摘要
目的观察半夏泻心汤对溃疡性结肠炎(Ulcerative colitis,UC)大鼠肠道粘膜中的Toll样受体(Toll-like receptors,TLRs)/核转录因子κB(Nuclear factor kappa-light-chain-enhancer of activated B cells,NF-κB)/髓样分化蛋白88(Myeloid differentiation factor SS,MyD88)信号通路及抑制因子基质金属蛋白酶(Matrix metalloproteinases,MMPs)MMP-1、MMP-2表达影响,并探讨其作用机制。方法建立葡聚糖硫酸钠盐(Dextran sulphate sodium salt,DSS)诱导产生UC模型,根据随机数字表法,将35只无特定病原体(Specific pathogen free,SPF)级大鼠进行分组,分为正常组、模型组、低浓度给药组(0.473 g·mL^(-1))、中浓度给药组(0.945 g·mL^(-1))和高浓度给药组(1.890 g·mL^(-1))。除正常组外,其余4组大鼠连续10天使用2.5%浓度的葡聚糖硫酸钠盐溶液每天2 mL进行灌胃,造模成功后,连续给药1周,观察大鼠疾病活动指数(Disease activity index,DAI)变化;采用苏木精-伊红(Hematoxylin-eosin,HE)染色法,确认结肠组织病变损伤程度;同时应用蛋白质免疫印迹试验(Western blot)法,检测TLR4、NF-κB、MyD88蛋白在各组大鼠结肠中的表达;实时荧光定量聚合酶链反应(Real-time fluorescent quantitative polymerase chain reaction,RT-PCR)技术则测定了大鼠结肠黏膜TLR4、NF-κB、IκB-α(人核因子κB抑制蛋白α,NF-κB inhibitorα)、MMP-1、MMP-2的mRNA的表达。结果造模后大鼠体质量、食欲、被毛、活动度、尿量较造模前均出现不同程度的下降,同时部分大鼠出现腹泻、便血的症状。模型组与正常组相比,大鼠结肠组织中TLR4、NF-κB、MyD88蛋白的表达,及TLR4、NF-κB、IκB-α、MMP-1、MMP-2的mRNA表达均显著升高(P<0.01);给予半夏泻心汤治疗后,UC大鼠体质量下降、腹泻、便血等症状明显减轻。低、中、高剂量的半夏泻心汤实验组中,大鼠结肠黏膜组织TLR4、NF-κB、MyD88蛋白的表达相较于模型组有不同程度地降低;MMP-1、MMP-2水平也有所下降。结论半夏泻心汤可能通过影响TLRs/NF-κB/MyD88信号通路及其相关因子的表达,改善溃疡性结肠炎大鼠结肠损伤程度,进而防治UC。
Objective This study was conducted to investigate the effects of Banxia Xiexin Decoction on the expression of TLRs/NF-κB/myD88 signaling pathway and the inhibitory factors MMP-1 and MMP-2 in the intestinal mucosa of rats with ulcerative colitis(UC),and to explore the mechanisms of their effects.Methods The UC model induced by dextran sulphate sodium salt(DSS)was established and 35 Specific pathogen Free(SPF)rats were divided into normal group,model group,low concentration group(0.473 g·mL^(-1)),medium concentration group(0.945 g·mL^(-1))and high concentration group(1.890 g·mL^(-1))according to random number table method.Except for normal group,rats in the other four groups were given 2.5%DSS solution 2 mL/day for 10 consecutive days.After modeling,rats were given DSS solution 2 mL/day for one week,and DAI changes were observed.HE staining was used to confirm the extent of lesion damage to the colonic tissue by observation.Meanwhile,the expression of TLR4,NF-κB and MyD88 in the colon of each group of rats was also detected by Western Blot.The mRNA expression of TLR4,NF-κB,IκB-α,MMP-1 and MMP-2 in rat colon mucosa was detected by Real-time fluorescent quantitative polymerase chain reaction(RT-PCR).Results After modeling,the weight,appetite,coat,activity and urine volume of the rats decreased to varying degrees compared with before modeling,and some of the rats showed symptoms of diarrhea and hematoia.The expression of TLR4,NF-κB and MyD88 protein,as well as mRNA expression of TLR4,NF-κB,IκB-α,MMP-1 and MMP-2 were significantly higher in the model group compared with the normal group(P<0.01).After treatment with Banxia Xiexin Decoction,weight loss,diarrhea,hematochezia and other symptoms of UC rats were significantly alleviated.The expressions of TLR4,NF-κB and MyD88 proteins in the colonic mucosa of rats in the low,medium and high doses of Banxia Xiexin Decoction were decreased to varying degrees compared to the model group.The levels of MMP-1 and MMP-2 have also declined.Conclusions Banxia Xiexin Decoction may affect the expression of TLRs/NF-κB/MyD88 signaling pathway and related factors,improve the degree of colonic injury in rats with ulcerative colitis,and prevent and treat UC.
作者
李室箕
张林
牟永旭
朱烔依
罗文晔
王杰
谢田炜
陈兰秋
Li Shiji;Zhang Lin;Mu Yongxu;Zhu Tongyi;Luo Wenye;Wang Jie;Xie Tianwei;Chen Lanqiu(Liaoning University of Traditional Chinese Medicine,Shenyang 110847,China;College of Traditional Chinese Medicine,Liaoning University of Traditional Chinese Medicine,Shenyang 110847,China)
出处
《世界科学技术-中医药现代化》
CSCD
北大核心
2023年第3期1133-1139,共7页
Modernization of Traditional Chinese Medicine and Materia Medica-World Science and Technology
基金
辽宁中医药大学创新学院大学生创新创业项目(X202010162063):半夏泻心汤对溃疡性结肠炎大鼠肠道粘膜中的TLRs/NF-κB/myD88信号通路及抑制因子MMP-1、MMP-2表达影响的实验研究,负责人:李室箕
作者简介
通讯作者:张林,博士,博士生导师,主要研究方向:方剂药效机制与配伍规律研究;牟永旭,本科,主要研究方向:方剂药效机制与配伍规律研究。
