[關鍵詞]
[摘要]
目的 考察復方扶芳藤合劑(CFM)對免疫抑制小鼠Th1/Th2細胞水平的調節(jié)作用及機制。方法 將SPF級Balb/c小鼠隨機分為對照組,單給CFM低、中、高劑量(3.75、7.50、15.00 g·kg-1)組,模型組,CFM低、中、高劑量(3.75、7.50、15.00 g·kg-1)組,模型組和CFM低、中、高劑量組連續(xù)3 d ip 80 mg·kg-1的環(huán)磷酰胺(CTX)建立免疫抑制模型,對照組和CFM單給藥組不造模,僅ip等體積0.9%氯化鈉溶液。造模結束后,每天ig給藥1次,連續(xù)給藥10 d,對照組和模型組ig等體積0.9%氯化鈉溶液。末次給藥24 h后取材,流式細胞術檢測小鼠外周血Th1、Th2細胞水平及其表面活化(CD38、CD69)和耗竭[CD95、程序性死亡受體1(PD-1)]分子的表達水平,脾臟CD3+CD4+T細胞中γ干擾素(IFN-γ)、白細胞介素(IL)-2、IL-10細胞因子水平。結果 與對照組比較,模型組外周血Th1/Th2細胞向Th1型偏移,Th1細胞上CD38、CD95、PD-1的表達顯著升高(P<0.001),Th2細胞上CD38、PD-1的表達顯著升高(P<0.05、0.001),脾臟CD3+CD4+T細胞中IFN-γ的表達顯著升高(P<0.001);與模型組比較,CFM 3.75 g·kg-1組外周血Th1/Th2趨于平衡穩(wěn)態(tài),Th1細胞中CD38、PD-1的表達、Th2細胞CD38、PD-1的表達均顯著下降(P<0.05、0.01、0.001);小鼠脾臟CD3+CD4+T細胞中IFN-γ和IL-2表達均顯著下降(P<0.05),IL-10的表達顯著上升(P<0.05、0.01、0.001)。單給CFM并不改變正常小鼠的Th1/Th2平衡及功能。結論 CFM改善免疫抑制小鼠外周血Th1/Th2細胞向Th1型偏移,對免疫抑制小鼠外周血Th1、Th2細胞的活化、耗竭具有正向調節(jié)作用,促進Th2細胞分泌IL-10,抑制Th1細胞分泌IFN-γ、IL-2。
[Key word]
[Abstract]
Objective To investigate the regulatory effect of Compound Fufangteng Mixture (CMF) on Th1/Th2 cell level in immunosuppressed mice. Methods Balb/c mice were randomly divided into control group, single CFM low, medium, and high dose (3.75, 7.50, 15.00 g·kg-1) groups, model group, CFM low, medium, and high dose groups, model group, and CFM low, medium, and high dose groups that were ip with 80 mg·kg-1 of CTX for three consecutive days to establish an immunosuppressive model. The control group and CFM single-dose group were not subjected to modeling and were ip with an equal volume of 0.9% sodium chloride solution. After modeling was completed, the mice were ig given the drug once daily for 10 consecutive days. The control group and model group were ig given an equal volume of 0.9% sodium chloride solution. After the last drug administration, the mice were sacrificed and the peripheral blood Th1/Th2 cell levels, the surface activation (CD38, CD69) and exhaustion [CD95, programmed death receptor 1 (PD-1)] molecule expression levels, and the levels of cytokines interferon gamma (IFN-γ), interleukin (IL)-2, and IL-10 in the spleen CD3+CD4+T cells were detected by flow cytometry. Results Compared with the control group, the Th1/Th2 cells in the peripheral blood of the model group shifted towards the Th1 type. The expressions of CD38, CD95, and PD-1 on Th1 cells were significantly increased (P < 0.001), and the expressions of CD38 and PD-1 on Th2 cells were significantly increased (P < 0.05, 0.001). The expression of IFN-γ in the spleen CD3+CD4+T cells was significantly increased (P < 0.001). Compared with the model group, the Th1/Th2 in the peripheral blood of the CFM 3.75 g·kg-1 group tended to be in a balanced and stable state. The expressions of CD38 and PD-1 on Th1 cells and the expressions of CD38 and PD-1 on Th2 cells were significantly decreased (P < 0.05, 0.01, 0.001). The expressions of IFN-γ and IL-2 in the spleen CD3+CD4+T cells of mice were significantly decreased (P < 0.05), and the expression of IL-10 was significantly increased (P < 0.05, 0.01, 0.001). The single administration of CFM did not change the Th1/Th2 balance and function of normal mice. Conclusion CFM improves the shift of Th1/Th2 cells in the peripheral blood of immunosuppressed mice towards the Th1 type. It has a positive regulatory effect on the activation and exhaustion of Th1 and Th2 cells in the peripheral blood of immunosuppressed mice, promotes the secretion of IL-10 by Th2 cells, and inhibits the secretion of IFN-γ and IL-2 by Th1 cells.
[中圖分類號]
R285.5
[基金項目]
國家自然科學基金資助項目(81860780);廣西自然科學基金資助項目(2020GXNSFAA297162)