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Article(id=1322912232216076363, tenantId=1146029695717560320, journalId=1255847919539208197, issueId=1322912162913596032, articleNumber=null, orderNo=null, doi=10.13193/j.issn.1673-7717.2025.01.010, 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=1793336454681, onlineDateStr=2026-10-30, pubDate=1736438400000, pubDateStr=2025-01-10, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1793336454681, onlineIssueDateStr=2026-10-30, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1793336454681, creator=13701087609, updateTime=1793336454681, updator=13701087609, issue=Issue{id=1322912162913596032, tenantId=1146029695717560320, journalId=1255847919539208197, year='2025', volume='43', issue='1', pageStart='1', pageEnd='258', issueExtLink='null', onlineDate='null', pubDate='1736438400000', pubDateStr='2025-01-10', beforeIssueId=null, nextIssueId=null, price=null, status=1, issueComplete=1, articleOrder=1, issueType=-1, specialIssue=0, createTime=1793336438157, creator='13701087609', updateTime=1793340633627, updator='13701087609', preIssue=null, nextIssue=null, articleTotal=null, ext={EN=IssueExt(id=1322929760048906989, tenantId=1146029695717560320, journalId=1255847919539208197, issueId=1322912162913596032, language=EN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=), CN=IssueExt(id=1322929760048906990, tenantId=1146029695717560320, journalId=1255847919539208197, issueId=1322912162913596032, language=CN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=)}, issueFiles=null, downloadFileDto=null}, startPage=55, endPage=59, ext={EN=ArticleExt(id=1322912232497094733, articleId=1322912232216076363, tenantId=1146029695717560320, journalId=1255847919539208197, language=EN, title=Mechanism of Wendan Decoction（温胆汤）-Containing Serum Regulating Neuronal Synaptic Plasticity to Improve Cognitive Impairment in Schizophrenia, columnId=1256183361521967297, journalTitle=Chinese Archives of Traditional Chinese Medicine, columnName=Target on National Project, runingTitle=null, highlight=null, articleAbstract=
Objective

To explore the molecular mechanism ofWendan Decoction（温胆汤）improving cognitive impairment in schizophrenia from the perspective of neuronal synaptic plasticity by using glutamate-induced HT22 cells as a cellularmodel of schizophrenia and detecting synaptic plasticity related proteins，postsynaptic density protein-95（PSD-95），growth associated protein-43（GAP-43），c-Jun NH2-terminal kinase-interacting protein-3（JIP-3）and connexin-43（Cx-43）.

Method

The SD ratswere randomly divided into four groups：Wendan Decoction 40 g/kg，30 g/kg and 20 g/kg groups and clozapine 0.02 g/kg group. Each drug group was given the corresponding dose of drug by gavage，once a day. After seven days，the ratswere be killed to collect blood and prepare the drug-containing serum. CCK8 assay was used to determine the concentration and time point of glutamate action on astrocytes. HT22 cells at 10 mmol/L glutamate for 24 h were used as a cellmodel to detect the effect of Wendan Decoction-containing serum on the apoptosis rate of HT22 cells by flow cytometry. RT-qPCR was used to detect the expressions of Psd95，Gap43，Jip3 and Cx43 mRNA in HT22 cells. The expressions of PSD-95，GAP-43，JIP-3 and Cx-43 proteins in HT22 cellswere detected by Western blot.

Results

According to the results of the effectof different concentrations of glutamate on the proliferation rate of HT22 cells，10 mmol/L glutamate HT22 cells for 24 h were determined as the cell experimental conditions. Compared with thatof the normal control group，the apoptosis rate of HT22 cells in the glutamate model group increased significantly. The mRNA and protein expressions of PSD-95，GAP-43，JIP-3 and Cx-43 were also significantly down-regulated（P<0.01）. Compared with that of the glutamate model group，the apoptosis rate of HT22 cells in Wendan Decoction-containing serum groups were significantly decreased and the expressions of PSD-95，GAP-43，JIP-3，Cx-43 mRNA and protein significantly up-regulated（P<0.05 or P<0.01）.

Conclusion

Wendan Decoction may be to reduce apoptosis in neurons and up-regulate the expressions of PSD-95，GAP-43，JIP-3 and Cx-43，and then regulate synaptic plasticity，in order to effectively improve cognitive deficits in schizophrenia.

, authors=Jinhua ZHU, Wei YANG, Linlin LI, Jing CHEN, Milan ZHANG, Bingyi LIU, Yongcuo REN QING, Jun ZHOU, Hongjiao WAN, authorsList=Jinhua ZHU, Wei YANG, Linlin LI, Jing CHEN, Milan ZHANG, Bingyi LIU, Yongcuo REN QING, Jun ZHOU, Hongjiao WAN, authorCompany=null, correspAuthors=Hongjiao WAN, 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=1322912235613462620, articleId=1322912232216076363, tenantId=1146029695717560320, journalId=1255847919539208197, language=CN, title=温胆汤含药血清调控神经元突触可塑性改善精神分裂症认知障碍的机制研究, columnId=1256183362172142382, journalTitle=中华中医药学刊, columnName=国家项目点击, runingTitle=null, highlight=null, articleAbstract=
目的

以谷氨酸作用的HT22细胞为精神分裂症细胞模型，通过检测突触可塑性相关蛋白突触后致密蛋白95（postsynaptic density protein-95，PSD-95）、生长相关蛋白43（growth associated protein-43，GAP-43）、JNK相互作用蛋白3（c-Jun NH2-terminal kinase-interacting protein-3，JIP-3）、连接蛋白43（connexin-43，Cx-43）的变化，并从神经元突触可塑性角度探讨温胆汤改善精神分裂症认知障碍的分子机制。

方法

将大鼠随机分成4组：温胆汤40、30、20 g/kg组和氯氮平0.02 g/kg组。各用药组灌胃相应的分组剂量药物，1次/d，7 d后取血，制备含药血清。采用CCK8实验确定谷氨酸对HT22细胞的作用浓度和时间点；以10 mmol/L谷氨酸作用24 h的HT22细胞为细胞模型，流式细胞术检测温胆汤含药血清对HT22细胞凋亡率的影响；RT-qPCR检测HT22细胞中Psd95、Gap43、Jip3和Cx43 mRNA表达；WB检测HT22细胞中PSD-95、GAP-43、JIP-3、Cx-43蛋白表达。

结果

根据不同浓度谷氨酸对HT22细胞增殖率影响的结果，确定10 mmol/L谷氨酸作用HT22细胞24 h为细胞实验条件。与正常对照组比较，谷氨酸模型组HT22细胞凋亡率显著上升；其PSD-95、GAP-43、JIP-3、Cx-43 mRNA和蛋白表达也明显下调（P<0.01）。与谷氨酸模型组比较，谷氨酸+温胆汤40、30、20 g/kg含药血清组HT22细胞凋亡率明显下降；并显著上调PSD-95、GAP-43、JIP-3、Cx-43 mRNA和蛋白的表达（P<0.05或P<0.01）。

结论

温胆汤可能是通过降低神经元细胞凋亡，上调其PSD-95、GAP-43、JIP-3、Cx-43表达水平，进而调控突触可塑性，以期达到有效改善精神分裂症认知缺陷的目的。

, authors=朱金华, 杨威, 李琳琳, 陈静, 张米兰, 刘冰怡, 仁青永措, 周军, 万红娇, authorsList=朱金华, 杨威, 李琳琳, 陈静, 张米兰, 刘冰怡, 仁青永措, 周军, 万红娇, authorCompany=null, correspAuthors=万红娇, authorNote=

朱金华（1977-），男，江西临川人，正高级实验师，硕士研究生导师，博士在读，研究方向：中医药与精神分裂症。

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万红娇（1964-），女，江西南昌人，教授，博士研究生导师，博士，研究方向：中医药与神经保护。E-mail：。
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朱金华（1977-），男，江西临川人，正高级实验师，硕士研究生导师，博士在读，研究方向：中医药与精神分裂症。

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朱金华（1977-），男，江西临川人，正高级实验师，硕士研究生导师，博士在读，研究方向：中医药与精神分裂症。

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articleTitle=Morphogenesis of the telencephalic commissure requires scaffold protein JNKinteracting protein 3（JIP3）, refAbstract=null), Reference(id=1322912583249953266, tenantId=1146029695717560320, journalId=1255847919539208197, articleId=1322912232216076363, doi=null, pmid=null, pmcid=null, year=2017, volume=23, issue=2, pageStart=208, pageEnd=211, url=null, language=null, rfNumber=[35], rfOrder=34, authorNames=王思茹, 石玉秀, journalName=解剖科学进展, refType=null, unstructuredReference=王思茹，石玉秀．突触相关蛋白与精神障碍性疾病［J］．解剖科学进展，2017，23（2）：208-211., articleTitle=突触相关蛋白与精神障碍性疾病, refAbstract=null), Reference(id=1322912583312867827, tenantId=1146029695717560320, journalId=1255847919539208197, articleId=1322912232216076363, doi=null, pmid=null, pmcid=null, year=2020, volume=11, issue=null, pageStart=888, pageEnd=null, url=null, language=null, rfNumber=[36], rfOrder=35, authorNames=YANG K L, ZHOU Y, ZHOU L Q, journalName=Front Neurol, refType=null, unstructuredReference=YANG K L，ZHOU Y，ZHOU L Q，et 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remark=江西中医药大学，江西　南昌　330004)])], figs=[ArticleFig(id=1322912573263315388, tenantId=1146029695717560320, journalId=1255847919539208197, articleId=1322912232216076363, language=EN, label=null, caption=null, figureFileSmall=UivCGKrsi/03ZzI/ePQAxQ==, figureFileBig=jDAp9s3C5uMfXy1mXZTEnA==, tableContent=null), ArticleFig(id=1322912573347201469, tenantId=1146029695717560320, journalId=1255847919539208197, articleId=1322912232216076363, language=CN, label=图1, caption=温胆汤含药血清对谷氨酸作用下HT22细胞凋亡的影响, figureFileSmall=UivCGKrsi/03ZzI/ePQAxQ==, figureFileBig=jDAp9s3C5uMfXy1mXZTEnA==, tableContent=null), ArticleFig(id=1322912573447864766, tenantId=1146029695717560320, journalId=1255847919539208197, articleId=1322912232216076363, language=EN, label=null, caption=null, figureFileSmall=HodC2+Kacn6kEaRirohK2w==, figureFileBig=pQ/2k8EqOoV4PU85RYKZEg==, tableContent=null), ArticleFig(id=1322912573510779327, tenantId=1146029695717560320, journalId=1255847919539208197, articleId=1322912232216076363, language=CN, label=图2, caption=温胆汤含药血清对谷氨酸作用下HT22细胞PSD-95、GAP-43、JIP-3、Cx-43蛋白表达的影响

注：A.正常对照组；B.谷氨酸模型组；C.氯氮平0.02 g/kg含药血清组；D.温胆汤40 g/kg含药血清组；E.温胆汤30 g/kg含药血清组；F.温胆汤20 g/kg含药血清组。

, figureFileSmall=HodC2+Kacn6kEaRirohK2w==, figureFileBig=pQ/2k8EqOoV4PU85RYKZEg==, tableContent=null), ArticleFig(id=1322912573603054016, tenantId=1146029695717560320, journalId=1255847919539208197, articleId=1322912232216076363, language=EN, label=null, caption=null, figureFileSmall=null, figureFileBig=null, tableContent=
引物名称引物序列引物长度/（nt）产物长度/（bp）退火温度/（℃）
Psd95 FTCCGGGAGGTGACCCATTC198360.9
Psd95 RTTTCCGGCGCATGACGTAG19
Gap43 FAAGGCAGGGGAAGATACCAC208759.1
Gap43 RTTGTTCAATCTTTTGGTCCTCAT23
Jip3 FGCGTCCCACCTCTCTGAATG2012960.5
Jip3 RCTGGAGCTGGACTGTAGCTG20
Cx43 FCTGAGTGCGGTCTACACCTG2012760.1
Cx43 RGAGCGAGAGACACCAAGGAC20
β actin FAGCCATGTACGTAGCCATCC2022759.6
β actin RTCTCAGCTGTGGTGGTGAAG20
), ArticleFig(id=1322912573670162881, tenantId=1146029695717560320, journalId=1255847919539208197, articleId=1322912232216076363, language=CN, label=表1, caption=

PCR引物序列

, figureFileSmall=null, figureFileBig=null, tableContent=
引物名称引物序列引物长度/（nt）产物长度/（bp）退火温度/（℃）
Psd95 FTCCGGGAGGTGACCCATTC198360.9
Psd95 RTTTCCGGCGCATGACGTAG19
Gap43 FAAGGCAGGGGAAGATACCAC208759.1
Gap43 RTTGTTCAATCTTTTGGTCCTCAT23
Jip3 FGCGTCCCACCTCTCTGAATG2012960.5
Jip3 RCTGGAGCTGGACTGTAGCTG20
Cx43 FCTGAGTGCGGTCTACACCTG2012760.1
Cx43 RGAGCGAGAGACACCAAGGAC20
β actin FAGCCATGTACGTAGCCATCC2022759.6
β actin RTCTCAGCTGTGGTGGTGAAG20
), ArticleFig(id=1322912573745660354, tenantId=1146029695717560320, journalId=1255847919539208197, articleId=1322912232216076363, language=EN, label=null, caption=null, figureFileSmall=null, figureFileBig=null, tableContent=
浓度/（mmol/L）n24 h48 h
05100.0±2.2100.0±4.1
1599.2±4.779.4±2.2**
2595.0±4.4*72.2±4.3**
5583.5±2.8**59.8±5.5**
10569.5±2.5**54.6±3.4**
20555.1±2.9**46.1±3.0**
), ArticleFig(id=1322912573816963523, tenantId=1146029695717560320, journalId=1255847919539208197, articleId=1322912232216076363, language=CN, label=表2, caption=

不同浓度谷氨酸对HT22细胞增殖率的影响（±s）

, figureFileSmall=null, figureFileBig=null, tableContent=
浓度/（mmol/L）n24 h48 h
05100.0±2.2100.0±4.1
1599.2±4.779.4±2.2**
2595.0±4.4*72.2±4.3**
5583.5±2.8**59.8±5.5**
10569.5±2.5**54.6±3.4**
20555.1±2.9**46.1±3.0**
), ArticleFig(id=1322912573888266692, tenantId=1146029695717560320, journalId=1255847919539208197, articleId=1322912232216076363, language=EN, label=null, caption=null, figureFileSmall=null, figureFileBig=null, tableContent=
组别n剂量/（g/kg）细胞凋亡率/（%）
正常对照组34.01±0.16**
谷氨酸模型组313.74±3.48
氯氮平0.02 g/kg含药血清组30.020.92±0.06**
温胆汤40 g/kg含药血清组3400.94±0.31**
温胆汤30 g/kg含药血清组3300.88±0.08**
温胆汤20 g/kg含药血清组3200.77±0.17**
), ArticleFig(id=1322912573972152773, tenantId=1146029695717560320, journalId=1255847919539208197, articleId=1322912232216076363, language=CN, label=表3, caption=

温胆汤含药血清对谷氨酸作用下HT22细胞凋亡的影响（±s）

, figureFileSmall=null, figureFileBig=null, tableContent=
组别n剂量/（g/kg）细胞凋亡率/（%）
正常对照组34.01±0.16**
谷氨酸模型组313.74±3.48
氯氮平0.02 g/kg含药血清组30.020.92±0.06**
温胆汤40 g/kg含药血清组3400.94±0.31**
温胆汤30 g/kg含药血清组3300.88±0.08**
温胆汤20 g/kg含药血清组3200.77±0.17**
), ArticleFig(id=1322912574077010374, tenantId=1146029695717560320, journalId=1255847919539208197, articleId=1322912232216076363, language=EN, label=null, caption=null, figureFileSmall=null, figureFileBig=null, tableContent=
组别n剂量/（g/kg）Psd95/2-ΔΔCtGap43/2-ΔΔCtJip3/2-ΔΔCtCx43/2-ΔΔCt
正常对照组31.000±0.000**1.000±0.000**1.000±0.000**1.000±0.000**
谷氨酸模型组30.443±0.0850.473±0.0250.420±0.0400.537±0.076
氯氮平0.02 g/kg含药血清组30.020.570±0.066*0.730±0.040**0.767±0.025**0.683±0.085*
温胆汤40 g/kg含药血清组3400.633±0.091**0.887±0.012**0.813±0.045**0.750±0.050**
温胆汤30 g/kg含药血清组3300.667±0.057**0.623±0.050**0.780±0.082**0.763±0.076**
温胆汤20 g/kg含药血清组3200.593±0.080*0.570±0.010**0.537±0.087*0.660±0.080*
), ArticleFig(id=1322912574152507847, tenantId=1146029695717560320, journalId=1255847919539208197, articleId=1322912232216076363, language=CN, label=表4, caption=

温胆汤含药血清对谷氨酸作用下海马神经元细胞Psd95、Gap43、Jip3、Cx43 mRNA表达的影响（±s）

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组别n剂量/（g/kg）Psd95/2-ΔΔCtGap43/2-ΔΔCtJip3/2-ΔΔCtCx43/2-ΔΔCt
正常对照组31.000±0.000**1.000±0.000**1.000±0.000**1.000±0.000**
谷氨酸模型组30.443±0.0850.473±0.0250.420±0.0400.537±0.076
氯氮平0.02 g/kg含药血清组30.020.570±0.066*0.730±0.040**0.767±0.025**0.683±0.085*
温胆汤40 g/kg含药血清组3400.633±0.091**0.887±0.012**0.813±0.045**0.750±0.050**
温胆汤30 g/kg含药血清组3300.667±0.057**0.623±0.050**0.780±0.082**0.763±0.076**
温胆汤20 g/kg含药血清组3200.593±0.080*0.570±0.010**0.537±0.087*0.660±0.080*
), ArticleFig(id=1322912574228005320, tenantId=1146029695717560320, journalId=1255847919539208197, articleId=1322912232216076363, language=EN, label=null, caption=null, figureFileSmall=null, figureFileBig=null, tableContent=
组别n剂量/（g/kg）PSD-95/GAPDHGAP-43/GAPDHJIP-3/GAPDHCx-43/GAPDH
正常对照组31.31±0.17**0.98±0.15**1.46±0.26**1.13±0.22**
谷氨酸模型组30.64±0.060.37±0.040.47±0.130.43±0.09
氯氮平0.02 g/kg含药血清组30.021.26±0.15**0.71±0.08**0.98±0.35**0.97±0.20**
温胆汤40 g/kg含药血清组3401.06±0.05**0.75±0.05**1.05±0.31**0.81±0.07*
温胆汤30 g/kg含药血清组3300.87±0.06*0.65±0.07**1.21±0.11**0.90±0.30*
温胆汤20 g/kg含药血清组3200.95±0.07**0.72±0.21**1.42±0.06**0.97±0.27**
), ArticleFig(id=1322912574328668617, tenantId=1146029695717560320, journalId=1255847919539208197, articleId=1322912232216076363, language=CN, label=表5, caption=

温胆汤含药血清对谷氨酸作用下HT22细胞PSD-95、GAP-43、JIP-3、Cx-43蛋白表达的影响（±s）

, figureFileSmall=null, figureFileBig=null, tableContent=
组别n剂量/（g/kg）PSD-95/GAPDHGAP-43/GAPDHJIP-3/GAPDHCx-43/GAPDH
正常对照组31.31±0.17**0.98±0.15**1.46±0.26**1.13±0.22**
谷氨酸模型组30.64±0.060.37±0.040.47±0.130.43±0.09
氯氮平0.02 g/kg含药血清组30.021.26±0.15**0.71±0.08**0.98±0.35**0.97±0.20**
温胆汤40 g/kg含药血清组3401.06±0.05**0.75±0.05**1.05±0.31**0.81±0.07*
温胆汤30 g/kg含药血清组3300.87±0.06*0.65±0.07**1.21±0.11**0.90±0.30*
温胆汤20 g/kg含药血清组3200.95±0.07**0.72±0.21**1.42±0.06**0.97±0.27**
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温胆汤含药血清调控神经元突触可塑性改善精神分裂症认知障碍的机制研究
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朱金华 , 杨威 , 李琳琳 , 陈静 , 张米兰 , 刘冰怡 , 仁青永措 , 周军 , 万红娇
中华中医药学刊 | 国家项目点击 2025,43(1): 55-59
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中华中医药学刊 |国家项目点击 2025 , 43 (1) : 55 -59
温胆汤含药血清调控神经元突触可塑性改善精神分裂症认知障碍的机制研究
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朱金华（1977-），男，江西临川人，正高级实验师，硕士研究生导师，博士在读，研究方向：中医药与精神分裂症。

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朱金华（1977-），男，江西临川人，正高级实验师，硕士研究生导师，博士在读，研究方向：中医药与精神分裂症。

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朱金华, 杨威, 李琳琳, 陈静, 张米兰, 刘冰怡, 仁青永措, 周军, 万红娇
作者信息
  • 江西中医药大学，江西　南昌　330004
通讯作者:
万红娇（1964-），女，江西南昌人，教授，博士研究生导师，博士，研究方向：中医药与神经保护。E-mail：。
作者简介:

朱金华（1977-），男，江西临川人，正高级实验师，硕士研究生导师，博士在读，研究方向：中医药与精神分裂症。

Mechanism of Wendan Decoction（温胆汤）-Containing Serum Regulating Neuronal Synaptic Plasticity to Improve Cognitive Impairment in Schizophrenia
Jinhua ZHU, Wei YANG, Linlin LI, Jing CHEN, Milan ZHANG, Bingyi LIU, Yongcuo REN QING, Jun ZHOU, Hongjiao WAN
Affiliations
  • Jiangxi University of Chinese Medicine, Nanchang 330004, Jiangxi, China
出版时间: 2025-01-10 doi: 10.13193/j.issn.1673-7717.2025.01.010
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摘要
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目的

以谷氨酸作用的HT22细胞为精神分裂症细胞模型，通过检测突触可塑性相关蛋白突触后致密蛋白95（postsynaptic density protein-95，PSD-95）、生长相关蛋白43（growth associated protein-43，GAP-43）、JNK相互作用蛋白3（c-Jun NH2-terminal kinase-interacting protein-3，JIP-3）、连接蛋白43（connexin-43，Cx-43）的变化，并从神经元突触可塑性角度探讨温胆汤改善精神分裂症认知障碍的分子机制。

方法

将大鼠随机分成4组：温胆汤40、30、20 g/kg组和氯氮平0.02 g/kg组。各用药组灌胃相应的分组剂量药物，1次/d，7 d后取血，制备含药血清。采用CCK8实验确定谷氨酸对HT22细胞的作用浓度和时间点；以10 mmol/L谷氨酸作用24 h的HT22细胞为细胞模型，流式细胞术检测温胆汤含药血清对HT22细胞凋亡率的影响；RT-qPCR检测HT22细胞中Psd95、Gap43、Jip3和Cx43 mRNA表达；WB检测HT22细胞中PSD-95、GAP-43、JIP-3、Cx-43蛋白表达。

结果

根据不同浓度谷氨酸对HT22细胞增殖率影响的结果，确定10 mmol/L谷氨酸作用HT22细胞24 h为细胞实验条件。与正常对照组比较，谷氨酸模型组HT22细胞凋亡率显著上升；其PSD-95、GAP-43、JIP-3、Cx-43 mRNA和蛋白表达也明显下调（P<0.01）。与谷氨酸模型组比较，谷氨酸+温胆汤40、30、20 g/kg含药血清组HT22细胞凋亡率明显下降；并显著上调PSD-95、GAP-43、JIP-3、Cx-43 mRNA和蛋白的表达（P<0.05或P<0.01）。

结论

温胆汤可能是通过降低神经元细胞凋亡，上调其PSD-95、GAP-43、JIP-3、Cx-43表达水平，进而调控突触可塑性，以期达到有效改善精神分裂症认知缺陷的目的。

温胆汤含药血清  /  精神分裂症  /  HT22细胞  /  突触可塑性
Objective

To explore the molecular mechanism ofWendan Decoction（温胆汤）improving cognitive impairment in schizophrenia from the perspective of neuronal synaptic plasticity by using glutamate-induced HT22 cells as a cellularmodel of schizophrenia and detecting synaptic plasticity related proteins，postsynaptic density protein-95（PSD-95），growth associated protein-43（GAP-43），c-Jun NH2-terminal kinase-interacting protein-3（JIP-3）and connexin-43（Cx-43）.

Method

The SD ratswere randomly divided into four groups：Wendan Decoction 40 g/kg，30 g/kg and 20 g/kg groups and clozapine 0.02 g/kg group. Each drug group was given the corresponding dose of drug by gavage，once a day. After seven days，the ratswere be killed to collect blood and prepare the drug-containing serum. CCK8 assay was used to determine the concentration and time point of glutamate action on astrocytes. HT22 cells at 10 mmol/L glutamate for 24 h were used as a cellmodel to detect the effect of Wendan Decoction-containing serum on the apoptosis rate of HT22 cells by flow cytometry. RT-qPCR was used to detect the expressions of Psd95，Gap43，Jip3 and Cx43 mRNA in HT22 cells. The expressions of PSD-95，GAP-43，JIP-3 and Cx-43 proteins in HT22 cellswere detected by Western blot.

Results

According to the results of the effectof different concentrations of glutamate on the proliferation rate of HT22 cells，10 mmol/L glutamate HT22 cells for 24 h were determined as the cell experimental conditions. Compared with thatof the normal control group，the apoptosis rate of HT22 cells in the glutamate model group increased significantly. The mRNA and protein expressions of PSD-95，GAP-43，JIP-3 and Cx-43 were also significantly down-regulated（P<0.01）. Compared with that of the glutamate model group，the apoptosis rate of HT22 cells in Wendan Decoction-containing serum groups were significantly decreased and the expressions of PSD-95，GAP-43，JIP-3，Cx-43 mRNA and protein significantly up-regulated（P<0.05 or P<0.01）.

Conclusion

Wendan Decoction may be to reduce apoptosis in neurons and up-regulate the expressions of PSD-95，GAP-43，JIP-3 and Cx-43，and then regulate synaptic plasticity，in order to effectively improve cognitive deficits in schizophrenia.

Wendan Decoction（温胆汤）-containing serum  /  schizophrenia  /  HT22 cells  /  synaptic plasticity
朱金华, 杨威, 李琳琳, 陈静, 张米兰, 刘冰怡, 仁青永措, 周军, 万红娇. 温胆汤含药血清调控神经元突触可塑性改善精神分裂症认知障碍的机制研究. 中华中医药学刊, 2025 , 43 (1) : 55 -59 . DOI: 10.13193/j.issn.1673-7717.2025.01.010
Jinhua ZHU, Wei YANG, Linlin LI, Jing CHEN, Milan ZHANG, Bingyi LIU, Yongcuo REN QING, Jun ZHOU, Hongjiao WAN. Mechanism of Wendan Decoction（温胆汤）-Containing Serum Regulating Neuronal Synaptic Plasticity to Improve Cognitive Impairment in Schizophrenia[J]. Chinese Archives of Traditional Chinese Medicine, 2025 , 43 (1) : 55 -59 . DOI: 10.13193/j.issn.1673-7717.2025.01.010
正文
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精神分裂症是最严重的精神疾病之一，其症状主要可以分为阴性症状、阳性症状和认知功能障碍三大类，世界上多达1%的人口受其影响，也是主要的致残因素之一[1-3]。该病的病因至今尚未明确，复发率和病死率高，在病因学和治疗上仍是一个巨大的挑战。认知功能障碍作为精神分裂症的核心症状之一，具有发生于精神分裂症其他症状之前、持续时间长、存在于绝大多数精神分裂症患者中的特征，并且认知已被确定为改善精神病患者功能结果的一个重要治疗目标[4]。大量文献表明，精神分裂症认知功能障碍与突触可塑性之间存在着紧密的联系[5-7]。突触可塑性反映了神经元突触连接强度的可调节能力，是学习和记忆的重要神经机制[8]。突触相关蛋白是突触可塑性的重要分子基础，其调控突触结构和功能的稳定，影响神经的生长与修复，维持神经系统正常的生理功能，与精神障碍疾病的关系十分密切[9]。
中医神志病中的“癫证”“狂证”与精神分裂症相类似。从中医角度分析癫狂的病因病机是由于多种因素引起机体阴阳与脏腑功能失调，出现气、火、痰、瘀等病理产物，最终引起神机逆乱。其中“痰”是至关重要的致病因素，痰浊内生或与其他病理产物相互作用，蒙蔽心神与清窍，出现各种精神错乱的症状 [10-11]。历代医家向来重视痰邪对癫狂的影响，治疗以治痰为主。温胆汤是经典的治痰名方，其功效清胆和胃，理气化痰，临床上通过对温胆汤进行化裁或联合其他抗精神病药物治疗精神分裂症，且取得了明显的治疗效果[12-14]。本课题组前期研究发现，温胆汤能够明显缓解精神分裂症模型大鼠刻板行为、增强学习记忆能力以及上调海马组织中突触相关蛋白突触可塑性相关蛋白突触后致密蛋白95（postsynaptic density protein-95，PSD-95）、生长相关蛋白43（growth associated protein-43，GAP-43）、JNK相互作用蛋白3（c-Jun NH2-terminal kinase-interacting protein-3，JIP-3）、连接蛋白43（connexin-43，Cx-43）的表达等，但具体的分子机制还需进一步研究[15]。因此，本研究在此基础上以谷氨酸作用于海马神经元细胞，通过流式细胞法、蛋白免疫印迹法等，在细胞水平上观察温胆汤含药血清对细胞凋亡的影响，检测突触相关蛋白PSD-95、GAP-43、JIP-3、Cx-43的表达，进一步探讨温胆汤是否通过调控突触可塑性来改善精神分裂症的认知功能障碍，并为临床运用温胆汤防治精神分裂症提供更多的实验依据。
1　材料和方法
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1.1　细胞株
HT22细胞（CL-0595，Procell）。
1.2　动物
SPF级SD大鼠40只，雄性，体质量（250±20）g，动物许可证号：SCXK（湘）2019-0004，由湖南斯莱克景达实验动物有限公司提供。本研究动物实验已获江西中医药大学实验动物伦理委员会审查（No. JZLLSC20230638）。大鼠饲养于屏障环境中，温度25℃，湿度50%，12 h昼夜节律，饮水和饲料充足，动物实验设施许可证编号：SYXK（赣）2022-0002。
1.3　试验药物
温胆汤原方：半夏6 g，茯苓4.5 g，竹茹6 g，枳实6 g，陈皮9 g，炙甘草3 g，生姜3片，红枣2粒，每剂含生药38 g，购买于江西中医药大学附属医院中药房。全方药材由江西中医药大学中药资源与民族药研究中心曹岚副教授鉴定合格。全方药物加水浸泡，一煎、二煎后，过滤取汁，使用旋转蒸发仪浓缩药物，制成浓度为2 g生药/mL的温胆汤汤剂，于4℃冰箱保存备用。
1.4　试剂
谷氨酸（批号：G8415，Merck公司），DMEM高糖培养基、CCK-8法细胞增殖检测试剂盒（批号：KGM12800S、KGA317，江苏凯基生物技术股份有限公司），FBS（批号：E600001-0500，BBI Life Sciences），Annexin V-FITC/PI Apoptosis Kit（AP101，联科生物），RNAiso Plus、PrimeScriptTM RT reagent Kit with gDNA Eraser（Perfect Real Time）、TB Green® Premix Ex TaqTMⅡ（Tli RNaseH Plus）（批号：9109、RR047A、RR820A，Takara公司），PSD-95、GAP-43、Cx-43、GAPDH一抗（批号：＃3409、＃8945、＃3512、＃3409，CST公司），JIP-3一抗（批号：sc-46663，SANTA CRUZ公司）。
1.5　仪器
BPN-80CW CO2培养箱（上海一恒科学仪器有限公司），MF53倒置荧光显微镜（广州市明美光电有限公司），2060R NovoCyteTM流式细胞仪［艾森生物（杭州）有限公司］，Power-PacTW 通用Bio-Rad型电泳仪、小型垂直电泳槽（Bio-Rad公司），NP80紫外分光光度仪（Nano Photometer公司），ND2000核酸蛋白测定仪（Thermo公司），CFX ConnectTM实时荧光PCR仪、Chemi DocTM XRS+超高灵敏度化学发光成像系统［伯乐生命医学产品（上海）有限公司］。
1.6　方法
1.6.1　含药血清制备
将40只SPF级SD大鼠适应性喂养5 d后，每组10只随机分为4组：温胆汤40 g/kg组、温胆汤30 g/kg组、温胆汤20 g/kg组、氯氮平0.02 g/kg组。氯氮平0.02 g/kg组灌胃氯氮平原药0.02 g/kg，温胆汤40、30、20 g/kg组分别灌胃温胆汤生药40、30、20 g/kg，1次/d。7 d后末次给药2 h麻醉大鼠，腹主动脉采血，离心取血清，灭活，过0.22 μm滤膜除菌，-80℃备用。
1.6.2　细胞复苏与传代
冻存细胞从液氮中取出，放37℃水浴快速融化，移至装有10 mL培养基的离心管中，1000 r/min离心5min，弃上清，加1 mL新鲜培养基重悬细胞，转移至装有DMEM完全培养基的培养瓶或培养皿中，37℃、5% CO2 培养。当细胞培养到80%～90%密度时需传代，先吸去原有培养基，用1×PBS洗2遍，加入适量0.25%胰酶消化，待细胞变圆后加入培养基终止消化，并收集细胞悬液于离心管，1000 r/min离心5 min，弃尽上清，加入培养基吹匀，将细胞悬液按照1∶2的比例加入准备好的培养瓶或培养皿中，置于细胞培养箱中培养。
1.6.3　CCK8实验检测不同浓度谷氨酸对HT22细胞增殖率的影响
将HT22细胞100 μL接种于96孔板中，细胞数为1×104个细胞/孔，37℃、5% CO2培养24 h。细胞状态良好，分组加入10 μL不同浓度的谷氨酸（终浓度为：0、1、2、5、10、20 mmol/L）作用24 h和48 h，每组设6复孔，每孔再加入10 μLCCK8试剂，培养箱中孵育1～4 h后，放振动器上轻轻混匀1 min，使用酶标仪在450 nm处检测每孔的吸光值。实验重复5次。数据分析：细胞存活率（%）＝［（As-Ab）/（Ac-Ab）］×100、抑制率（%）＝［（Ac-As）/（Ac-Ab）］×100；As＝实验孔吸光值（含有细胞，培养基，CCK8试剂和谷氨酸的孔的吸光值）、Ab＝空白孔吸光值（含有培养基和CCK8试剂的孔的吸光值）、Ac＝实验孔吸光值（含有细胞、培养基和CCK8试剂的孔的吸光值）。
1.6.4　细胞分组及药物干预
实验分组为6组，分设正常对照组、谷氨酸模型组、氯氮平0.02 g/kg含药血清组、温胆汤40 g/kg含药血清组、温胆汤30 g/kg含药血清组、温胆汤20 g/kg含药血清组。将对数期生长的HT22细胞均匀地接种于培养皿中，每皿细胞1×106个，放置培养箱培养，待细胞完全贴壁后进行加药处理，除正常对照组外，按照实验分组加入10 mmol/L的谷氨酸和对应的含药血清（各给药组加入相应大鼠含药血清，含药血清量为总培养体系的10%）处理24 h后，收集细胞做后续实验。
1.6.5　流式细胞术检测HT22细胞凋亡率
实验分组和细胞铺板、培养同1.6.4项，实验操作严格按照Annexin V-FITC/PI Apoptosis Kit说明书要求。收集1×106～3×106个细胞，加PBS 1500 r/min离心3 min，洗两次。取预冷的1×Binding Buffer重悬细胞。各管加入5 μL Annexin V-FITC和10 μL PI，轻微混匀后，室温避光孵育10 min，再向每管中加入预冷的1×Binding Buffer，混匀后上流式仪检测。
1.6.6　RT-qPCR检测HT22细胞Psd95、Gap43、Jip3、Cx43 mRNA的表达水平
实验分组和细胞铺板、培养同1.6.4项。收集各组细胞抽提RNA，提取后使用微量紫外分光光度仪测定RNA浓度及纯度，通过琼脂糖凝胶电泳来确定RNA质量情况。使用PrimeScriptTM RT reagent Kit with gDNA Eraser（Perfect Real Time）反转录试剂盒合成cDNA，以cDNA为模板，在荧光定量PCR仪上设定程序进行扩增检测。两步法PCR扩增标准程序：预变性95℃ 5 min循环数1，变性95℃ 10 s、退火＼延伸60℃ 30 s循环数40。根据相对定量的2-△△Ct法，以β actin为内参，计算出各组Psd95、Gap43、Jip3和Cx43的相对表达量。所有引物由通用生物系统（安徽）有限公司合成。详见表1。
1.6.7　WB检测HT22细胞PSD-95、GAP-43、JIP-3、Cx-43蛋白的表达水平
实验分组和细胞铺板、培养同1.6.4项。弃掉培养皿中液体，加入细胞裂解液，于冰上裂解30 min，在12 000 r/min离心10 min，取上清液，BCA测定蛋白浓度。蛋白变性、制胶、上样、电泳、转膜，脱脂牛奶封闭1 h，一抗（1∶1000）4℃过夜，洗膜5次，二抗（1∶2000）室温2 h，洗膜5次，加ECL曝光液曝光拍照。用“Quantity one”软件分析各抗体条带灰度值。
1.6.8　统计学分析
采用SPSS 19.0软件对实验数据进行统计分析，以均数±标准差（±s）来表示，多组间比较采用单因素方差分析（One-way ANOVA），组间两两比较用LSD检验；以P<0.05表示差异有统计学意义。
2　结果
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2.1　不同浓度谷氨酸对HT22细胞增殖率的影响
利用CCK8法在24 h、48 h时间点下检测HT22细胞增殖情况。如表2所示，2个时间点细胞增殖能力随着谷氨酸浓度的增加而减少，综合各时间点细胞的状态情况，综合选择10 mmol/mL谷氨酸浓度作用24 h进行后续实验。
2.2　温胆汤含药血清对谷氨酸作用下HT22细胞凋亡的影响
经流式细胞仪检测，与正常对照组比较，谷氨酸模型组HT22细胞凋亡率显著上升（P<0.01）；与谷氨酸模型组比较，温胆汤40、30、20 g/kg含药血清组与氯氮平0.02 g/kg含药血清组HT22细胞凋亡率明显下降（P<0.01）；与正常对照组比较，温胆汤40、30、20 g/kg含药血清组与氯氮平0.02 g/kg含药血清组HT22细胞凋亡率也明显下降（P<0.05）。见表3、插页Ⅹ图1。
2.3　温胆汤含药血清对谷氨酸作用下HT22细胞Psd95、Gap43、Jip3、Cx43 mRNA表达的影响
经RT-qPCR检测，与正常对照组比较，谷氨酸模型组HT22细胞Psd95、Gap43、Jip3、Cx43 mRNA的表达显著下调（P<0.01）；与谷氨酸模型组比较，温胆汤40、30、20 g/kg含药血清组与氯氮平0.02 g/kg含药血清组均能明显上调HT22细胞Psd95、Gap43、Jip3、Cx43 mRNA的表达（P<0.05或P<0.01）。见表4。
2.4　温胆汤含药血清对谷氨酸作用下HT22细胞PSD-95、GAP-43、JIP-3、Cx-43蛋白表达的影响
与正常对照组比较，谷氨酸模型组HT22细胞PSD-95、GAP-43、JIP-3、Cx-43蛋白表达显著下调（P<0.01）；与谷氨酸模型组比较，温胆汤40、30、20 g/kg含药血清组与氯氮平0.02 g/kg含药血清组HT22细胞PSD-95、GAP-43、JIP-3、Cx-43蛋白的表达均明显上调（P<0.05或P<0.01）。见表5、图2。
3　讨论
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精神分裂症是一类严重危害生命健康和造成沉重社会负担的精神障碍疾病，患者会出现思维、情感、记忆、行为等方面的异常，严重者甚至会自残或者自杀。谷氨酸假说是目前研究精神分裂症的热门领域，诸多研究也证实了谷氨酸功能的失调在精神分裂症病理生理中的作用[16-18]。谷氨酸是广泛分布于中枢神经系统的一种兴奋性神经递质，当谷氨酸释放过量会引起谷氨酸受体过度活化、钙稳态失衡等，继而产生兴奋毒性的效应，最终导致神经元的死亡以及功能的障碍[19-20]。鉴于谷氨酸的兴奋毒性作用，实验研究中常用谷氨酸来建立细胞损伤模型。其次，海马体是在学习与记忆巩固等功能中发挥关键作用的重要脑区，在海马体的神经回路中也发现了许多突触可塑性的实证[21-22]。广泛的尸检和神经影像学研究表明海马体是精神分裂症病理生理学中的关键部位。有学者研究发现精神分裂症患者海马体积减小以及神经元细胞显著减少[23]。因此，作为海马体基本结构和功能单位的海马神经元细胞是具有代表性和可行性的。本研究以谷氨酸作用于海马神经元细胞，经流式细胞仪检测发现海马神经元细胞凋亡率明显升高，与上述文献报道一致。而经温胆汤含药血清干预后发现凋亡率显著下降，说明温胆汤可能对维持神经元细胞的存活、保护神经等方面有一定的作用。
PSD-95是一种广泛分布于突触后致密区的支架蛋白，其通过不同结构域与多种受体、相关蛋白、黏附分子结合，在参与调控突触信号的传导、维持突触后膜结构的稳定性以及影响突触的数量和发育等过程扮演着重要的角色，通常作为突触后可塑性的标志性蛋白[24]。研究发现，在精神分裂症患者的海马和前额叶皮层中参与突触可塑性的关键蛋白PSD-95的表达明显降低[25-26]。GAP-43在轴突末端和突触前膜高度表达，与轴突延伸、再生以及突触连接与形成等过程密切相关，是研究神经系统生长发育、损伤修复和突触可塑性的神经元特异性蛋白 [27-28]。此外，作为神经元中蛋白激酶C（protein kinase C，PKC）的主要底物，其磷酸化状态可以影响突触传递效率，最终影响突触的功能可塑性[29]。FUNG S J等[30]发现精神分裂症患者存在突触末端可塑性的降低且检测到突触可塑标志物GAP-43的表达低于正常人。JIP-3是JNK相互作用蛋白（JNK-interacting proteins，JIPs）的家族成员之一，在体内多种组织中均有表达，但在脑组织中高度表达，对轴突发育产生重要的影响[31]。JIP-3作为支架蛋白能够介导多种货物蛋白的轴突转运，并且聚集于轴突末梢影响着轴突的生长[32]。孙涛[33]通过体内外实验证实JIP-3能够促进海马神经元的轴突生长。此外，有研究发现敲除JIP-3基因的小鼠在出生后死亡并伴随着大脑发育的缺陷[34]。Cx-43是缝隙连接中的重要组成部分，能够调控缝隙连接中通道的开放，参与邻近细胞间的信号传递[35]。研究表明，Cx-43可以调控突触数目和促进损伤突触的修复，在神经系统中发挥着保护神经及调控突触可塑性等重要作用[36]。本文结果显示，在10 mmol/L谷氨酸作用下，海马神经元细胞PSD-95、GAP-43、JIP-3、Cx-43mRNA和蛋白表达均明显下调；但经温胆汤含药血清干预后，海马神经元细胞PSD-95、GAP-43、JIP-3、Cx-43的mRNA和蛋白表达明显升高。说明温胆汤可能通过上调PSD-95、GAP-43、JIP-3、Cx-43的表达来调控突触可塑性。
综上所述，温胆汤可能是通过降低神经元细胞凋亡，上调其PSD-95、GAP-43、JIP-3、Cx-43表达水平，进而调控突触可塑性，以期达到有效改善精神分裂症认知缺陷的目的。这可能是温胆汤改善精神分裂症患者认知功能的机制之一。本研究只是从突触可塑性这一切入点，后续研究将从更多方面探讨温胆汤防治精神分裂症的作用机制。
基金
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  • 国家自然科学基金项目(82160839)
  • 江西省自然科学基金项目(20192BAB205113)
  • 江西省研究生创新专项资金项目(YC2023-B222)
  • 江西省教育厅科学技术研究项目(GJJ190642)
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参考文献 引证文献
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2025年第43卷第1期
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doi: 10.13193/j.issn.1673-7717.2025.01.010
  • 首发时间：2026-10-30
  • 出版时间：2025-01-10
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基金
国家自然科学基金项目(82160839)
江西省自然科学基金项目(20192BAB205113)
江西省研究生创新专项资金项目(YC2023-B222)
江西省教育厅科学技术研究项目(GJJ190642)
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    江西中医药大学，江西　南昌　330004

通讯作者:

万红娇（1964-），女，江西南昌人，教授，博士研究生导师，博士，研究方向：中医药与神经保护。E-mail：。
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