066203976627, language=EN, label=null, caption=null, figureFileSmall=q0VtbETcbqPzyNDHFfXBTw==, figureFileBig=q0ug75zOFBfX6QKG6NWjkQ==, tableContent=null), ArticleFig(id=1322923075389501454, tenantId=1146029695717560320, journalId=1255847919539208197, articleId=1322923066203976627, language=CN, label=图3, caption=肉苁蓉多糖对衰老小鼠肠道菌群的影响

注:A:各组小鼠的界、门、纲、目、科、属和种的肠道菌群分布。B:基于shannon指数的基于alpha多样性的箱形图。C:基于Weighted Unifrac Beta多样性的箱形图。D:LEfSe图。

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肉苁蓉多糖通过改善肠道菌群稳态产生抗衰老作用的研究
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张春生 1, 3 , 李冰 1 , 王鲁豫 1 , 周志华 1 , 张婷 1 , 朱莉 1 , 梁彦 1 , 刘爽 4 , 王鑫 5 , 李刚 1, 2
中华中医药学刊 | 国家项目点击 2025,43(3): 92-95
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中华中医药学刊 |国家项目点击 2025 , 43 (3) : 92 -95
肉苁蓉多糖通过改善肠道菌群稳态产生抗衰老作用的研究
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张春生(1999-),男,内蒙古赤峰人,硕士在读,研究方向:神经药理学,中蒙药药理学。

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张春生(1999-),男,内蒙古赤峰人,硕士在读,研究方向:神经药理学,中蒙药药理学。

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张春生1, 3, 李冰1, 王鲁豫1, 周志华1, 张婷1, 朱莉1, 梁彦1, 刘爽4, 王鑫5, 李刚1, 2
作者信息
  • 1.内蒙古医科大学药学院,内蒙古 呼和浩特 010100
  • 2.内蒙古医科大学民族医药创新中心,内蒙古 呼和浩特 010100
  • 3.呼和浩特市第三医院,内蒙古 呼和浩特 010100
  • 4.内蒙古医科大学第二附属医院,内蒙古 呼和浩特 010100
  • 5.内蒙古科技大学包头医学院,内蒙古 包头 014000
通讯作者:
李刚(1979-),男,内蒙古呼和浩特人,教授,硕士研究生导师,博士,研究方向:神经药理学,中蒙药药理学。E-mail:。
作者简介:

张春生(1999-),男,内蒙古赤峰人,硕士在读,研究方向:神经药理学,中蒙药药理学。

Study on Anti-Aging Effect of Cistanche deserticola Polysaccharide by Improving Homeostasis of Intestinal Flora
Chunsheng ZHANG1, 3, Bing LI1, luyu WANG1, Zhihua ZHOU1, Ting ZHANG1, Li ZHU1, Shuang LIU4, Xin WANG5, Gang LI1, 2
Affiliations
  • 1.School of Pharmacy,Inner Mongolia Medical University,Hohhot 010100,China
  • 2.National Medicine Innovation Center of Inner Mongolia Medical University,Hohhot 010100,China
  • 3.Hohhot Third Hospital,Hohhot 010100,China
  • 4.The Second Affiliated Hospital of Inner Mongolia Medical University,Hohhot 010100,China
  • 5.Baotou Medical College of Inner Mongolia University of Science and Technology,Baotou 014000,China
出版时间: 2025-03-10 doi: 10.13193/j.issn.1673-7717.2025.03.015
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目的

通过体内研究肉苁蓉多糖(Polysaccharides of Cistanche deserticola,CDPS)的抗衰老作用,探究产生这种抗衰老作用可能发生的机制。

方法

75只12月龄的昆明种(Kunming species,KM)小鼠按照完全随机的方法分为空白对照组(Wild Type group,WT)、肉苁蓉多糖低剂量组(CDPS.L)、肉苁蓉多糖中剂量组(CDPS.M)、肉苁蓉多糖高剂量组(CDPS.H),双歧杆菌四联活菌片组(Positive drug group,P),分别给予相应的药物或相同体积的蒸馏水灌胃直至小鼠出现明显死亡证候,其余未出现明显死亡证候的小鼠在给药后180 d统一取材。进行血清中的谷胱甘肽过氧化物酶(Glutathione peroxidase,GSH-Px),总超氧化物歧化酶(Total Superoxide Dismutase,T-SOD)和丙二醛(Malonaldehyde,MDA)的检测来观察肉苁蓉多糖的抗氧化作用;采用16SrRNA测序分析肠道菌群结构;通过小鼠海马和肠的病理切片HE染色观察对小鼠认知能力和对肠道功能的影响。

结果

与WT组小鼠相比,CDPS给药组小鼠的生存率均提高,且CDPS.M及CDPS.H组的效果优于P组,但是并不具有显著性;与WT组小鼠相比,CDPS给药组小鼠的GSH-Px活力、T-SOD活力均有明显提高,各给药组MDA水平较WT组小鼠明显下降,且CDPS.H组的GSH-PX活力较P组有明显改善;CDPS给药可以改善小鼠的肠道菌群稳态,使Akkermansia muciniphila、Alloprevotella和Prevotellaceae这三种益生菌的水平升高,且可以改善小鼠的肠道功能,与其他组相比小鼠肠道的肠绒毛长度较长,肠壁厚度也较厚。CDPS给药同时可以减少衰老引起的海马神经元死亡。

结论

CDPS具有抗衰老作用,这种作用可能是因为其具有抗氧化作用,及其可以通过改善肠道功能,进而影响肠道菌群稳态,使Akkermansia muciniphila、Alloprevotella和Prevotellaceae这三种益生菌的水平升高而实现的,并且CDPS减少小鼠海马神经元死亡的作用也可能是CDPS抗衰老作用机制之一。

肉苁蓉多糖  /  衰老  /  肠道菌群
Objective

By studying the anti-aging effect of Cistanche deserticola polysaccharides(CDPS)in vivo,it aimed to explore the possible mechanisms underlying this anti-aging effect.

Methods

Seventy-five Kunming species(KM)mice aged 12 months were randomly divided into a blank control group(WT),a low-dose group of Cistanche deserticola polysaccharides(CDPS. L),a medium dose group of Cistanche deserticola polysaccharides(CDPS. M),a high-dose group of Cistanche deserticola polysaccharides(CDPS. H),and a positive drug group(P). The mice were given corresponding drugs or the same volume of distilled water by gavage until obvious signs of death appeared. The other mice that did not show obvious signs of death were uniformly sampled 180 days after administration. The levels of glutathione peroxidase(GSH-Px),total superoxide dismutase(TSOD)and malondialdehyde(MDA)in serum were detected to observe the antioxidant effect of Cistanche deserticola polysaccharides.16SrRNA sequencing was used to analyze the structure of gut microbiota. The effects on cognitive ability and intestinal function of mice were observed through HE staining of pathological sections of the hippocampus and intestines.

Results

Compared with that o the WT group,the survival rate of the mice treated with CDPSwas improved,and the effect of CDPS. M and CDPS. H groups was better than that of Pgroup,but it was not significant. Compared with those of the WT group mice,the GSH-Px activity and T-SOD activity of the CDPSgroups were significantly increased,and the MDA levels in each treatment group were significantly reduced compared to that in the WT group,and the GSH-Px activity of CDPS. H group was significantly improved compared to that in Pgroup. CDPScan improve the intestinal microbiota homeostasis in mice,increase the levels of Akkermansia muciniphila,Alloprevotella and Prevotellaceae probiotics,and improve intestinal function. Compared with those of the other groups,the intestinal villus length and wall thickness of mice were longer. CDPSalso can reduce hippocampal neuronal death caused by aging.

Conclusion

CDPShas anti-aging effects,which may be due to its antioxidant properties and its ability to improve intestinal function,thereby affecting the homeostasis of gut microbiota,resulting in an increase in the levels of Akkermansia muciniphila,Alloprevotella and Prevotellaceae probiotics. Additionally,the effect of CDPSon reducing hippocampal neuronal death in mice may also be one of the anti-aging mechanisms of CDPS.

Cistanche deserticola polysaccharides  /  aging  /  intestinal microbiota
张春生, 李冰, 王鲁豫, 周志华, 张婷, 朱莉, 梁彦, 刘爽, 王鑫, 李刚. 肉苁蓉多糖通过改善肠道菌群稳态产生抗衰老作用的研究. 中华中医药学刊, 2025 , 43 (3) : 92 -95 . DOI: 10.13193/j.issn.1673-7717.2025.03.015
Chunsheng ZHANG, Bing LI, luyu WANG, Zhihua ZHOU, Ting ZHANG, Li ZHU, Shuang LIU, Xin WANG, Gang LI. Study on Anti-Aging Effect of Cistanche deserticola Polysaccharide by Improving Homeostasis of Intestinal Flora[J]. Chinese Archives of Traditional Chinese Medicine, 2025 , 43 (3) : 92 -95 . DOI: 10.13193/j.issn.1673-7717.2025.03.015
全球正在步入老龄化,我国的人口老龄化问题也日益严重,根据2020年第七次全国人口普查结果,60岁及以上人口占比达到了18.70%,其中65岁及以上人口占比13.50%。与2010年第六次全国人口普查相比,60岁及以上人口的比重上升5.44个百分点,65岁及以上人口的比重上升4.63个百分点[1]。人口老龄化问题已经十分严峻。
中药肉苁蓉是列当科植物肉苁蓉的干燥肉质茎,具有补阳、益精血、润肠通便等功效,肉苁蓉入药始载于《神农本草经》,是沙生名贵的滋补中药材[2-3]。在许多传统中医药古籍中被描述为具有抗衰老特性。作为肉苁蓉的主要有效成分,肉苁蓉多糖已被多项研究证明其具有抗衰老作用,在本课题组之前的一项研究中,我们利用D-半乳糖建立体外急性衰老细胞模型,结果表明各给药组与模型组比较,环磷酸腺苷(cAMP)/蛋白激酶A(PKA)/环磷酸腺苷反应成分结合蛋白(CREB)信号通路水平上调,提示肉苁蓉多糖改善D-半乳糖急性衰老模型的作用与调节cAMP/PKA/CREB信号通路有关[4]。在另一项研究中,使用D-半乳糖建立衰老大鼠模型肉苁蓉水提物给药,发现肉苁蓉给药组的SOD有上升趋势,MDA含量以及NO含量显著降低。因此认为肉苁蓉具有一定的抗衰老作用[5],并认为这种肉苁蓉水提物(成分主要为多糖)的抗衰老作用主要是通过抗氧化作用所实现的。
在本研究通过小鼠长期灌服肉苁蓉多糖直至自然死亡或180 d统一取材,通过分析小鼠的生存率,检测血清的GSHPx,T-SOD和MDA的水平来观察肉苁蓉多糖的抗衰老作用,并且观察脑和盲肠的病理变化以及肠道菌群的改变来验证其可能的作用机制。
健康12月龄雄性KM小鼠75只,体质量为45~55 g,购买于内蒙古医科大学实验动物中心,许可证编号:[SCXK(蒙)2020-0001]。饲养于内蒙古医科大学实验动物中心SPF级动物房,实验环境温度20~25℃,相对湿度40%~60%。
肉苁蓉多糖(将清洗后的肉苁蓉约1.0 kg在40℃烘箱中风干,粉碎成粗粉。粉末用热乙醇提取3 h,残渣用纱布过滤除去滤液,然后用8倍量水稀释,在90℃下依次回流2 h、1.5 h和1 h,每个时间点离心分离上清液,与滤液结合。滤液在减压下浓缩,冷却至室温,缓慢加入3倍量95%乙醇,4℃静置24 h,然后以6000 r/min离心20 min,重复水提和乙醇沉淀三次后收集沉淀物。将沉淀物在水中复溶、脱蛋白、冷冻干燥,得到肉苁蓉多糖。紫外分光光度法测定多糖含量在90%以上);双歧杆菌四联活菌片(杭州远大生物制药有限公司,生产批号20210911,规格:0.5 g/片)于-4℃冰箱内保存,现用现配;GSH-Px测定试剂盒(A005-1-2,南京建成生物工程研究所);T-SOD测定试剂盒(A001-1-1,南京建成生物工程研究所);MDA测定试剂盒(A003-1-2,南京建成生物工程研究所)。
Molecular Devices多功能酶标仪(SpectraMax i3x,美谷分子仪器有限公司);紫外分光光度计(TU-1901,上海精密仪器仪表有限公司);电子天平(PL-203,梅特勒-托利多仪器上海有限公司);移液枪[Rainin Instrument公司(美国),10、100、200、1000μL];超低温离心机(TGL-16,长沙平凡仪器仪表有限公司);低温冰箱(三洋公司,日本);十万分之一电子天平(Mettler Toledo公司,美国);w H-1微型旋涡混合器(上海沪西分析仪器厂);SB2200-T超声波清洗器(上海必能信超声有限公司);GraphPad Prism 9.0.0(GraphPad Software,La Jolla,加利福尼亚,美国)。
12月龄雄性KM小鼠,适应性饲养1周后,按照体质量分层随机的方法分为空白对照组(Wild Type group,WT)15只,肉苁蓉多糖低剂量组(CDPS.L)(2.5 mg/kg)15只,肉苁蓉多糖中剂量组(CDPS.M)(5 mg/kg)15只,肉苁蓉多糖高剂量组(CDPS.H)(10 mg/kg)15只,双歧杆菌四联活菌片组(Positive drug group,P)(双歧杆菌四联活菌片0.5 g/kg)15只。每天分别给予相应的药物或相同体积的蒸馏水灌胃直至小鼠出现明显死亡证候(皮毛蓬松、精神委顿、眼目不睁、活动明显减少等)。其余未出现明显死亡证候的小鼠在给药后180 d统一采用碳酸气体麻醉处死,具体方法为将小鼠放置在密闭的装置中,注入大量的二氧化碳,使其失去知觉并最终死亡。确保最小限度的痛苦和不适。
使用生化检测试剂盒测量GSH-PX、SOD和MDA水平。使用比色法评估血清GSH-PX水平。羟胺法用于测量血清T-SOD水平,TBA法用于评估血清MDA水平。
所有小鼠的无菌粪便样本-80℃保存,每组随机选取3只进行肠道菌群分析。16S rRNA基因的V4区序列用Illumina MiSeq(北京诺新世股份有限公司,中国,北京)测序,并用QIIME开放平台进行分析,以确定肠道微生物区系。
实验结果以均数±标准差()表示,使用Graphpad Prism 9.0.0进行数据分析及作图,运用单因素方差分析(One way-ANOVA)进行组间差异分析,采用Kaplan-Meier法、COX比例风险模型进行生存分析,P<0.05表示差异有统计学意义,P<0.01表示差异有显著性统计学意义,所有统计结果重复3次。
各实验组小鼠每天分别给予相应药物或相同体积的蒸馏水灌胃,直至出现明显死亡证候后取材,其余未出现明显死亡证候的小鼠在给药180 d后统一取材。获得了插页ⅩⅩ图1A所示的生存曲线图,但是各组间均无显著性差异。接下来我们观察了各组动物的生存率,如插页ⅩⅩ图1B所示,截至给药后第180天,WT组、CDPS.L组、CDPS.M组、CDPS.H组和P组的生存率分别为20%、26.26%、40%、53.33%和33.33%。可见CDPS给药组的生存率高于空白组,且CDPS.M组和CDPS.H组生存率也高于P组。
在插页ⅩⅩ图1C、D、E中,我们测定了各组小鼠血清中GSH-PX、T-SOD和MDA水平。可见与WT组相比,CDPS.H组显著升高了血清中GSH-Px活力(P<0.000 1),且这种升高作用明显优于P组(P<0.05);与WT组相比,CDPS.M组、CDPS.H组和P组均显著升高了血清中T-SOD活力(P<0.01、P<0.001、P<0.05);给药组均可使血清中MDA含量降低,且CDPS.L、CDPS.H和P组有显著地降低血清MDA水平的作用(P<0.05、P<0.01、P<0.001)。
我们的结果表明各给药组小鼠的生存率均高于WT组,但并无统计学意义,在血清生化指标的检测中,我们观察到各给药组的血清氧化应激水平均显著低于WT组,且CDPS.H组在血清GSH-Px水平上优于P组,证明CDPS在一定程度上抗氧化能力优于双歧杆菌四联活菌片。
接下来我们用各组小鼠的无菌粪便进行了16S rRNA高通量测序,基于Illumina Nova测序平台测序,构建PCR-free文库,然后进行双末端测序。通过对Reads拼接,平均每样品测得87 435条tags,经过质控平均得到87 019条有效数据,质控有效数据量达66 976,质控有效率达76.60%。以97%的一致性将序列聚类成为操作性分类单元(Operational Taxonomic Units,OTUs),共得到3117个OTUs,然后对OTUs序列与Silva138数据库进行物种注释。注释结果中,共有1315(42.19%)个OTU注释到属水平。根据物种注释情况,做出基于OTU的Venn图(插页ⅩⅪ图2A),可见各组小鼠的肠道菌群都有不同程度的差异。
比较各组小鼠肠道菌群在门水平上的结构(插页ⅩⅪ图2 B-C),可见Verrucomicrobiota在CDPS.H组和P组中相较于其他三组比例有明显的提高,在插页ⅩⅫ图3A-C中,由基于shannon指数Alpha多样性箱形图和基于Weighted Unifrac Beta多样性箱形图分别观察各组小鼠肠道菌群的α多样性指数组内差异分析和β多样性指数组间分析可见,CDPS.H组小鼠的肠道菌群的组内物种多样性降低,组间样本相似性升高,这反映了Verrucomicrobiota所占比例的升高。由线性判别分析图(Linear discriminant analysis Effect Size,LEfSe)可见(插页ⅩⅫ图3D),在CDPS.H组小鼠的肠道菌群中找到了具有统计学意义的Biomarker两种,分别为Alloprevotella和Prevotellaceae,有研究表明,两者均被认为是产生丁酸的有益菌种[6-7]。
结果表明,肉苁蓉多糖高剂量给药改善了肠道菌群稳态,使Akkermansia muciniphila、Alloprevotella和Prevotellaceae的水平显著提高。
自然衰老会导致肠道组织结构发生改变。随着年龄的增长,小鼠肠道的消化和吸收能力下降,肠黏膜屏障受损,具体表现为肠绒毛数量减少、长度变短[8]。在小鼠取材后,用4%的中性甲醛固定小鼠的脑和肠道,石蜡包埋,用HE染色的方法制作小鼠的大脑海马区和盲肠的病理切片,观察各组实验小鼠的肠道和海马神经元的变化。我们发现CDPS.H组和P组小鼠肠道的肠绒毛长度较其他组明显增加,肠壁厚度也增厚(插页ⅩⅩⅢ图4),逆转了小鼠衰老所引起的肠绒毛长度变短,且CDPS.H组相较P组表现出更明显的治疗作用。
因为我们之前的研究中发现CDPS给药可以通过恢复肠道菌群-脑轴减轻D-半乳糖诱导的衰老模型小鼠的认知能力下降[9],因此,我们接下来观察了各组小鼠海马区的病理切片(插页ⅩⅩⅣ图5),海马组HE染色结果显示,WT组小鼠海马神经元排列紊乱,海马CA1区和DG区正常神经元数目减少,可见大量明显的细胞核固缩及老化细胞(蓝色圆圈标记处)。相较于WT组小鼠,CDPS给药组及P组小鼠的海马神经元排列比较紧密,且细胞核固缩神经元相对较少。
结果表明,CDPS给药可以改善小鼠的肠道功能,改善小鼠海马神经元的核固缩及老化。因此,我们认为CDPS给药小鼠生存率较高的原因可能是因为其可以通过改善小鼠的肠道菌群结构引起小鼠肠道功能的改善,并且减少小鼠的海马神经元死亡,改善小鼠的认知能力,最终引起小鼠寿命得到延长。
衰老是生物体内所有细胞、组织、器官和整体普遍存在的现象,是由自然趋势所造成的内生性变化。肉苁蓉的抗衰老作用已经得到了多次证实,但是其作用机制还不是特别明确,有研究表明多糖可以改善衰老小鼠的学习记忆能力,其机制可能与脑内海马区环磷酸腺苷(cAMP)反应元件结合蛋白环磷腺苷效应元件结合蛋白(CREB)表达水平的上调有关[10]。我们把研究重点放在了改善肠道菌群和肠屏障作用上,并且也观察了其对海马神经元的改善作用。
在健康的有机体中,肠道微生物及其代谢物具有多种调节机体的作用,进而形成抵御传染源的保护屏障[11]。人们对肠道菌群的认识正在逐渐积累,但是更多的研究重点放在了与某些疾病的关系上,如阿尔茨海默病、糖尿病、肠道炎、类风湿关节炎和肿瘤等[12-16]。近来有研究表明可以通过调节肠道菌群稳态进而产生抗衰老作用[17-19],但相关研究依然不多。在本项研究中,我们通过对不同组动物粪便进行16s rRNA测序,发现了不同的给药策略可以引起小鼠肠道菌群的显著变化,这为CDPS的药效的产生机制提供了依据。
在这项研究中,我们发现CDPS给药可以改善小鼠海马区的神经元凋亡,这与我们之前在D-半乳糖诱导的衰老小鼠模型中的结果一致[10],我们今后会对CDPS对脑病理及认知记忆能力的影响做进一步的研究。
Akkermansia muciniphila是一种椭圆形细菌,定植于胃肠道的黏液层。使用黏蛋白作为碳和氮元素的唯一来源[20]。这种黏蛋白降解剂受到位于肠上皮附近的黏液层中营养物质的影响[21]。由于这种独特的功能,Akkermansia muciniphila被认为最有希望成为下一代治疗性益生菌。已经有研究表明,Akkermansia muciniphila可以刺激黏膜微生物网络[22-24],改善肠道屏障功能,抑制肠道炎症。同时该肠道菌也被证实在抗肿瘤方面有一定的疗效[25]。并且已有研究通过粪便微生物移植(Fecal Microbiota Transplantation,FMT)的方法,将富含Akkermansia muciniphila的粪便移植到早衰小鼠,发现接受治疗小鼠的平均寿命也比未接受治疗的长了近15%(前者为160 d,后者为141 d)[26]。综上所述,我们认为高剂量肉苁蓉多糖给药可以改善小鼠肠道菌群结构,使肠道益生菌Akkermansia muciniphila水平显著升高,并且这种升高效果优于双歧杆菌四联活菌片(插页ⅩⅫ图3A所示,CDPS.H组为该菌为20.90%,P组为8.612%)。高剂量的肉苁蓉多糖给药可以通过对肠道菌群的这一影响间接地延长自然衰老小鼠的寿命,产生抗衰老的作用。
本课题组前期已经研究了CDPS用于D-半乳糖诱导的小鼠衰老模型,产生了抗衰老改善认知功能的作用,在前期研究的基础上,我们将CDPS给药拓展到自然衰老小鼠模型,结合不同给药策略,观察小鼠肠道菌群的差异及肠道病理变化,探讨CDPS的抗衰老作用的可能机制。我们认为CDPS给药产生的抗衰老作用可能与其改善肠道菌群稳态,使肠道中Akkermansia muciniphila、Alloprevotella和Prevotellaceae这三种菌群的上调有关。
  • 国家自然科学基金项目(82260777)
  • 内蒙古自治区自然科学基金项目(2023QN08014)
  • 内蒙古自治区蒙医药协同创新中心科学研究项目(MYYXTZD202102)
  • 内蒙古自治区高等学校科学研究项目(NJZY22646)
  • 内蒙古自治区高等学校科学技术研究项目(NJZY22613)
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2025年第43卷第3期
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doi: 10.13193/j.issn.1673-7717.2025.03.015
  • 首发时间:2026-10-30
  • 出版时间:2025-03-10
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基金
国家自然科学基金项目(82260777)
内蒙古自治区自然科学基金项目(2023QN08014)
内蒙古自治区蒙医药协同创新中心科学研究项目(MYYXTZD202102)
内蒙古自治区高等学校科学研究项目(NJZY22646)
内蒙古自治区高等学校科学技术研究项目(NJZY22613)
作者信息
    1.内蒙古医科大学药学院,内蒙古 呼和浩特 010100
    2.内蒙古医科大学民族医药创新中心,内蒙古 呼和浩特 010100
    3.呼和浩特市第三医院,内蒙古 呼和浩特 010100
    4.内蒙古医科大学第二附属医院,内蒙古 呼和浩特 010100
    5.内蒙古科技大学包头医学院,内蒙古 包头 014000

通讯作者:

李刚(1979-),男,内蒙古呼和浩特人,教授,硕士研究生导师,博士,研究方向:神经药理学,中蒙药药理学。E-mail:。
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https://castjournals.cast.org.cn/joweb/zhzyyxk/CN/10.13193/j.issn.1673-7717.2025.03.015
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2种不同金属材料的力学参数
科
Family
属数
Number of
genus
种数
Number of
species
占总种数比例
Percentage of
total species (%)
属
Genus
种数
Number of
species
占总种数比例
Percentage of total
species (%)
鹅膏菌科Amanitaceae 2 11 5.26 鹅膏菌属 Amanita 10 4.78
小菇科 Mycenaceae 2 12 5.74 丝盖伞属 Inocybe 5 2.39
多孔菌科 Polyporaceae 8 14 6.70 蜡蘑属 Laccaria 5 2.39
红菇科 Russulaceae 3 23 11.00 小皮伞属 Marasmius 6 2.87
小菇属 Mycena 11 5.26
光柄菇属 Pluteus 5 2.39
红菇属 Russula 17 8.13
栓菌属 Trametes 5 2.39
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