文献类型：期刊文献

中文题名：基于NLRP3/ASC/Caspase-1通路探讨真武汤改善脾肾阳虚型糖尿病肾病炎症损伤的机制

英文题名：Mechanism of Zhenwu Decoction in improving renal inflammatory injury in diabetes with spleen-kidney yang deficiency based on NLRP3/ASC/Caspase-1 pathway

作者：赵哲[1,2];陈光顺[1];白敏[1,2];靳玉秋[1];田萌媛[1];张延英[1,2];刘臻华[1];宋冰[1,2]

第一作者：赵哲

机构：[1]甘肃中医药大学,兰州730000;[2]甘肃省实验动物行业技术中心,兰州730000

第一机构：甘肃中医药大学

年份：2023

卷号：31

期号：7

起止页码：853

中文期刊名：中国实验动物学报

外文期刊名：Acta Laboratorium Animalis Scientia Sinica

收录：CSTPCD;;北大核心:【北大核心2020】；CSCD:【CSCD_E2023_2024】；

基金：国家自然科学基金(82160865);甘肃省优秀研究生“创新之星”基金(2022CXZX-769);甘肃省自然科学基金(23JRRA1227)。

语种：中文

中文关键词：真武汤;脾肾阳虚;糖尿病肾病;NLRP3/ASC/Caspase-1通路;炎症损伤

外文关键词：Zhenwu Decoction;spleen and kidney yang deficiency;diabetic nephropathy;NLRP3/ASC/Caspase-1 pathway;inflammatory injury

摘要：目的研究真武汤对脾肾阳虚型糖尿病肾病(diabetic nephropathy,DN)小鼠肾组织炎症损伤的改善作用及可能机制。方法除空白对照组(db/m小鼠),其余各组均使用灌服生大黄溶液和氢化可的松制备db/db小鼠脾肾阳虚证模型,造模成功后随机分为模型对照组、厄贝沙坦组(25 mg/kg)、真武汤高、中、低剂量(33.8、16.9、8.45 g/kg)组,每组15只。除空白对照组和模型对照组给予生理盐水灌胃,其余各组给予相应药物灌胃,连续灌胃8周。测定各组小鼠中医证候评分;检测各组小鼠空腹血糖(fasting blood glucose,FBG)、尿肌酐(urine creatinine,Ucr)以及24 h尿蛋白(24 hour urinary protein,24 h UTP);酶免疫吸附测定法(ELISA)检测肾组织中睾酮(testosterone,T)、三碘甲状腺原氨酸(triiodothyronine,T3)、四碘甲状腺原氨酸(tetraiodothyronine,T4)、雌二醇(estradiol,E2)、白介素-1β(interleukin-1,IL-1β)、白介素-18(interleukin-18,IL-18)含量;苏木精-伊红染色法(hematoxylin-eosin staining,HE)观察各组肾组织病理学改变;以实时荧光定量PCR(Real-time quantitative PCR,RT-qPCR)和蛋白免疫印迹法(Western Blot)检测各组小鼠肾组织中NLRP3、ASC、Caspase-1 mRNA和蛋白表达水平。结果与空白对照组相比,模型对照组小鼠出现少食、倦卧、便溏下降等脾肾阳虚症状,FBG、24 h UTP、IL-1β、IL-18、E2含量均显著升高(P<0.05),T、T3、T4、Ucr含量显著降低(P<0.05),肾组织病理显示肾小球肥大,肾小球系膜和基底增厚明显,且肾组织NLRP3、ASC、Caspase-1 mRNA及蛋白表达水平显著上升(P<0.05);与模型对照组相比,真武汤高、中、低剂量组小鼠精神状态改善、活动增加、便质成型,T、T3、T4、Ucr含量明显增加(P<0.05),24 h UTP、IL-1β、IL-18、E2含量均显著降低(P<0.05);各给药组小鼠肾组织病理损伤明显改善,肾组织中NLRP3、ASC、Caspase-1 mRNA及蛋白表达水平显著降低(P<0.05)。结论真武汤能够显著改善脾肾阳虚型db/db小鼠一般状态、肾功能,降低炎症反应、减轻肾病理损伤,其作用机制可能与真武汤抑制肾组织中NLRP3/ASC/Caspase-1通路表达有关。
Objective To study the effect and possible mechanism of Zhenwu Decoction on renal inflammatory injury in mice with spleen-kidney yang deficiency-type diabetic nephropathy.Methods Mice(db/m mice)were treated with rhubarb solution and hydrocortisone to induce a db/db mouse spleen and kidney yang deficiency syndrome model.Mice in the blank control group were untreated.After successful modeling,the db/db mice were divided randomly into a model control group,irbesartan group(25 mg/kg),and high-,medium-,and low-dose Zhenwu Decoction groups(33.8,16.9,8.45 g/kg,respectively)(n=15 per group).Mice in all groups except the blank control and model control groups were given the corresponding drugs by gavage once a day for 8 weeks.Mice in the blank control and model control groups received the same volume of normal saline.Traditional Chinese medicine syndrome scores were measured in each group.Fasting blood glucose(FBG),urinary creatinine(Ucr),and 24 h urinary protein(24 h UTP)were measured.Testosterone(T),triiodothyronine(T3),tetraiodothyronine(T4),estradiol(E2),interleukin(IL)-1β,and IL-18 levels in renal tissues were measured by enzyme-linked immunosorbent assay.Histopathological changes in the kidneys were observed by hematoxylin/eosin staining.NOD-like receptor thermal protein domain associated protein 3(NLRP3),Apoptosis-associated speckle-like protein(ASC),Cysteinyl aspartate specific proteinase-1(Caspase-1)mRNA and protein expression levels in renal tissues were detected by Real-time quantitative PCR(RT-qPCR)and Western Blot.Results Mice in the model control group showed symptoms of spleen-kidney yang deficiency,such as anorexia,fatigue,and decreased,loose stools compared with the blank control group,while FBG,24 h UTP,IL-1β,IL-18,and E2 were significantly increased(P<0.05)and T,T3,T4,and Ucr were significantly decreased(P<0.05).Renal pathology showed glomerular hypertrophy,and glomerular mesangial and basal thickening.Protein and mRNA levels of NLRP3,ASC,and caspase-1 in renal tissue were significantly increased(P<0.05).In contrast,mice in the high-,medium-,and low-dose Zhenwu Decoction groups had increased body weight,activity,and stool molding compared with the model control group,as well as significantly increased levels of T,T3,T4,and Ucr(P<0.05)and significantly decreased levels of 24 h UTP,IL-1β,IL-18,and E2(P<0.05).In addition,pathological damage to the renal tissue was significantly improved and the protein and mRNA expression levels of NLRP3,ASC,and Caspase-1 were significantly decreased in all treatment groups(P<0.05).Conclusions Zhenwu Decoction can improve the general condition and renal function,and reduce the inflammatory reaction and renal pathological injury in db/db mice with spleen-kidney yang deficiency,via a mechanism possibly related to inhibition of the NLRP3/ASC/Caspase-1 pathway.