Li Li, Qi Wei, Wang Zhao-Hui, Zhi Ding-Ming, Jiang Hai-Lin, Zheng Li-Wen, Wang Fu-Chun
School of Acupuncture-moxibustion and Tuina, Changchun University of Chinese Medicine, Changchun 130117, China; School of Pharmacy and Medicine, Tonghua Normal University, Tonghua 134000, Jilin Province.
School of Acupuncture-moxibustion and Tuina, Changchun University of Chinese Medicine, Changchun 130117, China;Orthopedics Department, Bao'an Authentic Traditional Chinese Medicine Therapy Hospital, Shenzhen 518101, Guangdong Province.
Zhen Ci Yan Jiu. 2021 Mar 25;46(3):173-9. doi: 10.13702/j.1000-0607.200963.
To observe the effect of electroacupuncture preconditioning at "Zusanli"(ST36,Lower Confluent point) and "Zhongwan"(CV12,Front-Mu point) combination on oxidative stress and inflammation-related indicators, Toll-like receptor 4 (TLR4), myeloid differentiation factor 88 (MyD88) and inhibitor-α of nuclear transcription factor κB (IκB-α) in serum and gastric tissue of rats with stress gastric ulcer(SGU),so as to explore its mechanisms underlying prevention of SGU.
A total of 36 Wistar rats were randomly divided into blank control, model, positive drug and He-Sea-Front-Mu point combination groups (=9 in each group). A rat model of SGU was established by restraint water-immersion stress method. Ten days before mode-ling, rats in the He-Sea-Front-Mu point combination group received electroacupuncture (2 Hz, 0.6 mA)at ST36 and CV12 for 10 min once every other day for 10 days, and those in the positive drug group was treated by gavage of omeprazole (20 mg/kg) once every other day for 10 days. The morphology of the gastric mucosa was observed by naked eyes and hematoxylin-eosin staining, and the ulcer index (UI) and lesion score were calculated. TBA and colorimetric methods, ELISA and Western blot were used to detect malondialdehyde (MDA), myeloperoxidase (MPO), glutathione peroxidase (GSH-Px), tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) levels and the relative expressions of TLR4, MyD88, and IκB-α protein, separately.
The gastric mucosa of rats in the blank control group was smooth and intact, the cells were arranged neatly, and there was no telangiec-tasia, hyperemia and inflammatory cell infiltration. The gastric mucosal epithelial structure of rats in the model group was destroyed, and a large number of mucosal epithelial cell death and inflammatory cell infiltration were seen. The degree of gastric mucosal injury and inflammatory cell infiltration in the positive drug group and the combined point group was less than that in the model group. Compared with the blank control group, the UI and lesion score of rats in the model group were significantly increased (<0.05), the levels of MDA and MPO in the serum and gastric tissues were significantly increased (<0.05), GSH-Px was significantly reduced (<0.05), the contents of TNF-α and IL-6 in serum were markedly increased (<0.05), the expression levels of TLR4 and MyD88 proteins in gastric tissue were significantly increased (<0.05), IκB-α was significantly reduced (<0.05). After intervention and in comparison with the model group showed that, the UI and lesion score, the levels of MDA and MPO, contents of serum TNF-α and IL-6, expression levels of TLR4 and MyD88 proteins in positive drug and He-Sea-Front-Mu point combination groups were significantly decreased (<0.05), while GSH-Px and IκB-α were significantly increased (<0.05); There were no significant differences in the above indicators between the positive drug and the He-Sea-Front -Mu point combination groups (except TNF-α).
Electroacupuncture preconditioning at ST36 and CV12 can prevent SGU, which may be related to its effects in anti-oxidant, anti-inflammatory and regulating TLR4/MyD88/IκB signaling pathway.
观察电针“足三里”(ST36,下合穴)与“中脘”(CV12,募穴)配伍预处理对应激性胃溃疡(SGU)大鼠血清及胃组织氧化应激和炎症相关指标、Toll样受体4(TLR4)、髓样分化因子88(MyD88)及核转录因子κB抑制因子-α(IκB-α)的影响,探讨其预防SGU的作用机制。
将36只Wistar大鼠随机分为空白对照组、模型组、阳性药物组和合募配穴组,每组9只。采用束缚水浸应激法制备SGU大鼠模型。造模前10天,合募配穴组大鼠隔日接受电针双侧“足三里”和“中脘”(2 Hz,0.6 mA)10分钟,共10次;阳性药物组大鼠隔日灌胃奥美拉唑(20 mg/kg),共10次。肉眼及苏木精-伊红染色观察胃黏膜形态,计算溃疡指数(UI)和损伤评分。采用硫代巴比妥酸法、比色法、酶联免疫吸附测定法和蛋白质免疫印迹法分别检测丙二醛(MDA)、髓过氧化物酶(MPO)、谷胱甘肽过氧化物酶(GSH-Px)、肿瘤坏死因子-α(TNF-α)和白细胞介素-6(IL-6)水平以及TLR4、MyD88和IκB-α蛋白的相对表达量。
空白对照组大鼠胃黏膜光滑完整,细胞排列整齐,无毛细血管扩张、充血及炎症细胞浸润。模型组大鼠胃黏膜上皮结构破坏,可见大量黏膜上皮细胞死亡及炎症细胞浸润。阳性药物组和合募配穴组大鼠胃黏膜损伤程度及炎症细胞浸润程度均轻于模型组。与空白对照组比较,模型组大鼠UI及损伤评分显著升高(P<0.05),血清及胃组织MDA、MPO水平显著升高(P<0.05),GSH-Px显著降低(P<0.05),血清TNF-α、IL-6含量明显升高(P<0.05),胃组织TLR4、MyD88蛋白表达水平显著升高(P<0.05),IκB-α显著降低(P<0.05)。与模型组比较,干预后阳性药物组和合募配穴组大鼠UI及损伤评分、MDA及MPO水平、血清TNF-α及IL-6含量、胃组织TLR4及MyD88蛋白表达水平均显著降低(P<0.05),GSH-Px及IκB-α显著升高(P<0.05);阳性药物组和合募配穴组上述指标比较,除TNF-α外,差异均无统计学意义。
电针“足三里”与“中脘”预处理可预防SGU,其机制可能与抗氧化、抗炎及调节TLR4/MyD88/IκB信号通路有关。