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中国人民解放军总医院老年心血管病研究所
中国科技出版传媒股份有限公司
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中华老年多器官疾病杂志编辑委员会
100853, 北京市复兴路28号
电话:010-66936756
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E-mail: zhlndqg@mode301.cn
创刊人 王士雯
总编辑 范利
副总编辑 陈韵岱
执行主编 叶大训
编辑部主任 王雪萍
ISSN 1671-5403
CN 11-4786
创刊时间 2002年
出版周期 月刊
邮发代号 82-408
友情链接
向家培,雷玉华.栀子苷抑制高糖诱导的乳鼠心脏成纤维细胞向肌成纤维细胞的表型转化[J].中华老年多器官疾病杂志,2019,18(9):699~704
栀子苷抑制高糖诱导的乳鼠心脏成纤维细胞向肌成纤维细胞的表型转化
Geniposide suppresses phenotypic transformation of neonatal rat cardiac fibroblasts into myofibroblasts induced by high glucose
投稿时间:2019-04-22  
DOI:10.11915/j.issn.1671-5403.2019.09.151
中文关键词:  栀子苷;高糖;心肌成纤维细胞;糖尿病心肌病
英文关键词:geniposide; high glucose; cardiac fibroblast; diabetic cardiomyopathy
基金项目:
作者单位E-mail
向家培 恩施土家族苗族自治州中心医院心血管内科,恩施 445000  
雷玉华 恩施土家族苗族自治州中心医院心血管内科,恩施 445000 huayulei_0319@126.com 
摘要点击次数: 60
全文下载次数: 101
中文摘要:
      目的 探讨栀子苷(GE)对高糖诱导过程中体外培养的乳鼠心脏成纤维细胞(CF)表型转化及胶原合成的影响及机制。方法 提取新生大鼠原代CF,给予高糖刺激(葡萄糖浓度为33.3mmol/L)和GE干预,24h后,应用逆转录聚合酶链反应(RT-PCR)测定CF细胞内Ⅰ型胶原(Col Ⅰ)、Col Ⅲ及结缔组织生长因子(CTGF)的mRNA表达水平;试剂盒检测氧化应激相关指标,包括超氧化物歧化酶(SOD)、还原型烟酰胺腺嘌呤二核苷酸磷酸(NADPH)活性及丙二醛(MDA)的产量;Western blot检测转化生长因子β(TGF-β)、乙酰化的Smad3(ac-Smad3)、α-平滑肌肌动蛋白(α-SMA)及沉默信息调节因子1(SIRT1)蛋白质的表达水平。给予SIRT1抑制剂EX-527后,采用免疫荧光共染SIRT1及α-SMA。再次检测氧化应激及ac-Smad3信号通路相关蛋白的表达。采用SPSS 22.0统计软件进行统计学分析。结果 不同浓度的GE处理高糖刺激后的大鼠CF 24h后,Col Ⅰ、Ⅲ及CTGF mRNA表达有所降低,且随GE浓度的增加,抑制作用增强,在1~100μmol/L之间呈现浓度依赖性。GE可明显降低高糖诱导过程中NADPH的活性及MDA的含量,增强SOD活性。Western blot结果显示,高糖诱导下,TGF-β、ac-Smad3及α-SMA 水平明显升高,而GE能明显抑制这3种蛋白的升高。同时,GE明显逆转了高糖条件下SIRT1的下调。免疫荧光结果显示,应用SIRT1抑制剂EX-527后,GE对高糖条件下α-SMA及氧化应激的抑制作用消失。结论 GE能够抑制高糖诱导的大鼠CF表型转化及胶原合成,其机制可能与SIRT1介导的TGF-β/ac-Smad3信号通路及氧化应激有关。
英文摘要:
      Objective To investigate the effect of geniposide (GE) on phenotypic transformation of neonatal rat cardiac fibroblasts (CF) as well as on collagen synthesis under the induction of high glucose (HG), and explore the underlying mechanisms. Methods Primary CF were extracted from neonatal rats, and then treated in presence or absence of 33.3mmol/L HG or GE for 24h. The mRNA levels of collagen Ⅰ (Col I), collagen Ⅲ (Col Ⅲ) and connective tissue growth factor (CTGF)were detected by reverse transcription-polymerase chain reaction (RT-PCR). The markers of oxidative stress, including superoxide dismutase (SOD), reduced nicotinamide adenine dinucleotide phosphate (NADPH) and malondialdehyde (MDA) were measured by corresponding test kits. The protein levels of transforming growth factor-beta (TGF-β), acetylated-Smad3 (ac-Smad3), α-smooth muscle actin (SMA) and silent information regulator 1 (SIRT1) were detected using Western blotting. After SIRT1 inhibitor, EX-527 was added, the expression of SIRT1 and α-SMA, activities and production of oxidative stress markers, and protein levels of ac-Smad3 signal pathway related proteins were observed and measured to further explore the protective mechanisms of GE. SPSS 22.0 statistics was used to analyze the data. Results Treatment of GE at different concentrations (1-100μmol/L) for 24h resulted in decreased mRNA levels of Col Ⅰ, Col Ⅲ and CTGF in a dose-dependent manner. Meantime, GE significantly reduced the activity of NADPH, decreased the production of MDA and enhanced the activity of SOD in the process of HG induction. Western blotting indicated that HG treatment induced up-regulation of TGF-β, ac-Smad3 and α-SMA, while GE obviously inhibited the elevations and reversed the down-regulation of SIRT1 induced by HG. Immunofluorescence assay showed that the inhibitor EX-527 could block the inhibitory effect of GE on HG-induced α-SMA and oxidative stress. Conclusion GE exerts inhibitory effect on high HG-induced phenotypic transformation of CF as well as collagen synthesis, which may be associated with TGF-β/ac-Smad3 signal pathway and oxidative stress mediated by SIRT1.
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