四君子汤激活TLR-2/MyD88信号通路促进小肠黏膜上皮细胞损伤修复  

Sijunzi Decoction Promote the Repair of Intestinal Epithelial Cell Injury via Activating TLR-2/My D88 Signaling Pathway

作　　者：宋厚盼[1] 李如意[1,2] 黄惠勇[1] 蔡雄[1] 袁振仪[1] 刘平安[1,2] 周驰[3] 

机构地区：[1]湖南中医药大学中医诊断学湖南省重点实验室,湖南长沙410208 [2]湖南中医药大学药学院,湖南长沙410208 [3]广州中医药大学第一附属医院三骨科,广东广州510405

出　　处：《中药材》2016年第9期2081-2085,共5页Journal of Chinese Medicinal Materials

基　　金：湖南省教育厅优秀青年项目(15B179);湖南中医药大学科技创新团队项目(201503);湖南中医药大学博士科研启动项目(1001-017);湖南中医药大学中医诊断学国家重点学科开放基金项目(2015ZYZD19;2015ZYZD16)

摘　　要：目的:探讨四君子汤促进胃肠黏膜损伤修复的作用及机制。方法:采用血清药理学方法制备四君子汤含药血清;tips划痕法建立IEC-6细胞迁移模型,观察四君子汤含药血清对ICE-6细胞迁移的影响;实时细胞分析仪(RTCA)检测四君子汤含药血清对IEC-6细胞增殖的影响;荧光定量PCR(qRT-PCR)法检测TLR-2和My D88 mRNA表达;Western blot法检测TLR-2与My D88蛋白表达。结果:中(10%)、高(20%)浓度四君子汤含药血清可促进IEC-6细胞迁移(P<0.01);细胞损伤后12 h,中(10%)、高(20%)浓度四君子汤含药血清可促进IEC-6细胞增殖(P<0.01);细胞损伤后24 h和36 h,低(5%)、中(10%)、高(20%)浓度四君子汤含药血清均可促进IEC-6细胞增殖(P<0.05,P<0.01);中(10%)、高(20%)浓度四君子汤含药血清可上调TLR-2及My D88 mRNA和蛋白表达(P<0.01)。结论:四君子汤可加速胃肠黏膜损伤修复过程,其机制可能与激活TLR-2/My D88信号通路,促进黏膜上皮细胞迁移和增殖有关。Objective： To study the effect and mechanism of Sijunzi decoction（ SJZD） containing serum on the repairing of gastrointestinal mucosal damage. Methods： SJZD containing serum was prepared by serum pharmacological method. Cell migration model was established by tips scratch method,Real-time-cell-analyzer（ RTCA） was used to mensurate IEC-6 cell proliferation,the mRNA and protein expression of TLR-2 and My D88 mRNA were detected by qRT-PCR and Western blot analysis,respectively. Results： Medium dose（ 10%） and high dose（ 20%） of SJZD containing serum stimulated IEC-6 cell migration at 8 h after cell damage; medium dose（ 10%）and high dose（ 20%） of SJZD containing serum increased IEC-6 cell proliferation at 12 h after cell damage; low dose（ 5%）,medium dose（ 10%） and high dose（ 20%） of SJZD containing serum enhanced IEC-6 proliferation both at 24 h and 36 h after cell damage. Medium dose（ 10%） and high dose（ 20%） of SJZD containing serum upregulated TLR-2 and My D88 mRNA and protein expression,respectively. Conclusion： Sijunzi decoction can repair the injury of gastrointestinal mucosal barrier,the mechanism may be related to its effect on activating TLR-2 / My D88 signaling pathway,and promoting IEC-6 cell migration and proliferation.

关 键 词：四君子汤 胃肠黏膜损伤 细胞迁移 细胞增殖 TLR-2/My D88信号通路 

分 类 号：R285.5[医药卫生—中药学]