摘要
目的探讨丹参酮ⅡA(TⅡA)对miR-148-5p过激活NF-kB通路引起缺氧/复氧(H/R)大鼠心肌细胞损伤和炎性反应的影响。方法CCK-8检测TⅡA对心肌细胞增殖和活力;RT-qPCR检测miR-148-5p、Bcl-2、Bax和casepase 3的表达;ELISA检测TNF-ɑ、IL-17、IL-4、IL-10的表达;Westen Blotting和免疫荧光检测NF-kB和凋亡的表达。结果高剂量组的TⅡA在72 h对心肌细胞增殖最佳;与正常组相比,H/R组Bax、casepase3、TNF-ɑ、IL-17、p-p50、p-p65表达升高(P<0.05),Bcl-2、IL-4、IL-10、表达降低(P<0.05);与H/R组相比,H/R+miR-148-5p组Bax、casepase 3、TNF-ɑ、IL-17、IKBKG、p-p50、p-p65表达显著升高(P<0.05),Bcl-2、IL-4、IL-10、表达显著降低(P<0.05);与H/R+miR-148-5p组相比,TⅡA干预后,Bax、casepase 3、TNF-ɑ、IL-17、IKBKG、p-p50、p-p65表达显著降低(P<0.05),Bcl-2、IL-4、IL-10表达显著升高(P<0.05)。结论TⅡA可以保护心肌细胞,可能与其通过调控miR-148-5p,抑制NF-kB通路过激活,抑制细胞凋亡,下调促炎因子、上调抑炎因子,减轻炎症反应有关。
Objective To investigate the influence of tanshinoneⅡA(TⅡA)on injury and inflammatory response of cardiomyocytes induced by miR-148-5p over-activating NF-kB signaling pathway in hypoxia/reoxygenation(H/R)rats Methods The effect of TIIA on cardiomyocyte proliferation and activity was detected by using CCK-8.The expressions of miR-148-5p,Bcl-2,Bax and casepase3 were detected by using RT-qPCR.The expressions of TNF-ɑ,IL-17,IL-4 and IL-10 were detected by using ELISA.The expression of NF-kB was detected by using Western blotting assay and immunofluorescence.Results TⅡA in a high dose had the best effect on cardiomyocyte proliferation at 72 h.Compared with normal group,the expressions of Bax,casepase3,TNF-ɑ,IL-17,p-p50 and p-p65 increased(P<0.05),and expressions of Bcl-2,IL-4 and IL-10 decreased(P<0.05)in H/R group.Compared with H/R group,the expressions of Bax,casepase3,TNF-ɑ,IL-17,IKBKG,p-p50 and p-p65 increased significantly(P<0.05),and expressions of Bcl-2,IL-4 and IL-10 decreased significantly(P<0.05)in H/R+miR-148-5p group.Compared with H/R+miR-148-5p group,the expressions of Bax,casepase3,TNF-ɑ,IL-17,IKBKG,p-p50 and p-p65 decreased significantly(P<0.05),and expressions of Bcl-2,IL-4 and IL-10 increased significantly(P<0.05)after TⅡA intervention.Conclusion TⅡA can protect cardiomyocytes may through regulating miR-148-5p,inhibiting over-activating of NF-kB signaling pathway,inhibiting apoptosis,down-regulating pro-inflammatory factors,up-regulating anti-inflammatory factors,and relieving inflammatory response.
作者
张亚文
唐海沁
黄荣彩
Zhang Yawen;Tang Haiqin;Huang Rongcai(Department of Cardiovascular Medicine,First Affiliated Hospital of Anhui Medical University,Hefei 230032,China;不详)
出处
《中国循证心血管医学杂志》
2020年第12期1447-1449,1457,共4页
Chinese Journal of Evidence-Based Cardiovascular Medicine
基金
国家中医药管理局国家中医临床研究基地业务建设科研专项课题(JDZX2015133)。
