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Article(id=1322926329980657880, tenantId=1146029695717560320, journalId=1255847919539208197, issueId=1322926217711726961, articleNumber=null, orderNo=null, doi=10.13193/j.issn.1673-7717.2025.08.018, 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=1793339815850, onlineDateStr=2026-10-30, pubDate=1754755200000, pubDateStr=2025-08-10, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1793339815850, onlineIssueDateStr=2026-10-30, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1793339815850, creator=13701087609, updateTime=1793339815850, updator=13701087609, issue=Issue{id=1322926217711726961, tenantId=1146029695717560320, journalId=1255847919539208197, year='2025', volume='43', issue='8', pageStart='1', pageEnd='258', issueExtLink='null', onlineDate='null', pubDate='1754755200000', pubDateStr='2025-08-10', beforeIssueId=null, nextIssueId=null, price=null, status=1, issueComplete=1, articleOrder=1, issueType=-1, specialIssue=0, createTime=1793339789083, creator='13701087609', updateTime=1793340517761, updator='13701087609', preIssue=null, nextIssue=null, articleTotal=null, ext={EN=IssueExt(id=1322929274071679978, tenantId=1146029695717560320, journalId=1255847919539208197, issueId=1322926217711726961, language=EN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=), CN=IssueExt(id=1322929274071679979, tenantId=1146029695717560320, journalId=1255847919539208197, issueId=1322926217711726961, language=CN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=)}, issueFiles=null, downloadFileDto=null}, startPage=83, endPage=87, ext={EN=ArticleExt(id=1322926330190373081, articleId=1322926329980657880, tenantId=1146029695717560320, journalId=1255847919539208197, language=EN, title=Effect of Kangxian Yixin Decoction（抗纤益心方）on Mitochondrial Ridge Remode And Its Structural Proteins Mic10，Mic60 and Mic19 in Mice with Dilated Cardiomyopathy, columnId=1256183361521967297, journalTitle=Chinese Archives of Traditional Chinese Medicine, columnName=Target on National Project, runingTitle=null, highlight=null, articleAbstract=
Objective

To observe the effects of Kangxian Yixin Decoction（抗纤益心方，KYD）on myocardial mitochondrial ridge and its structural proteins mitochondrial inner membrane ridge structure protein 10（Mic10），mitochondrial inner membrane ridge structure protein 60（Mic60）and mitochondrial inner membrane ridge structure protein 19（Mic19）in mice with dilatedcardiomyopathy（DCM），and explore the mechanism of protecting DCM mitochondrial ridge structure and improving mitochondrial energy metabolism.

Methods

According to random number table method，40 cTnTR141W transgenic DCM mice were divided into model group，KYD low-dose group，KYD medium-dose group，KYD high-dose group and captopril group，with 8 mice in each group，and 8 C57BL/6J mice in normal group.Left ventricular end-systolic diameter（LVESD），left ventricular end-diastolic diameter（LVEDD），ejection fraction（EF）and fraction shorting（FS）were detected by ultrasonography.Myocardial pathological morphology was observed by HE staining，and mitochondrial ultrastructure was observed by electron microscopy.Enzyme-linked immunosorbent assay（ELISA）was used to detect the contents of NT-proBNP and adenosine triphosphate（ATP）in serum of mice.The mRNA and protein expressions of Mic10，Mic60 and Mic19 were detected by Real-time PCR and Western blotting，respectively.

Result

Compared with those in the normal group，in the model group，the levels of LVESD and LVEDD were increased，the levels of FS and EF were decreased（P<0.05），myocardial pathological injury was observed，apoptotic number was increased（P<0.01），NT-proBNP content was increased，ATP content was decreased（P<0.01），mitochondrial ridge structure was damaged and disordered.The mRNA and protein expressions of mitochondrial ridge structure proteins Mic10，Mic60 and Mic19 were decreased（P<0.01）. Compared with those in the model group，in the KYD group，the levels of LVESD and LVEDD were decreased，the levels of FS and EF were increased（P<0.05），the myocardial injury was alleviated，the NT-proBNP content was decreased（P<0.01），the apoptosis was decreased，the ATP content was increased（P<0.05），the mitochondrial ridge damage and arrangement disorder were improved and the mRNA and protein expressions of mitochondrial ridge structure proteins Mic10，Mic60 and Mic19 were increased（P<0.01）.

Conclusion

KYD can improve DCM mice mitochondrial ridge arrangement disorder，alleviate myocardial cell apoptosis，and improve cardiac function，which may be related to increasing the expression of mitochondrial ridge structural proteins Mic10，Mic60 and Mic19 and improving mitochondrial energy metabolism.

, authors=Hongbo CHANG1, Zhentao WANG2, Hong WU1, Chuiyi ZENG2, authorsList=Hongbo CHANG, Zhentao WANG, Hong WU, Chuiyi ZENG, authorCompany=null, correspAuthors=Zhentao WANG, 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=1322926335747825904, articleId=1322926329980657880, tenantId=1146029695717560320, journalId=1255847919539208197, language=CN, title=抗纤益心方对扩张型心肌病小鼠线粒体嵴重构及其结构蛋白Mic10、Mic60、Mic19的影响, columnId=1256183362172142382, journalTitle=中华中医药学刊, columnName=国家项目点击, runingTitle=null, highlight=null, articleAbstract=
目的

观察抗纤益心方对扩张型心肌病（Dilated cardiomyopathy，DCM）小鼠心肌线粒体内膜嵴重构及其线粒体内膜嵴结构蛋白10（Mitochondrial inner membrane ridge structure protein 10，Mic10）、线粒体内膜嵴结构蛋白60（Mitochondrial inner membrane ridge structure protein 60，Mic60）、线粒体内膜嵴结构蛋白19（Mitochondrial inner membrane ridge structure protein 19，Mic19）的影响，探讨其保护DCM线粒体嵴结构，改善线粒体能量代谢的作用机制。

方法

按照随机数字表法将40只cTnTR141W转基因DCM小鼠分为模型组、抗纤益心方低剂量组、抗纤益心方中剂量组、抗纤益心方高剂量组、卡托普利组，每组8只，另设C57BL/6J小鼠为正常组8只，各组小鼠灌胃8周后，小动物超声仪检测左心室收缩末期内径（Left ventricular end-systolic diameter，LVESD）、左心室舒张末期内径（Left ventricular end-diastolic diameter，LVEDD）、射血分数（Ejection fraction，EF）、缩短率（Fraction shorting，FS），HE染色观察心肌病理形态，电镜观察线粒体超微结构，酶联免疫吸附测定法（Enzyme linked immunosorbent assay，ELISA）检测小鼠血清氨基端前脑钠素（N-terminal brain natriuretic peptide precursor，NT-proBNP）、三磷酸腺苷（Adenosine triphosphate，ATP）含量，TUNEL法检测心肌组织凋亡情况，实时荧光定量聚合酶链式反应（Real-time PCR，RT-PCR）和蛋白免疫印迹法（Western Blotting，WB）分别检测心肌线粒体嵴Mic10、Mic60、Mic19 mRNA和蛋白表达。

结果

与正常组比较，模型组小鼠NT-proBNP含量增加（P<0.01），LVESD和LVEDD升高、FS和EF降低（P<0.05），心肌病理损伤，细胞凋亡数增加（P<0.01），ATP含量降低（P<0.01），线粒体嵴结构受损、排列紊乱，线粒体嵴结构蛋白Mic10、Mic60、Mic19 mRNA和蛋白表达均降低（P<0.01）；与模型组比较，抗纤益心方组NT-proBNP含量降低（P<0.05），LVESD和LVEDD降低、FS和EF增加（P<0.05），心肌损伤减轻，细胞凋亡减少，ATP含量增加（P<0.05），线粒体嵴受损及排列紊乱改善，线粒体嵴Mic10、Mic60、Mic19 mRNA和蛋白表达均增加（P<0.01）。

结论

抗纤益心方能够改善DCM小鼠线粒体嵴排列紊乱，减轻心肌细胞凋亡，提高心脏功能，其作用机制可能与增加线粒体嵴结构蛋白Mic10、Mic60、Mic19表达，改善线粒体能量代谢有关。

, authors=常红波1, 王振涛2, 吴鸿1, 曾垂义2, authorsList=常红波, 王振涛, 吴鸿, 曾垂义, authorCompany=null, correspAuthors=王振涛, authorNote=

常红波（1987-），男，河南开封人，讲师、主治医师，硕士研究生导师，博士，研究方向：中医药防治心血管疾病的临床与基础。

, correspAuthorsNote=
王振涛（1962-），男，河南滑县人，教授，博士研究生导师，博士，研究方向：中医药防治心血管疾病的临床与基础。E-mail：。
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常红波（1987-），男，河南开封人，讲师、主治医师，硕士研究生导师，博士，研究方向：中医药防治心血管疾病的临床与基础。

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常红波（1987-），男，河南开封人，讲师、主治医师，硕士研究生导师，博士，研究方向：中医药防治心血管疾病的临床与基础。

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注：A正常组；B模型组；C抗纤益心方高剂量组；D抗纤益心方中剂量组；E抗纤益心方低剂量组；F卡托普利组。

, figureFileSmall=1D88yXSj6grYEVpFskZlyA==, figureFileBig=REMqmHTwKoaUamEdJx/fMQ==, tableContent=null), ArticleFig(id=1322926344023187747, tenantId=1146029695717560320, journalId=1255847919539208197, articleId=1322926329980657880, language=EN, label=null, caption=null, figureFileSmall=null, figureFileBig=null, tableContent=
组别n剂量/（g/kg）LVESD/（mm）LVEDD/（mm）FS/（%）LVEF/（%）
正常组8-2.74±0.231.81±0.2433.94±1.0771.32±2.09
模型组8-3.92±0.27▲3.34±0.25▲14.80±1.13▲38.14±1.57▲
抗纤益心方高剂量组85.43.27±0.24#2.52±0.46#22.93±1.51#54.23±1.86#
抗纤益心方中剂量组82.73.49±0.25#2.76±0.25#20.92±1.43#50.60±1.37#
抗纤益心方低剂量组81.353.76±0.193.17±0.27#15.69±1.31#40.06±1.21#
卡托普利组80.013.39±0.12#2.61±0.23#23.01±1.16#54.36±1.63#
), ArticleFig(id=1322926344119656740, tenantId=1146029695717560320, journalId=1255847919539208197, articleId=1322926329980657880, language=CN, label=表1, caption=

各组小鼠心脏超声结果比较（）

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组别n剂量/（g/kg）LVESD/（mm）LVEDD/（mm）FS/（%）LVEF/（%）
正常组8-2.74±0.231.81±0.2433.94±1.0771.32±2.09
模型组8-3.92±0.27▲3.34±0.25▲14.80±1.13▲38.14±1.57▲
抗纤益心方高剂量组85.43.27±0.24#2.52±0.46#22.93±1.51#54.23±1.86#
抗纤益心方中剂量组82.73.49±0.25#2.76±0.25#20.92±1.43#50.60±1.37#
抗纤益心方低剂量组81.353.76±0.193.17±0.27#15.69±1.31#40.06±1.21#
卡托普利组80.013.39±0.12#2.61±0.23#23.01±1.16#54.36±1.63#
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组别n剂量/（g·kg）NT-proBNP/（pg/mL）ATP/（nmol/L）
正常组6-87.00±0.501 801.00±127.12
模型组6-204.80±16.98▲▲1 044.80±118.99▲▲
抗纤益心方高剂量组65.4118.15±4.47##1 536.60±76.07##
抗纤益心方中剂量组62.7164.41±7.28#1 404.00±94.88#
抗纤益心方低剂量组61.35195.20±14.31#1 121.80±72.07#
卡托普利组60.01112.40±9.74#1 568.40±54.49#
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各组小鼠血清NT-proBNP、ATP含量结果比较（）

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组别n剂量/（g·kg）NT-proBNP/（pg/mL）ATP/（nmol/L）
正常组6-87.00±0.501 801.00±127.12
模型组6-204.80±16.98▲▲1 044.80±118.99▲▲
抗纤益心方高剂量组65.4118.15±4.47##1 536.60±76.07##
抗纤益心方中剂量组62.7164.41±7.28#1 404.00±94.88#
抗纤益心方低剂量组61.35195.20±14.31#1 121.80±72.07#
卡托普利组60.01112.40±9.74#1 568.40±54.49#
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组别n剂量/（g/kg）凋亡/（%）
正常组3-7.50±1.44
模型组3-32.73±3.79▲▲
抗纤益心方高剂量组35.415.60±1.80##
抗纤益心方中剂量组32.719.03±1.72##
抗纤益心方低剂量组31.3523.03±1.46#
卡托普利组30.0114.30±1.25##
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各组小鼠心肌细胞凋亡情况结果比较（）

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组别n剂量/（g/kg）凋亡/（%）
正常组3-7.50±1.44
模型组3-32.73±3.79▲▲
抗纤益心方高剂量组35.415.60±1.80##
抗纤益心方中剂量组32.719.03±1.72##
抗纤益心方低剂量组31.3523.03±1.46#
卡托普利组30.0114.30±1.25##
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组别n剂量/（g/kg）Mic10Mic60Mic19
正常组3-1.00±0.00　　1.00±0.001.00±0.00
模型组3-0.42±0.07▲▲0.39±0.11▲▲0.39±0.07▲▲
抗纤益心方高剂量组35.41.03±0.21##0.87±0.09##1.09±0.15##
抗纤益心方中剂量组32.70.86±0.10#0.67±0.10#0.76±0.06#
抗纤益心方低剂量组31.350.52±0.96#0.41±0.05#0.47±0.09#
卡托普利组30.010.98±0.15#1.04±0.11#0.99±0.16#
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各组小鼠心肌组织Mic10、Mic60、Mic19 mRNA表达水平（）

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组别n剂量/（g/kg）Mic10Mic60Mic19
正常组3-1.00±0.00　　1.00±0.001.00±0.00
模型组3-0.42±0.07▲▲0.39±0.11▲▲0.39±0.07▲▲
抗纤益心方高剂量组35.41.03±0.21##0.87±0.09##1.09±0.15##
抗纤益心方中剂量组32.70.86±0.10#0.67±0.10#0.76±0.06#
抗纤益心方低剂量组31.350.52±0.96#0.41±0.05#0.47±0.09#
卡托普利组30.010.98±0.15#1.04±0.11#0.99±0.16#
), ArticleFig(id=1322926344648139051, tenantId=1146029695717560320, journalId=1255847919539208197, articleId=1322926329980657880, language=EN, label=null, caption=null, figureFileSmall=null, figureFileBig=null, tableContent=
组别n剂量/（g/kg）Mic10Mic60Mic19
正常组3-1.00±0.051.00±0.031.00±0.02
模型组3-0.31±0.04▲▲0.37±0.05▲▲0.31±0.07▲▲
抗纤益心方高剂量组35.40.76±0.07#1.08±0.13##1.05±0.08#
抗纤益心方中剂量组32.70.63±0.09#0.71±0.03#0.68±0.06#
抗纤益心方低剂量组31.350.36±0.05#0.48±0.09#0.38±0.07#
卡托普利组30.010.66±0.12#1.13±0.14#0.97±0.06#
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各组小鼠心肌组织Mic10、Mic60、Mic19蛋白表达水平（）

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组别n剂量/（g/kg）Mic10Mic60Mic19
正常组3-1.00±0.051.00±0.031.00±0.02
模型组3-0.31±0.04▲▲0.37±0.05▲▲0.31±0.07▲▲
抗纤益心方高剂量组35.40.76±0.07#1.08±0.13##1.05±0.08#
抗纤益心方中剂量组32.70.63±0.09#0.71±0.03#0.68±0.06#
抗纤益心方低剂量组31.350.36±0.05#0.48±0.09#0.38±0.07#
卡托普利组30.010.66±0.12#1.13±0.14#0.97±0.06#
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抗纤益心方对扩张型心肌病小鼠线粒体嵴重构及其结构蛋白Mic10、Mic60、Mic19的影响
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常红波 1 , 王振涛 2 , 吴鸿 1 , 曾垂义 2
中华中医药学刊 | 国家项目点击 2025,43(8): 83-87
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中华中医药学刊 |国家项目点击 2025 , 43 (8) : 83 -87
抗纤益心方对扩张型心肌病小鼠线粒体嵴重构及其结构蛋白Mic10、Mic60、Mic19的影响
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常红波1, 王振涛2 , 吴鸿1, 曾垂义2
作者信息
  • 1.河南中医药大学，河南 郑州 450046
  • 2.河南省中医院，河南 郑州 450002
通讯作者:
王振涛（1962-），男，河南滑县人，教授，博士研究生导师，博士，研究方向：中医药防治心血管疾病的临床与基础。E-mail：。
作者简介:

常红波（1987-），男，河南开封人，讲师、主治医师，硕士研究生导师，博士，研究方向：中医药防治心血管疾病的临床与基础。

Effect of Kangxian Yixin Decoction（抗纤益心方）on Mitochondrial Ridge Remode And Its Structural Proteins Mic10，Mic60 and Mic19 in Mice with Dilated Cardiomyopathy
Hongbo CHANG1, Zhentao WANG2 , Hong WU1, Chuiyi ZENG2
Affiliations
  • 1.Henan University of Chinese Medicine，Zhengzhou 450046，Henan，China
  • 2.Henan Province Hospital of Traditional Chinese Medicine，Zhengzhou 450002，Henan，China
出版时间: 2025-08-10 doi: 10.13193/j.issn.1673-7717.2025.08.018
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摘要
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目的

观察抗纤益心方对扩张型心肌病（Dilated cardiomyopathy，DCM）小鼠心肌线粒体内膜嵴重构及其线粒体内膜嵴结构蛋白10（Mitochondrial inner membrane ridge structure protein 10，Mic10）、线粒体内膜嵴结构蛋白60（Mitochondrial inner membrane ridge structure protein 60，Mic60）、线粒体内膜嵴结构蛋白19（Mitochondrial inner membrane ridge structure protein 19，Mic19）的影响，探讨其保护DCM线粒体嵴结构，改善线粒体能量代谢的作用机制。

方法

按照随机数字表法将40只cTnTR141W转基因DCM小鼠分为模型组、抗纤益心方低剂量组、抗纤益心方中剂量组、抗纤益心方高剂量组、卡托普利组，每组8只，另设C57BL/6J小鼠为正常组8只，各组小鼠灌胃8周后，小动物超声仪检测左心室收缩末期内径（Left ventricular end-systolic diameter，LVESD）、左心室舒张末期内径（Left ventricular end-diastolic diameter，LVEDD）、射血分数（Ejection fraction，EF）、缩短率（Fraction shorting，FS），HE染色观察心肌病理形态，电镜观察线粒体超微结构，酶联免疫吸附测定法（Enzyme linked immunosorbent assay，ELISA）检测小鼠血清氨基端前脑钠素（N-terminal brain natriuretic peptide precursor，NT-proBNP）、三磷酸腺苷（Adenosine triphosphate，ATP）含量，TUNEL法检测心肌组织凋亡情况，实时荧光定量聚合酶链式反应（Real-time PCR，RT-PCR）和蛋白免疫印迹法（Western Blotting，WB）分别检测心肌线粒体嵴Mic10、Mic60、Mic19 mRNA和蛋白表达。

结果

与正常组比较，模型组小鼠NT-proBNP含量增加（P<0.01），LVESD和LVEDD升高、FS和EF降低（P<0.05），心肌病理损伤，细胞凋亡数增加（P<0.01），ATP含量降低（P<0.01），线粒体嵴结构受损、排列紊乱，线粒体嵴结构蛋白Mic10、Mic60、Mic19 mRNA和蛋白表达均降低（P<0.01）；与模型组比较，抗纤益心方组NT-proBNP含量降低（P<0.05），LVESD和LVEDD降低、FS和EF增加（P<0.05），心肌损伤减轻，细胞凋亡减少，ATP含量增加（P<0.05），线粒体嵴受损及排列紊乱改善，线粒体嵴Mic10、Mic60、Mic19 mRNA和蛋白表达均增加（P<0.01）。

结论

抗纤益心方能够改善DCM小鼠线粒体嵴排列紊乱，减轻心肌细胞凋亡，提高心脏功能，其作用机制可能与增加线粒体嵴结构蛋白Mic10、Mic60、Mic19表达，改善线粒体能量代谢有关。

抗纤益心方  /  扩张型心肌病  /  线粒体  /  Mic10  /  Mic60  /  Mic19
Objective

To observe the effects of Kangxian Yixin Decoction（抗纤益心方，KYD）on myocardial mitochondrial ridge and its structural proteins mitochondrial inner membrane ridge structure protein 10（Mic10），mitochondrial inner membrane ridge structure protein 60（Mic60）and mitochondrial inner membrane ridge structure protein 19（Mic19）in mice with dilatedcardiomyopathy（DCM），and explore the mechanism of protecting DCM mitochondrial ridge structure and improving mitochondrial energy metabolism.

Methods

According to random number table method，40 cTnTR141W transgenic DCM mice were divided into model group，KYD low-dose group，KYD medium-dose group，KYD high-dose group and captopril group，with 8 mice in each group，and 8 C57BL/6J mice in normal group.Left ventricular end-systolic diameter（LVESD），left ventricular end-diastolic diameter（LVEDD），ejection fraction（EF）and fraction shorting（FS）were detected by ultrasonography.Myocardial pathological morphology was observed by HE staining，and mitochondrial ultrastructure was observed by electron microscopy.Enzyme-linked immunosorbent assay（ELISA）was used to detect the contents of NT-proBNP and adenosine triphosphate（ATP）in serum of mice.The mRNA and protein expressions of Mic10，Mic60 and Mic19 were detected by Real-time PCR and Western blotting，respectively.

Result

Compared with those in the normal group，in the model group，the levels of LVESD and LVEDD were increased，the levels of FS and EF were decreased（P<0.05），myocardial pathological injury was observed，apoptotic number was increased（P<0.01），NT-proBNP content was increased，ATP content was decreased（P<0.01），mitochondrial ridge structure was damaged and disordered.The mRNA and protein expressions of mitochondrial ridge structure proteins Mic10，Mic60 and Mic19 were decreased（P<0.01）. Compared with those in the model group，in the KYD group，the levels of LVESD and LVEDD were decreased，the levels of FS and EF were increased（P<0.05），the myocardial injury was alleviated，the NT-proBNP content was decreased（P<0.01），the apoptosis was decreased，the ATP content was increased（P<0.05），the mitochondrial ridge damage and arrangement disorder were improved and the mRNA and protein expressions of mitochondrial ridge structure proteins Mic10，Mic60 and Mic19 were increased（P<0.01）.

Conclusion

KYD can improve DCM mice mitochondrial ridge arrangement disorder，alleviate myocardial cell apoptosis，and improve cardiac function，which may be related to increasing the expression of mitochondrial ridge structural proteins Mic10，Mic60 and Mic19 and improving mitochondrial energy metabolism.

Kangxian Yixin Decoction（抗纤益心方）  /  dilated cardiomyopathy  /  mitochondrial ridge  /  Mic10  /  Mic60  /  Mic19
常红波, 王振涛, 吴鸿, 曾垂义. 抗纤益心方对扩张型心肌病小鼠线粒体嵴重构及其结构蛋白Mic10、Mic60、Mic19的影响. 中华中医药学刊, 2025 , 43 (8) : 83 -87 . DOI: 10.13193/j.issn.1673-7717.2025.08.018
Hongbo CHANG, Zhentao WANG, Hong WU, Chuiyi ZENG. Effect of Kangxian Yixin Decoction（抗纤益心方）on Mitochondrial Ridge Remode And Its Structural Proteins Mic10，Mic60 and Mic19 in Mice with Dilated Cardiomyopathy[J]. Chinese Archives of Traditional Chinese Medicine, 2025 , 43 (8) : 83 -87 . DOI: 10.13193/j.issn.1673-7717.2025.08.018
正文
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扩张型心肌病（Dilated cardiomyopathy，DCM）是引起心力衰竭的重要病因，主要特征心肌收缩功能障碍、心脏扩大，病情呈进行性加重，其病因及病理机制尚不完全明确[1-2]，目前缺乏有效的治疗手段[3]，预后较差。研究表明，线粒体能量代谢在DCM发展过程中具有重要作用[4-5]。线粒体是含有双层膜结构的动态细胞器，其内膜深入基质处突起形成线粒体嵴是线粒体氧化磷酸化的关键位点，亦是产生ATP供能基地[6]。线粒体嵴形态及重构调控细胞多种功能[7]，DCM发生时，细胞内活性氧增加，嵴连接被重构以促进凋亡膜间隙蛋白的释放，ATP产能不足，线粒体嵴损伤，嵴形态重构发生，能量代谢障碍，引发心肌重构以及细胞凋亡[8]，促使DCM发展。Mic10、Mic60、Mic19为线粒体嵴结构关键蛋白，在维持线粒体嵴结构形成和功能中发挥重要作用[9]。因此，调控线粒体嵴结构蛋白与功能可能成为防治DCM的潜在作用靶点。
中医学认为DCM可归属“心胀”“心衰”“喘证”等范畴，为本虚标实证，本团队王振涛教授长期致力于DCM研究，认为宗气下陷为DCM核心病机，标实责之瘀血、水湿等。“益气升陷”为针对DCM“宗气下陷”的病机所立治法，根据治法拟抗纤益心方，具有益气升陷、活血化瘀功效，临床取得较好效果[10-11]，前期研究发现，抗纤益心方能通过减轻细胞凋亡、抑制心肌重构、提高心功能，从而延缓DCM进程，且对线粒体结构和功能及能量代谢具有一定的改善作用[12]。然而，抗纤益心方调节线粒体结构与功能的潜在机制尚未阐明。本研究通过建立DCM小鼠模型，以观察抗纤益心方对DCM小鼠心功能、线粒体超微结构及线粒体嵴结构蛋白Mic10、Mic60、Mic19表达的影响及作用机制，为抗纤益心方治疗DCM提供科学的实验依据。
1　材料
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1.1　动物
cTnTR141W转基因DCM小鼠体质量（25±2）g，SPF级雄性，购自中国医学科学院实验动物研究所，许可证号SCXK（京）2021-0065，合格证编号1103261911000021。SPF级雄性，体质量（25±2）g，C57BL/6J小鼠，购自北京维通利华实验动物技术有限公司，许可证号SCXK（京）2021-0018，合格证编号1102861914000028。饲养于河南省中医院中心实验室动物中心，室内温度20～24℃，相对湿度40%～60%，12 h/12 h交替照明。该实验由河南省中医院动物实验伦理委员会评审通过，编号：PZ-HNSZYY-2022-060。
1.2　药物
抗纤益心方组成：黄芪30 g，红参12 g，丹参15 g，益母草15 g，茯苓15 g，白术15 g，泽兰15 g，麦冬12 g，升麻9 g，由四川新绿药业科技发展有限公司提供，药物质量控制采用GMP认证（证书编号：SC20150037），1.0 g颗粒相当于生药8.0 g。卡托普利（上海信宜天平药业有限公司生产，国药准字H31021832，批号：187280，25 mg/片）。
1.3　试剂
NT-proBNP试剂盒（武汉伊莱瑞特生物科技股份有限公司，货号：E-EL-0053c），ATP（酶联生物科技有限公司，货号：批号202304）；伊红染色液和苏木精染色液（北京索莱宝科技有限公司，批号：G1120-3）；BCA蛋白定量试剂盒（上海碧云天生物技术有限公司，批号：P0010）；PVDF膜（美国Millipore公司，批号：IPVH0010）；SDS-PAGE凝胶（西安晶彩生物公司，批号：JCPE022）；mRNA提取试剂盒、实时荧光定量聚合酶链式反应（Real-timePCR）反转录试剂盒（日本Takara公司，批号分别为FB26 KB3904，AK51598A）。Mic10、Mic60、Mic19及COXⅣ抗体（武汉三鹰生物公司，批号：10179-1-AP、12494-1-AP、25625-1-AP、66110-1-lg）。
1.4　仪器
Mini-PROTEAN SDS-PAGE凝胶电泳系统、Mini Trans-Blot蛋白湿转仪、ChemiDoc XRS+凝胶成像系统（美国Bio-Rad公司）；JEM-1400透射电镜（日本电子株式会社）。
2　方法
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2.1　分组与给药
将40只cTnTR141W转基因DCM小鼠随机分为模型组，抗纤益心方高、中、低剂量组，卡托普利组。小鼠灌胃剂量根据临床成人服用剂量换算得到，小鼠每公斤体质量用药量为成人公斤体质量的9倍。抗纤益心方高、中、低剂量浓度分别是5.4、2.7、1.35 g/kg，卡托普利0.01 g/kg，正常组和模型组小鼠每日给予灌胃对应剂量的生理盐水，连续干预8周。
2.2　指标检测
2.2.1　超声心动仪检测小鼠心功能
在小鼠乳头肌水平处用小动物超声心动仪进行二维定向M 型示踪，测量LVEDD、LVESD、EF、FS。检测由专业超声人员进行操作，每次检测需连续3个心动周期。
2.2.2　HE染色检测小鼠心肌组织病理形态
麻醉后立即取心脏组织于4%多聚甲醛固定，在24 h后开始常规脱水，然后依次性按照说明书操作步骤进行石蜡包埋，切片，脱洗，染色，封片过夜，每个标本5个视野选取，显微镜观察并拍照。
2.2.3　电镜观察心肌线粒体超微结构
取材时于心脏左心室心尖部位剪取一小块心肌组织，约1 mm3大小的组织标本，迅速放入2.5%戊二醛EP管子中固定，每个标本切成3～4个，4 h，换取新鲜2.5%戊二醛固定液，漂洗，锇酸1%加入，丙酮浸染，半薄切片定位、超薄切片，然后依次饱和醋酸铀溶液，柠檬酸铅溶液，电镜观察拍照。
2.2.4　ELISA检测小鼠血清NT-proBNP、ATP含量
收集各组血清，分别将100μL上清液加入到NT-proBNP、ATP抗体酶标板中。在室温条件下孵育，2 h后开始洗涤，于每孔添加酶标抗体，孵育1 h室温下，循环进行洗涤，每孔添加底物和显色剂孵育20 min。加入终止溶液，在酶标仪450 nm和570 nm处读取吸光度值，计算NT-proBNP、ATP含量。
2.2.5　TUNEL法检测心肌组织细胞凋亡
将石蜡切片样本依次放入二甲苯Ⅰ、二甲苯Ⅱ、不同浓度乙醇。用滤纸吸干切片样本周围液体，滴加蛋白酶K溶液孵育。参照TUNEL细胞凋亡检测试剂盒说明书进行操作，荧光显微镜下观察并拍照。
2.2.6　RT-PCR检测小鼠心肌Mic10、Mic60、Mic19 mRNA表达
从心肌组织中提取总RNA，用Takara反转录试剂盒，配制反转录体系总体积20μL进行反转录反应，进行扩增，反转录cDNA，按照试剂盒说明配制反应液，经过95℃预变性，95℃变性，60℃退火/延伸，再加热至95℃，循环数为40。引物由上海生工生物工程技术服务有限公司合成。引物序列：Mic10上游ATGAGAAGTGGGATCGCTGC，下游TTGCACTCCTCATAGGCACG，bp：157；Mic60上游TACACGTTGCTGTTGAGGCT，下游GATGGATTAGAGGCGATCAT，bp：142；Mic19上游GGTAACCTACCATCGTCCGA， 下 游 TCTGTAGGCGCATGCAACTC，bp：115；GAPDH 上游ATCGGCTCCTTTGAACCTCG，下游CTTGGTGTCATCCCACTCCC，bp：155。根据2-ΔΔCt定量法计算各组mRNA表达水平。
2.2.7　WB检测小鼠心肌线粒体嵴结构蛋白Mic10、Mic60、Mic19表达
从-80℃取冻存心肌组织约100 mg，RIPA裂解，匀浆，离心，蛋白的浓度测定BCA法，蛋白上样缓冲液加入，制胶，上样和Maker，凝胶电泳，转模，4℃一抗孵育，清洗，孵育二抗，显影，成像，结果使用Image J软件分析条带的光密度值，以目的蛋白/COXⅣ比值表示目的蛋白相对表达量。
2.3　统计学分析
本实验所得实验数据分析均采用SPSS 21.0统计软件，实验数据符合正态分布以均数±标准差（）表示，多组间比较用单因素方差分析，方差齐时采用LSD法检验，方差不齐时采用Tamhane′s法检验。P<0.05为差异有统计学意义。
3　结果
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3.1　抗纤益心方对DCM小鼠心脏功能的影响
模型组小鼠与正常组比较，LVESD和LVEDD增加（P<0.05），FS和EF降低（P<0.05）；抗纤益心方组与模型组比较，LVESD和LVEDD降低（P<0.05），FS和EF增加（P<0.05）。见插页ⅩⅩⅣ图1，表1。
3.2　抗纤益心方对DCM小鼠心肌病理损伤的影响
与正常组比较，模型组心肌受损明显，细胞核排列不规则，肌纤维排列、走形紊乱，胶原沉积，局部心肌纤维化严重；与模型组比较，抗纤益心方各剂量组心肌受损减轻，心肌细胞形态与纤维排列顺序明显改善，胶原沉积减少。见插页ⅩⅩⅤ图2。
3.3　抗纤益心方对DCM小鼠心肌线粒体嵴形态结构的影响
正常组线粒体双层膜完整清晰、线粒体嵴排列整齐；与正常组比较，模型组线粒体双层膜模糊或消失、线粒体嵴破裂、排列紊乱；与模型组比较，抗纤益心方剂量组线粒体形态部分恢复正常，线粒体双层膜结构清晰，仍有少量的受损线粒体嵴，嵴排列较整齐。见插页ⅩⅩⅥ图3。
3.4　抗纤益心方对DCM小鼠血清NT-proBNP、ATP的影响
与正常组比较，模型组NT-proBNP含量增加（P<0.01），ATP含量减少（P<0.01）。与模型组比较，抗纤益心方各剂量组NT-proBNP降低，ATP含量增加（P<0.01，P<0.05）。见表2。
3.5　抗纤益心方对DCM小鼠心肌细胞凋亡的影响
与正常组比较，模型组细胞凋亡增加；与模型组比较，抗纤益心方剂量组细胞凋亡减少。见插页ⅩⅩⅦ图4，表3。
3.6　抗纤益心方对DCM小鼠心肌Mic10、Mic60、Mic19 mRNA表达的影响
与正常组比较，模型组Mic10、Mic60、Mic19 mRNA表达降低（P<0.01）；与模型组比较，抗纤益心方组Mic10、Mic60、Mic19 mRNA表达增加（P<0.05）。见表4。
3.7　抗纤益心方对DCM小鼠心肌Mic10、Mic60、Mic19蛋白表达的影响
与正常组比较，模型组Mic10、Mic60、Mic19蛋白表达降低（P<0.01）；与模型组比较，抗纤益心方组Mic10、Mic60、Mic19蛋白表达增加（P<0.05）。见图5，表5。
4　讨论
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扩张型心肌病属于临床常见心肌病之一，是导致心力衰竭的重要原因。我国最近一项报道显示其52个月病死率为42.64%[2]，其发病率有逐渐上升的趋势。能量代谢障碍是DCM发生发展主要病理过程，而线粒体结构和功能损伤是引起能量代谢障碍，导致DCM的重要因素[13]。临床上通过应用改善心肌能量代谢药物辅酶Q10、磷酸肌酸钠、曲美他嗪等可以起到缓解作用，但效果有限[14-15]，因此开发改善线粒体心肌能量代谢药物尤为迫切需要。
中医药治疗DCM具有一定优势，我们团队王振涛教授长期从事DCM研究，根据四十余年临床实践，提出宗气下陷为DCM核心病机，瘀血贯穿于本病始终，故临床治疗以益气升陷活血法为基本治法，根据治法拟定抗纤益心方治疗DCM，疗效获得患者满意。方中黄芪为君药，具有益气升陷功效，发挥直补宗气下陷，直达病所的作用；宗气下陷与脾胃之气密切相关，故臣药用红参补脾胃之气，白术、茯苓健运脾胃，以恢复脾胃功能；益母草、丹参、泽兰活血化瘀利水以解决标实之症；麦冬具有滋心阴而不助湿，为佐药，且可制约补气药的燥性；升麻具有升陷功效，作为使药升提引经作用。前期研究表明，抗纤益心方能够改善DCM小鼠线粒体结构异常，提高线粒体功能作用[12]，为进一步探讨抗纤益心方调节线粒体的作用机制提供实验依据。
本实验利用cTnTR141W转基因小鼠成功建立DCM模型，探讨抗纤益心方对调节线粒体嵴作用机制。研究表明，不同因素导致DCM出现心肌病理性损伤，活性氧增加[16]，细胞凋亡增加，线粒体嵴破裂、排列紊乱，ATP产生减少，线粒体功能异常，能量代谢障碍[17-18]，从而导致DCM心力衰竭。在这一过程中，线粒体嵴结构受损与重构是导致线粒体功能下降加重DCM心力衰竭的重要病理机制。Mic10、Mic60、Mic19是线粒体嵴结构关键蛋白，三者分布于线粒体内膜[19-20]，在维持线粒体的正常功能及嵴形态上具有重要作用[21-22]。研究表明，线粒体对不同生理条件的适应高度依赖于线粒体超微结构的调节，特别是在线粒体嵴区室水平上[23]，嵴数量和形状的变化决定了线粒体呼吸能力和细胞活力[24-25]。在DCM演变过程中，心肌损伤，线粒体结构与功能出现失调，线粒体嵴结构蛋白Mic10、Mic60、Mic19丢失，线粒体嵴与内膜直接物理接触丧失[26]，线粒体内膜边界堆积，嵴受到不可修复的损害，线粒体无法维持其正常的生理功能[27]，细胞内重要的生命活动都会受到不同程度的抑制[28-29]。综合以上因素，通过调节线粒体嵴结构蛋白表达，是维持线粒体功能及嵴形态的正常、改善心肌能量代谢、提高心脏功能的潜在作用靶点。研究报道，线粒体嵴结构蛋白Mic10或Mic60或Mic19缺失，线粒体嵴与内膜的连接中断，嵴形态改变，线粒体结构与功能损伤。而通过调节Mic10、Mic60或Mic19表达，可保护线粒体结构和功能，改善能量代谢[30]。
本实验研究显示，模型组NT-proBNP水平明显增加，心脏超声显示心脏FS和EF降低、心功能下降，心肌组织病理心肌出现间质水肿、心肌纤维排列紊乱，纤维胶原沉积，电镜下显示线粒体结构损伤、线粒体双层膜模糊或消失、线粒体嵴数量减少、线粒体嵴破裂、排列紊乱，TUNEL结果显示细胞凋亡率增多，ATP含量降低，同时线粒体嵴结构蛋白Mic10、Mic60、Mic19的mRNA和蛋白表达均明显下调，表明DCM心功能下降，心肌纤维排列紊乱，心肌细胞凋亡增加可能与线粒体嵴相关。通过与模型组比较，抗纤益心方高、中、低剂量干预后，DCM小鼠NT-proBNP水平明显降低，心脏FS和EF增加，心肌组织病理心肌出现间质水肿和纤维胶原沉积减少，心肌损伤减轻，电镜下线粒体形态部分恢复正常、有少量的受损线粒体嵴、嵴排列较整齐，细胞凋亡程度减轻，且线粒体嵴结构蛋白Mic10、Mic60、Mic19的mRNA和蛋白表达明显增加，提示抗纤益心方可能通过上调线粒体嵴结构蛋白表达，保护线粒体结构与功能，改善能量代谢，提高心脏功能。
综上所述，抗纤益心方能够调节DCM心肌线粒体嵴结构蛋白Mic10、Mic60、Mic19表达，促进ATP产能，改善线粒体嵴形态与重构，减少细胞凋亡，提高心脏功能，从而改善DCM。此研究为从调节线粒体能量代谢角度改善DCM药物开发提供一定参考。但DCM发病机制复杂，本研究只表明了抗纤益心方改善DCM机制可能与调节线粒体嵴结构蛋白Mic10、Mic60、Mic19表达，改善线粒体嵴形态与重构及能量代谢有关。未能全面阐述治疗DCM机制，今后仍需进一步研究，以期为中医药改善线粒体嵴结构与功能，治疗DCM提供更加完善的科学实验依据。
基金
收起
  • 国家自然科学基金青年科学基金项目(82205082)
  • 河南省科技研发计划（优势学科培育类）项目(232301420066)
  • 河南省高等学校重点科研项目(24A360001)
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2025年第43卷第8期
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doi: 10.13193/j.issn.1673-7717.2025.08.018
  • 首发时间：2026-10-30
  • 出版时间：2025-08-10
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基金
国家自然科学基金青年科学基金项目(82205082)
河南省科技研发计划（优势学科培育类）项目(232301420066)
河南省高等学校重点科研项目(24A360001)
作者信息
    1.河南中医药大学，河南 郑州 450046
    2.河南省中医院，河南 郑州 450002

通讯作者:

王振涛（1962-），男，河南滑县人，教授，博士研究生导师，博士，研究方向：中医药防治心血管疾病的临床与基础。E-mail：。
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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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