复方生地合剂从TLR-NF-κB通路调节MRL/lpr小鼠Th17/Treg平衡

Compound Shengdi Mixture regulated the balance of Th17/Treg in MRL/lpr mice through the TLR-NF-κB pathway

目的:探讨复方生地合剂从TLR-NF-κB信号通路调节MRL/lpr小鼠Th17/Treg细胞平衡的作用。方法:将MRL/lpr狼疮模型小鼠从随机分为模型组、中药组、西药组,每组10只。中药组给予复方生地合剂灌胃治疗,西药组给予强的松灌胃治疗,模型组以0.9%氯化钠溶液灌胃治疗,干预8周。采用酶联免疫方法检测外周血细胞因子IL-17、TGF-β、IFN-α和TNF-α的水平,采用流式细胞术检测Th17细胞和Treg细胞水平,采用定量聚合酶链反应测定脾组织中TLR7、TLR9、MyD88、NF-κB、RORγt和Foxp3 mRNA表达。结果:两给药组脾组织TLR7、TLR9、MyD88、NF-κB、RORγt mRNA表达水平均低于模型组(P<0.05,P<0.01),西药组脾组织Foxp3 mRNA表达水平高于模型组(P<0.05)。中药组TNF-α、IFN-α、IL-17A水平均低于模型组(P<0.05),TGF-β1水平高于模型组(P<0.05)。西药组IFN-α、IL-17A水平均低于模型组(P<0.05,P<0.01),TGF-β1水平高于模型组(P<0.05)。两给药组Th17细胞/CD_4^+T细胞比例低于模型组(P<0.05,P<0.01),Treg细胞/CD_4^+T细胞比例高于模型组(P<0.05),Th17细胞/Treg细胞比例均显著低于模型组(P<0.01)。结论:复方生地合剂可能通过抑制MRL/lpr小鼠TLR-NF-κB通路信号传导调节Th17/Treg细胞平衡。

Objective:To investigate the effects of compound Shengdi Mixture on regulating the balance of Th17/Treg cells in the MRL/lpr mice through TLR-NF-κB signaling pathway.Methods:MRL/lpr lupus model mice were randomly divided into model group,Chinese medicine group and western medicine group,ten mice in each group.The Chinese medicine group was given the compound Shengdi Mixture for intragas tric administration,and the western medicine group was given the treatment with Prednisone.The model group was replaced with physiological saline.After 8 weeks,the peripheral blood and spleen were taken and treated.The expression of cytokines IL-17,TGF-β,IFN-αand TNF-αin peripheral blood was detected by enzymelinked immunosorbent assay(ELISA).The levels of Th17 cells and Treg cells were detected by FCM.The expression of TLR7,TLR9,MyD88,NF-κB,RORyt and Foxp3 mRNA in spleen was determined by quantitative polymerase chain reaction.Results:The expression levels of TLR7,TLR9,MyD88,NF-κB and RORyt mRNA were all lower than those of the model group(P<0.05,P<0.01),and the expression levels of Foxp3 mRNA in western medicine group were higher than those of the model group(P<0.05).Levels of TNF-α,IFN-a and IL-17 A in the Chinese medicine group were lower than those in the model group(P<0.05),and the level of TGF-β1 was higher than that in the model group(P<0.05).Levels of IFN-a and IL-17 A in the western medicine group were lower than those in the model group(P<0.05,P<0.01),and the level of TGF-β1 was higher than that in the model group(P<0.05).The proportion of Th17 cell/CD4+T cells in the Chinese medicine group and the western medicine group was lower than that in the model group(P<0.05,P<0.01),the proportion of Treg cell/CD4+T cells was higher than that in the model group(P<0.05),and the proportion of Th17 cell/Treg cell was significantly lower than that in the model group(P<0.01).Conclusion:The compound Shengdi Mixture may regulate the balance of Th17/Treg cells by inhibiting the signal transduction of the TLR-NF-κB pathway in MRL/lpr mice.