刺五加注射液通过抑制TLR4/NF-κB信号通路减轻糖尿病肾病小鼠肾损伤的研究  

Study on Acanthopanax senticosus Injection on alleviating renal injury in diabetic kidney disease mice by inhibiting TLR4/NF-κB signaling pathway

作　　者：郭金明[1] 付永涛 索大伟 江旗琳 白敏[1] 郑聪聪 王莉平[1] GUO Jinming;FU Yongtao;SUO Dawei;JIANG Qilin;BAI Min;ZHENG Congcong;WANG Liping(Handan Central Hospital,Handan 056001,China)

机构地区：[1]邯郸市中心医院,邯郸056001

出　　处：《天津中医药》2025年第1期100-107,共8页Tianjin Journal of Traditional Chinese Medicine

基　　金：河北省医学科学研究课题计划(20210854)。

摘　　要：[目的]研究刺五加注射液(ASI)调控Toll样受体4/核因子-κB(TLR4/NF-κB)信号通路对糖尿病肾病(DKD)小鼠肾损伤的影响。[方法]采用高脂饮食结合腹腔注射链脲佐菌素的方法构建DKD小鼠动物模型,设正常对照组、模型组、ASI(75 mg/kg)组、ASI(75 mg/kg)+瑞沙托维(TAK-242,TLR4抑制剂,0.5 mg/kg)组和ASI(75 mg/kg)+脂多糖(LPS,TLR4激动剂,10 mg/kg)组,每组10只。持续给药8周后,检测小鼠空腹血糖(FBG)和肾功能指标[24 h尿蛋白量(24 h UTP)、尿酸(UA)、血尿素氮(BUN)和血清肌酐(Scr)],称量体质量、肾脏质量并计算肾脏系数,酶联免疫吸附法(ELISA)检测肾组织炎症因子[白细胞介素(IL)-1β、IL-6、肿瘤坏死因子-α(TNF-α)]含量,苏木精-伊红(HE)染色法观察肾组织病理改变,马森(Masson)染色和末端标记法(TUNEL)检查肾纤维化和肾细胞凋亡,逆转录实时定量聚合酶链反应(RT-PCR)法检测肾组织TLR4、NF-κB p65 mRNA表达,蛋白免疫印迹(Western blot)法检测肾组织TLR4、NF-κB p65、B细胞淋巴瘤-2(Bcl-2)、Bcl-2相关X蛋白(Bax)、半胱氨酸蛋白酶-3(Caspase-3)、活化型Caspase-3(Cleaved Caspase-3)蛋白表达。[结果]与模型组相比,ASI组和ASI+TAK-242组小鼠FBG、24 h UTP、UA、BUN、Scr、肾脏质量、肾脏系数、IL-1β、IL-6、TNF-α均显著降低,体质量显著升高(P<0.05);肾组织病理改变、纤维化及肾细胞凋亡状况均明显改善,肾组织损伤评分、胶原容积分数(CVF)及细胞凋亡指数(AI)均显著降低(P<0.05);TLR4、NF-κB p65 mRNA和蛋白表达量显著降低,Bax/Bcl-2、Cleaved Caspase-3/Caspase-3蛋白表达比值显著降低(P<0.05)。TAK-242能够明显增强ASI对DKD小鼠FBG、肾功能、体质量、肾脏系数、肾组织病变、纤维化、细胞凋亡及TLR4/NF-κB信号通路相关mRNA和蛋白表达的作用;LPS则明显逆转ASI对DKD小鼠的上述作用。[结论]ASI具有减轻DKD小鼠肾损伤并改善其肾功能的作用,其机制可能与下调TLR4/NF-κB信号通�[Objective]To investigate the effect of Acanthopanth senticosus Injection(ASI)regulating toll-like receptor 4/nuclear factor-κB(TLR4/NF-κB)signaling pathway on renal injury in diabetic kidney disease(DKD)mice.[Methods]The DKD mice model was established by high-fat diet combined with intrabitoneal injection of streptozotocin.And set up the normal control group,model group,ASI(75 mg/kg)group,ASI(75 mg/kg)+ressatovi(TAK-242,TLR4 inhibitor,0.5 mg/kg)group and ASI(75 mg/kg)+lipopolysaccharide(LPS,TLR4 agonist,10 mg/kg)group,with 10 mice in each group.After continuous administration for 8 weeks,the fasting blood glucose(FBG)and renal function indexes[24 h urinary protein(24 h UTP),uric acid(UA),blood urea nitrogen(BUN),serum creatinine(Scr)]of the mice were detected;the body mass and renal mass were weighed,and the renal index was calculated.The content of inflammatory factors[interleukin-1β(IL-1β),IL-6,tumor necrosis factor-α(TNF-α)]in renal tissue were detected by ELISA.The pathological changes of renal tissue were observed through HE staining.The renal fibrosis and renal cells apoptosis were detected through masson and TUNEL staining.The expression of TLR4,NF-κB p65 mRNA in renal tissue were detected by RT-PCR.The expression of TLR4,NF-κB p65,B-cell lymphoma-2(Bcl-2),Bcl-2 associated X protein(Bax),Caspase-3,Cleaved Caspase-3 proteins were detected by Western blot.[Results]Compared with the model group,the FBG,24 h UTP,UA,BUN,Scr,renal mass,renal index,IL-1β,IL-6,TNF-αwere significantly decreased in ASI group and ASI+TAK-242 group,while the body mass was significantly increased(P<0.05).The pathological changes and fibrosis of renal tissue,apoptosis of renal cells were significantly improved;and the renal tissue injury score,collagen volume fraction(CVF)and apoptosis index(AI)were significantly decreased(P<0.05).The mRNA and protein expressions of TLR4,NF-κB p65 were significantly decreased,and the protein expression ratio of Bax/Bcl-2 and Cleaved Caspase-3/Caspase-3 were significantly decreased(P<0.05).TA

关 键 词：糖尿病肾病 刺五加注射液 肾损伤 TLR4/NF-κB信号通路 炎症 凋亡 

分 类 号：R587.2[医药卫生—内分泌]