摘要
目的:探讨黄芪甲苷(astragalosideⅣ,AS-Ⅳ)调控肌醇依赖性激酶1α(inositol requiring enzyme 1α,IRE-1α)信号通路,减轻足细胞内质网应激,改善糖尿病肾病(diabetic nephropathy,DN)足细胞损伤。方法:(1)培养大鼠足细胞,将其分为正常组(5.5 mmol/L葡萄糖)、高糖组(30 mmol/L葡萄糖)、AS-Ⅳ组(20μmmol/L黄芪甲苷)、IRE-1α抑制剂组(20μmol/L STF-083010),干预24 h后收集细胞,采用RT-PCR检测内质网应激指标葡萄糖调节蛋白78(glucose-regulated protein 78,GRP78)、IRE-1α、肿瘤坏死因子相关受体2(tumor necrosis factor receptor-associated factor 2,TRAF-2)、IKK-α基因表达。(2)40只SPF级雄性SD大鼠,随机分为正常组10只和造模组30只。喂养1周后,采用链脲佐菌素(STZ)55 mg/kg腹腔注射,连续3 d,造模成功后随机分为DN组、AS-Ⅳ组(AS-Ⅳ40 mg·kg^(-1)·d^(-1)灌胃)、IRE-1α抑制剂组(STF-083010 10 mg·kg^(-1)·d^(-1)灌胃),干预8周后收集各组大鼠24 h尿液,ELISA测定尿蛋白定量,收集各组大鼠肾组织,行HE染色,免疫组化及RT-PCR检测GRP78、IRE-1α、TRAF-2、IKK-α表达。结果:(1)与正常组比较,高糖组足细胞GRP78、IRE-1α、TRAF-2、IKK-α mRNA表达升高(P<0.05);与高糖组相比,AS-Ⅳ组和IRE-1α抑制剂组表达下降(P<0.05);(2)与正常组比较,DN组大鼠24 h尿蛋白定量升高(P<0.05),病理HE染色可见肾小球肥大、系膜增生,基底膜增厚;与DN组比较,AS-Ⅳ组、IRE-1α抑制剂组大鼠24 h尿蛋白定量及病理改变减轻(P<0.05);(3)免疫组化结果显示,与正常组比较,DN组大鼠肾组织GRP78、IRE-1α表达增高(P<0.05);RT-PCR结果显示,与正常组比较,DN组GRP78、IRE-1α、TRAF-2、IKK-α mRNA表达增高(P<0.05);与DN组比较,AS-Ⅳ组、IRE-1α抑制剂组上述表达下降(P<0.05)。结论:AS-Ⅳ通过调控IRE-1α信号通路,减轻内质网应激,改善糖尿病肾病足细胞损伤。
Objective:Explore the mechanism of Astragaloside Ⅳ(AS-Ⅳ) alleviates podocyte endoplasmic reticulum stress in diabetic nephropathy by regulating IRE-1α signaling pathway.Methods:(1) Cultivate rat podocytes and divide them into normal group(5.5 mmol/L glucose),high glucose group(30 mmol/L glucose),AS-Ⅳ group(20 μmol/L astragaloside Ⅳ),and IRE-1 α inhibitor group(20 μmol/L STF-083010),after 24 hours of intervention,cells were collected,and RT-PCR was used to detect expression of glucose regulated protein 78(GRP78),IRE-1α,TRAF-2 and IKK-α.(2) 40 SPF grade male SD rats were randomly divided into normal group and model group.After one week of feeding,55 mg/kg streptozotocin(STZ) was injected intraperitoneally for 3 consecutive days to create DN model.After successful modeling,the rats were randomly divided into DN group,AS-Ⅳ group(AS-Ⅳ 40 mg·kg^(-1)·d^(-1) gavage),and IRE-1 α inhibitor group(STF-083010 10 mg·kg^(-1)·d^(-1) gavage).After 8 weeks of intervention,collected 24-hour urine and measure urinary protein quantification using ELISA.Collected kidney tissue from rats in each group,perform HE staining,Immunohistochemistry and RT-PCR detected GRP78,IRE-1α,TRAF-2 and IKK-α expression.Results:(1) Compared with the normal group,GRP78,IRE-1α,TRAF-2,IKK-α mRNA expression increased in the high glucose group(P<0.05).Compared with high glucose group,the expression decreased in AS-Ⅳ group and IRE-1αinhibitor group(P<0.05).(2) Compared with the normal group,the DN group showed an increase in 24-hour urine protein quantification(P<0.05).HE staining showed glomerular hypertrophy and mesangial widening in the DN group.Compared with DN group,the 24-hour urine protein quantification and pathological changes were reduced in AS-Ⅳ group and IRE-1αinhibitor group(P<0.05).(3) Immunohistochemical results showed that the expression of GRP78 and IRE-1αin DN group increased compared with the normal group(P<0.05).RT-PCR results showed that compared with the normal group,GRP78,IRE-1 α,TRAF-2 and IKK-α mRNA
作者
王蕊花
孙大林
王金晶
刘文媛
张紫媛
胡雅玲
方敬爱
WANG Ruihua;SUN Dalin;WANG Jinjing(Department of Nephrology,First Hospital of Shanxi Medical University,Taiyuan 030001)
出处
《中国中西医结合肾病杂志》
2023年第11期957-960,I0001,共5页
Chinese Journal of Integrated Traditional and Western Nephrology
基金
山西省中医药管理局科研项目(No.2023ZYYB030)。
