电针“内关-中脘”对功能性消化不良大鼠胃动力及相关脑肠肽的影响

Effects of electroacupuncture‘Neiguan(PC 6)-Zhongwan(RN 12)’on gastric motility and related brain gut peptides in functional dyspepsia rats

目的:观察电针“内关”“中脘”及“内关-中脘”对功能性消化不良(FD)大鼠胃动力和相关脑肠肽的影响。方法:将56只SPF级雄性SD幼鼠随机分为正常组(16只)和造模组(40只),造模组采用0.1%蔗糖碘乙酰胺溶液灌胃叠加改良小平台法制备FD大鼠模型。正常组和造模组各随机选8只进行模型评价,确认模型制备成功后,将造模组剩余32只大鼠随机分为模型组、内关组、中脘组、内关-中脘组,每组8只。上述干预组分别予以电针干预双侧“内关”“中脘”以及双侧“内关”和“中脘”,参数为连续波、频率2 Hz、电流强度1 mA,30 min/d,持续7 d。干预结束后测量各组大鼠胃排空率;HE染色观察大鼠胃窦组织病理形态学改变,ELISA检测血清中5-羟色胺(5-HT)含量,Western Blot法检测血清中胃泌素(GAS)、胃动素(MTL)、降钙素基因相关肽(CGRP)以及胃窦组织中P物质(SP)、缩胆囊素(CCK)、饥饿激素(Ghrelin)蛋白表达量。结果:光镜下可见各组胃窦组织黏膜层完整,腺体排列有序,胃黏膜未见明显炎症浸润。与模型组比较,中脘组、内关-中脘组大鼠胃排空率显著上升(P<0.01),血清5-HT含量显著降低(P<0.05);内关组、中脘组及内关-中脘组GAS、MTL、Ghrelin蛋白表达量显著升高(P<0.05,P<0.01),SP蛋白表达量显著降低(P<0.05,P<0.01);内关-中脘组CGRP蛋白表达量显著降低(P<0.05),CCK蛋白表达量显著升高(P<0.05)。内关-中脘组GAS、MTL蛋白表达量显著高于中脘组、内关组(P<0.01,P<0.05),内关-中脘组SP蛋白表达量显著低于内关组(P<0.05);内关-中脘组Ghrelin蛋白表达量显著高于中脘组(P<0.01),中脘组MTL、Ghrelin蛋白表达量显著高于内关组(P<0.01,P<0.05)。结论:电针“内关-中脘”能够改善FD大鼠胃动力障碍,缓解大鼠消化不良症状,降低CCK、CGRP等脑肠肽水平,且明显优于单用“中脘”“内关”,可能与两穴配伍协同调节脑肠轴的功能,从而影响脑肠肽的分泌有关。

Objective:To observe the effects of electroacupuncture Neiguan(PC 6),Zhongwan(RN 12)and Neiguan-Zhongwan on gastric motile and braingutpeptide in functional dyspepsia(FD)rats.Methods:A total of 56 SPF grade male SD rats were randomly divided into normal group(n=16)and modeling group(n=40).The FD rat model was prepared by using 0.1%sucrose iodoacetamide solution and modified small platform method.After confirming the successful preparation of the model,the remaining 32 rats in the modeling group were randomly divided into model group,Neiguan group,Zhongwan group and Neiguan-Zhongwan group,with 8 rats in each group.The above intervention groups were respectively subjected to electroacupuncture intervention of bilateral Neiguan(PC 6)and Zhongwan(RN 12)and Neiguan(PC 6),and Zhongwan(RN 12),with continuous wave,frequency 2Hz,current intensity 1 mA,30 min/d for 7 days.The gastric emptying rate was measured after the intervention.The histopathologic changes of gastric antrum were observed by HE staining,and the content of 5-HT in serum was detected by ELISA.The expression levels of gastrin(GAS),motilin(MTL),calcitonin gene-related peptide(CGRP)in serum and substance P(SP),cholecystokinin(CCK)and Ghrelin in antrum tissues were detected by Western Blot.Results:Under light microscope,the mucosa of gastric antrum tissue was intact,the glands were arranged in order,and no obvious inflammatory infiltration was observed in gastric mucosa.Compared with model group,gastric emptying rate in Zhongwan group and Neiguan-Zhongwan group was significantly increased(P<0.01),and serum 5-HT content was significantly decreased(P<0.05).The protein expressions of GAS,MTL and Ghrelin in Neiguan group,Zhongwan group and Neiguan-Zhongwan group were significantly increased(P<0.05,P<0.01),while the protein expression of SP was significantly decreased(P<0.05.P<0.01).The expression level of CGRP protein in Neiguan-Zhongwan group was significantly decreased(P<0.05),and the expression level of CCK protein was significantly increased(P<0.05).The protein expression levels of GAS and MTL in Neiguan-Zhongwan group were significantly higher than those in Zhongwan and Neiguan groups(P<0.01,P<0.05),and the protein expression levels of SP in Neiguan-Zhongwan group were significantly lower than those in Neiguan group(P<0.05).The expression of Ghrelin protein in Neiguan-Zhongwan group was significantly higher than that in Zhongwan group(P<0.01),the expression of MTL and Ghrelin protein in Zhongwan group was significantly higher than that in Neiguan group(P<0.01,P<0.05).Conclusion:Electroacupuncture Neiguan-Zhongwan can improve gastric motility disorder in FD rats,relieve dyspeptic symptoms in rats,and reduce the level of brain and intestinal peptides such as CCK and CGRP,which is obviously better than Zhongwan and Neiguan alone.which may be related to the coordination of the two points to regulate the function of brain and intestinal axis,thus affecting the secretion of brain and intestinal peptides.