甲泼尼龙对高糖腹膜透析液诱导人腹膜间皮细胞表达纤维化相关细胞因子的影响  

Effect of methylprednisolone on expression of fibrosis-related cytokines in human peritoneal mesothelial cells induced by high glucose peritoneal dialysate

作　　者：蔡家驹 戴卫波[1] 吴惠妃[1] 叶秋明[1] 萧俊祺 CAI Jiaju;DAI Weibo;WU Huifei;YE Qiuming;XIAO Junqi(Department of Clinical Pharmacy,Zhongshan Hospital of Traditional Chinese Medicine,Zhongshan 528400,China)

机构地区：[1]中山市中医院临床药学室,广东中山528400

出　　处：《山东医药》2021年第22期37-41,共5页Shandong Medical Journal

基　　金：中山市医学科研项目(2019A020479)。

摘　　要：目的观察甲泼尼龙(MP)对高糖腹膜透析液(PDS)诱导人腹膜间皮细胞(HMPCs)纤维化相关细胞因子表达的影响。方法体外培养HPMCs,随机分为空白组(10%FBS培养液)、模型组(4.25%PDS和10%FBS培养液)和MP低、中、高剂量组(含MP浓度为0.1、0.2、0.4μg/mL的4.25%PDS和10%FBS培养液)。采用CCK-8法检测细胞存活率;MTT法检测各组细胞增殖活性;流式细胞仪检测细胞内ROS水平;RT-PCR法检测IL-6、α-SMA、HA的mRNA表达;Westernblotting法检测核转录因子(NF-κB)、转化生长因子β1(TGF-β1)、结缔组织生长因子(CTGF)、血管内皮生长因子(VEGF)、α-平滑肌肌动蛋白(α-SMA)、透明质酸(HA)、Smads蛋白表达。结果MP浓度在0.032~0.256μg/mL时,HPMCs存活率升高(P<0.05);MP对HPMCs的半抑制率(IC_(50))为0.842μg/mL。与空白组相比,模型组增殖活性减弱(P<0.05),ROS、IL-6、NF-κB、TGF-β1、CTGF、VEGF、α-SMA、HA、p-Smad2/3、Smad7的表达增强(P均<0.05);与模型组相比,ROS在MP中、高剂量组的表达减少(P均<0.05),IL-6、NF-κB、TGF-β1、CTGF、VEGF、α-SMA、HA、p-Smad2/3、Smad7在MP三个剂量组表达下调(P均<0.05)。结论MP通过减少高糖PDS诱导HPMCs纤维化相关细胞因子的表达,发挥抗PF的作用。Objective To investigate the effect of methylprednisolone(MP)on the expression of fibrosis-related cytokines in human peritoneal mesothelial cells(HPMCs)induced by high glucose peritoneal dialysate(PDS).Methods HPMCs were cultured in vitro and randomly divided into five groups:the blank group which contained 10% FBS only,the model group which contained 4.25% PDS and 10% FBS,and low-,medium-and high-dose MP groups which contained 0.1,0.2 and 0.4μg/mL MP as well as 4.25%PDS and 10%FBS.The cytotoxicity of MP was detected by CCK-8;the activity of HPMCs was detected by MTT;the level of ROS was determined by flow cytometry;the mRNA expression levels of IL-6,α-SMA and HA were detected by RT-PCR;the protein expression levels of nuclear factor-κB(NF-κB),transforming growth factor-β1(TGF-β1),vascular endothelial growth factor(VEGF),connective tissue growth factor(CTGF),Smad 2/3,p-smad 2/3 and Smad7 were detected by Western blotting.Results Survivals of HPMCs increased significantly at the concentrations of MP from 0.032-0.256μg/mL(P<0.05).The IC_(50) of MP was 0.842μg/mL.Compared with the blank group,the HPMCs activity of the model group decreased,and the expression levels of ROS,IL-6,NF-κB,TGF-β1,CTGF,VEGF,α-SMA,HA,p-Smad2/3,and Smad7 increased significantly(all P<0.05).Compared with the model group,the expression levels of IL-6,NF-κB,TGF-β1,CTGF,VEGF,α-SMA,HA,p-Smad2/3,and Smad7 decreased significantly in the low-,medium-,and high-dose MP groups,while the expression of ROS decreased only in the medium-and high-dose PM groups(all P<0.05).Conclusion MP plays a role of anti-PF via decreasing the expression of fibrosis-related cytokines in HPMCs induced by high glucose PDS.

关 键 词：肾脏疾病 腹膜透析 甲泼尼龙 人腹膜间皮细胞 腹膜纤维化 

分 类 号：R692[医药卫生—泌尿科学]