大黄泄浊方调控ROS/TXNIP/NLRP3通路对5/6肾切除大鼠肾间质纤维化的影响  

Effect of Dahuang Xiezhuo Prescription on Renal Interstitial Fibrosis of Rats with 5/6 Nephrectomy by Regulating ROS/TXNIP/NLRP3 Pathway

作　　者：曹雯萱 辛鑫 檀淼[3] 高飞[4] 杨凤文[2] 陈素枝[2] 任美芳[2] 檀金川[2] CAO Wenxuan;XIN Xin;TAN Miao;GAO Fei;YANG Fengwen;CHEN Suzhi;REN Meifang;TAN Jinchuan(Hebei University of Chinese Medicine,Shijiazhuang 050200,China;Hebei Provincial Hospital of Traditional Chinese Medicine,Shijiazhuang 050011,China;The Fourth Hospital of Hebei Medical University,Shijiazhuang 050000,China;Dongzhimen Hospital,Beijing University of Chinese Medicine,Beijing 100700,China)

机构地区：[1]河北中医学院,石家庄050200 [2]河北省中医院,石家庄050011 [3]河北医科大学第四医院,石家庄050000 [4]北京中医药大学东直门医院,北京100700

出　　处：《中国实验方剂学杂志》2022年第21期81-89,共9页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：河北省中医药管理局科研计划项目(2021030)。

摘　　要：目的:通过观察大黄泄浊方对5/6肾切除大鼠肾脏病理变化,肾组织活性氧(ROS)/硫氧还蛋白互作蛋白(TXNIP)/NOD样受体蛋白3(NLRP3)通路表达的变化,研究其保护肾功能,延缓肾间质纤维化的作用机制及可能性。方法:90只健康SD雄性大鼠随机分为假手术组、模型组、大黄泄浊方低、中、高剂量(6.825、13.65、27.30 g·kg^(-1))组和尿毒清颗粒(2.60 g·kg^(-1))组。除假手术组外其余5组均采用5/6肾切除术复制CRF大鼠模型。造模成功后,各给药组分别给予相应剂量的药物混悬液灌胃,每日1次,连续8周。给药结束后,检测大鼠血清中肌酐(SCr)、尿素氮(BUN)水平和24 h尿蛋白定量(UTP)水平;蛋白免疫印迹法(Western blot)检测硫氧还蛋白(TRX)、TXNIP和NLRP3蛋白的表达情况;免疫组化法检测NLRP3、TRX、TXNIP、接头蛋白凋亡相关斑点样蛋白(ASC)、转化生长因子-β(TGF-β)、Ⅳ型胶原蛋白(CollagenⅣ)、α-平滑肌肌动蛋白(α-SMA)和调纤连蛋白(FN)的表达。结果:与假手术组比较,模型组大鼠血SCr、BUN、24 h UTP水平明显增高(P<0.05),TRX、TXNIP、NLRP3、ASC、TGF-β、CollagenⅣ、α-SMA和FN蛋白显著增高(P<0.01),肾间质发生显著纤维化;与模型组比较,大黄泄浊方各剂量组和尿毒清颗粒组血SCr、BUN、24 h UTP水平均有不同程度降低(P<0.05),TRX、TXNIP、NLRP3、ASC、TGF-β、CollagenⅣ、α-SMA和FN蛋白显著降低(P<0.01),肾间质纤维化不同程度改善。结论:大黄泄浊方可保护慢性肾衰竭大鼠肾功能、延缓肾间质纤维化。Objective:To observe the effect of Dahuang Xiezhuo prescription on the changes in rena pathology and reactive oxygen species (ROS)/thioredoxin-interacting protein (TXNIP)/NOD-like receptor protein 3(NLRP3)pathway expression in the kidney tissues of rats with 5/6 nephrectomy,and to explore the mechanism of Dahuang Xiezhuo prescription in protecting renal function and delaying renal interstitial fibrosis and the possibility.Method:Ninety healthy male SD rats were randomly divided into a sham operation group,a model group,low,medium,and high-dose(6.825,13.65,27.30 g·kg^(-1))Dahuang Xiezhuo prescription groups,and a Niaoduqing granule group(2.60 g·kg^(-1)).Except the sham operation group,5/6 nephrectomy was used to replicate the rat model of chronic renal failure(CRF).After modeling,each administration group was given the corresponding dose of drug suspension by intragastric administration,once a day for consecutive 8weeks.After administration,serum creatinine(SCr)and urea nitrogen(BUN)levels and 24 h urinary protein quantification(UTP)levels were detected.Western blot assay was used to detect the protein expressions of thioredoxin(TRX),TXNIP,and NLRP3.The protein expressions of TRX,TXNIP,NLRP3,apoptosisassociated speck-like protein containing a CARD(ASC),transformation growth factor-β(TGF-β),CollagenⅣ,α-smooth muscle actin(α-SMA),and fibronectin(FN)were detected by immunohistochemistry.Result:As compared with the sham operation group,serum levels of SCr,BUN,and UTP in the model group were increased(P<0.05),TRX,TXNIP,NLRP3,ASC,TGF-β,CollagenⅣ,α-SMA,and FN proteins were increased(P<0.01),and renal interstitial fibrosis significantly occurred.As compared with the model group,the levels of SCr,24 h BUN,and UTP in the low,medium,and high-dose Dahuang Xiezhuo prescription groups and the Niaoduqing granule group were decreased to varying degrees(P<0.05),TRX,TXNIP,NLRP3,ASC,TGF-β,CollagenⅣ,α-SMA,and FN were decreased(P<0.01),and renal interstitial fibrosis was improved to varying degrees.Conclusion:Dahuang X

关 键 词：大黄泄浊方 慢性肾衰竭 肾间质纤维化 氧化应激 硫氧还蛋白相互作用蛋白(TXNIP) NOD样受体蛋白(3 NLRP3)炎性小体 

分 类 号：R2-0[医药卫生—中医学] R33R289R692