润燥灵方对干燥综合征模型小鼠血清Th1/Th2的影响

Impact of Runzaoling Decoction(润燥灵方) on Serum Th1/Th2 in Model Mice with Sjogren's Syndrome

目的探讨润燥灵方治疗干燥综合征的可能作用机制。方法将90只C57bl/6j小鼠随机分为空白组、模型组、强的松组及润燥灵低、中、高剂量组,每组15只。除空白组外,其余各组采用免疫诱导法建立干燥综合征小鼠模型。造模成功后润燥灵高、中、低剂量组分别给予润燥灵方水煎剂36、18、9 g/(kg·d),强的松组给予强的松0.2 g/(kg·d),空白组和模型组灌服0.2 ml生理盐水,每日1次。连续给药4周后,ELISA法检测血清干扰素γ(IFN-γ)和白细胞介素4(IL-4)水平,PCR法检测颌下腺T-bet和GATA-3转录因子的表达量,流式细胞仪检测血清Th1和Th2细胞的数量。结果模型组IFN-γ、IL-4、T-bet表达及T-bet/GATA-3、CD3+CD8-/IFN-γ、CD3+CD8-/IL-4及IFN-γ/IL-4明显高于空白组,润燥灵高、中剂量均能够降低IFN-γ、T-bet表达及CD3+CD8-/IFN-γ、CD3+CD8-/IL-4,以润燥灵高剂量组作用最明显(P<0.05或P<0.01)。结论润燥灵方能够下调干燥综合征模型小鼠血清Th1/Th2,可能是其治疗干燥综合征的作用机制之一。

Objective To explore the possible mechanism of Runzaoling Decoction（ 润 燥 灵 方） in treating Sjogren＇s syndrome（ SS）. Methods Ninety C57 mice were randomized into a blank group,a model group,a prednisone group,and low,medium and high dose Runzaoling Decoction groups,with 15 mice in each group. Each group was established SS mice models by immune induction method except the blank group. After the models were successfully established,the high,medium and low dose Runzaoling Decoction groups were given Runzaoling Decoction water decoction 36,18,or 9 g/（ kg·d）,respectively; the prednisone group was given prednisone 0. 2 g/（ kg·d）,the blank group and the model group given 0. 2 ml of normal saline,once every day. After 4 weeks of successive administration,the levels of serum interferonγ（ IFN-γ） and interleukin 4（ IL-4） were detected with Enzyme-Linked Immuno Sorbent Assay（ ELISA） method. The expressions of salivary gland T-bet and GATA-3 transcription factor were detected with polymerase chain reaction（ PCR） method. The number of serum Th1 and Th2 cells were calculated with flow cytometry. Results Levels of IFN-γ and IL-4,expression of T-bet,as well as T-bet/GATA-3,CD3＋CD8-/IFN-γ,CD3＋CD8-/IL-4 and IFN-γ/IL-4 in the model group were much higher than those in the blank group. Level of IFN-γ,T-bet expression,as well as CD3＋CD8-/IFN-γand CD3＋CD8-/IL-4 were lower in the High and medium dose Runzaoling Decoction groups,especially in the high dose Runzaoling Decoction group（ P 0. 05 or P 0. 01）. Conclusion Runzaoling Decoction might down-regulate serum levels of Th1/Th2 in SS model mice,which might be one of the mechanisms of treating SS.