文献类型：期刊文献

中文题名：真武汤调控ROCK/IKK/NF-κB通路关键分子表达改善脾肾阳虚型DN小鼠肾脏炎症损伤的机制研究

英文题名：Mechanism of Zhenwu Decoction in improving renal inflammatory injury in mice with DN of spleen-kidney Yang deficiency syndrome by regulating ROCK/IKK/NF-κB pathway

作者：靳玉秋[1];陈光顺[1];白敏[1];赵哲[1];陈彦旭[1];田萌媛[1];陈家涟[1];王庆胜[1];刘臻华[1]

第一作者：靳玉秋

机构：[1]甘肃中医药大学,甘肃兰州730000

第一机构：甘肃中医药大学

年份：2023

卷号：48

期号：18

起止页码：5041

中文期刊名：中国中药杂志

外文期刊名：China Journal of Chinese Materia Medica

收录：CSTPCD;;Scopus;北大核心:【北大核心2020】；CSCD:【CSCD2023_2024】；PubMed;

基金：国家自然科学基金地区科学基金项目(82160865)。

语种：中文

中文关键词：糖尿病肾病;脾肾阳虚型;真武汤;ROCK/IKK/NF-κB通路;炎症

外文关键词：diabetic nephropathy;spleen-kidney Yang deficiency syndrome;Zhenwu Decoction;ROCK/IKK/NF-κB pathway;inflammation

摘要：基于Rho相关卷曲螺旋激酶(Rho-associated coiled helix kinase,ROCK)/核因子-κB上游激酶(IκB kinase,IKK)/核因子-κB(nuclear factor kappaB,NF-κB)通路探讨真武汤对脾肾阳虚型糖尿病肾病(diabetic nephropathy,DN)小鼠的干预作用及机制。95只7周龄db/db雄性小鼠和25只7周龄db/m雄性小鼠适应性喂养1周,采用灌服大黄水煎液联合氢化可的松法复制脾肾阳虚型DN小鼠模型,随后进行模型评价,造模成功后随机分为模型组,真武汤高、中、低剂量组(33.8、16.9、8.45 g·kg^(-1)·d^(-1)),厄贝沙坦组(25 mg·kg^(-1)·d^(-1)),每组至少15只,持续干预8周。干预结束后,检测体质量、饮食量;测定血清肌酐(serum creatinine,Scr)、尿素氮(blood urea nitrogen,BUN)、空腹血糖(fasting blood glucose,FBG)、尿微量白蛋白(urinary albumin,uALb)、尿肌酐(urine creatinine,Ucr)含量,计算尿微量白蛋白与尿肌酐比值(ACR)、24 h尿蛋白(urinary protein,UTP);HE染色、Masson染色评价肾脏病理形态;采用蛋白免疫印迹(Western blot)法检测ROCK/IKK/NF-κB通路关键分子蛋白水平;采用酶联免疫吸附测定法(enzyme-linked immunosorbent assay,ELISA)检测白细胞介素-1β(interleukin-1β,IL-1β)、白细胞介素-6(interleukin-6,IL-6)、白细胞介素-8(interleukin-8,IL-8)、白细胞介素^(-1)0(interleukin-10,IL-10)、肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)含量。与空白组相比,模型组小鼠BUN、uALb、SCr含量上升,24 h UTP、ACR上升,Ucr含量下降(P<0.05);肾小球增大,基底膜变厚且系膜基质出现增生,炎性细胞浸润,胶原纤维沉积;肾脏中ROCK1、ROCK2、IKK、NF-κB以及磷酸化后的核因子-κB上游激酶(p-IκB kinase,p-IKK)、磷酸化后的核因子-κB(p-nuclear factor-kappaB,p-NF-κB)、磷酸化后的NF-κB抑制蛋白(p-inhibitor of NF-κB,p-IκB)蛋白表达上升(P<0.05),NF-κB抑制蛋白(inhibitor of NF-κB,IκB)蛋白表达下降(P<0.05);炎症因子IL-1β、IL-6、IL-8、TNF-α含量升高(P<0.05),IL-10含量降低(P<0.05)。与模型组相比,各组小鼠BUN、uALb、SCr含量下降,24 h UTP、ACR均下降,Ucr含量升高(P<0.05);肾脏病理状态与模型组相比均有不同程度改善;真武汤高、中剂量组及厄贝沙坦组小鼠肾脏中ROCK1、ROCK2、IKK、NF-κB、p-IKK、p-NF-κB、p-IκB蛋白表达下降(P<0.05),IκB蛋白表达上升(P<0.05),血清炎症因子IL-1β、IL-6、IL-8、TNF-α含量降低(P<0.05),IL-10含量升高(P<0.05)。真武汤能够显著改善脾肾阳虚型DN小鼠肾功能及肾脏病理损伤,其具体机制与真武汤下调肾脏中ROCK/IKK/NF-κB通路关键分子表达抑制炎症反应发生有关。
To investigate the intervention effect and mechanism of Zhenwu Decoction on diabetic nephro pathy(DN)mice of spleen-kidney Yang deficiency syndrome based on the Rho-associated coiled-coil kinase(ROCK)/IκB kinase(IKK)/nuclear factor-κB(NF-κB)pathway.Ninety-five 7-week-old db/db male mice and 257-week-old db/m male mice were fed adaptively for one week.The DN model of spleen-kidney yang deficiency syndrome was induced by Dahuang Decoction combined with hydrocortisone by gavage,and then the model was evaluated.After modeling,they were randomly divided into a model group,high-dose,medium-dose,and low-dose Zhenwu Decoction groups(33.8,16.9,and 8.45 g·kg^(-1)·d^(-1)),and an irbesartan group(25 mg·kg^(-1)·d^(-1)),with at least 15 animals in each group.The intervention lasted for eight weeks.After the intervention,body weight and food intake were measured.Serum creatinine(Scr),blood urea nitrogen(BUN),fasting blood glucose(FBG),urinary albumin(uALb),and urine creatinine(Ucr)were determined.The uALb/Ucr ratio(ACR)and 24 h urinary protein(UTP)were calculated.Renal pathological morphology was evaluated by HE staining/Masson staining.The levels of key molecular proteins in the ROCK/IKK/NF-κB pathway were detected by Western blot.Enzyme-linked immunosorbent assay(ELISA)was used to detect interleukin-1β(IL-1β),interleukin-6(IL-6),interleukin-8(IL-8),interleukin-10(IL-10),and tumor necrosis factor-α(TNF-α).Compared with the blank group,the model group showed increased content of BUN,uALb,and SCr,increased values of 24 h UTP and ACR,decreased content of Ucr(P<0.05),enlarged glomeruli,thickened basement membrane,mesangial matrix proliferation,inflammatory cell infiltration,and collagen fiber deposition.The protein expression of ROCK1,ROCK2,IKK,NF-κB,phosphorylated IKK(p-IKK),phosphorylated NF-κB(p-NF-κB),and phosphorylated inhibitor of NF-κB(p-IκB)increased(P<0.05),while the protein expression of inhibitor of NF-κB(IκB)decreased(P<0.05).The levels of inflammatory factors IL-1β,IL-6,IL-8,and TNF-αincreased(P<0.05),while the level of IL-10 decreased(P<0.05).Compared with the model group,the groups with drug treatment showed decreased levels of BUN,uALb,SCr,24 h UTP,and ACR,increased levels of Ucr(P<0.05),and improved renal pathological status to varying degrees.The high-and medium-dose Zhenwu Decoction groups and the irbesartan group showed reduced protein expression of ROCK1,ROCK2,IKK,NF-κB,p-IKK,p-NF-κB,and p-IκB in the kidneys(P<0.05),increased protein expression of IκB(P<0.05),decreased levels of serum inflammatory factors IL-1β,IL-6,IL-8,and TNF-α(P<0.05),and increased level of IL-10(P<0.05).Zhenwu Decoction can significantly improve renal function and renal pathological damage in DN mice of spleen-kidney Yang deficiency syndrome,and its specific mechanism may be related to the inhibition of inflammatory response by down-regulating the expression of key molecules in the ROCK/IKK/NF-κB pathway in the kidneys.