文献类型：期刊文献

中文题名：四神丸对脾肾阳虚型溃疡性结肠炎模型大鼠结肠组织Toll样受体4及其负性调控因子IRAK-M表达的影响

英文题名：Effect of Sishenwan on Toll-like Receptor 4 and IRAK-M Expression in Colonic Tissue of Rats with Ulcerative Colitis of Spleen-kidney Yang Deficiency Type

作者：王爱华[1];何兰娟[2];朱向东[2]

第一作者：王爱华

机构：[1]甘肃省中医院,兰州730050;[2]甘肃中医药大学,兰州730000

第一机构：甘肃省中医院,兰州730050

年份：2019

卷号：25

期号：14

起止页码：70

中文期刊名：中国实验方剂学杂志

外文期刊名：Chinese Journal of Experimental Traditional Medical Formulae

收录：CSTPCD;;北大核心:【北大核心2017】；CSCD:【CSCD2019_2020】；

基金：国家自然科学基金项目(81360541)

语种：中文

中文关键词：Toll样受体4;白细胞介素1受体相关激酶M;溃疡性结肠炎;四神丸;作用机制

外文关键词：Toll-like receptor 4 ( TLR4 );interleukin-1 receptor-associated kinase-M ( IRAK-M);ulcerative colitis;Sishenwan;mechanism

摘要：目的:观察Toll样受体4(TLR4)及其负性调控因子白细胞介素-1受体相关激酶M(IRAK-M)在实验性溃疡性结肠炎(UC)模型大鼠结肠黏膜中的表达,并探讨四神丸干预UC的作用机制。方法:将90只Wistar大鼠随机分成6组,即空白组,模型组,柳氮磺胺嘧啶组(0. 36 g·kg^-1),四神丸低、中、高剂量组(2. 5,5,10 g·kg^-1),每组15只。以三硝基苯磺酸/乙醇溶液法制备UC大鼠模型,苏木素-伊红(HE)染色观察大鼠结肠组织病理学改变;采用放射免疫法测定血清游离三碘甲腺原氨酸(FT3),血清游离甲状腺素(FT4),免疫球蛋白(Ig) E,白细胞介素(IL)-2的含量;采用黄嘌呤氧化法测定大鼠血清超氧化物歧化酶(SOD)活性;采用硫代巴比妥酸(TBA)比色法测定大鼠血清丙二醛(MDA)的活性。采用实时荧光定量PCR(Real-time PCR),免疫组化和蛋白免疫印迹法(Western blot)分别检测TLR4,IRAK-M的mRNA及蛋白表达。结果:与空白组比较,模型组大鼠肠黏膜损伤评分显著增高(P <0. 01),大鼠血清Ig E,MDA含量显著升高(P <0. 01),FT3,FT4,IL-2,SOD表达水平显著下降(P <0. 01),TLR4 mRNA和蛋白表达显著升高(P <0. 01),IRAK-M mRNA和蛋白表达显著降低(P <0. 01);与模型组比较,各治疗组鼠肠黏膜损伤评分均显著降低(P <0. 01),Ig E,MDA含量明显下降(P <0. 05,P <0. 01),FT3,FT4,IL-2,SOD水平明显升高(P <0. 05,P <0. 01),TLR4 mRNA和蛋白表达明显降低(P <0. 05,P <0. 01),IRAK-M mRNA和蛋白表达水平显著升高(P <0. 01)。结论:UC的发病机制与TLR4及其负性调控因子IRAK-M的表达失衡有关,且四神丸可能通过抑制TLR4mRNA和蛋白的表达,促进其负性调控因子IRAK-M的表达,起到有效治疗UC的作用。
Objective: To observe the Toll-like receptor 4 ( TLR4 ) and its negative regulating factorInterleukin-1 receptor-associated kinase-M ( IRAK-M) in colonic mucosa of rats with experimental ulcerative colitis ( UC),and to discuss the mechanism of the Chinese medicine Sishenwan. Method: The 90 Wistar rats were randomly divide into six groups,blank group,model group,sulfasalazine group ( 0.36 g·kg ^- 1 ),Sishenwan low,medium and high-dose group ( 2.5,5,10 g·kg ^- 1 ),15 cases in each group. A rat model of UC was prepared by using a solution of trinitrobenzenesulfonic acid / ethano. The histopathological changes of colon were observed by hematoxylin-eosin ( HE) staining. The contents of serum free triiodothyroid acid ( FT3 ),serum free thyroxine ( FT4 ),immunoglobulin ( Ig) E and interleukin ( IL)-2 were determined by radioimmunoassay. The activity of superoxide dismutase ( SOD) in rat serum was determined by xanthine oxidation method. The activity of malondialdehyde ( MDA) in serum of rats was determined by thiobarbituric acid ( TBA) colorimetry. Result: Compared with blank group,intestinal mucosal injury score of rats in model group was significantly increased ( P < 0.01),serum IgE and MDA contents were significantly increased ( P < 0.01). The expression levels of FT3 ,FT4 , IL-2 and SOD were significantly decreased ( P < 0.01). The TLR4 mRNA and protein expression in model group increased significantly ( P < 0.01). The expression of IRAK-M mRNA and protein decreased significantly ( P < 0.01). Compared with model group,scores of each treatment group were significantly decreased ( P < 0.01),IgE and MDA contents were significantly decreased ( P < 0.05,P < 0.01),FT3 ,FT4 ,IL-2,and SOD contents were significantly increased ( P < 0.05,P < 0.01). The expression of TLR4 mRNA and proteinin each treatment group was significantly reduced ( P < 0.05,P < 0.01),IRAK-M mRNA and protein expression level increased ( P < 0.01). Conclusion: The unbalanced expressions of TLR4 and its negative regulating factor IRAK-M are connected with the pathogenesis of UC. Sishenwan can cure UC and control the expression of TLR4 and promote the expression of IRAK-M.