Lu Weifeng, He Ying, Yu Tao, Huang Keli, Liu Shengzhong
Sichuan Academy of Medical Sciences & Sichuan Provincial People's Hospital, Operation room, Chengdu, China.
Sichuan Academy of Medical Sciences & Sichuan Provincial People's Hospital, Psychosomatic Medicine Center, Chengdu, China.
Turk Gogus Kalp Damar Cerrahisi Derg. 2019 Jan 24;27(2):178-184. doi: 10.5606/tgkdc.dergisi.2019.17294. eCollection 2019 Apr.
This study aims to investigate the changes in miR-30a expression and myocardial autophagy following osthole treatment in a myocardial ischemia/reperfusion injury model.
Thirty male Wistar rats (weighing 170 to 220 g, aged 8 weeks) were randomly divided into three groups as control (sham) group, ischemia/reperfusion (model) group, and ischemia/ reperfusion + osthole post-treatment (osthole) group. Masson"s trichrome staining was used to detect myocardial collagen changes. Apoptotic cardiomyocytes in the ischemic area were labeled in situ by terminal deoxynucleotidyl transferase-mediated dUTP (2'-deoxyuridine 5'-triphosphate) nick end labeling assay. Levels of autophagy markers light chain 3 beta (LC3b) and Beclin-1 in myocardial tissue were detected by western blotting. Expression of miR-30a was detected by quantitative reverse transcriptionpolymerase chain reaction.
Compared with the sham group, ischemia/reperfusion significantly increased collagen contents. Osthole significantly inhibited the ischemia/reperfusion-increased collagen contents. Osthole inhibited the ischemia/reperfusion-increased myocardial fibrosis, myocardial swelling, necrosis, and myocardial atrophy. Osthole also significantly inhibited the ischemia/reperfusionincreased apoptosis of myocardial cells. Moreover, the conversion of LC3b-I to LC3b-II and the Beclin-1 expression were significantly inhibited by ischemia/reperfusion. Osthole treatment significantly increased the conversion of LC3b-I to LC3b-II and Beclin-1 expression in ischemia/reperfusion rats. Finally, the expression of miR-30a was significantly increased in ischemia/reperfusion rats, while Osthole suppressed the expression of miR-30a.
Osthole promoted autophagy, thereby alleviating myocardial ischemia/reperfusion injury. Osthole protects the myocardium during autophagy by down-regulating miR-30a expression.
本研究旨在探讨在心肌缺血/再灌注损伤模型中蛇床子素治疗后miR-30a表达及心肌自噬的变化。
将30只雄性Wistar大鼠（体重170至220克，8周龄）随机分为三组，即对照组（假手术组）、缺血/再灌注（模型）组和缺血/再灌注+蛇床子素后处理（蛇床子素）组。采用Masson三色染色法检测心肌胶原变化。通过末端脱氧核苷酸转移酶介导的dUTP缺口末端标记法对缺血区域的凋亡心肌细胞进行原位标记。采用蛋白质免疫印迹法检测心肌组织中自噬标志物轻链3β（LC3b）和Beclin-1的水平。采用定量逆转录聚合酶链反应检测miR-30a的表达。
与假手术组相比，缺血/再灌注显著增加了胶原含量。蛇床子素显著抑制了缺血/再灌注增加的胶原含量。蛇床子素抑制了缺血/再灌注增加的心肌纤维化、心肌肿胀、坏死和心肌萎缩。蛇床子素还显著抑制了缺血/再灌注增加的心肌细胞凋亡。此外，缺血/再灌注显著抑制了LC3b-I向LC3b-II的转化及Beclin-1的表达。蛇床子素治疗显著增加了缺血/再灌注大鼠中LC3b-I向LC3b-II的转化及Beclin-1的表达。最后，缺血/再灌注大鼠中miR-30a的表达显著增加，而蛇床子素抑制了miR-30a的表达。
蛇床子素促进自噬，从而减轻心肌缺血/再灌注损伤。蛇床子素通过下调miR-30a表达在自噬过程中保护心肌。
