p38 MAPK通路调控自噬-内质网应激途径介导草酸钙肾结石形成的机制研究  

Mechanism of calcium oxalate kidney stone formation mediated by autophagy and endoplasmic reticulum stress pathway regulated by p38 MAPK pathway

作　　者：谢亚彬 王飞 王康扬 林师帅 XIE Yabin;WANG Fei;WANG Kangyang;LIN Shishuai(Department of Urology,Wenchang People s Hospital,Wenchang 571000,China)

机构地区：[1]文昌市人民医院泌尿外科,海南文昌571000

出　　处：《现代泌尿外科杂志》2024年第1期73-80,共8页Journal of Modern Urology

基　　金：海南省卫生健康行业科研项目(No.22A200166)。

摘　　要：目的 探究p38 MAPK通路对大鼠草酸钙(CaOx)肾结石形成的影响及其作用机制,为肾结石的治疗选择提供新的思路。方法 将40只大鼠分为对照组、SB203580组、CaOx组、SB203580+CaOx组,每组10只,CaOx组和SB203580+CaOx组以1%乙二醇与1%氯化铵配制的混合液灌胃建立CaOx肾结石模型,对照组和SB203580组以饮用水灌胃;造模后,SB203580组和SB203580+CaOx组腹腔注入剂量为5 mg/kg的SB203580,1次/d,连续注射14 d,对照组和CaOx组腹腔注入等体积生理盐水。称量大鼠肾脏质量并计算肾脏系数,全自动生化分析仪检测血清中血清尿素氮(BUN)和血肌酐(SCr)水平,酶联免疫吸附法(ELISA)测定尿液中中性粒细胞明胶酶相关脂质运载蛋白(NGAL)和肾损伤分子-1(KIM-1)水平,钙盐染色(Von Kossa)观察黑色晶体沉积及组织损伤情况,原位末端标记法(TUNEL)观察肾小管细胞凋亡情况,免疫组化染色检测自噬标记物表达,实时荧光定量聚合酶链反应(RT-qPCR)与蛋白质印迹(Western blotting)测定自噬及内质网应激途径相关分子表达。结果 与CaOx组比较,SB203580+CaOx组大鼠造模后体质量较高(P<0.05),肾脏质量、肾脏系数、BUN、SCr、NGAL、KIM-1水平均较低(P<0.05),肾脏组织病理损伤减轻且黑色结晶明显减少,TUNEL阳性细胞比例、LC3B与Beclin-1阳性表达面积占比、LC3B、Beclin-1、CHOP、GRP78 mRNA表达、LC3-Ⅱ/LC3-Ⅰ蛋白比值、Beclin-1、CHOP、GRP78蛋白表达均下调(P<0.05),p62 mRNA和蛋白表达上调(P<0.05)。结论 p38 MAPK通路参与大鼠CaOx肾结石形成,抑制该通路能够减少肾结石形成,该作用可能与其调控自噬-内质网应激有关。Objective To explore the effects and mechanism of p38 mitogen activated protein kinase(MAPK) pathway on the formation of calcium oxalate(CaOx) kidney stones in rats,so as to provide new ideas for the treatment of kidney stones.Methods A total of 40 rats were divided into control,SB203580,CaOx and SB203580+CaOx groups,with 10 rats in each group.Intragastric administration of a mixture of 1% ethylene glycol and 1% ammonium chloride was given to the CaOx and SB203580+CaOx groups to construct CaOx models,while intragastric administration of drinking water was given to the control and SB203580 groups.After molding,SB203580 and SB203580+CaOx groups were injected with 5 mg/kg SB203580 peritoneally once a day for 14 days,while the control and CaOx groups were injected with equal volume of normal saline.The renal mass of rats was measured and the renal coefficient was calculated;the serum levels of blood urea nitrogen(BUN) and serum creatinine(SCr) were measured with an automated biochemical analyzer;the urinary levels of neutrophil gelatinase-associated lipid carrier protein(NGAL) and kidney injury molecule-1(KIM-1) were determined with enzyme-linked immunosorbent assay(ELISA);the crystal deposition and tissue damage in renal tissues were observed with Von Kossa staining;the apoptosis of renal tubule cells was observed with TUNEL;the expressions of autophagy markers in kidney tissues were detected with immunohistochemical staining;the molecular expressions of autophagy-endoplasmic reticulum stress related pathways in renal tissues were determined with RT-qPCR and Western blot.Results Compared with the CaOx group,the SB203580+CaOx group had increased body mass after molding(P<0.05);decreased kidney mass,kidney coefficient,BUN,SCr,NGAL and KIM-1 levels(P<0.05);alleviated pathological damage of kidney tissues;significantly reduced black crystal;down-regulated proportion of positive TUNEL cells,positive expression area of LC3B and Beclin-1,mRNA expressions of LC3B,Beclin-1,CHOP and GRP78,protein ratio of LC3-Ⅱ/LC3-Ⅰ,and

关 键 词：肾结石 草酸钙 P38丝裂原激活蛋白激酶 自噬 内质网应激 结石形成 肾结石动物模型 

分 类 号：R692.4[医药卫生—泌尿科学]