小青龙汤通过TLR4/MyD88/NF-κB信号通路抑制LPS诱导慢性肾小球肾炎的机制研究  

Xiaoqinglong Decoction inhibits LPS-induced chronic glomerulonephritis through TLR4/MyD88/NF-κB signaling pathway

作　　者：柴希 储开博 赖永健 柴溥苒 张育敏 何丽清 CHAI Xi;CHU Kai-bo;LAI Yong-jian;CHAI Pu-ran;ZHANG Yu-min;HE Li-qing(Shanxi University of Traditional Chinese Medicine,Jinzhong 030619,China)

机构地区：[1]山西中医药大学,山西晋中030619

出　　处：《时珍国医国药》2023年第10期2333-2337,共5页Lishizhen Medicine and Materia Medica Research

基　　金：国家自然科学基金面上项目(82274365);山西省自然科学基金面上项目(20210302123228);山西中医药大学科技创新团队(2022TD1011)。

摘　　要：目的探究小青龙汤通过TLR4/MyD88/NF-κB信号通路抑制LPS诱导的慢性肾小球肾炎的机制。方法24只健康Wistar大鼠随机分为对照组、模型组、小青龙汤组。对照组切片在DMEM基础培养基中培养80min,LPS模型组将10μg/mL浓度的LPS加入基础培养基中,切片在基础培养基中培养30min,转入含有LPS的基础培养基中培养50min。小青龙汤组:将含药血清溶解于基础培养基中,继续培养LPS模型组。造模成功后,取出切片进行指标检测。观察肾组织形态学变化;用ELISA检测肾组织中IL-1β、IL-4、TNF-α等含量;RT-qPCR和Western blot检测肾组织TLR4、MyD88、NF-κB mRNA及蛋白表达水平。结果与对照组相比,模型组大鼠肾组织中可见明显的炎性细胞浸润,偶见肾小球体积增大,增生的肾小球内细胞数量增多,肾小管的管腔扩张且内有明显的透明管型,肾组织中IL-1β、TNF-α、IL-6和IL-8的含量显著升高,IL-4显著下降(P<0.01);Western blot检测结果显示,与对照组比较,模型组大鼠肾组织TLR4、MyD88和P-NF-κB-p65蛋白表达升高,NF-κBp65蛋白表达降低(P<0.01);RT-qPCR显示,与对照组比较,模型组大鼠肾组织中TLR4,MyD88,NF-κB p65 mRNA表达水平有不同程度增高(P<0.01)。结论小青龙汤通过TLR4/MyD88/NF-κB信号通路改善LPS诱导的慢性肾小球肾炎大鼠肾组织的炎症损伤。Objective To investigate the inhibitory effect of Xiaoqinglong Decoction on LPS-induced chronic glomerulonephritis through TLR4/MyD88/NF-κB signaling pathway.Methods Twenty-four healthy Wistar rats were randomly divided into control group,model group and Xiaoqinglong Decoction group.The slices in the control group were cultured in DMEM basal medium for 80 minutes,and the slices in the LPS model group were cultured in basal medium with 10μg/mL LPS for 30 minutes,and then transferred to basal medium containing LPS for 50 minutes.Xiaoqinglong Decoction group:the drug-containing serum was dis-solved in the basal medium,and the LPS model group was further cultured.After the model was successfully established,the slices were taken out for index detection.The morphological changes of renal tissue were observed.The contents of IL-1β,IL-4 and TNF-αin renal tissue were detected by ELISA.Rt-qpcr and Western blot were used to detect the mRNA and protein ex-pression levels of TLR4,MyD88,and NF-κB in kidney tissue.Results Compared with the control group,the renal tissue of the model group showed obvious inflammatory cell infiltration,occasionally increased glomerular volume,increased number of cells in the proliferative glomerulus,dilated lumen of the renal tubules with obvious transparent cast,and significantly increased IL-1β,TNF-α,IL-6 and IL-8 contents in the renal tissue.The level of IL-4 was significantly decreased(P<0.01).Western blot results showed that compared with the control group,the protein expressions of TLR4,MyD88 and p-NF-κB-p65 in renal tissue of rats in the model group were increased,and the protein expression of NF-κBp65 was decreased(P<0.01).Rt-qpcr showed that compared with the control group,the expression levels of TLR4,MyD88 and NF-κB p65 mRNA in the kidney tissue of the model group were increased to varying degrees(P<0.01).Conclusion Xiaoqinglong Decoction can improve renal inflammatory injury in rats with LPS-induced chronic glomerulonephritis through TLR4/MyD88/NF-κB signaling pathway.

关 键 词：小青龙汤 Toll样受体4(TLR4)/髓样分化因子88(MyD88)/核转录因子-κB(NF-κB)信号通路 慢性肾小球肾炎 炎症反应 

分 类 号：R285.5[医药卫生—中药学]