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Title: [Effect of acupuncture on intestinal flora and TLR4 in brain and intestinal tissues in rats with stress gastric ulcer].
Author: Zhao JY, Wang TN, Wang LJ, Ting X, Wu YQ, Zhou ZX, Yang YC, Shao YX, Ma HF.
Journal: Zhongguo Zhen Jiu; 2021 Apr 12; 41(4):413-9. PubMed ID: 33909363.
Abstract:
OBJECTIVE: To observe the effect of acupuncture at "Guanyuan" (CV 4) and "Xiajuxu" (ST 39) on intestinal flora and Toll-like receptors-4 (TLR4) in brain and intestinal tissue in rats with stress gastric ulcer (SGU), and to explore the possible mechanism of acupuncture for SGU. METHODS: Thirty-one male SD rats were randomly divided into a blank group (n=7), a model group (n=8), an acupuncture group (n=8) and a drug group (n=8). The rats in the model group, acupuncture group and drug group were treated with modified restraint plus water-immersion stress method to establish SGU model. After modeling, the rats in the acupuncture group were treated with acupuncture at "Guanyuan" (CV 4) and "Xiajuxu" (ST 39), 20 min each time, and the needles were twirled for 30 s every 5 min. The rats in the drug group were treated with intragastric administration of 2 mL omeprazole enteric-coated tablets (20 mg/mL). Both the treatments were given once a day for 5 days. After the intervention, the gastric mucosal damage index was calculated by Guth method; the morphological changes of gastric mucosa were observed by HE staining; the diversity of intestinal flora was detected by 16S rDNA high-throughput sequencing; TLR4 contents in brain and intestinal tissues were determined by ELISA. RESULTS: Compared with the blank group, the gastric mucosal damage index was significantly increased in the model group (P<0.01); the morphological changes of gastric mucosa were obvious; the Observed Species index and Shannon index of α diversity index of intestinal flora were decreased (P<0.05); the β diversity showed that the spatial distance between the model group and the blank group was far; the TLR4 contents in the brain and intestinal tissue were increased (P<0.05, P<0.01). Compared with the model group, the gastric mucosal damage index was decreased in the acupuncture group and the drug group (P<0.05); the morphology of gastric mucosa was improved; the Observed Species index and Shannon index of α diversity index of intestinal flora in the acupuncture group was increased (P<0.05), and the Shannon index in the drug group was increased (P<0.05); the β diversity showed that the spatial distance between the acupuncture group and the blank group was close; the TLR4 contents in the brain and intestinal tissues of the acupuncture group were decreased (P<0.01). Compared with the drug group, the contents of TLR4 in the intestinal tissue of the acupuncture group were decreased (P<0.05). CONCLUSION: Acupuncture at "Guanyuan" (CV 4) and "Xiajuxu" (ST 39) could alleviate SGU in rats, and its mechanism may be related to increasing the diversity of intestinal flora, promoting the disorder of intestinal flora to normal, and reducing the overexpression of TLR4 in brain and intestinal tissues. 目的：观察针刺“关元”“下巨虚”对应激性胃溃疡（SGU）模型大鼠肠道菌群及脑和肠组织内Toll样受体4（TLR4）含量的影响，探讨针刺治疗SGU可能的作用机制。方法：将31只雄性SD大鼠随机分为空白组（7只）、模型组（8只）、针刺组（8只）和药物组（8只）。除空白组外，其余3组采用束缚-浸水应激法的改良办法制备大鼠SGU模型。造模后，针刺组于“关元”“下巨虚”行针刺干预，每次20 min，每5分钟捻转行针30 s；药物组予浓度20 mg/mL奥美拉唑肠溶片溶液2 mL灌胃。均每日1次，共干预5 d。干预结束后，应用Guth法计算胃黏膜损伤指数，HE染色法观察胃黏膜形态学改变，16S rDNA高通量测序法检测肠道菌群结构多样性，ELISA法测定脑和肠组织内TLR4的含量。结果：与空白组比较，模型组大鼠胃黏膜损伤指数明显升高（P<0.01）；胃黏膜形态改变明显；肠道菌群α多样性指数Observed Species与Shannon降低（P<0.05），β多样性显示模型组与空白组空间距离较远；脑和肠组织内TLR4含量升高（P<0.05，P<0.01）。与模型组比较，针刺组和药物组大鼠胃黏膜损伤指数降低（P<0.05）,胃黏膜形态改善；针刺组大鼠肠道菌群α多样性指数Observed Species与Shannon升高（P<0.05），药物组大鼠Shannon指数升高（P<0.05）；β多样性显示针刺组与空白组空间距离较近；针刺组大鼠脑及肠组织内TLR4含量降低（P<0.01）。与药物组比较，针刺组大鼠肠组织内TLR4含量降低（P<0.05）。结论：针刺“关元”“下巨虚”能够缓解大鼠应激性胃溃疡，其机制可能与增加肠道菌群多样性，促使紊乱的肠道菌群趋于正常，降低脑和肠组织内TLR4的过度表达有关。.

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