基于TLR4/ERK通路探讨电针对功能性消化不良大鼠肠黏膜通透性的影响  

The Effect of Electroacupuncture on Intestinal Permeability in Rats with Functional Dyspepsia Based on TLR4/ERK Pathway

作　　者：康朝霞 崔冬梅 KANG Zhaoxia;CUI Dongmei(Guizhou University of Traditional Chinese Medicine,Guiyang Guizhou 550025,China)

机构地区：[1]贵州中医药大学,贵州贵阳550025

出　　处：《中医药导报》2024年第5期29-35,共7页Guiding Journal of Traditional Chinese Medicine and Pharmacy

基　　金：国家自然科学基金项目(82060897);贵州省基础研究(自然学科)项目(黔科合基础-ZK[2022]一般466);贵州省中医药管理局中医药、民族医药科学技术研究项目(QZYY-2022-007);贵州省教育厅高等学校科学研究项目青年项目(黔科技[2022]211);贵州中医药大学博士启动基金(贵中医博士启动[2020]27号)。

摘　　要：目的:基于TLR4/ERK通路探讨电针对功能性消化不良(FD)大鼠肠黏膜通透性的影响。方法:选用50只SPF级8周龄SD雄性大鼠,按随机数字表法分为空白组(10只)、造模组(40只),造模组采用郭氏夹尾刺激法+不规则饮食法+冰生理盐水灌胃法的复合造模方法建立FD模型。选取造模成功的大鼠30只随机分为模型组、电针组、TAK242组,每组10只。电针组针刺“足三里“”太冲”穴后在足三里穴接电针仪干预,连续波,频率2 Hz,强度1 mA,每次干预30 min,1次/d,共14 d;TAK242组予以TLR4抑制剂TAK242(0.5 mg/kg)尾静脉注射,1次/d,共14 d。干预结束后比较各组大鼠胃内残留率、小肠推进率;ELISA检测血清中二胺氧化酶(DAO)浓度;生化检测方法检测脂多糖(LPS)、D-乳酸(D-LA)的浓度;免疫组化法检测MBP蛋白的表达;免疫印迹法检测TLR4、MyD88、MEK、ERK、p-MEK、p-ERK1/2蛋白的表达。结果:模型组大鼠胃内残留率较空白组明显上升(P<0.01);与模型组比较,电针组、TAK242组胃内残留率均下降(P<0.01);电针组胃内残留率低于TAK242组(P<0.05)。模型组大鼠小肠推进率较空白组显著下降(P<0.01);电针组、TAK242组小肠推进率较模型组明显增加(P<0.01);电针组小肠推进率高于TAK242组(P<0.05)。与空白组比较,模型组大鼠十二指肠TLR4、MyD88蛋白相对表达量显著增加(P<0.01);与模型组比较,电针组、TAK242组十二指肠组织TLR4、MyD88蛋白相对表达量显著降低(P<0.01);TAK242组TLR4蛋白相对表达量低于电针组(P<0.05),TAK242组MyD88蛋白相对表达量与电针组比较,差异无统计学意义(P>0.05)。与空白组比较,模型组大鼠十二指肠组织MEK、ERK、p-MEK、p-ERK1/2蛋白相对表达量显著上调(P<0.01);电针组、TAK242组大鼠十二指肠组织MEK、ERK、p-MEK、p-ERK1/2蛋白相对表达量均较模型组降低(P<0.01);电针组MEK、ERK、p-MEK、p-ERK1/2蛋白相对表达量低于TAK242组(P<0.05)。模型组大鼠十二指肠组织MBObjective:To explore the effect of electroacupuncture on intestinal mucosal permeability in rats with functional dyspepsia(FD)through the TLR4/ERK pathway.Methods:A total of 50 SPF-grade 8-week-old male SD rats were randomly divided into blank group(n=10)and modeling group(n=40).The FD model was established in the modeling group using a combination of Guo's tail clamping stimulation method,irregular diet method and ice physiological saline gavage method.A total of 30 successfully modeled rats were randomly divided into model group,electroacupuncture group and TAK242 group,with 10 rats in each group.The electroacupuncture group received electroacupuncture intervention after needling"Zusanli"(ST 36)and"Taichong"(LR 3),with continuous waves,a frequency of 2 Hz,intensity of 1 mA,30 minutes per session,once daily for 14 days.The TAK242 group was intravenously injected with the TLR4 inhibitor TAK242(0.5 mg/kg)once daily for 14 days.After the intervention,the residual rate in the stomach and the propulsion rate in the small intestine were compared among the groups.The serum concentration of diamine oxidase(DAO)was measured by ELISA.The concentrations of lipopolysaccharide(LPS)and D-lactic acid(D-LA)were measured by biochemical methods.The MBP protein expression was measured by immunohistochemical.The protein expressions of TLR4,MyD88,MEK,ERK,p-MEK and p-ERK1/2 in the duodenal tissue were detected by Western blotting.Results:The gastric residue rate significantly increased in model group compared to the blank group(P<0.01).After intervention,both the electroacupuncture group and the TAK242 group showed a significant decrease in gastric residue rate compared to the model group(P<0.01).The electroacupuncture group showed lower gastric residue rate than TAK242 group(P<0.05).The small intestinal propulsion rate significantly decreased in model group compared to the blank group(P<0.01).After intervention,both the electroacupuncture group and the TAK242 group showed a significant increase in small intestinal propulsion rate co

关 键 词：功能性消化不良 电针 肠黏膜通透性 TLR4 大鼠 

分 类 号：R285.5[医药卫生—中药学]