积雪草苷调节JAK/STAT信号通路对糖尿病肾病大鼠炎症反应的影响

Effect of asiaticoside on inflammatory response in diabetic nephropathy rats by regulating JAK/STAT signaling pathway

目的探讨积雪草苷(ASI)调节Janus激酶(JAK)/细胞信号转导转录因子(STAT)信号通路对糖尿病肾病(DN)大鼠炎症反应的影响。方法30只SD大鼠随机取6只作为对照组(NC组),其余大鼠使用链脲佐菌素(STZ)构建DN大鼠模型,并随机均分为DN组、ASI组(45 mg/kg)、JAK2/STAT3信号通路激活剂Coumermycin A1(C-A1)组(100μg/kg)及ASI+C-A1组(45 mg/kg ASI+100μg/kg C-A1),NC组、DN组给予等量生理盐水,1次/d,连续2周;药物处理结束12 h,使用全自动生化分析仪测定肌酐、白蛋白和尿素水平,ELISA法检测血清白细胞介素-1β(IL-1β)、白细胞介素-6(IL-6)、肿瘤坏死因子(TNF-α)、丙二醛(MDA)及超氧化物歧化酶(SOD)水平,HE染色检测肾脏组织病理变化,TUNEL染色检测肾脏细胞凋亡情况,Western blot检测JAK2/STAT3通路相关蛋白表达。结果与NC组相比,DN组大鼠出现肾小球萎缩、肾小球类囊体严重增生、肾小管上皮分离、基底膜暴露、炎性细胞浸润和间质性水肿等症状;DN组较NC组IL-1β、IL-6、TNF-α、MDA水平、肌酐、白蛋白、尿素、细胞凋亡率、p-JAK2/JAK2、p-STAT3/STAT3蛋白水平显著升高(P<0.05),SOD水平显著降低(P<0.05);与DN组相比,ASI组肾组织病变严重程度有所缓解,炎性细胞浸润现象减轻,IL-1β、IL-6、TNF-α、肌酐、白蛋白、尿素、MDA、细胞凋亡率、p-JAK2/JAK2、p-STAT3/STAT3蛋白水平降低,SOD水平升高(P<0.05);C-A1减弱了ASI对DN大鼠炎症损伤的改善效果。结论ASI可能通过下调JAK2/STAT3信号通路抑制DN大鼠炎症反应。

Objective To investigate the effect of asiaticoside(ASI)on inflammatory response in diabetic nephropathy(DN)rats by regulating Janus kinase(JAK)/signal transducer and activator of transcription(STAT)signaling pathway.Methods Six SD rats were randomly selected as the control group(NC group),and the remaining rats were used to construct a DN rat model using streptozotocin(STZ).The successfully modeled rats were randomly divided into DN group,ASI group(45 mg/kg),JAK2/STAT3 signaling pathway activator Coumermycin A1(C-A1)group(100μg/kg),and ASI+C-A1 group(45 mg/kg ASI+100μg/kg C-A1).The NC group and DN group were given the same amount of physiological saline once a day for two consecutive weeks.At 12h after drug treatment,the levels of creatinine,albumin,and urea were measured by automated biochemical analyzer.The levels of serum interleukin-1β(IL-1β),interleukin-6(IL-6),tumor necrosis factor(TNF)-α,malondialdehyde(MDA),and superoxide dismutase(SOD)were detected by ELISA method.Pathological changes in renal tissue were detected by HE staining.The renal cell apoptosis was detected by TUNEL staining.The expression of JAK2/STAT3 pathway proteins was detected by Western blot.Results Compared with the NC group,the DN group exhibited symptoms such as glomerular atrophy,severe hyperplasia of glomerular thylakoids,separation of renal tubular epithelial,exposure of basement membrane,inflammatory cell infiltration,and interstitial edema.Compared with NC group,the levels of IL-1β,IL-6,TNF-α,MDA levels,creatinine,albumin,urea,cell apoptosis rate,p-JAK2/JAK2,p-STAT3/STAT3 protein in the DN group were significantly higher than those in NC group(P<0.05),and the SOD level was significantly lower than that in NC group(P<0.05).Compared with the DN group,the renal tissue lesions,inflammatory cell infiltration,IL-1β,IL-6,TNF-α,creatinine,albumin,urea,MDA,cell apoptosis rate,p-JAK2/JAK2,p-STAT3/STAT3 protein levels decreased in the ASI group,and the SOD level increased in the ASI group(P<0.05).C-A1 attenuated the improvement effect of ASI on inflammatory damage in DN rats.Conclusion ASI may inhibit inflammatory response in DN rats by down-regulating the JAK2/STAT3 signaling pathway.