摘要
目的观察参黛清肠汤对溃疡性结肠炎(ulcerative colitis,UC)小鼠肠道黏膜的保护作用,并探讨其机制。方法55只C57BL/6小鼠以2.5%葡聚糖硫酸钠(dextran sodium sulfate,DSS)灌胃3个周期诱导建立UC模型,建模成功后随机分为5组,另10只C57BL/6小鼠记为正常组。美沙拉嗪组予以200 mg/kg小鼠体质量的美沙拉嗪溶于1 mL/100 g小鼠体质量生理盐水中灌胃,参黛清肠汤低、中、高剂量组分别予以15、30、60 mg/kg参黛清肠汤同法灌胃,模型组和正常组均予以等量生理盐水灌胃,1次/d,给药2周。干预后评价各组疾病活动指数(disease activity index,DAI)和肠道黏膜损伤指数(colonic mucosal damage index,CMDI);苏木素-伊红(HE)染色观察肠道黏膜病理改变;酶联免疫法检测肠道黏膜组织白细胞介素-1β(IL-1β)、肿瘤坏死因子-α(TNF-α)含量;实时反转录荧光聚合酶链反应(RT-qPCR)和蛋白质免疫印迹法(Western blotting,WB)分别检测肠道黏膜组织Ras同源基因家族A(RhoA)、Rho相关卷曲螺旋形成蛋白激酶1(ROCK-1)、Ras相关的C3肉毒素底物1(Rac-1)、p21激活激酶1(PAK-1)、核转录因子-κB(NF-κB)mRNA和蛋白表达。结果HE染色证实UC小鼠建模成功;模型组DAI和CMDI评分,肠道黏膜组织IL-1β、TNF-α含量,肠道黏膜组织RhoA、ROCK-1、NF-κB mRNA及蛋白表达均高于正常组(P<0.05),美沙拉嗪组和参黛清肠汤3剂量组均低于模型组(P<0.05),参黛清肠汤低剂量组均高于美沙拉嗪组(P<0.05),参黛清肠汤高剂量组均低于美沙拉嗪组(P<0.05),模型组肠道黏膜组织Rac-1、PAK-1 mRNA及蛋白表达均低于正常组(P<0.05),美沙拉嗪组和参黛清肠汤3剂量组均高于模型组(P<0.05),参黛清肠汤低剂量组均低于美沙拉嗪组(P<0.05),参黛清肠汤高剂量组均高于美沙拉嗪组(P<0.05),且参黛清肠汤中剂量组与美沙拉嗪组差异均无统计学意义(P>0.05)。模型组肠道黏膜组织病理改变严重,美沙拉嗪组和参黛清肠汤3剂量组�
Objective To observe the protective effect of Shendai Qingchang Decoction(参黛清肠汤)on intestinal mucosa in mice with ulcerative colitis(UC)and explore its mechanism.Methods 55 C57BL/6 mice were induced to establish UC model by intragastric administration of 2.5%dextran sodium sulfate(DSS)for 3 cycles.After successful modeling,5 groups were randomly divided into 5 groups,and the other 10 C57BL/6 mice were recorded as normal group.Mesalazine group was given 200 mg/kg weight of mesalazine dissolved in 1 mL/100 g weight of normal saline by gavage,and the low,medium and high dose groups of Shendai Qingchang decoction were given 15,30 and 60 mg/kg Shendai Qingchang Decoction by gavage respectively,while the model group and the normal group were given the same amount of normal saline by gavage,once a day for 2 weeks.Disease activity index(DAI)and colonic mucosal damage index(CMDI)were evaluated after intervention.Hematoxylin eosin(HE)staining was used to observe the pathological changes of intestinal mucosa.Contents of interleukin-1β(IL-1β)and tumor necrosis factor-α(TNF-α)in intestinal mucosa by enzyme-linked immunosorbent assay were detected.The expressions of Ras homologous gene family A(RhoA),Rho associated coiled coil forming protein kinase 1(ROCK-1),Ras associated C3 botulinum toxin substrate 1(Rac-1),p21 activated kinase 1(PAK-1)and nuclear transcription factor-κB(NF-κB)messenger ribonucleic acid(mRNA)and proteins were detected by real-time quantitative polymerase chain reaction(RT-qPCR)and Western blotting(WB).Results HE staining confirmed the successful modeling of UC mice.DAI and CMDI scores,IL-1βand TNF-αcontents in intestinal mucosa,the expressions of RhoA,Rock-1,NF-κB mRNA and proteins in intestinal mucosa of the model group were higher than those in the normal group(P<0.05),of which the mesalazine group and Shendai Qingchang Decoction three dose groups were lower than those in the model group(P<0.05),and those in the low dose group of Shendai Qingchang Decoction were higher than those of me
作者
杜明民
郎晓猛
张晓利
崔建从
刘建平
DU Mingmin;LANG Xiaomeng;ZHANG Xiaoli;CUI Jiancong;LIU Jianping(Hebei Hospital of Traditional Chinese Medicine,Shijiazhuang 050000,Hebei,China)
出处
《辽宁中医药大学学报》
CAS
2022年第9期19-24,F0003,共7页
Journal of Liaoning University of Traditional Chinese Medicine
基金
河北省中医药管理局中医药类科研计划项目(2020098)。
