浏览全部资源
扫码关注微信
泻肺利水合剂对异丙肾上腺素损伤大鼠心肌细胞线粒体钙超载的改善作用及机制
Effect and mechanism of
Xiefei Lishui Mixture on mitochondrial overload of myocardial cells injured by isoproterenol- 北京中医药 2025年44卷第2期 页码：149-156
- 作者机构：
首都医科大学附属北京中医医院心血管科，北京 100010
- 作者简介：
梁立新，女，33岁，博士研究生，主治医师。研究方向：中西医结合诊治心血管疾病、急诊危重症。
刘红旭，E-mail：lhx_@263.net
- 基金信息：中医药循证能力建设项目-专科专病循证能力提升(2019XZZX-XXG001);北京市医管局扬帆计划重点医学专业（中医心血管病）(ZYLX201817)
DOI：10.16025/j.1674-1307.2025.02.007
中图分类号：收稿日期：2024-02-20，
纸质出版日期：2025-02-25
- 稿件说明：
移动端阅览
梁立新，魏鹏路，刘红旭，等.泻肺利水合剂对异丙肾上腺素损伤大鼠心肌细胞线粒体钙超载的改善作用及机制［J］.北京中医药，2025，44（2）：149-156.
LIANG Lixin,WEI Penglu,LIU Hongxu,et al.Effect and mechanism of Xiefei Lishui Mixture on mitochondrial overload of myocardial cells injured by isoproterenol[J]. Beijing Journal of Traditional Chinese Medicine,2025,44(02):149-156.
梁立新，魏鹏路，刘红旭，等.泻肺利水合剂对异丙肾上腺素损伤大鼠心肌细胞线粒体钙超载的改善作用及机制［J］.北京中医药，2025，44（2）：149-156. DOI： 10.16025/j.1674-1307.2025.02.007.
LIANG Lixin,WEI Penglu,LIU Hongxu,et al.Effect and mechanism of Xiefei Lishui Mixture on mitochondrial overload of myocardial cells injured by isoproterenol[J]. Beijing Journal of Traditional Chinese Medicine,2025,44(02):149-156. DOI： 10.16025/j.1674-1307.2025.02.007.
摘要
目的
2
研究泻肺利水合剂对异丙肾上腺素（isoproterenol，ISO）损伤大鼠心肌细胞（H9c2细胞）线粒体钙超载的改善作用及机制。
方法
2
体外培养H9c2细胞，取部分H9c2细胞随机分为正常组、模型组、中药组、卡托普利组，除正常组外，各组均先使用含ISO的培养基培养48 h，正常组、模型组用20%生理盐水含药血清、中药组用20%泻肺利水含药血清、卡托普利组用20%卡托普利含药血清培养基再培养48 h。取部分H9c2细胞随机分为转染对照组（si-NC组）、转染中药对照组（si-NC+中药组）、转染卡托普利对照组（si-NC+卡托普利组）、转染模型组（si-GRP75组）、转染中药模型组（si-GRP75+中药组）、转染卡托普利模型组（si-GRP75+卡托普利组），将转染后的各组细胞均先用含ISO的培养基培养48 h，si-NC组、si-GRP75组加入20%生理盐水含药血清培养基，si-NC+中药组、si-GRP75+中药组加入20%泻肺利水含药血清培养基，si-NC+卡托普利组、si-GRP75+卡托普利组加入20%卡托普利含药血清培养基，再培养48 h。CCK-8法检测细胞活力，荧光定量PCR检测肌醇-1，4，5三磷酸受体（IP3R）、葡萄糖调节蛋白75（GRP75）、电压依赖性阴离子通道蛋白1（VDAC1）、线粒体融合蛋白2（MFN2） mRNA表达，Western blotting法检测IP3R、GRP75、VDAC1、MFN2蛋白表达，Rhod2-AM Ca
2+
荧光探针法检测线粒体内Ca
2+
含量。
结果
2
与正常组比较，模型组IP3R、GRP75、VDAC1 mRNA、蛋白表达及线粒体Ca
2+
含量高，MFN2 mRNA和蛋白表达低（
P
<
0.05）；与模型组比较，中药组、卡托普利组IP3R、GRP75、VDAC1 mRNA、蛋白表达及线粒体Ca
2+
含量低，MFN2 mRNA和蛋白表达高（
P
<
0.05）；与si-NC组比较，si-NC+中药组、si-GRP75组细胞存活率高，IP3R、GRP75、VDAC1 mRNA和蛋白表达低，线粒体Ca
2+
含量低，MFN2 mRNA和蛋白表达高（
P
<
0.05）；与si-NC+中药组比较，si-GRP75+中药组细胞存活率高，IP3R、GRP75、VDAC1 mRNA、蛋白表达及线粒体Ca
2+
含量低，MFN2 mRNA、蛋白表达高（
P
<
0.05）；si-GRP75组、si-GRP75+中药组、si-GRP75+卡托普利组细胞存活率、线粒体Ca
2+
含量差异无统计学意义（
P
>
0.05）。
结论
2
泻肺利水合剂可能通过调控IP3R/GRP75/VDAC1通路减轻ISO损伤大鼠心肌细胞线粒体钙超载。
Abstract
Objective
2
To investigate the protective effect of
Xiefei Lishui Mixture
containing serum on isoproterenol （ISO） injured rat cardiomyocytes （H9c2） and its mechanism.
Method
2
H9c2 cells were cultured in vitro to construct a cell model of ISO injury. First， the cells were divided into 4 groups： normal group， model group， Chinese medicine group and captopril group. The GRP75 gene was knocked down by plasmid transfection， and then the cells were divided into 6 groups： si-NC group， si-NC+ Chinese medicine group， si-NC+ captopril group， si-GRP75 group， si-GRP75+ Chinese medicine group and si-GRP75+ captopril group， which were cultured with corresponding serum. The cell viability of each group was detected dy CCK-8 method， the mRNA levels of IP3R， GRP75， VDAC1 and MFN2 were detected dy real-time fluorescence quantitative PCR， the protein expressions of IP3R， GRP75， VDAC1 and MFN2 were detected dy Western blot， and the mitochondrial Ca
2+
content was detected dy Rhod2-AM Ca
2+
fluorescent prode.
Result
2
Compared to the normal group， the model group exhibited increased mRNA and protein expressions of IP3R， GRP75， VDAC1， and mitochondrial Ca
2+
content （
P
<
0.05）. Conversely， the mRNA and protein expressions of MFN2 were decreased in the model group （
P
<
0.05）. In comparison to the model group， both traditional Chinese medicine and captopril groups showed reduced mRNA and protein expressions of IP3R， GRP75， VDAC1 as well as mitochondrial Ca
2+
content； meanwhile， they exhibited increased mRNA and protein expressions of MFN2 （
P
<
0.05）. Furthermore， compared t
o the si-NC group， both si-NC+traditional Chinese medicine group and si-GRP75 group demonstrated improved cell survival rate along with decreased levels of IP3R， GRP75 mRNA/protein expressions as well as mitochondrial Ca
2+
content； additionally， they displayed increased levels of MFN2 mRNA/protein expressions （
P
<
0.05）. Notably， the si-GRP75+traditional Chinese medicine group had a higher cell survival rate than the si-NC+traditional Chinese medicine group； it also exhibited lower levels of IP3R/GRP75/VDAC1 mRNA/protein expressions along with reduced mitochondrial Ca
2+
content while showing elevated levels of MFN2 mRNA/protein expressions （
P
<
0.05）. No significant differences were observed in terms of cell survival rate or mitochondrial Ca
2+
content among the si-GRP75 group，si-GRP75 + traditional Chinese medicine group or si-GRP75+captopril group（
P
>
0.05）.
Conclusion
2
Xiefei Lishui Mixture
can improve the survival rate of ISO injured rat H9c2 cells and reduce mitochondrial Ca
2+
overload， and its mechanism may relate to down regulation the IP3R/GRP75/VDAC1 pathway.
关键词
Keywords
references
中华医学会心血管病学分会心力衰竭学组 , 中国医师协会心力衰竭专业委员会 , 中华心血管病杂志编辑委员会 . 中国心力衰竭诊断和治疗指南2018 [J ] . 中华心力衰竭和心肌病杂志（中英文） , 2018 , 46 ( 10 ): 760 - 789 .
李超 , 王巍 , 杨雪卿 . 泻肺利水合剂对慢性心衰大鼠心肌氧化应激和线粒体自噬的影响 [J ] . 贵州医科大学学报 , 2020 , 45 ( 9 ): 1036 - 1041 .
来晓磊 , 尚菊菊 , 刘红旭 , 等 . 泻肺利水合剂联合西医常规方法治疗慢性缺血性心力衰竭气虚血瘀证疗效的真实世界、多中心、队列研究 [J ] . 中医杂志 , 2022 , 63 ( 6 ): 544 - 550 .
陈嘉兴 , 佟彤 , 杨志海 , 等 . 泻肺利水合剂对肥大心肌细胞线粒体MAO、COX活性的影响 [J ] . 中国生化药物杂志 , 2016 , 36 ( 9 ): 24 - 26 .
王亚君 , 常莉 , 姚天明 . 羌活醇对异丙肾上腺素诱导的H9c2心肌细胞损伤的保护作用 [J ] . 中国临床药理学杂志 , 2021 , 37 ( 15 ): 2015 - 2018 .
邱雨美 , 尹史帝 , 董伟 , 等 . 薯蓣皂苷对异丙肾上腺素诱导H9c2心肌细胞肥大的保护作用 [J ] . 中药药理与临床 , 2022 , 38 ( 2 ): 94 - 99 .
王华 , 刘宇佳 , 杨杰孚 . 心力衰竭流行病学 [J ] . 临床心血管病杂志 , 2023 , 39 ( 4 ): 243 - 247 .
SAVARESE G , BECHER PM , LUND LH , et al . Global burden of heart failure: a comprehensive and updated review of epidemiology [J ] . Cardiovasc Res , 2023 , 118 ( 17 ): 3272 - 3287 .
LI Y , SUN XL , QIU H , et al . Long-term outcomes and independent predictors of mortality in patients presenting to emergency departments with acute heart failure in Beijing: a multicenter cohort study with a 5-year follow-up [J ] . Chin Med J (Engl) , 2021 , 134 ( 15 ): 1803 - 1811 .
王丽 , 王峰 , 翟兴荣 . 中药复方治疗射血分数保留的心衰研究进展 [J ] . 中国中医急症 , 2023 , 32 ( 2 ): 373 - 376 .
孔吉武 , 李应宏 , 杨旺胜 , 等 . 中药复方治疗心力衰竭的研究进展分析 [J ] . 中国社区医师 , 2023 , 39 ( 1 ): 6 - 8 .
汤同娟 , 王翔 , 周鹏 , 等 . 基于Nrf2/BNIP3信号通路探讨苓桂术甘汤含药血清对心肌细胞线粒体氧化应激的影响 [J ] . 中国中药杂志 , 2022 , 47 ( 12 ): 3303 - 3311
黄凯 , 高佳明 , 贺爽 , 等 . 线粒体作为中药治疗心力衰竭靶点的研究进展 [J ] . 中国中药杂志 , 2020 , 45 ( 9 ): 2082 - 2090 .
中华中医药学会慢性心力衰竭中医诊疗指南项目组 . 慢性心力衰竭中医诊疗指南（2022年） [J ] . 中医杂志 , 2023 , 64 ( 7 ): 743 - 756 .
陈可冀 , 吴宗贵 , 朱明军 , 等 . 慢性心力衰竭中西医结合诊疗专家共识 [J ] . 中国中西医结合杂志 , 2016 , 36 ( 2 ): 133 - 141 .
许心如 , 魏执真 , 许信国 , 等 . 心衰合剂治疗充血性心力衰竭30例临床观察 [J ] . 中医杂志 , 1983 , 25 ( 11 ): 25 - 26 .
连妍洁 , 蒋燕君 , 佟彤 , 等 . 基于数据挖掘探析刘红旭治疗慢性心力衰竭用药规律 [J ] . 北京中医药 , 2022 , 41 ( 1 ): 82 - 87 .
来晓磊 , 邢文龙 , 刘红旭 , 等 . 泻肺利水法治疗心力衰竭的研究进展 [J ] . 北京中医药 , 2018 , 37 ( 6 ): 507 - 509 .
符竣杰 , 杨雪卿 , 李俊平 , 等 . 心衰合剂对慢性心力衰竭患者认知功能及焦虑抑郁状态的影响 [J ] . 北京中医药 , 2022 , 41 ( 12 ): 1368 - 1371 .
XU HX , CUI SM , ZhANG YM , et al . Mitochondrial Ca 2+ regulation in the etiology of heart failure: physiological and pathophysiological implications [J ] . Acta Pharmacol Sin , 2020 , 41 ( 10 ): 1301 - 1309 .
LI YE , SOWERS JR , HETZ C , et al . Cell death regulation by MAMs: from molecular mechanisms to therapeutic implications in cardiovascular diseases [J ] . Cell Death Dis , 2022 , 13 ( 5 ): 504 .
WANG P , XU S , XU J , et al . Elevated MCU expression by CaMKIIδB limits pathological cardiac remodeling [J ] . Circulation , 2022 , 145 ( 14 ): 1067 - 1083 .
VANCE JE . Phospholipid synthesis in a membrane fraction associated with mitochondria [J ] . J Biol Chem , 1990 , 265 ( 13 ): 7248 - 7256 .
MARCHI S , PATERGNANI S , PINTON P . The endoplasmic reticulum-mitochondria connection: one touch, multiple functions [J ] . Biochim Biophys Acta , 2014 , 1837 ( 4 ): 461 - 469 .
FILADI R , THEUREY P , PIZZO P . The endoplasmic reticulum-mitochondria coupling in health and disease: Molecules, functions and significance [J ] . Cell Calcium , 2017 , 62 ( 3 ): 1 - 15 .
YU J , LI Y , LIU X , et al . Mitochondrial dynamics modulation as a critical contribution for Shenmai Injection in attenuating hypoxia /reoxygenation injury [J ] . J Ethnopharmacol , 2019 , 237 ( 10 ): 9 - 19 ．
PANG LZ , JU AC , ZHENG XJ , et al . YiQiFuMai powder injection attenuates coronary artery ligation-induced myocardial remodeling and heart failure through modulating MAPKs signaling pathway [J ] . J Ethnopharmacol , 2017 , 8 ( 202 ): 67 - 77 .
SAUTULLI G , XIE W , REIKEN SR , et al . Mitochondrial calcium overload is a key determinant in heart failure [J ] . Proc Natl Acad Sci U S A , 2015 , 112 ( 36 ): 11389 - 11394 .
LU M , LENG B , HE X , et al . Calcium sensing receptor-related pathway contributes to cardiac injury and the mechanism of astragaloside IV on cardioprotection [J ] . Front Pharmacol , 2018 , 10 ( 11 ): 1 - 15 .
冯翠娜 , 赵子晖 , 胥磊 , 等 . 卡托普利改善大鼠心力衰竭的分子机制 [J ] . 临床和实验医学杂志 , 2020 , 19 ( 10 ): 1048 - 1053 .
EL-SAYED SF , ABDELHAMID AM , ZEINELABDEEN SG , et al . Melatonin enhances captopril mediated cardioprotective effects and improves mitochondrial dynamics in male Wistar rats with chronic heart failure [J ] . Sci Rep , 2024 , 14 ( 1 ): 575 .
0
浏览量
0
下载量
0
CSCD
- Meta-XML下载
- BibTeX下载
- Endnote下载
- RefMan 下载
- NoteExpress下载
- NoteFirst下载
关联资源
相关文章
相关作者
相关机构
- 技术支持由北京北大方正电子有限公司提供 京ICP备18046208号-2
京公网安备 11010102006710号 - 本系统建议在Chrome、 IE9+ 以上版本浏览器阅读本站内容，360浏览器请切换至极速模式
- Cookies帮助我们提供服务并提供个性化体验。使用本网站，即表示您同意我们使用Cookies
- 总访问量：200207 今日访问量：1