二苯乙烯苷调控Klotho蛋白抑制CKD小鼠肾纤维化  

Inhibition of tetrahydroxystilbene glucoside on renal fibrosis in mice with chronic kidney disease via regulating soluble Klotho protein expression

作　　者：张冬[1] 李宗英 高原[1] 肖娜[1] ZHANG Dong;LI Zongying;GAO Yuan;XIAO Na(Department of Nephrology,Cangzhou Central Hospital,Cangzhou 061001,China)

机构地区：[1]沧州市中心医院肾内一科,河北沧州061001

出　　处：《生物技术》2024年第4期497-503,共7页Biotechnology

基　　金：沧州市重点研发计划指导项目(213106065)。

摘　　要：[目的]探究二苯乙烯苷(TSG)调控Klotho蛋白对慢性肾脏病(CKD)小鼠肾纤维化的影响。[方法] 30只C57BL/6J小鼠随机分为对照组(Control)、模型组(CKD)、模型+二苯乙烯苷处理组(CKD+T),取肾组织检测各组小鼠肾功能[24 h尿蛋白(UP)、24 h尿白蛋白(UA),血肌酐(SCr),尿素氮(BUN)]、氧化应激指标[肾超氧化物歧化酶(SOD),丙二醛(MDA),尿8-羟基脱氧鸟苷(8-ohdg)],RT-PCR检测Klotho mRNA表达,Western Blot检测肾组织纤维化相关因子[中性粒细胞明胶酶相关脂质运载蛋白(NGAL),纤连蛋白(FN),结缔组织生长因子(CTGF)]及Klotho蛋白表达。[结果]与Control组相比,CKD组、CKD+T组的24 h UP、24 h UA、ACR、SCr、BUN、尿8-ohdg、肾MDA、NGAL、FN、CTGF蛋白表达水平均升高,肾SOD、Klotho mRNA及蛋白表达水平降低,且CKD+T组的24 h UP、24 h UA、ACR、SCr、BUN、肾MDA、尿8-ohdg含量、肾小管损伤评分、胶原纤维阳性面积(14.1±2.7 vs 6.4±1.5)%、NGAL、FN、CTGF蛋白表达水平低于CKD组,肾SOD、Klotho mRNA及蛋白表达量(1.21±0.23 vs 1.46±0.29,1.18±0.16 vs 1.37±0.21)高于CKD组(P<0.05)。[结论] TSG可改善CKD小鼠肾功能,抑制氧化应激反应和延缓肾纤维化进程,可能是通过上调Klotho蛋白表达来起作用的。[Objective]To explore the effect of tetrahydroxystilbene glucoside(TSG) on renal fibrosis in mice with chronic kidney disease(CKD) via regulating soluble Klotho protein expression.[Method]Thirty C57BL/6J mice were randomized to one of three interventions: 1) control(Control group),2) CKD model construction(CKD group) and 3) CKD model construction+TSG treatment(CKD+T group).Renal tissues were collected to measure renal function parameters[24 hours urinary protein(UP),24 hours urinary albumin(UA),serum creatinine(SCr),blood urea nitrogen(BUN),urinary albumin/creatinine ratio(ACR)] and oxidative stress indexes[superoxide dismutase(SOD),malondialdehyde(MDA),8-hydroxydeoxyguanosine(8-ohdg)] of mice in each group.Klotho mRNA expression was detected by real time polymerase chain reaction(RT-PCR),and the protein expression levels of fibrosis-related factors[neutrophil gelatinase-associated lipocalin(NGAL),fibronectin(FN),connective tissue growth factor(CTGF)] and Klotho protein expression were tested by Western Blot.[Result] Compared with control group, CKD group and CKD+T group were found to have an elevation in the values of 24 hours UP,24 hours UA,ACR,SCr, BUN,urinary 8-ohdg and renal MDA as well as the protein expression of NGAL,FN,and CTGF,and also reported a decrease in renal SOD plus the mRNA and protein expression of Klotho.In addition, the values of 24 hours UP,24 hours UA,ACR,SCr, BUN,renal MDA and urinary 8-ohdg content, the renal tubular injury score, the collagen fiber positive area(14.1±2.7 vs 6.4±1.5)%,along with the protein expression of NGAL,FN,and CTGF were lower in CKD+T group than in CKD group, and the renal SOD,Klotho mRNA expression(1.21±0.23 vs 1.46±0.29) and Klotho protein expression(1.18±0.16 vs 1.37±0.21) were higher in CKD+T group than in CKD group, with statistical difference(all P<0.05).[Conclusion] TSG can inhibit the expression of fiber-related factors and delay the process of renal fibrosis in CKD mice, which may be related to the down-regulation of soluble Klotho protein expression.

关 键 词：二苯乙烯苷 KLOTHO 慢性肾脏病 肾纤维化 肾小管损伤 氧化应激 血肌酐 尿素氮 衰老 

分 类 号：R692.5[医药卫生—泌尿科学]