ACSL4抑制剂对草酸钙结石致小鼠肾损伤及间质纤维化的影响  

Effect of ACSL4 inhibitor on calcium oxalate stone-induced renal injury and interstitial fibrosis in mice

作　　者：李磊[1] 叶泽华 夏煜琦 李柏均 陈礼嘉 周向军[1] 阮远[1] 饶婷[1] 余伟民[1] 程帆[1] LI Lei;YE Zehua;XIA Yuqi;LI Bojun;CHEN Lijia;ZHOU Xiangjun;RUAN Yuan;RAO Ting;YU Weimin;CHENG Fan(Department of Urology,Renmin Hospital of Wuhan University,Wuhan,Hubei 430060,China)

机构地区：[1]武汉大学人民医院泌尿外科,湖北武汉430060

出　　处：《中华实用诊断与治疗杂志》2023年第10期999-1003,共5页Journal of Chinese Practical Diagnosis and Therapy

基　　金：国家自然科学基金(82170775,82100806)。

摘　　要：目的探讨ACSL4抑制剂调控铁死亡在草酸钙结石所致小鼠肾损伤及间质纤维化中的肾保护作用及可能机制。方法15只雄性C57BL/6小鼠随机分为模型组、ACSL4抑制剂组、对照组各5只,其中模型组腹腔注射乙醛酸水溶液(100 mg/kg)构建草酸钙肾结石小鼠模型,ACSL4抑制剂组腹腔注射乙醛酸水溶液(100 mg/kg)并饮用含Abemaciclib(30 mg/kg)无菌水,对照组腹腔注射等量生理盐水,3组小鼠均连续处理12 d。12 d后检测3组小鼠血清肌酐、尿素水平。取3组小鼠左侧肾脏组织,行组织病理检查Von-Kossa染色、HE染色、Masson染色,观察肾脏晶体沉积、组织损伤及间质纤维化情况;行免疫组织化学检查,观察肾组织ACSL4、α-SMA表达情况;采用Western blot法检测肾组织ACSL4、collagenⅠ、vimentin、α-SMA、SLC7A11、GPX4蛋白相对表达量。结果(1)模型组血清肌酐[(68.91±14.18)μmol/L]、尿素[(7.49±1.92)mmol/L]水平均高于对照组[(30.22±6.80)μmol/L、(3.76±0.38)mmol/L]、ACSL4抑制剂组[(44.10±4.42)μmol/L、(4.61±0.76)mmol/L](P<0.05),ACSL4抑制剂组均高于对照组(P<0.05)。(2)对照组肾组织未见草酸钙晶体沉积,肾组织结构基本正常,未见明显炎症细胞浸润及胶原纤维沉积;模型组肾组织大量黑色草酸钙晶体沉积,肾小管上皮细胞肿胀、脱落,细胞核染色质浓缩,管腔扩张、变形,周围炎症细胞浸润,肾间质胶原纤维大量沉积;与模型组相比,ACSL4抑制剂组肾组织草酸钙晶体沉积减少,肾小管上皮细胞肿胀减轻,周围炎症细胞浸润减少,肾间质胶原沉积减少,纤维化程度减轻。(3)对照组肾组织ACSL4和α-SMA呈弱阳性表达;模型组肾组织ACSL4和α-SMA阳性表达较对照组明显增多;ACSL4抑制剂组肾组织ACSL4和α-SMA阳性表达较模型组明显减少。(4)模型组肾组织ACSL4、collagenⅠ、vimentin、α-SMA蛋白相对表达量(0.93±0.01、1.19±0.06、1.19±0.08、1.20±0.08)均高于对照组(0.33±0.01、0.53±0.0Objective To investigate the renoprotective effects and potential mechanisms of ACSL4 inhibitor-mediated regulation of ferroptosis in renal injury and interstitial fibrosis induced by calcium oxalate stone in mice.Methods Fifteen male C57BL/6 mice were randomly divided into model group,ACSL4 inhibitor group and control group,with 5 mice in each group.In model group,the mice received intraperitoneal injection of aqueous glyoxylate(100 mg/kg)to establish calcium oxalate stone mice models.In ACSL4 inhibitor group,the mice were given intraperitoneal injection of aqueous glyoxylate(100 mg/kg)and were provided with sterile water containing Abemaciclib(30 mg/kg).Control group received intraperitoneal injection of an equivalent volume of normal saline.All three groups were treated continuously for 12 days.After 12 days,the serum creatinine and urea levels were compared among three groups.The left kidneys of three groups underwent histopathological examinations,including Von-Kossa staining,HE staining and Masson staining to assess crystal deposition,tissue injury and fibrosis.Im munohistochemistry was employed to evaluate ACSL4 andα-SMA expressions in renal tissues,while Western blot analysis was used to detect the relative expressions of ACSL4,collagenⅠ,vimentin,α-SMA,SLC7A11 and GPX4 proteins.Results(1)The levels of serum creatinine and urea were higher in model group[(68.91±14.18)μmol/L,(7.49±1.92)mmol/L]than those in control group[(30.22±6.80)μmol/L,(3.76±0.38)mmol/L]and ACSL4 inhibitor group[(44.100±4.42)μmol/L,(4,61±76)mmol/L](P<0.05),and higher in ACSL4 inhibitor group that those in control group(P<0.05),(2)The renal tissues from control group revealed an absence of calcium oxalate crystal deposition,maintaining a primarily normal renal tissue structure,no discernible inflammatory cell infiltration or collagen fiber deposition.In contrast,model group exhibited extensive deposition of black calcium oxalate crystals in renal tissues,marked swelling or detachment of renal tubular epithelial cells,chromati

关 键 词：肾损伤 肾间质纤维化 ACSL4 Abemaciclib 铁死亡 草酸钙结石 

分 类 号：R692.4[医药卫生—泌尿科学]