Deng Duo-Xi, Tan Jie, Zhang Hong, Huang Gui-Lan, Li Shuai, Guo Kui-Kui, Wu Yun-Yun, Yang Bi-Ying, Xiao Yao, Peng Liang, Xie Hui
College of Acupuncture-moxibustion and Massage, Hunan University of Traditional Chinese Medicine, Changsha 410208, China.
Department of Rehabilitation, Chenzhou First People's Hospital, Chenzhou 423000, Hunan Province, China.
Zhen Ci Yan Jiu. 2018 Aug 25;43(8):485-91. doi: 10.13702/j.1000-0607.170513.
To observe the effect of electroacupuncture (EA) stimulation of "Shangjuxu" (ST 37, Lower Confluent point) and "Tianshu" (ST 25, Front-Mu point) on visceral pain and expression of colonic tryptase(Try), proteinase-activated receptor 2(PAR-2),transient receptor potential channel vanilloid subfamily member 1 (TPRV 1),substance P (SP) and calcitonin gene-related peptide (CGRP) in rats with irritable bowel syndrome (IBS), so as to explore its mechanisms underlying improvement of IBS.
Forty male Sprague Dawley rats were equally randomized into normal control (control), model, medication and EA groups (=10 in each). The IBS model was established by chronic acute combining stress (CACS, water deprivation, fasting, tail clamping, forced swimming in ice water, restraint, etc.) for 21 days. Rats of the medication group were treated by gavage of Pinaverium Bromide (1 mg/mL, 15 mg/kg), once daily for 14 d. EA (10 Hz/50 Hz, 0.2-0.3 mA) was applied to bilateral ST 37 and ST 25 for 30 min, once daily for 14 d. The muscular withdrawal reflex (AWR) of both abdomen and buttock was detected by colorectal distension (CRD) with a water-filled balloon for examining the visceral hypersensitivity. The number of mast cells in the colonic tissue was counted after toluidine blue stain. The immunoactivity of colonic Try was determined by immunochemistry and the expression of colonic PAR-2, TRVP 1, SP and CGRP proteins detected by Western blot.
After modeling, the body weight was significantly decreased in IBS rats of the model, medication and EA groups compared with their own individual pre-treatment and with the control group (<0.01), and markedly higher in both medication and EA groups than in the model group (<0.05, <0.01). The intra-colonic volume thresholds for inducing abdominal and hip AWR were significantly lower in the model group than in the normal control group (<0.01), and obviously higher in both medication and EA groups than in the model group (<0.05,<0.01). The AWR scores of intra-colorectal balloon at volumes of 0.5, 1.0 and 1.5 mL of water were significantly higher in the model group than in the control group (<0.01), and considerably lower in the EA and medication groups than in the model group (<0.01). The number of colonic MC and the expression levels of colonic Try, PAR-2, TRPV 1, SP and CGRP proteins were significantly higher in the model group than in the control group (<0.01), and obviously decreased in both medication and EA groups relevant to the model group (<0.01). Comparison between the medication and EA groups showed that the decreased expression levels of colonic PAR-2, TRPV 1, SP and CGRP proteins were significantly lower in the EA group than in the medication group (<0.05), but no significant differences were found between the two groups in intra-colonic volumes for inducing AWR, AWR scores, body weight, and colonic MC number and Try immunoactivity levels (>0.05)..
EA of ST 37 and ST 25 can relieve visceral hypersensitivity in IBS rats, which may be associated with its effects in down-regulating the number of MC and the expression of PAR-2, TRVP 1, SP, CGRP and Try proteins in the colonic tissue.
观察电针刺激“上巨虚”(ST 37,下合穴)和“天枢”(ST 25,募穴)对肠易激综合征(IBS)大鼠内脏痛及结肠组织中类胰蛋白酶(Try)、蛋白酶激活受体2(PAR - 2)、瞬时受体电位香草酸受体1(TRPV 1)、P物质(SP)和降钙素基因相关肽(CGRP)表达的影响,以探讨其改善IBS的作用机制。
将40只雄性Sprague Dawley大鼠随机分为正常对照组、模型组、药物组和电针组,每组10只。采用慢性急性复合应激(CACS,包括禁水、禁食、夹尾、冰水强迫游泳、束缚等)方法建立IBS模型,持续21天。药物组大鼠给予溴丙胺太林灌胃(1 mg/mL,15 mg/kg),每日1次,共14天。电针组采用频率为10 Hz/50 Hz、强度为0.2~0.3 mA的电针刺激双侧ST 37和ST 25,每次30分钟,每日1次,共14天。通过向结直肠内充水球囊扩张(CRD)检测腹部和臀部的肌肉收缩反射(AWR),以评估内脏高敏感性。甲苯胺蓝染色后计数结肠组织中肥大细胞的数量。采用免疫化学方法测定结肠组织中Try的免疫活性,采用蛋白质印迹法检测结肠组织中PAR - 2、TRVP 1、SP和CGRP蛋白的表达。
造模后,模型组、药物组和电针组IBS大鼠的体重较各自造模前及正常对照组均显著下降(P<0.01),且药物组和电针组大鼠体重显著高于模型组(P<0.05,P<0.01)。模型组大鼠诱发腹部和臀部AWR的结肠内体积阈值显著低于正常对照组(P<0.01),药物组和电针组显著高于模型组(P<0.05,P<0.01)。模型组大鼠在结肠内注入0.5、1.0和1.5 mL水时的AWR评分显著高于正常对照组(P<0.01),电针组和药物组显著低于模型组(P<0.01)。模型组大鼠结肠肥大细胞数量及结肠组织中Try、PAR - 2、TRPV 1、SP和CGRP蛋白的表达水平显著高于正常对照组(P<0.01),药物组和电针组与模型组相比均显著降低(P<0.01)。药物组与电针组比较,电针组结肠组织中PAR - 2、TRPV 1、SP和CGRP蛋白表达水平的降低幅度显著低于药物组(P<0.05),但两组在诱发AWR的结肠内体积、AWR评分、体重、结肠肥大细胞数量及Try免疫活性水平方面差异无统计学意义(P>0.05)。
电针ST 37和ST 25可缓解IBS大鼠的内脏高敏感性,其机制可能与下调结肠组织中肥大细胞数量及PAR - 2、TRVP 1、SP、CGRP和Try蛋白的表达有关。