基于滤泡辅助性T细胞通路研究蜈蚣败毒饮治疗银屑病的机制  

Mechanism of Wugong Baidu Decoction in Treating Psoriasis Based onGene Transcription of Follicular Helper T Cell Pathway

作　　者：郑毅[1] 杨素清[2] 张琼 安月鹏[2] ZHENG Yi;YANG Suqing;ZHANG Qiong;AN Yuepeng(Heilongjiang University of Chinese Medicine,Harbin Heilongjiang China 150040;The First Affiliated Hospital of Heilongjiang University of Chinese Medicine,Harbin Heilongjiang China 150040)

机构地区：[1]黑龙江中医药大学,黑龙江哈尔滨150040 [2]黑龙江中医药大学附属第一医院,黑龙江哈尔滨150040

出　　处：《中医学报》2023年第4期787-793,共7页Acta Chinese Medicine

基　　金：国家自然科学基金项目(81904201,81973846);国家重点研发计划“中医药现代化研究”重点专项课题项目(2018YFC1705301);黑龙江省博士后资助经费项目(LBH-Z20198);黑龙江省普通本科高等学校青年创新人才培养计划项目(UNPYSCT-2020229)。

摘　　要：目的:在基因转录水平上研究中药蜈蚣败毒饮对银屑病小鼠皮损组织滤泡辅助性T细胞(follicular helper T cell,Tfh)通路中关键刺激、转录、分泌因子的调控作用。方法:将30只SPF级4~6周龄健康C57BL/6小鼠随机分为空白组、模型组、甲氨蝶呤组(3 mg·L^(-1))、蜈蚣败毒饮低剂量组(0.08 g·mL^(-1))、蜈蚣败毒饮中剂量组(0.16 g·mL^(-1))、蜈蚣败毒饮高剂量组(0.32 g·mL^(-1)),每组5只。除空白组外,其余组小鼠建立银屑病皮损模型。建模完成后进行药物干预,甲氨蝶呤组小鼠于第1—4天采用3 mg·L^(-1)甲氨蝶呤混悬液进行灌胃,第5~8天采用9 g·L^(-1)氯化钠注射液进行灌胃;蜈蚣败毒饮低、中、高剂量组小鼠灌胃给予不同浓度的蜈蚣败毒饮溶液;空白组、模型组小鼠灌胃给予9 g·L^(-1)氯化钠注射液,灌胃体积均为5 mL·kg-1,每日2次,持续8 d。采用鼠银屑病皮损面积和严重程度指数(mice psoriasis area and severity index,MPASI)评分法评估小鼠的皮损指数,并计算灌胃前后MPASI分值的变化量(△MPASI);RT-PCR法检测IL-6、IL-21、Bcl-6、STAT3、CXCR5、ICOS、PD-1、CD40L的mRNA表达水平;Pearson相关性分析考察△MPASI与皮损组织Tfh通路各因子转录表达量的相关性。结果:与空白组比较,造模小鼠皮损指数及IL-6 mRNA、IL-21 mRNA、Bcl-6 mRNA、STAT3 mRNA、CXCR5 mRNA、ICOS mRNA、CD40L mRNA表达水平显著升高(P<0.01),PD-1 mRNA表达水平显著降低(P<0.01)。与模型组比较,各给药组小鼠皮损指数显著降低(P<0.05),△MPASI显著升高(P<0.05);甲氨蝶呤组及蜈蚣败毒饮中、高剂量组小鼠皮损组织Tfh细胞通路关键因子IL-6 mRNA、IL-21 mRNA、Bcl-6 mRNA、STAT3 mRNA、CXCR5 mRNA、ICOS mRNA、CD40L mRNA表达水平显著降低(P<0.05),PD-1 mRNA表达水平显著升高(P<0.05)。Pearson分析显示,银屑病小鼠皮损组织Tfh细胞通路中IL-6 mRNA、IL-21 mRNA、Bcl-6 mRNA、STAT3 mRNA、CXCR5 mRNA、ICOS mRNA、CD40L mRNA表达量与△MObjective:To study the regulatory effect of Wugong Baidu Decoction on the key stimulation,transcription,and secretion factors in the follicular helper T cell(Tfh)pathway in psoriasis mouse skin lesions at the gene transcription level.Methods:30 healthy C57BL/6 mice aged 4-6 weeks in SPF grade were randomly divided into blank group,model group,methotrexate group(3 mg·L^(-1)),Wugong Baidu Decoction low-dose group(0.08 g·mL^(-1)),medium-dose group(0.16 g·mL^(-1)),and high-dose group(0.32 g·mL^(-1)),5 rats in each group.Except for the blank group,the mice in other groups were established with psoriasis skin lesion model on the back.Drug intervention was performed after the modeling was completed.Mice in the methotrexate group were intragastrically administered with 3 mg·L^(-1)methotrexate suspension on days 1-4,and 9 g·L^(-1)sodium chloride injection on days 5-8 by gavage.The mice in the low,medium,and high dose groups of Wugong Baidu Decoction were intragastrically given different concentrations of Wugong Baidu Decoction solution.Mice in the blank group and the model group were given 9 g·L^(-1)sodium chloride injection by intragastric administration,with a volume of 5 mL·kg-1,twice a day for 8 days.The mouse psoriasis area and severity index(MPASI)scoring method was used to evaluate the skin lesion index of mice,and the change of MPASI score before and after gavage was calculated(△MPASI).RT-PCR method was used to detect the mRNA expression levels of IL-6,IL-21,Bcl-6,STAT3,CXCR5,ICOS,PD-1,and CD40L.Pearson correlation analysis was used to investigate the correlation between△MPASI and the transcriptional expression of each factor of the Tfh pathway in skin lesion tissue.Results:Compared with the blank group,the skin lesion index and the expression levels of IL-6 mRNA,IL-21 mRNA,Bcl-6 mRNA,STAT3 mRNA,CXCR5 mRNA,ICOS mRNA,and CD40L mRNA in the modeled mice were significantly increased(P<0.01),the expression level of PD-1 mRNA was significantly decreased(P<0.01).Compared with the model group,the skin lesion i

关 键 词：银屑病 蜈蚣败毒饮 滤泡辅助性T细胞通路 小鼠 

分 类 号：R285.5[医药卫生—中药学]