摘要
目的探讨苦参碱(MT)调节白细胞介素-6(IL-6)/信号转导和转录激活因子3(STAT3)/核转录因子κB(NF-κB)通路对炎症性肠病大鼠辅助性T细胞17(Th17)/调节性T细胞(Treg)平衡的影响。方法按照随机数字表法将SD大鼠分为空白对照组(CK组)、Model组,低、中、高剂量MT组(MT-L组,50 mg/kg;MT-M组,100 mg/kg;MT-H组,200 mg/kg),美沙拉嗪组(MSLM组,0.42 g/kg)、MT-H+rIL-6(IL-6激活剂)组(200 mg/kg+0.05 mg/kg),每组18只。除CK组外,其余组大鼠均采用50 g/L三硝基苯磺酸(20 mg/kg)缓冲液与500 mL/L乙醇按照1∶1比例混匀后灌肠以构建炎症性肠病大鼠模型,建模成功后,分别进行给药处理,每天1次,持续7周。分别在给药1、3、5、7周时对大鼠进行体质量的测量;比较各组大鼠结肠长度的变化;HE染色检测大鼠结肠组织病理损伤;ELISA法检测大鼠血清中肿瘤坏死因子-α(TNF-α)、白细胞介素(IL)-17(IL-17)、IL-10水平;流式细胞术检测大鼠外周血中Th17、Treg细胞比例;Western blotting检测大鼠结肠组织中维甲酸相关孤核受体γt(RORγt)、叉头框蛋白P3(Foxp3)、IL-6、p-STAT3、p-NF-κB p65蛋白表达。结果与CK组比较,Model组大鼠结肠组织病理损伤严重,体质量(3、5、7周时)、IL-10水平、Treg细胞比例、Foxp3蛋白表达降低,结肠变短,TNF-α、IL-17水平、Th17细胞比例、Th17/Treg比值、RORγt、IL-6、p-STAT3、p-NF-κB p65蛋白表达增高(P<0.05);与Model组比较,MT-L组、MT-M组、MT-H组、MSLM组对应指标变化趋势与上述相反(P<0.05);rIL-6减弱了高剂量MT对炎症性肠病大鼠Th17/Treg平衡的促进作用。结论MT可能通过抑制IL-6/STAT3/NF-κB信号通路,促进炎症性肠病大鼠Th17/Treg平衡。
Objective To investigate the influence of matrine(MT)on the balance of T helper cell 17(Th17)/regulatory T cells(Treg)in rats with inflammatory bowel disease by regulating interleukin-6(IL-6)/signal transducer and activator of transcription 3(STAT3)/nuclear transcription factor-κB(NF-κB)pathway.Methods SD rats were grouped into control check group(CK group),model group,low-dose MT group(MT-L group,50 mg/kg),medium-dose MT group(MT-M group,100 mg/kg),high-dose MT group(MT-H group,200 mg/kg),mesalazine group(MSLM group,0.42 g/kg),and MT-H+rIL-6(IL-6 activator)group(200 mg/kg+0.05 mg/kg)according to the random number table method,with 18 in each group.Except for the CK group,the rats in other groups all received with 5%trinitrobenzenesulfonic acid(20 mg/kg)buffer solution mixed with 50%ethanol at a ratio of 1∶1 and then enema to construct a rat model of inflammatory bowel disease.After the successful modeling,they were treated with drug administration once a day for 7 weeks.The body weight of rats was measured at 1,3,5,and 7 weeks of administration;the changes of colon length of rats in each group were compared;HE staining was used to detect the pathological damage of rat colon tissue;the levels of tumor necrosis factor-α(TNF-α),interleukin(IL)-17 and IL-10 in serum of rats were detected by ELISA;the proportions of Th17 and Treg cells in peripheral blood of rats were detected by flow cytometry;Western blottingt was performed to detect the protein expression of retinoic acid-related orphan receptorγt(RORγt),forkhead box protein P3(Foxp3),IL-6,p-STAT3,and p-NF-κB p65 in rat colon tissue.Results Compared with the CK group,the colon tissue of the model group was severely damaged by pathology,and the body weight(at 3,5,and 7 weeks),the level of IL-10,the proportion of Treg cell,and the expression of Foxp3 protein were decreased,the colon length shortened,the levels of TNF-α,IL-17,the proportions of Th17 cell,Th17/Treg ratio,and the protein expression of RORγt,IL-6,p-STAT3,and p-NF-κB p65 increased(P<0.05).Compared with the model group,the corresponding indicators of the MT-L group,MT-M group,MT-H group,and MSLM group had the opposite trends(P<0.05);rIL-6 attenuated the promoting effect of high-dose MT on Th17/Treg balance in inflammatory bowel disease rats.Conclusion MT may promote Th17/Treg balance in inflammatory bowel disease rats by inhibiting IL-6/STAT3/NF-κB signaling pathway.
作者
吴茸
王栋
王晶敏
张玮宇
陶庆松
WU Rong;WANG Dong;WANG Jingmin;ZHANG Weiyu;TAO Qingsong(Department of General Surgery,Zhongda Hospital Affiliated to Southeast University,Nanjing 210000,China)
出处
《西安交通大学学报（医学版）》
CAS
CSCD
北大核心
2023年第5期809-816,共8页
Journal of Xi’an Jiaotong University（Medical Sciences）
基金
国家自然科学基金青年项目资助(No.81500421)。
作者简介
通信作者:陶庆松.E-mail:2479881882@qq.com。
