心阳片含药血清调控MLK3介导的铁死亡对心肌细胞的保护作用及机制研究  

Protective Effect and Mechanism of Xinyang Tablet(心阳片)-containing Serum Regulating MLK3-mediated Ferroptosis on Cardiomyocytes

作　　者：王俊岩 于忠杨 郭依宁 邓波 王陵军[2,3] 杨忠奇 冼绍祥[2,3] 黄育生 WANG Junyan;YU Zhongyang;GUO Yining;DENG Bo;WANG Lingjun;YANG Zhongqi;XIAN Shaoxiang;HUANG Yusheng(School of Pharmaceutical Sciences,Guangzhou University of Chinese Medicine,Guangzhou 510006,Guangdong,China;The First Affiliated Hospital of Guangzhou University of Chinese Medicine,Guangzhou 510405,Guangdong,China;Lingnan Medical Research Center,Guangzhou University of Chinese Medicine,Guangzhou 510405,Guangdong,China;Qingdao No.6 People’s Hospial,Qingdao 266033,Shandong,China)

机构地区：[1]广州中医药大学中药学院,广东广州510006 [2]广州中医药大学附属医院,广东广州510405 [3]广州中医药大学岭南医学研究中心,广东广州510405 [4]青岛市第六人民医院,山东青岛266033

出　　处：《中华中医药学刊》2022年第2期32-35,I0013,I0014,共6页Chinese Archives of Traditional Chinese Medicine

基　　金：国家自然科学基金(81804048,81973776,81973777,81673796);中国博士后科学基金面上项目(2021M700966);广州市科技计划(202102020123);广州市慢性心力衰竭中医药防治重点实验室项目(201705030006)。

摘　　要：目的探讨心阳片含药血清调控MLK3介导的铁死亡对心肌细胞的保护作用及机制。方法20只SD大鼠随机分为对照组和含药血清组,含药血清组按照人与动物体表面积等效剂量换算,给予心阳片0.24 g/kg进行灌胃,每日2次,对照组给予等量生理盐水灌胃,连续灌胃7 d后制备血清。将心肌细胞HL-1分为空白组(Control)、FIN56组、FIN56+URMC-099组(FIN56+U-099)、FIN56+心阳片含药血清组(FIN56+XYP)。收集上述各组细胞,ROS荧光染色检测HL-1细胞活性氧水平,测定HL-1细胞MDA、T-SOD和GSH表达水平,RT-PCR检测各组细胞ANP和BNP mRNA水平,Western Blot法检测各组细胞MLK3、p53、GPX4和FTH1蛋白表达水平。结果与空白组相比,FIN56组细胞ROS、MDA表达水平显著升高(P<0.01),T-SOD和GSH表达水平降低(P<0.01)。ANP和BNP mRNA表达水平显著升高(P<0.01),MLK3和p53蛋白表达水平显著升高(P<0.01),GPX4和FTH1蛋白表达水平显著降低(P<0.01);与FIN56组比较,给予URMC-099和心阳片含药血清能降低ROS、MDA表达水平(P<0.01),升高T-SOD和GSH表达水平(P<0.01)。显著抑制ANP和BNP mRNA的表达(P<0.01)以及MLK3和p53蛋白表达(P<0.01),激活GPX4和FTH1蛋白表达(P<0.01)。结论心阳片含药血清可能通过调控MLK3,抑制心肌细胞铁死亡改善心肌损伤。Objective To explore the protective effect and mechanism of Xinyang Tablet(心阳片)-containing serum regulating MLK3-mediated ferroptosis on cardiomyocytes.Methods Twenty SD rats were randomly divided into control group and Xinyang Tablet-containing serum group.According to the equivalent dose of human and animal body surface area,Xinyang Tablet(0.24 g/kg)was given by gavage,twice a day.The control group was given the same amount of normal saline,and the serum was prepared after continuous gavage for 7 days.HL-1 was divided into control group,FIN56 group,FIN56+URMC-099 group(FIN56+U-099)and FIN56+Xinyang Tablet-containing serum group(FIN56+XYP).ROS fluorescence staining detected the level of reactive oxygen species and the expression levels of MDA,T-SOD and GSH in HL-1 cells.RT-PCR was used to detect the levels of ANP and BNP mRNA,and Western Blot method was used to detect the protein expression levels of MLK3,p53,GPX4 and FTH1.Results Compared with those of the control group,the expression levels of ROS and MDA in the FIN56 group were increased significantly(P<0.01).The expression levels of T-SOD and GSH were decreased(P<0.01)and the expressions of ANP and BNP mRNA significantly increased(P<0.01).MLK3 and p53 protein expression levels were increased(P<0.01)and the protein expression levels of GPX4 and FTH1 significantly decreased(P<0.01).Compared with the FIN56 group,FIN56+U-099 and FIN56+XYP group can significantly reduce the expression levels of ROS and MDA(P<0.01),increase the expression levels of T-SOD and GSH(P<0.01),inhibit the expressions of ANP and BNP mRNA(P<0.01)and expressions of MLK3 and p53 protein(P<0.01)and activate the expressions of GPX4 and FTH1 protein(P<0.01).Conclusion Xinyang Tablet-containing serum may improve myocardial injury by regulating MLK3 and inhibiting ferroptosis of myocardial cells.

关 键 词：心阳片 铁死亡 慢性心力衰竭 MLK3 

分 类 号：R285.5[医药卫生—中药学]