小檗碱对心肌缺血再灌注损伤大鼠线粒体自噬及对PINK1/Parkin通路的影响

目的 探讨小檗碱对心肌缺血再灌注损伤大鼠线粒体自噬及PTEN诱导激酶1(PTEN induced putative kinase 1,PINK1)/帕金森病蛋白(Parkin)通路的影响.方法 建立心肌缺血再灌注损伤大鼠模型,随机分组为模型组、小檗碱低、高剂量(75、150 mg/kg)组,自噬抑制剂三甲基腺嘌呤(3-...

Effects of berberine on mitochondrial autophagy and PINK1/Parkin pathway in rats with myocardial ischemia-reperfusion

Objective To investigate the effects of berberine on mitochondrial autophagy and phosphatase and tensin homology deleted on chromosome ten (PTEN)-induced putative kinase 1 (PINK1)/Parkin pathway in rats with myocardial ischemia-reperfusion (MIR). Methods MIR rat model was established and randomly divided into model group, low-dose berberine group (75 mg/kg), high-dose berberine group (150 mg/kg), autophagy inhibitor trimethyladenine (3-MA, 100 mmol/L), berberine + autophagy inhibitor group (150 mg/kg + 100 mmol/L), with 12 rats in each group, and another 12 rats were set as Sham operation group. After grouping and treatment, left ventricular function was detected by echocardiography, left ventricular end systolic diameter (LVESD), left ventricular end diastolic diameter (LVEDD), left ventricular ejection fraction (LVEF) and left ventricular short axis shortening (FS) were recorded. The infarct size was detected by 2,3,5-triphenyltetrazolium chloride (TTC) staining, and the levels of creatine kinase isoenzyme (CK-MB) and troponin I (cTnI) in serum were detected by enzyme-linked immunosorbent assay (ELISA). HE staining was used to observe the pathological changes of myocardium. The ultrastructure and mitochondrial autophagy of cardiomyocytes were observed by transmission electron microscopy (TEM), and mitochondrial damage score was analyzed. The protein expressions of PINK1, Parkin, microtubule associated protein 1 light chain 3B (LC3B), mitochondrial autophagy receptor p62 and ubiquitin specific protease 30 (USP30) were detected by Western blotting. Results Compared with those in the Sham operation group, the pathological damage of myocardial tissue in the model group was serious, and the swelling and vacuolation of mitochondria were more serious, the mitochondrial injury score, myocardial infarction area, LVEDD, LVESD, levels of CK-MB and cTnI, the protein expression levels of PINK1, Parkin, LC3B and p62 were higher (P<0.05), but LVEF, the protein expression levels of FS and USP30 were lower (P<0.05). Compared with those in the model group, the pathological damage of myocardial tissue and mitochondria in 3-MA group was aggravated, LVEF, FS, the protein expression levels of PINK1, Parkin, LC3B and p62 were lower (P<0.05), the mitochondrial injury score, myocardial infarction area, LVEDD, LVESD, levels of CK-MB and cTnI and the protein expression level of USP30 were higher (P<0.05). The pathological damage of myocardial tissue and mitochondria in rats in low and high dose berberine groups was alleviated, LVEF, FS, the protein expression levels of PINK1, Parkin, LC3B, p62 and USP30 were higher (P<0.05), the mitochondrial damage score, myocardial infarction area, LVEDD, LVESD, levels of CK-MB and cTnI were lower (P < 0.05). The changes of the above indexes in berberine + 3-MA group were contrary to those in high-dose berberine group, and the difference was statistically significant (P<0.05). Conclusion Berberine may promote mitochondrial autophagy by activating PINK1/Parkin/p62/LC3B pathway, increase USP30 expression, reduce abnormal autophagy and alleviate MIR injury.