摘要
目的基于内质网应激途径探究XBP1抑制剂在缺血再灌注肾损伤中的作用及机制。方法将36只成年健康雄性SD大鼠随机分为3组,每组12只,分别记为假手术组(sham组)、模型组(model组)和XBP1抑制剂干预组(STF080310组)。sham组大鼠暴露双侧肾动脉,不夹闭动脉;model组大鼠夹闭双侧肾动脉45 min后恢复灌流,制备缺血再灌注肾损伤模型;STF080310组在建模前1 h给予3 mg/100 g STF080310(浓度:3 g/L)腹腔注射,其余处理同model组。造模成功后24 h取大鼠尾静脉血,随后处死大鼠取肾脏组织。采用全自动生化仪检测大鼠血清尿素氮(blood urea nitrogen,BUN)、肌酐(serum creatinine,sCr)等肾脏过滤能力指标水平;采用酶联免疫吸附测定(enzyme-linked immunosorbent assay,ELISA)法检测大鼠血清肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)、白介素-1β(interleukin-1β,IL-1β)、白介素-6(interleukin-6,IL-6)等氧化应激指标水平;采用HE染色观察大鼠肾脏组织形态学改变,并计算肾小管损伤评分;采用TUNEL法检测肾脏细胞凋亡指数;采用RT-qPCR法和Western blot法分别检测大鼠肾组织中XBP1及内质网应激相关蛋白Caspase-12、JNK1、CHOP、GRP78蛋白及mRNA表达。结果与sham组大鼠比较,model组和STF080310组大鼠血清BUN、sCr水平及血清TNF-α、IL-1β、IL-6等氧化应激指标水平均明显升高(P<0.05),肾小管损伤评分及肾脏细胞凋亡指数均明显升高(P<0.05),肾脏组织中XBP1 mRNA及蛋白表达水平明显升高(P<0.05);与model组大鼠比较,STF080310组大鼠血清BUN、sCr水平及血清TNF-α、IL-1β、IL-6水平明显降低(P<0.05),肾小管损伤评分及肾脏细胞凋亡指数均明显降低(P<0.05);与sham组大鼠比较,model组和STF080310组大鼠肾脏组织中Caspase-12、JNK1、CHOP、GRP78 mRNA及蛋白表达水平均明显升高(P<0.05);与model组大鼠比较,STF080310组大鼠肾脏组织中Caspase-12、JNK1、CHOP等内质网应激相关蛋白mRNA及蛋白表达水平则明显降低(P<0.05),GRP78 mRNA及蛋白表达水平则明显升高(P<0.05)。结论XBP1抑制剂通过采用预防性给药的方式,可对内质网应激相关凋亡蛋白表达产生抑制作用,由此降低血清炎症因子水平,达到减轻大鼠肾脏缺血再灌注损伤的目的。
Objective To explore the role of XBP1 inhibitor in renal ischemia-reperfusion injury based on the endoplasmic reticulum stress pathway and its mechanism.Methods Thirty-six adult healthy male SD rats were randomly divided into 3 groups(n=12):sham operation group(sham group),model group and XBP1 inhibitor intervention group(STF080310 group).Rats in the sham group were exposed to bilateral renal arteries without clipping.The model group was prepared by clamping bilateral renal arteries for 45 min,and the STF080310 group was given intraperitoneal injection of 3 mg/100 g STF080310(concentration:3 g/L)1 h before modeling.The other treatments were the same as those in the model group.The rat tail venous blood was collected 24 h after successful modeling,and then the rats were sacrificed and kidney tissue was collected.Blood urea nitrogen(BUN),serum creatinine(sCr)and other indexes of renal filtration capacity were detected by automatic biochemical analyzer.Serum levels of tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β),interleukin-6(IL-6)and other oxidative stress indexes were detected by enzyme-linked immunosorbent assay(ELISA).HE staining was used to observe the changes of renal histomorphology,and the renal tubular injury score was calculated.The apoptosis index of renal cells was detected by TUNEL assay.The protein and mRNA expressions of XBP1 and endoplasmic reticulum stress-related proteins Caspase-12,JNK1,CHOP and GRP78 in renal tissues of rats were detected by Western blot and RT-qPCR.Results Compared with sham group,the levels of serum BUN,sCr and oxidative stress indexes such as TNF-α,IL-1βand IL-6 in model group and STF080310 group were significantly higher(P<0.05),and the renal tubular injury score and renal cell apoptosis index were significantly higher(P<0.05).The mRNA and protein expression levels of XBP1 in kidney tissue were significantly higher(P<0.05).Compared with model group,the levels of BUN and sCr in serum,TNF-α,IL-1βand IL-6 in serum of STF080310 group were significantly lower(P<0.05),and the renal tubular injury score and renal cell apoptosis index were significantly lower(P<0.05).Compared with sham group,the mRNA and protein expression levels of Caspase-12,JNK1,CHOP and GRP78 in kidney tissues of model group and STF080310 group were significantly higher(P<0.05).Compared with model group,the mRNA and protein expression levels of endoplasmic reticulum stress-related proteins such as Caspase-12,JNK1 and CHOP were significantly lower in STF080310 group(P<0.05),while the mRNA and protein expression levels of GRP78 were significantly higher in STF080310 group(P<0.05).Conclusion XBP1 inhibitor can inhibit the expression of endoplasmic reticulum stress-related apoptotic proteins by prophylactic administration,thereby reducing the level of serum inflammatory factors and alleviating renal ischemia-reperfusion injury in rats.
作者
李海洋
李涛
LI Hai-yang;LI Tao(Department of Psychiatry,The First Affiliated Hospital of Harbin Medical University,Harbin 150001,China;Central Operating Room,The First Affiliated Hospital of Harbin Medical University,Harbin 150001,China)
出处
《哈尔滨医科大学学报》
CAS
2022年第6期525-530,540,共7页
Journal of Harbin Medical University
