中国全科医学2016，Vol.19Issue(36)：4489-4494,6.DOI:10.3969/j.issn.1007-9572.2016.36.017
自噬在尿毒症小鼠心肌细胞损伤中的作用研究
Effect of Autophagy on Cardiac Myocytes Injury in Uremic Cardiomyopathy Mice
摘要
Abstract
Objective To explore the effect and mechanism of autophagy on cardiac myocytes injury in uremic cardiomyopathy mice.Methods From August to December 2015, fifty male C57BL/6-PAX6 mice in a SPF grade were selected in our study.The mice were randomly divided into control group ( 6 ) , sham operation group ( 8 ) , test groups 〔5/6 nephrectomy (NC) group (6), 5/6NC+saline group (10), 5/6NC+wortmannin group (10) and 5/6NC+Beclin-1 group (10)〕 .The control group did not receive any intervention in the study .The sham operation group only received surgery and anesthesia procedures simultaneous with the test groups , but the kidneys were not removed .5/6 mice nephrectomy model were <br> established in the test groups.Six weeks after model establishment , 5/6 NC group did not receive intervention.5/6NC+saline group was given 0.9%sodium chloride solution 1 ml intraperitoneal injection, 1 time every other day.5/6NC+wortmannin group was given wortmannin 1 mg/kg intraperitoneal injection , 1 time/day.5/6NC+Beclin -1 group was given adenovirus carrying Beclin-1 1 ×109 pfu intramuscular injection , 1 time/once.12 weeks after model establishment , mice heart function was measured by ultrasonic cardiograph , and renal function was measured by ELISA .In addition , we analyzed the expression of autophagy and its associated protein in mice myocardium using Western blotting and immunohistochemical staining after sacrificed.Results 12 weeks after model establishment , compared with control group and sham operation group , LVEDD significantly increased , LVFS significantly increased and serum creatinine and urea nitrogen level significantly increased in the 5/6NC group and 5/6NC+saline group (P<0.05) .Compared with 5/6 NC+saline group, LVEDD significantly decreased, LVFS significantly increased , serum creatinine and urea nitrogen level significantly decreased in the 5/6NC+wartmannin group ( P<0.05 ) .Compared with control group and 5/6 NC +saline group , LVEDD significantly increased , LVFS significantly decreased , serum creatinine and urea nitrogen level significantly increased in the 5/6 NC +Beclin -1 group ( P <0.05 ) . Immunohistochemical staining showed that the expression of Atg 5, Cathepsin -D and Rab 7 in the myocardial cells of 5/6NC group were significantly higher than that in control group (P<0.05) .There were significant differences in the rate of Atg5, Cathepsin-D and Rab7 positive cells between the control group and 5/6NC group ( P<0.05) .The expressions of autophagy -related protein LC3, Beclin-1 and AKT in the 5/6 NC+saline group and 5/6NC+Beclin-1 group were significantly higher than those in the control group (P<0.05) .The expressions of LC3, Beclin-1 and AKT in 5/6 NC+wortmannin penicillin group were higher than those in control group , but lower than those in 5/6NC+saline group, 5/6NC+Beclin-1 group ( P<0.05) .The expression of LC3, Beclin-1 and AKT in the 5/6 NC+Beclin-1 group were higher than those in 5/6 NC+saline group and 5/6 NC+wartmannin group ( P<0.05 ) .Conclusion Autophagy was closely related to cardiomyocytes injury in uremic mice.Enhancement or inhibition of autophagy could aggravate or relieve cardiac and renal function in uremic mice .Beclin-1 gene and PI3K/AKT signal proteins played an important role in regulation cardiomyocytes autophagy .关键词
心肌疾病/尿毒症/自噬/自噬相关蛋白/PI3K/AKT信号蛋白Key words
Cardiomyopathies/Uremia/Autophagy/Autophagy associated protein/PI3K/AKT signal protein分类
医药卫生引用本文复制引用
龚建光,金娟,赵黎,林波,梁世凯,何强,李一文..自噬在尿毒症小鼠心肌细胞损伤中的作用研究[J].中国全科医学,2016,19(36):4489-4494,6.基金项目
浙江省自然科学基金资助项目(Y2080371）；浙江省中医药管理局项目(2016ZA023） （）
