基于TLR4/MyD88/NF-κB信号通路探讨大补脾汤联合奥沙利铂对胃癌荷瘤小鼠肠道屏障损伤的影响  

Effect of Dabupi Decoction(大补脾汤))Combined with Oxaliplatin on Intestinal Barrier Injury in Tumor-Bearing Mice with Gastric Cancer Based on TLR4/MyD88/NF-kB Signaling Pathway

作　　者：龚红霞 董焕成 苏韫 曹旺杰[1,2,3,4] 和建政 刘永琦 张晗 曾元丁[1,2,3] 李从艺 康倩[1,2,3] GONG Hongxia;DONG Huancheng;SU Yun;CAO Wangjie;HE Jianzheng;LIU Yongqi;ZHANG Han;ZENG Yuanding;LI Congyi;KANG Qian(Basic Medical College of Gansu University,of Chinese Medicine,Lanzhou73000;Provincial Key Laboratory of Molecular Medicine and Traditional Chinese Medicine Prevention and Treatment in Gansu Clleges and Universities,Lanzhou 730000;Key Laboratory of Dunhuang Ministry of Medicine and Transformation,Lanzhou 730000;Key Laboratory of Diagnosis and Treatment of Gastrointestinal Tumors of the National Health Commission,Lanzhou 730000;Key Laboratory of Traditional Chinese Medicine for Prevention and Treatment of Chronic Diseases in Gansu Province,Lanzhou 730000)

机构地区：[1]甘肃中医药大学基础医学院,兰州730000 [2]甘肃省高校重大疾病分子医学与中医药防治研究省级重点实验室,兰州730000 [3]敦煌医学与转化教育部重点实验室,兰州730000 [4]国家卫生健康委胃肠肿瘤诊治重点实验室,兰州730000 [5]甘肃省中医药防治慢性疾病重点实验室,兰州730000

出　　处：《中药药理与临床》2023年第8期31-36,共6页Pharmacology and Clinics of Chinese Materia Medica

基　　金：敦煌医学与转化教育部重点实验室开放基金(编号:DHYX19-08);中国医学科学院中央级公益性科研院所基本科研业务费专项资金资助(编号:2019PT320005);甘肃省高效中(藏)药化学与质量研究重点实验室开放基金(编号:zzy-2022-06);甘肃省“双一流”科研重点项目(编号:GSSYLXM-05)。

摘　　要：目的:探讨大补脾汤联合奥沙利铂通过影响TLR4/MyD88/NF-κB信号通路及下游相关分子的表达,对胃癌荷瘤小鼠肠道屏障损伤的影响。方法:建立MFC胃癌荷瘤小鼠模型,随机分为模型对照组、奥沙利铂0.01 g/kg组、奥沙利铂加大补脾汤5.4、10.8、21.6 g/kg组,另选10只健康小鼠作为正常对照组;各组小鼠经灌胃或腹腔注射给予相应药物或生理盐水,连续给药14 d。末次给药后,眼球取血收集血清并取其肠道组织样本。HE染色观察小肠组织形态的变化,ELISA法检测血清中肿瘤坏死因子α(TNF-α)及D-乳酸的含量,RT-qPCR和WB法分别检测小肠组织中Toll样受体4(Toll-like receptor 4,TLR4)、髓样分化因子88(myeloiddifferentiationfactor88,MyD88)、核转录因子κB(nuclear factor kappa-B,NF-κB)、闭合蛋白(Occludin)的mRNA和蛋白表达。结果:与正常对照组比较,模型对照组病理显示小鼠小肠绒毛稀疏、断裂,各给药组上述病理改变均有不同程度的改善,模型对照组小鼠血清中TNF-α和D-乳酸含量显著升高(P<0.01),小肠组织中Tlr4、Myd88和Nfκb p65 mRNA及蛋白的表达明显上调,Occludin mRNA及蛋白的表达明显下调(P<0.05或P<0.01);与模型对照组比较,奥沙利铂加大补脾汤各组血清中TNF-α、D-乳酸含量均显著降低(P<0.01);小肠组织中Tlr4、Myd88和Nfκb p65 mRNA及蛋白表达明显下调,Occludin mRNA及蛋白表达明显上调(P<0.05或P<0.01)。结论:大补脾汤联合奥沙利铂通过TLR4/MyD88/NF-κB信号通路可减轻对胃癌荷瘤小鼠肠道屏障的损伤。Objective:To investigate the effect of Dabupi Decoction(大补脾汤)combined with oxaliplatin on intestinal barrier injury in tumorbearing mice with gastric cancer by affecting the Toll-like receptor 4(TLR4)/myeloid differentiation factor 88(MyD88)/nuclear factor kappa-B(NF-kB)signaling pathway and downstream related molecules.Methods:An MFC tumor-bearing mouse model with gastric cancer was established and the model mice were randomly divided into a model control group,an oxaliplatin(0.01 g/kg)group,and oxaliplatin combined with Dabupi Decoction(21.58,10.79,and 5.40 g/kg)groups,with 10 mice in each group.Additionally,10 healthy mice were selected and assigned to the normal control group.The mice in each group were administered the corresponding drugs or saline solution by gavage or intraperitoneal injection for 14 consecutive days.After the final administration,blood samples were collected from the eyeballs to obtain serum and intestinal tissue samples were collected.HE staining was performed to observe the morphological changes in the small intestine tissues.ELISA was used to measure the levels of tumor necrosis factor-α(TNF-α)and D-lactic acid in the serum.RT-qPCR and Western blot were performed to detect the protein and mRNA expression of TLR4,MyD88,NF-kB,and Occludin in the intestinal tissues.Results:Compared with the normal control group,the model control group showed sparse and fractured villi in the smallintestine,which were improved to varying degrees in the treatment groups.The levels of TNF-αand D-lactic acid in the serum of the model control group significantly increased(P<0.01).The mRNA and protein expression levels of Tlr4,Myd88,and Nfkb p65 in the small intestine tssues were significantly up-regulated and Occludin was down-regulated(P<0.05 or P<0.01).Compared with the model control group,the combined treatment of oxaliplatin and Dabupi Decoction significantly reduced the levels of TNF-αand D-lactic acid in the serum(P<0.01),down-regulated the relative mRNA and protein expression levels of Tlr4,

关 键 词：大补脾汤 胃癌 肠道屏障损伤 Toll样受体4/髓样分化因子88/核转录因子-κB信号通路 

分 类 号：R285.5[医药卫生—中药学]