PKA调控TRPV1敏化介导CGRP表达探讨疏肝健脾方防治肠易激综合征研究  

Experiment on Mechanism of Shugan Jianpi Decoction in Treating IBS by Observing Expressions of PKA,TRPV1 and CGRP

作　　者：方健松 马媛萍 刘畅[1] 张涛[1] 

机构地区：[1]广西中医药大学附属瑞康医院,广西南宁530011

出　　处：《中华中医药学刊》2017年第3期601-604,I0009,I0010,共6页Chinese Archives of Traditional Chinese Medicine

基　　金：国家自然科学基金项目(81460724)

摘　　要：目的:通过观察肠易激综合征(irritable bowels syndrome,IBS)大鼠结肠黏膜组织形态、超微结构及蛋白激酶A(protein kinase A,PKA)、辣椒素受体(transient receptor potential vanilloid 1,TRPV1)、降钙素基因相关肽(caleitonin gene-relatedpeptide,CGRP)的表达变化,探讨疏肝健脾方治疗IBS可能机制。方法:72只清洁级雄性大鼠按体质量随机分为正常组,模型组,疏肝健脾方组(低、中、高剂量)和对照组;用辣素灌胃结合束缚应激刺激法复制IBS大鼠模型。正常组予同步饲养;疏肝健脾方组分别按低、中、高不同剂量给药;对照组予匹维溴胺;模型组予等剂量生理盐水;连续灌胃2周。末次给药后,处死所有大鼠,取结肠组织并应用HE染色法和醛品红-桔黄G法分别检测病理组织学改变及肥大细胞阳性表达情况;采用Western-blot技术和荧光定剂量PCR检测大鼠结肠PKA、TRPV1、CGRP的表达情况。结果:光镜下见各组黏膜及黏膜上皮绒毛完整,排列规则,可见少许炎症细胞,未见黏膜下血管扩张及弥漫性炎症浸润改变,腺体结构、排列正常;正常组除外,各组大鼠结肠组织黏膜间质或黏膜下层可见类圆形紫红至深紫色染色点。模型组大鼠结肠组织中PKA、TRPV1、CGRP基因和蛋白表达明显高于正常组(P"0.05)。高剂量组PKA、TRPV1、CGRP以及中剂量组CGRP基因表达和高剂量组PKA、CGRP蛋白表达均低于模型组(P"0.05),其余低、中剂量各组PKA、TRPV1及CGRP表达与模型组比较,差异均无统计学意义(P#0.05)。结论:疏肝健脾方可能通过PKA调节IBS大鼠TRPV1、CGRP的变化,是其发挥疗效的作用机制之一。Objective： The mechanism of Shugan Jianpi Decoction treating irritable bowel syndrome was investigated by observing the colonic morphology, ultrastructure and the expression of PKA, TRPV1 and CGRP. Methods： Seventy - two clean grade male SD rats according to weight were randomly divided into normal group, blank group, control group and intervention group （ low - dosage, middle - dosage, high - dosage）. Except the normal group, the other groups were model- ing by capsaicin and restraint stress. After the modeling, the normal group was given normal feeding. The intervention group and the control group were separately given different dosages of Shugan Jianpi Decoction and pinaverium bromide suspension liquid by garage. And the blank group was given the same volume saline for two weeks. All rats were killed after five weeks. The colon was detected by HE and fuchsin - aldehyde orange G reagent staining for pathology. The ex- pressions of PKA,TRPV1 and CGRP were detected by SYBR green real -time PCR and Western -blot. Results： Under the light microscope, the mucosal epithelium and the villi in each group were intact. Sub mucosal vessels were not dilated with some inflammation cells around. The arrangement of glandular and its structure were normal. Except the normal group, the colons of other groups were stained purple dot or dark purple dot. The expressions of PKA gene, TRPVI gene, CGRP gene and PKA protein, CGRP protein of high - dosage and the CGRP gene of middle - dosage were lower than those of the blank group （ P 〈 0.05 ）. Compared with the blank group, the expressions of gene and protein of low -dosage and other middle- dosage had no statistical significance（ P 〉0.05 ）. Conclusion： The mechanism of Shugan Jianpi De- coction treating rritable bowel syndrome may be associated with down - regulating the expressions of PKA, TRPV1 and CGRP.

关 键 词：肠易激综合征 疏肝健脾 PKA TRPV1 CGRP 

分 类 号：R574.4[医药卫生—消化系统]