NGF/PI3K/TRPV1通路介导针刺“上巨虚”对慢性内脏痛觉过敏的肠易激综合征大鼠内脏疼痛的调节。

Cai-Yi Chen, Zhi Yu, Feng Ji, Ling-Ling He, Xuan Wang, Bin Xu
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引用次数: 0

摘要

目的:观察手法针刺“上巨虚”对肠易激综合征(IBS)慢性内脏痛觉过敏大鼠神经生长因子(NGF)/磷脂酰肌醇3激酶(PI3K)/瞬时受体电位香草素1(TRPV1)信号通路的影响,探讨其治疗IBS慢性内脏痛觉敏的潜在机制。方法:SD幼犬出生后8天,通过结肠扩张刺激建立IBS慢性内脏痛觉过敏模型,持续2周,并在刺激后喂养至8周龄。然后将验证成功的成年大鼠随机分为模型组、上巨虚组和非穴位组,每组6只,选择6只未建模的大鼠作为正常组。模型评价次日,上巨虚组大鼠针刺右ST37,非穴位组大鼠分别针刺右疑病症非经穴和非穴位,时间均为15min,手法捻转,时间为30s,间隔5min,每天1次,共7d。采用腹部戒断反射(AWR)评分评估大鼠慢性内脏疼痛的程度。采用Western blot和实时荧光定量PCR方法检测NGF、原肌球蛋白受体激酶A(TrkA)、PI3K和TRPV1的结肠蛋白和mRNA表达。用免疫组织化学方法检测大鼠结肠组织中PI3K和TRPV1蛋白的阳性表达。结果:与正常组相比,结肠组织中20、40、60和80mm Hg 4个压力水平对应的AWR评分,NGF、TrkA、PI3K和TRPV1的mRNA和蛋白表达,结肠组织中PI3K和TRPV1的阳性表达显著增加(PPC结论:手针ST37可减轻大鼠IBS慢性内脏痛觉过敏,其镇痛作用可能与调节NGF/PI3K/TRPV1信号通路有关。
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NGF/PI3K/TRPV1 pathway mediates the regulation of visceral pain by acupuncture at "Shangjuxu" (ST37) in irritable bowel syndrome rats with chronic visceral hyperalgesia.

Objectives: To investigate the effect of manual acupuncture at "Shangjuxu"(ST37) on nerve growth factor(NGF)/phosphatidylinositol 3-kinase(PI3K)/transient receptor potential vanilloid 1(TRPV1) signaling pathway in rats with chronic visceral hyperalgesia of irritable bowel syndrome (IBS), so as to explore its underlying mechanism in treating IBS chronic visceral hyperalgesia.

Methods: IBS chronic visceral hyperalgesia model was established by colorectal dilation stimulation for 2 weeks for SD pups at 8 d after birth, which were fed until 8-week age after the stimulation. Then the verified successfully modeled adult rats were randomly divided into model, Shangjuxu, and non-acupoint groups, with 6 rats in each group, and 6 unmodeled rats were selected as normal group. On the next day of model evaluation, rats in the Shangjuxu group received acupuncture at right ST37 while rats in the non-acupoint group received acupuncture at the non-meridian and non-acupoint point in the right hypochondrium, both for 15 min, with manual twisting of mild reinforcing and reducing performed for 30 s at an interval of 5 min, once a day, for a total of 7 d. Abdominal withdrawal reflex(AWR) score was used to evaluate the degree of chronic visceral pain in rats. Western blot and real-time fluorescence quantitative PCR were used to detect the colonic protein and mRNA expressions of NGF, tropomyosin receptor kinase A (TrkA), PI3K and TRPV1. The positive expressions of PI3K and TRPV1 proteins in the colon of rats were detected by immunohistochemistry method.

Results: Compared with the normal group, AWR scores corresponding to 4 pressure levels of 20, 40, 60 and 80 mm Hg, mRNA and protein expressions of NGF, TrkA, PI3K and TRPV1 in colon tissue, and positive expressions of PI3K and TRPV1 in colon tissue were significantly increased(P<0.05) in the model group. After intervention, compared with the model group, rats in the Shangjuxu group had reduced AWR scores corresponding to 4 pressure levels of 20, 40, 60 and 80 mm Hg, lower colonic mRNA and protein expressions of NGF, TrkA, PI3K and TRPV1, and decreased positive expressions of PI3K and TRPV1 in colon tissue(P<0.05), while there were no significant differences in the above indexes of the non-acupoint group.

Conclusions: Manual acupuncture at ST37 can alleviate IBS chronic visceral hyperalgesia in rat and its analgesic effect may be related to regulating NGF/PI3K/TRPV1 signaling pathway.

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