摘要
目的探讨栀子苷对溃疡性结肠炎模型大鼠的抗炎作用及腺苷酸激活蛋白激酶(AMPK)/肌球蛋白轻链激酶(MLCK)信号通路的影响。方法90只SD大鼠按随机数字表法分成对照组、模型组、柳氮磺胺吡啶组(300mg/kg)、栀子苷低、高剂量组(40、80mg/kg),每组18只。2,4,6-三硝基苯磺酸(TNBS)诱导制备溃疡性结肠炎模型大鼠,建模成功后,各组大鼠给予相应药物灌胃4周,对照组及模型组给予等体积生理盐水。测定各组大鼠结肠黏膜损伤指数、疾病活动指数(DAI)评分、大体形态损伤评分,逆转录-聚合酶链反应(RT-PCR)法及蛋白免疫印迹(Western blot)法测定大鼠结肠组织AMPK、MLCK mRNA和蛋白水平,酶联免疫吸附试验(ELISA)法测定结肠组织白细胞介素-2(IL-2)、白细胞介素-6(IL-6)、肿瘤坏死因子-α(TNF-α)水平。结果与对照组比较,模型组结肠黏膜损伤指数[(3.69±0.23)分比(0.00±0.00)分,P<0.05]、DAI评分[(2.69±0.25)分比(0.00±0.00)分,P<0.05]、大体形态损伤评分[(2.26±0.21)分比(0.00±0.00)分,P<0.05]、结肠组织MLCK mRNA[(3.89±0.26)比(0.89±0.16),P<0.05]和蛋白表达[(1.15±0.12)比(0.31±0.07),P<0.05]、IL-2[(654.74±24.54)ng/g比(269.69±15.89)ng/g,P<0.05]、IL-6[(788.99±15.47)ng/g比(242.96±15.78)ng/g,P<0.05]、TNF-α[(610.89±18.14)ng/g比(154.99±18.47)ng/g,P<0.05]水平升高,AMPK mRNA[(1.25±0.85)比(5.36±0.19),P<0.05]和蛋白表达[(0.30±0.06)比(1.33±0.10),P<0.05]水平降低;与模型组比较,柳氮磺胺吡啶组、栀子苷低剂量组、栀子苷高剂量组结肠黏膜损伤指数[(1.01±0.10)分、(2.42±0.18)分、(0.99±0.09)分比(3.69±0.23)分,P均<0.05]、DAI评分[(0.95±0.23)分、(2.04±0.30)分、(1.00±0.22)分比(2.69±0.25)分,P均<0.05]、大体形态损伤评分[(1.01±0.27)分、(1.61±0.31)分、(1.03±0.25)分比(2.26±0.21)分,P均<0.05]、结肠组织MLCK mRNA[(1.29±0.18)、(2.87±0.15)、(1.31±0.18)比(3.89±0.26),P均<0.05]和蛋白[(0.78±0.11)、(0.95±0.12)、(0.79±0.10)�
Objective To investigate gardenoside anti-inflammatory activity in a rat model of ulcerative colitis and the underlying molecular events.Methods A total of 90 Sprague-Dawley rats were randomly divided into control,model,and model plus sulfasalazine(300mg/kg),model plus a low dose gardenoside(40mg/kg),and model plus a high dose gardenoside groups(80mg/kg;n=18 in each group).Rats were treated with 2,4,6-trinitrobenzenesulfonic acid(TNBS)to establish ulcerative colitis model.Thereafter,the rats were subjected to different treatments by oral gavage,while the control rats were given equal amount of saline daily for 4 weeks.At the end of the experiments,the intestinal mucosal damage index,disease activity index(DAI)score and gross morphological damage score were assessed.Levels of adenosine monophosphate-activated protein kinase(AMPK)and myosin light chain kinase(MLCK)mRNA and protein in rat intestinal mucosa were measured using qRT-PCR and western blot,while levels of interleukin-2(IL-2),IL-6,and tumor necrosis factor-α(TNF-α)in the tissues were detected using ELISA.Results Compared with those of colon tissues from the control group,the colonic mucosal injury index[(3.69±0.23)vs.(0.00±0.00);P<0.05],DAI score[(2.69±0.25)vs.(0.00±0.00);P<0.05],and gross morphological damage score[(2.26±0.21)vs.(0.00±0.00);P<0.05],level of MLCK mRNA[(3.89±0.26)vs.(0.89±0.16);P<0.05]and MLCK protein[(1.15±0.12)vs.(0.31±0.07);P<0.05],IL-2[(654.74±24.54)vs.(269.69±15.89)ng/g;P<0.05],IL-6[(788.99±15.47)vs.(242.96±15.78)ng/g;P<0.05],TNF-α[(610.89±18.14)vs.(154.99±18.47)ng/g;P<0.05]in the colon of the model group were all increased,whereas levels of AMPK mRNA[(1.25±0.85)vs.(5.36±0.19);P<0.05]and protein[(0.30±0.06)vs.(1.33±0.10);P<0.05]were decreased.Furthermore,compare with those of the model group,the colonic mucosal injury index[(1.01±0.10),(2.42±0.18),and(0.99±0.09)vs.(3.69±0.23);P<0.05],DAI score[(0.95±0.23),(2.04±0.30),and(1.00±0.22)vs.(2.69±0.25);P<0.05]and gross morphological damage score[(1.01±0.27),(1.
作者
莫霏霏
周晶晶
金玲玲
余光
Mo Fei-fei;Zhou Jing-jing;Jin Ling-ling;Yu Guang(Department of Gastroenterology,Taizhou Municipal Hospital,Taizhou,Zhejiang 318000,China)
出处
《浙江中西医结合杂志》
2021年第5期418-424,共7页
Zhejiang Journal of Integrated Traditional Chinese and Western Medicine
