大黄泄浊方对慢性肾衰竭大鼠炎症及SOCS3/TLR4通路的影响  

Effect of Dahuang Xiezhuo Prescription on Inflammation and SOCS3/TLR4 Pathway in Rats with Chronic Renal Failure

作　　者：辛鑫 吴振华 檀淼[3] 高飞[4] 杨凤文[2] 陈素枝[2] 张倩 檀金川[1,2] XIN Xin;WU Zhenhua;TAN Miao;GAO Fei;YANG Fengwen;CHEN Suzhi;ZHANG Qian;TAN Jinchuan(Hebei University of Chinese Medicine,Shijiazhuang 050200,China;Hebei Province Hospital of Traditional Chinese Medicine,Shijiazhuang 050011,China;The Fourth Hospital of Hebei Medical University,Shijiazhuang 050000,China;Dongzhimen Hospital Affiliated to 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》2023年第7期95-104,共10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:观察大黄泄浊方对慢性肾衰竭(CRF)大鼠信号传导抑制因子3(SOCS3)/Toll样受体4(TLR4)通路的干预作用,探讨其减轻肾组织炎症反应的分子机制。方法:取雄性SD大鼠90只,随机留取15只为假手术组,剩余75只为造模组,行5/6肾切除术复制CRF大鼠模型,模型复制成功后,随机分为模型组,大黄泄浊方低、中、高剂量(6.825、13.65、27.3 g·kg^(-1))组、尿毒清颗粒组(2.6 g·kg^(-1)),分别予相应剂量的药物灌胃,连续灌胃8周。给药结束后,苏木素-伊红(HE)染色及马松(Masson)染色观察大鼠肾脏组织病理形态改变;检测大鼠血肌酐(SCr)、血尿素氮(BUN)、血尿酸(UA)水平;酶联免疫吸附测定法(ELISA)检测大鼠血清中白细胞介素-6(IL-6)、肿瘤坏死因子-α(TNF-α)、C反应蛋白(CRP)含量;实时荧光定量聚合酶链式反应(Real-time PCR)检测肾组织中SOCS3和TLR4 m RNA的表达;蛋白免疫印迹法(Western blot)检测信号传导抑制因子3(SOCS3)、TLR4、核转录因子-κB(NF-κB)、髓样分化因子88(My D88)蛋白的表达;免疫组化法检测NF-κB、My D88、NOD样受体蛋白3(NLRP3)、黑色素瘤缺乏因子2(AIM2)蛋白的表达。结果:与假手术组比较,模型组大鼠肾组织发生明显的炎症反应,血SCr、BUN、UTP、IL-6、TNF-α、CRP水平明显升高(P<0.05),肾组织中SOCS3蛋白及SOCS3 m RNA含量明显下调,TLR4、NF-κB、My D88、NLRP3、AIM2蛋白及TLR4 m RNA含量均明显升高(P<0.05);与模型组比较,大黄泄浊方和尿毒清颗粒可明显减轻肾组织中的炎症反应,降低血SCr、BUN、UTP、IL-6、TNF-α、CRP的表达水平(P<0.05),肾组织中SOCS3蛋白及SOCS3 m RNA含量显著上升,TLR4、NF-κB、My D88、NLRP3、AIM2蛋白及TLR4 m RNA含量均明显下降(P<0.05)。结论:大黄泄浊方可减少炎症因子的释放和表达,抑制炎症反应,改善肾功能,其机制可能与调控SOCS3/TLR4信号通路有关。Objective:To observe the intervention effect of Dahuang Xiezhuo prescription(DHXZ)on inflammation and suppressor of cytokine signaling 3(SOCS3)/Toll-like receptor 4(TLR4)pathway in rats with chronic renal failure(CRF),and to explore its molecular mechanism in alleviating renal inflammatory response.Method:The 90 male SD rats,15 were randomly selected as sham group,and the remaining 75 were used as modeling group to replicate CRF rat model by 5/6 nephrectomy.After successful modeling,the rats were randomly divided into model group,DHXZ low-,medium-,high-dose groups(6.825,13.65,27.3 g·kg^(-1))and Niaoduqing Granules group(2.6 g·kg^(-1)).The drug intervention groups received corresponding drugs by gavage for 8 consecutive weeks.After administration,hematoxylin-eosin(HE)staining and Masson staining were used to observe the morphological changes of rat renal tissue,and blood creatinine(SCr),blood urea nitrogen(BUN)and blood uric acid(UA)were tested.Enzyme-linked immunosorbent assay(ELISA)was performed to detect the serum contents of interleukin-6(IL-6),tumor necrosis factor-α(TNF-α)and C-reactive protein(CRP).The m RNA expressions of SOCS3 and TLR4 in renal tissue were detected by Real-time fluorescence quantitative polymerase chain reaction(Real-time PCR),and the protein expressions of SOCS3,TLR4,nuclear transcription factor(NF-κB)and myeloid differentiation factor(My D88)were detected by Western blot.Immunohistochemistry was used to determine the protein expressions of NF-κB,My D88,NOD-like receptor protein 3(NLRP3)and melanoma deficiency factor 2(AIM2).Result:Compared with the sham group,the model group had a significant inflammatory response in renal tissue,and an increase in blood SCr,BUN,UTP,IL-6,TNF-αand CRP(P<0.05).The protein and m RNA expressions of SOCS3 in renal tissue of rats in the model group were lower while the protein expressions of TLR4,NF-κB,My D88,NLRP3 and AIM2 and the m RNA expression of TLR4 were higher than those in the sham group(P<0.05).Compared with the model group,DHXZ and Niaoduqi

关 键 词：大黄泄浊方 慢性肾衰竭 炎症 肾间质纤维化 信号传导抑制因子3 TOLL样受体4 

分 类 号：R2-0[医药卫生—中医学] R33R289R692