摘要
目的:探讨电针对溃疡性结肠炎(UC)大鼠的保护作用以及对Toll样受体4(TLR4)/髓样分化因子88(MyD88)/核因子-κB(NF-κB)信号通路的影响。方法:SD大鼠随机分为空白组、模型组、柳氮磺吡啶组、低强度电针组、高强度电针组,每组8只。采用2,4,6-三硝基苯磺酸灌肠复制UC模型。两个电针组电针"天枢""关元""足三里"("天枢""足三里"双侧交替取穴),低强度电针组电流强度1 mA,高强度电针组电流强度5 mA,每次15 min,每日1次,柳氮磺砒啶组大鼠每天给予柳氮磺砒啶混悬液灌胃1次,每次3 mL,均治疗15 d。HE染色法观察结肠病理情况,评估组织学损伤指数(TDI);ELISA法检测血清白细胞介素(IL)-4、IL-10、IL-17及前列腺素E2(PGE2)水平;采用免疫组织化学法及荧光定量PCR法检测结肠组织TLR4、MyD88、NF-κB蛋白及mRNA的表达水平。结果:与空白组比较,模型组大鼠体质量、血清IL-4和IL-10水平明显降低(P<0.05),TDI、血清IL-17和PGE2水平、结肠组织TLR4、MyD88、NF-κB蛋白及mRNA表达明显升高(P<0.05)。与模型组比较,柳氮磺砒啶组及电针各组大鼠的体质量、血清IL-4和IL-10水平升高(P<0.05),TDI、血清IL-17和PGE2水平、结肠组织TLR4、MyD88、NF-κB蛋白及mRNA表达明显降低(P<0.05)。与柳氮磺砒啶组比较,低强度电针组大鼠TDI、结肠组织TLR4、MyD88、NF-κB的蛋白及mRNA表达明显升高(P<0.05);高强度电针组大鼠体质量明显升高(P<0.05),TDI、结肠组织TLR4、MyD88、NF-κB的蛋白及mRNA表达明显降低(P<0.05)。与低强度电针组比较,高强度电针组大鼠的体质量升高,TLR4、MyD88、NF-κB蛋白和mRNA表达均明显降低(P<0.05)。结论:电针可能通过抑制TLR4/MyD88/NF-κB信号通路并减少致炎细胞因子的释放达到保护UC大鼠结肠黏膜组织的目的,且高强度电针效果更优。
Objective To investigate the protective effect of electroacupuncture(EA) on rats with ulcerative colitis(UC) and its effect on the Toll-like receptor 4(TLR4)/myeloid differentiation factor 88(MyD88)/nuclear factor-kappa B(NF-κB) signaling pathway. Methods Sprague-Dawley rats were randomly divided into blank group, model group, salicylazosulfapyridine(SASP) group, low-intensity EA group, and high-intensity EA group, with 8 rats in each group. Enema with 2,4,6-trinitrobenzenesulfonic acid was performed to establish a model of UC. The rats in the two EA groups were given EA at"Tianshu"(ST25), "Guanyuan"(CV4), and "Zusanli"(ST36) for 15 min each time, once a day, with a current intensity of 1 mA for the low-intensity EA group and 5 mA for the high-intensity EA group(among them, "Tianshu" "Zusanli" bilateral alternate acupoints);the rats in the SASP group were given SASP suspension 3 mL every day by gavage. The course of treatment was 15 days for all groups. HE staining was used to observe the pathology of the colon and determine tissue damage index(TDI);ELISA was used to measure the serum levels of interleukin-4(IL-4), interleukin-10(IL-10), interleukin-17(IL-17), and prostaglandin E2(PGE2);immunohistochemical staining and real-time PCR were used to measure the protein and mRNA expression of TLR4, MyD88, and NF-κB in colonic tissue. Results Compared with the blank group, the model group had significant reductions in body weight, serum IL-4, and IL-10(P<0.05) and significant increases in colonic mucosa TID, the serum levels of IL-17 and PGE2 and the protein and mRNA expression of TLR4, MyD88, and NF-κB in colonic tissue(P<0.05). Compared with the model group, the SASP group and the EA groups had significantly higher body weight and serum levels of IL-4 and IL-10(P<0.05), as well as significantly lower colonic TDI, serum levels of IL-17 and PGE2, and integrated optical density and mRNA expression of TLR4, MyD88, and NF-κB in colonic tissue(P<0.05). Compared with the SASP group, the low-intensity EA group had significantly higher colonic TDI and protein and mRNA expression of TLR4, MyD88, and NF-κB in colonic tissue(P<0.05), and compared with the SASP group, the high-intensity EA group had a significantly higher body weight(P<0.05) and lower colonic TDI and protein and mRNA expression of TLR4, MyD88, and NF-κB in colonic tissue(P<0.05). Compared with the low-intensity EA group, the high-intensity EA group had significantly higher body weight(P<0.05), and lower colonic TDI and protein and mRNA expression of TLR4, MyD88, and NF-kB in colonic tissue(P<0.05). Conclusion Electroacupuncture exerts a protective effect on the colonic mucosa in rats with UC possibly by inhibiting the TLR4/MyD88/NF-κB signaling pathway and reducing the release of inflammatory cytokines, and high-intensity EA may have a better effect than low-intensity EA.
作者
乔翠霞
赵刚
张立泽
邸爱婷
刘鹏林
王莹莹
曾莉
QIAO Cui-xia;ZHAO Gang;ZHANG Li-ze;DI Ai-ting;LIU Peng-lin;WANG Ying-ying;ZENG Li(First Clinical Medical School,Nanjing University of Chinese Medicine,Nanjing 210046,China;Department of Traditional Chinese Medicine,The Affiliated Hospital of Qingdao University,Qingdao 266003,Shandong Province)
出处
《针刺研究》
CAS
CSCD
北大核心
2020年第3期180-187,201,共9页
Acupuncture Research
基金
国家自然科学基金项目(No.81673982)
山东省中医药科技发展项目(No.2017-190)。
关键词
电针
TLR4/MyD88/NF-κB信号通路
炎性细胞因子
溃疡性结肠炎
Electroacupuncture
Toll-like receptor 4/myeloid differentiation factor 88/nuclear factor-kappa B signaling pathway
Inflammatory cytokine
Ulcerative colitis
