铁死亡抑制剂预培养的大鼠心肌细胞抵抗素诱导后细胞肥大程度及铁死亡相关蛋白表达观察  

Effects of resistin on cell hypertrophy and expression of ferroptosis-related protein in rat cardiomyocytes precultured with forrostatin-1

作　　者：朱贲贲 海日汗 王巍嵩[4] 杨鹏杰 ZHU Benben;Hairihan;WANG Weisong;YANG Pengjie(Department of Pharmacy,Inner Mongolia Autonomous Region Cancer Hospital,Hohhot 010020,China;不详)

机构地区：[1]内蒙古自治区肿瘤医院药剂科,呼和浩特010020 [2]内蒙古医科大学附属人民医院药剂科 [3]内蒙古医科大学麻醉系 [4]内蒙古医科大学附属医院药学部 [5]内蒙古自治区肿瘤医院胸外科 [6]内蒙古医科大学附属人民医院胸外科

出　　处：《山东医药》2023年第6期24-27,共4页Shandong Medical Journal

基　　金：内蒙古医科大学联合项目(YKD2022LH005);内蒙古自治区自然科学基金项目(2020MS08107;2020MS08187)。

摘　　要：目的观察铁死亡抑制剂Fer-1预培养的大鼠心肌细胞系H9c2抵抗素诱导后细胞肥大程度及铁死亡相关蛋白的表达情况。方法将H9c2细胞饥饿培养16 h,随机分为1、2、3、4组。1组用含1μmmol/L的Fer-1的培养基培养16 h,后换为含50 ng/mL抵抗素的培养基培养32 h;2组用含1μmmol/L的Fer-1培养基培养16 h,后换为0.1%FBS培养基培养32 h;3组用含50 ng/mL抵抗素的培养基培养48 h;4组(正常对照组)用0.1%FBS培养基培养48 h。取各组细胞,采用Image J软件测量细胞表面积,采用BCA法测算单细胞蛋白含量,采用RT-qPCR法检测心肌细胞肥大相关胚胎基因ANP、BNP和β-MHC;采用WESTERN Blotting法检测各组铁死亡相关蛋白谷胱甘肽过氧化物酶4(Glutathione Peroxidase 4,GPX4)、酰基辅酶A合成酶长链家族成员4(Acyl-CoA synthetase long-chain family member,ACSL4)及环氧合酶2(COX2)。结果与4组比较,3组H9c2细胞表面积增大,单细胞蛋白含量高,ANF、BNF及β-MHC基因相对表达量高,GPX4蛋白相对表达量低、ACSL4及COX2蛋白相对表达量高(P均<0.05)。与3组相比,1组细胞表面积小,单细胞蛋白含量低,ANF、BNF及β-MHC基因相对表达量低(P均<0.05);与3组相比,1组细胞GPX4蛋白相对表达量高、ACSL4及COX2蛋白相对表达量低(P均<0.05)。结论Fer-1能抑制抵抗素诱导的H9c2细胞肥大。Fer-1可能通过促进铁死亡相关蛋白GPX4表达、抑制ACSL4及COX2蛋白表达,抑制抵抗素诱导的H9c2肥大。Objective To observe the degree of hypertrophy and the expression of ferroptosis-related proteins in H9C2 resistin-induced rat cardiomyocytes pre-cultured with ferroptosis inhibitor ferrostatin-1(Fer-1).Methods H9c2 cells were starved for 16 h and randomly divided into four groups.H9c2 cells in the group 1 were cultured in the medium containing 1μmmol/L Fer-1 for 16 h,and then in the medium containing 50 ng/mL resistin for 32 h.H9c2 cells in the group 2 were cultured in Fer-1 medium containing 1μmmol/L for 16 h and then in 0.1%FBS medium for 32 h.H9c2 cells in the group 3 were cultured in medium containing 50 ng/mL resistin for 48 h.H9c2 cells in the group 4(normal con⁃trol group)were cultured with 0.1%FBS for 48 h.After cells in each group were cultured,and the cell surface area was measured by ImageJ software,the single cell protein content was measured by BCA method,and the cardiomyocyte hy⁃pertrophy-related genes ANP,BNP andβ-MHC were detected by RT-qPCR;ferroptosis-related proteins glutathione peroxi⁃dase 4(GPX4),acyl-CoA synthetase long-chain family member 4(ACSL4)and cyclooxygenase 2(Cox2)were detected by Western blotting.Results Compared with the group 4,the surface area of H9c2 cells in the group 3 increased significantly,the content of single-cell protein increased significantly,the expression levels of embryonic genes ANF,BNF,andβ-MHC in the group 3 were significantly up-regulated,the expression of GPX4 protein was significantly down-regulated,and the protein expression levels of ACSL4 and COX2 were significantly up-regulated(all P<0.05).Compared with the group 3,the surface area of H9c2 cells in the group 1 decreased significantly,the content of single-cell proteins decreased significantly,and the expression levels of embryonic genes ANF,BNF,andβ-MHC in the group 1 were significantly re⁃duced,the expression of GPX4 protein was significantly up-regulated,and the expression levels of ACSL4 and COX2 pro⁃teins were significantly down-regulated(all P<0.05).Conclusions Fer-1 could inhibit H9C

关 键 词：铁死亡 抵抗素 心肌肥厚 心肌细胞肥大 谷胱甘肽过氧化物酶4 酰基辅酶A合成酶长链家族成员4 环氧合酶2 

分 类 号：R544[医药卫生—心血管疾病]