Wu Yang Yang, Liu Ming Jiang, Yin Shao Jie, Wang An Yuan, Li Jin Gui
School of Veterinary Medicine, Yangzhou University, Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonosis, Yangzhou 225009, Jiangsu Province, China.
Zhen Ci Yan Jiu. 2020 Jan 25;45(1):8-13. doi: 10.13702/j.1000-0607.1806066.
To investigate the effect of manual acupuncture (MA) and electroacupuncture (EA) on histopathological changes, and levels of oxidative-stress related cytokines and key proteins of the endoplasmic reticulum stress (ERS) pathway in ulcerative colitis (UC) rats, so as to reveal their mechanisms underlying improvement of UC.
Twenty-eight male SD rats were randomly divided into control, model, EA and MA groups (＝ 7 rats per group). The UC model was established by enema of mixture solution of 5% 2, 4, 6-trinitrobenzene sulfonic acid (TNBS, 100 mg/kg). Rats of the control group received intra-rectal perfusion of normal saline. After modeling, the left "Quchi"(LI11) and "Zusanli"(ST36) were stimulated with EA (2-4 mA，8 Hz/25 Hz) or MA for 20 min, once every other day for consecutive 2 weeks. The rats in the control and model group were just anesthetized and fixed. At the end of experiments, the colon tissue was collected for observing histopathological changes with H．E. staining. The contents of oxidative stress-related factors as superoxide dismutase (SOD), catalase (CAT), malondialdehyde (MDA), reduced glutathione (GSH) and total antioxidant capacity (T-AOC) were detected by ELISA, and the expression levels of key proteins of ERS as phosphorylated inhibitor of nuclear factor kappa B kinase α (p-IκBα), phosphorylated p65 (p-p65), glucose regulated protein 78 (GRP78), phosphorylated protein kinase R-like endoplasmic reticulum kinase (p-PERK) and phosphorylated eukaryotic translation initiation factor 2α (p-eIF2α) by using Western blot.
After modeling, the colon tissues showed severe swelling, disordered arrangement of intestinal mucosal cells, hemorrhage with infiltration of inflammatory cell and partial loss of colon villus, which was relatively milder in the EA and MA groups. The colonic lesion score was remarkably increased in the model group in contrast to the control group (<0.01), and obviously reduced in both EA and MA groups relevant to the model group (<0.01). The levels of SOD, CAT, GSH and T-AOC were all significantly decreased (<0.01), and the content of MDA, and expression levels of p-IκBα， p-p65 and GRP78, p-PERK and p-eIF2α proteins were all significantly increased in the model group relevant to the control group (<0.01). After the treatment, modeling-induced down-regulation of SOD, CAT and GSH in both EA and MA groups, and T-AOC in the EA group, and up-regulation of levels of MDA, p-IκBα， p-p65, GRP78, p-PERK and p-eIF2α in both groups were reversed (<0.01, <0.05).
Both EA and MA treatment can obviously alleviate colonic inflammation in UC rats via inhibiting oxidative stress and ERS.
探讨手针（MA）和电针（EA）对溃疡性结肠炎（UC）大鼠组织病理学变化、氧化应激相关细胞因子水平及内质网应激（ERS）通路关键蛋白的影响，以揭示其改善UC的作用机制。
将28只雄性SD大鼠随机分为对照组、模型组、电针组和手针组（每组7只）。采用5% 2,4,6-三硝基苯磺酸（TNBS，100 mg/kg）混合溶液灌肠建立UC模型。对照组大鼠直肠内灌注生理盐水。造模后，用电针（2 - 4 mA，8 Hz/25 Hz）或手针刺激左侧“曲池”（LI11）和“足三里”（ST36）20分钟，隔日1次，连续2周。对照组和模型组大鼠仅进行麻醉和固定。实验结束时，采集结肠组织，用苏木精-伊红（H.E.）染色观察组织病理学变化。采用酶联免疫吸附测定（ELISA）法检测超氧化物歧化酶（SOD）、过氧化氢酶（CAT）、丙二醛（MDA）、还原型谷胱甘肽（GSH）和总抗氧化能力（T-AOC）等氧化应激相关因子的含量，并用蛋白质免疫印迹法检测ERS关键蛋白磷酸化核因子κB激酶α（p-IκBα）、磷酸化p65（p-p65）、葡萄糖调节蛋白78（GRP78）、磷酸化蛋白激酶R样内质网激酶（p-PERK）和磷酸化真核翻译起始因子2α（p-eIF2α）的表达水平。
造模后，结肠组织出现严重肿胀、肠黏膜细胞排列紊乱、出血伴炎症细胞浸润及部分结肠绒毛缺失，电针组和手针组病变相对较轻。与对照组相比，模型组结肠病变评分显著升高（<0.01），与模型组相比，电针组和手针组结肠病变评分均明显降低（<0.01）。与对照组相比，模型组SOD、CAT、GSH和T-AOC水平均显著降低（<0.01），MDA含量及p-IκBα、p-p65、GRP78、p-PERK和p-eIF2α蛋白表达水平均显著升高（<0.01）。治疗后，电针组和手针组造模所致的SOD、CAT和GSH水平降低以及电针组T-AOC水平降低，以及两组MDA、p-IκBα、p-p65、GRP78、p-PERK和p-eIF2α水平升高均得到逆转（<0.01，<0.05）。
电针和手针治疗均可通过抑制氧化应激和ERS明显减轻UC大鼠的结肠炎症。
