Cui Xia Qiao1, Gang Zhao2, Li Ze Zhang2, Ai Ting Di2, Peng Lin Liu2, Ying Ying Wang2, Li Zeng3. 1. 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. 2. Department of Traditional Chinese Medicine, The Affiliated Hospital of Qingdao University, Qingdao 266003, Shandong Province. 3. First Clinical Medical School, Nanjing University of Chinese Medicine, Nanjing 210046, China.
Abstract
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.
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.