Autophagy Involved In The Myocardial Fibrosis Of The P-selectin Gene Knockout Mice

Objective:To investigate the effect of P-selectin on angiotensin Ⅱ-induced myocardial fibrosis and the possible relationship with autophagy by establishing of the angiotensin Ⅱ-induced myocardial fibrosis model of P-selectin gene knockout mice.Methods:1. Select40weight matching18-20g wild C57BL/6mice (wild type) and P-selectin gene knockout mice (P-selectin-/-) of the same genetic background, divide into four groups randomly,(n=10), wild C57BL/6mice+angiotensin Ⅱ (1000ng/min*kg,7d)(positive control), P-selectin-/-+angiotensin Ⅱ (1000ng/min*kg,7d)(experimental group), wild C57BL/6mice+PBS (negative control), P-selectin-/-+PBS (negative control).Establish cardiac fibrosis model through Angiotensin Ⅱ micro pump infusion (7days), determine the success of the angiotensin Ⅱ infusion by checking blood pressure3days before and4-7days after the surgery.7days later, take off the heart by infusing heparin saline.2. Observe cardiac fibrosis by Masson’s staining calculate the proportion of the fibrosis areas in the overall area of the heart slices with NIS-ELEMENT software, analyze the difference between each group quantitatively.3.Western blot was used to detect the expression of LC3and Beclinl, the autophagy-related proteins.Results:1. Masson’staining of the Myocardial tissue results:the degree of myocardial fibrosis was significantly increased in the positive control group and experimental group compared with the negative control group (P<0.05). Compared with the positive control group,the degree of myocardial fibrosis in the experimental group significantly decreased (P<0.05).2. The results of Western blot:The expression of LC3and Beclinl in the positive control group and experimental group were significantly higher than that of the negative control group (P<0.05).Compared with the the positive control group, the expression of LC3and Beclinl in the experimental group decreased significantly(P<0.05).Conclusions:1. Ang Ⅱ can induce the myocardial fibrosis.2. P-selectin gene knockout can inhibit the angiotensin Ⅱ-induced myocardial fibrosis.3.P-selectin can aggravate myocardial fibrosis through promoting autophagy.