摘要
目的探讨甘草酸铵对过敏性紫癜性肾炎(HSPN)小鼠肾功能的影响及可能的作用机制。方法2020年7月—2021年1月在邯郸市中心医院实验室进行实验,C57BL/6雄性小鼠80只,随机选取12只作为空白对照组,其余小鼠建立HSPN模型。将造模成功的60只小鼠随机分为模型组、环磷酰胺组(22 mg/kg)及甘草酸铵低、中、高剂量组(25、50、100 mg/kg),每组12只。环磷酰胺组、甘草酸铵各剂量组小鼠腹腔注射相应剂量的药物,1次/d,空白对照组和模型组给予等量生理盐水,持续4周。给药结束后,检测小鼠24 h尿蛋白水平和尿红细胞计数,血清白介素-23(IL-23)、IL-17、免疫球蛋白A(IgA)、尿素氮(BUN)、血肌酐(SCr)、循环免疫复合物(CIC)水平,外周血单个核细胞中Th17、Treg细胞占比及肾脏组织中IL-23、信号传导和转录激活因子3(STAT3)及其磷酸化蛋白表达水平;苏木素—伊红(HE)染色观察肾脏组织的形态学变化。结果与空白对照组比较,模型组小鼠24 h尿蛋白水平、尿红细胞计数,血清IL-23、IL-17、IgA、CIC、BUN、SCr水平,外周血单个核细胞中Th17细胞占比、Th17/Treg细胞比值,肾脏组织中IL-23、磷酸化STAT3(p-STAT3)/STAT3蛋白表达升高(P<0.05),Treg细胞占比显著降低(P<0.05),肾脏组织出现严重的病理损伤。与模型组比较,环磷酰胺组及甘草酸铵中、高剂量组小鼠24 h尿蛋白水平、尿红细胞计数,血清IL-23、IL-17、IgA、CIC、BUN、SCr水平,外周血单个核细胞中Th17细胞占比、Th17/Treg细胞比值,肾脏组织中IL-23、p-STAT3/STAT3蛋白表达降低(F/P=45.928/0.000、52.976/0.000、64.629/0.000、151.382/0.000、277.542/0.000、55.985/0.000、203.736/0.000、261.569/0.000、214.406/0.000、226.046/0.000、228.611/0.000、148.613/0.000),Treg细胞占比显著升高(F/P=24.160/0.000),肾脏组织病理损伤明显减轻。与环磷酰胺组比较,甘草酸铵高剂量组小鼠上述指标差异无统计学意义(P>0.05)。结论甘草酸铵可减轻HSPN小鼠肾损伤,其作用机制可能与抑制IL-23/STAT3通路的活化,调节Th17/Treg细胞免疫平衡有关。
Objective To investigate the effect of ammonium glycyrrhizinate on renal function in mice with Henoch-Schonlein purpura nephritis(HSPN)and its possible mechanism.Methods The experiment was conducted in the Central Laboratory of Handan Central Hospital from July 2020 to January 2021.There were 80 C57BL/6 male mice,12 of which were randomly selected as the blank control group,and the other mice established HSPN model.Sixty successful mice were randomly divided into model group,cyclophosphamide group(22 mg/kg)and ammonium glycyrrhizate low,medium and high dose groups(25,50 and 100 mg/kg),with 12 mice in each group.Mice in cyclophosphamide group and ammonium glycyrrhizinate groups were intraperitoneally injected with corresponding doses of drugs,once a day.The blank control group and model group were given the same amount of normal saline for 4 weeks.After administration,the levels of 24 h urinary protein,urinary red blood cell count,serum interleukin 23(IL-23),IL-17,immunoglobulin A(IGA),urea nitrogen(BUN),serum creatinine(SCR),circulating immune complex(CIC),the proportion of Th17 and Treg cells in peripheral blood mononuclear cells,IL-23 and signal transduction and transcription activator 3(STAT3)in renal tissue were measured And its phosphorylated protein expression level;Hematoxylin eosin(he)staining was used to observe the morphological changes of renal tissue.Results Compared with the blank control group,the 24-hour urinary protein level,urinary red blood cell count,serum IL-23,IL-17,IgA,CIC,bun and SCR levels,the proportion of Th17 cells and Th17/Treg cells in peripheral blood mononuclear cells,the expression of IL-23,phosphorylated STAT3(P-STAT3)/STAT3 protein in kidney tissue increased(P<0.05),and the proportion of Treg cells decreased significantly(P<0.05)Compared with the model group,the 24-hour urinary protein level,urinary red blood cell count,serum IL-23,IL-17,IgA,CIC,bun and SCR levels,the proportion of Th17 cells and the ratio of Th17/Treg cells in peripheral blood mononuclear cells,and the expression of IL-23 and P STAT3/STAT3 protein in renal tissue decreased in cyclophosphamide group and ammonium glycyrrhizate medium and high dose groups(F/P=45.928/0.000,52.976/0.000,64.629/0.000,151.382/0.000,277.542/0.000,55.985/0.000,203.736/0.000,261.569/0.000,214.406/0.000,226.046/0.000,228.611/0.000,148.613/0.000),the proportion of Treg cells increased significantly(F/P=24.160/0.000)Compared with cyclophosphamide group,there was no significant difference in the above indexes in Ammonium Glycyrrhizinate high-dose group(P>0.05).Conclusion Ammonium glycyrrhizinate can reduce renal injury in HSPN mice,and its mechanism may be related to inhibiting the activation of IL-23/STAT3 pathway and regulating the immune balance of Th17/Treg cells.
作者
赵利平
梁卫章
张俊民
代宝春
姜红
刘秀珍
Zhao Liping;Liang Weizhang;Zhang Junmin;Dai Baochun;Jiang Hong;Liu Xiuzhen(The First Department of Pediatrics,Handan Central Hospital,Hebei Province,Handan 056000,China)
出处
《疑难病杂志》
CAS
2022年第1期74-79,共6页
Chinese Journal of Difficult and Complicated Cases
基金
河北省医学科学研究课题计划项目(20191844)。
关键词
过敏性紫癜性肾炎
肾损伤
甘草酸铵
白介素-23/信号传导和转录激活因子3
小鼠
Henoch-Schonlein purpura nephritis
Kidney injury
Ammonium glycyrrhizinate
Interleukin 23/signal transducer and activator of transcription 3
Mice
