艾沙利酮抑制妊娠加重梗阻性肾病大鼠淋巴管生成及肾间质纤维化  

Esaxerenone inhibits lymphangiogenesis and renal interstitial fibrosis in rats with pregnancy aggravated obstructive nephropathy

作　　者：牛洁琪 张淑琛 许畅 王洪双 方芳 高兰君 王香婷 王筝 NIU Jieqi;ZHANG Shuchen;XU Chang;WANG Hongshuang;FANGFang;GAO Lanjun;WANG Xiangting;WANG Zheng(College of Integrated Chinese and Western Medicine,Hebei University of Chinese Medicine,Shijiazhuang 050200,China;Hebei Key Laboratory of Integrative Medicine on Liver-Kidney Pattern,Shijiazhuang 050091,China;Graduate School,Hebei University of Chinese Medicine,Shijiazhuang 050091,China;College of Basic Medicine,Hebei University of Chinese Medicine,Shijiazhuang 050200,China)

机构地区：[1]河北中医药大学中西医结合学院,河北石家庄050200 [2]河北省中西医结合肝肾病证研究重点实验室,河北石家庄050091 [3]河北中医药大学研究生学院,河北石家庄050091 [4]河北中医药大学基础医学院,河北石家庄050200

出　　处：《中国病理生理杂志》2024年第9期1700-1710,共11页Chinese Journal of Pathophysiology

基　　金：河北省自然科学基金资助项目(No.H2024423013)。

摘　　要：目的:探讨艾沙利酮抑制妊娠加重梗阻性肾病大鼠淋巴管生成的机制及对肾脏的保护作用。方法:未经产雌性Wistar大鼠40只随机分为假手术组、假手术+妊娠组、模型组和艾沙利酮组,每组10只。将模型组与艾沙利酮组大鼠采用单侧输尿管结扎(unilateral ureteral obstruction,UUO)手术造成肾脏损伤,其余两组仅游离输尿管但不结扎。UUO术后第9周将假手术+妊娠组、模型组和艾沙利酮组的雌鼠进行阴道涂片,将处于动情前期或动情期的雌鼠与雄鼠按2∶1比例合笼,并密切观察,将发现阴栓且阴道涂片中含有大量精子细胞确定为妊娠第1天,建立梗阻性肾病妊娠大鼠模型。艾沙利酮组在UUO术后第2天开始给药,给予艾沙利酮1 mg·kg^(−1)·d^(−1)治疗。妊娠后第18天代谢笼留取24 h尿液,次日处死母鼠,留取血清标本,摘取梗阻对侧肾脏。采用常规生化法检测尿素氮(blood urea nitrogen,BUN),计算内生肌酐清除率(creatinine clearance rate,Ccr)。苏木精-伊红(HE)、马松(Masson)和天狼星红(Sirius red)染色观察大鼠肾脏组织病理形态学改变。ELISA法测定血清中醛固酮含量。免疫组化、real-time PCR和Western blot检测盐皮质激素受体(mineralocorticoid receptor,MR)活化、淋巴管生成以及相关信号通路、肾纤维化相关指标的表达。结果:肾功能结果显示模型组大鼠BUN升高,Ccr下降(P<0.01)。肾脏病理显示模型组大鼠肾小管扩张明显,有大量胶原纤维沉积及炎症细胞浸润。ELISA结果显示模型组血清醛固酮含量显著升高(P<0.01)。免疫组化结果显示模型组大鼠肾脏MR活化后入核数量增多。Western blot和real-time PCR结果显示模型组大鼠中性粒细胞明胶酶相关脂质运载蛋白(neutrophil gelatinase-associated lipocalin,NGAL)表达显著升高(P<0.01)。免疫组化结果显示模型组大鼠肾脏血管内皮生长因子C(vascular endothelial growth factor-C,VEGF-C)和VEGF受体3(VEGF recepAIM:To explore the mechanisms behind the inhibition of lymphangiogenesis in pregnant rats with obstructive nephropathy and assess the protective effects on kidney function.METHODS:Forty nulliparous female Wistar rats were randomly assigned to four groups:sham operation,sham operation+pregnancy,model,and Esaxerenone groups,with 10 rats in each group.Renal injury was induced in the model and Esaxerenone groups via unilateral ureteral obstruction(UUO).The other two groups underwent ureteral dissociation without ligation.Nine weeks post-UUO,female rats in the sham operation+pregnancy,model,and Esaxerenone groups were mated with male rats(2∶1 ratio)to establish a rat model of obstructive nephropathy during pregnancy.Starting the day after UUO,rats in the Esaxerenone group received Esaxerenone at 1 mg·kg^(−1)·d^(−1).On the 18th day of pregnancy,24-hour urine was collected using metabolic cages.The following day,the rats were sacrificed,serum samples collected,and the contralateral kidney removed.Blood urea nitrogen(BUN)was measured using standard biochemical methods,and endogenous creatinine clearance rate(Ccr)was calculated.Kidney tissue pathology was assessed using HE,Masson,and Sirius red staining.Serum aldosterone levels were determined via ELISA.Immunohistochemistry,real-time PCR,and Western blot were employed to assess mineralocorticoid receptor(MR)activation,lymphangiogenesis,signaling pathways,and fibrosis-related markers.RESULTS:Renal function tests revealed increased BUN levels and decreased Ccr in the model group(P<0.01).Pathological examination showed dilated renal tubules,significant collagen deposition,and inflammatory cell infiltration in the model group.ELISA results indicated a significant increase in serum aldosterone levels in the model group(P<0.01).Immunohistochemistry showed enhanced nuclear translocation of MR in the kidneys of the model group post-activation.Western blot and real-time PCR demonstrated a marked increase in neutrophil gelatinase-associated lipocalin(NGAL)expression in the

关 键 词：淋巴管生成 肾间质纤维化 艾沙利酮 梗阻性肾病 醛固酮 

分 类 号：R363.2[医药卫生—病理学] R692[医药卫生—基础医学] R714