粪便菌群移植通过调节Th17/Treg平衡发挥对帕金森小鼠的保护作用

Protective Effect of Fecal Flora Transplantation on Parkinson's Mice by Regulating Th17/Treg Balance

目的:探讨粪菌移植对Th17/Treg平衡的调控及帕金森病(PD)小鼠的神经保护作用。方法:将18只小鼠均分为正常对照组、粪菌移植对照组及粪菌移植组,后两组采用—方法建立PD模型;粪菌移植组小鼠在建立PD模型后、以正常对照组小鼠的粪便菌悬液灌胃,粪菌移植对照组和正常对照组予生理盐水灌胃;爬杆实验评价小鼠的行动能力,牵引实验评价小鼠肌肉力量及平衡能力;Real-time PCR、Western blot检测小鼠黑质致密部(SNpc)RORγt、Foxp3 m RNA及蛋白表达;免疫荧光双染检测小鼠SNpc中Th17细胞(RORγt+CD4+)及Treg细胞(Foxp3+CD4+)的表达;ELISA检测小鼠纹状体中DA、5-HT的表达以及外周血与SNpc中IL-17A、IL-22、TGF-β、IL-10的表达。结果:与粪菌移植对照组比较,粪菌移植组小鼠爬杆实验中返回到杆底部的时间明显缩短(P<0.01),牵引实验中得分明显增加(P<0.01),小鼠SNpc中RORγt m RNA和蛋白的表达明显下调(P<0.01),粪菌移植组小鼠Foxp3 m RNA及蛋白表达明显上调(P<0.01),小鼠纹状体内DA及5-HT的含量明显升高(P<0.01),小鼠外周血与SNpc中IL-17A与IL-22的含量明显下调(P<0.01),IL-10与TGF-β明显上调(P<0.01),小鼠SNpc中Th17细胞(RORγt+CD4+)的数量有所下调,而Treg细胞(Foxp3+CD4+)的数量有所上调。结论:粪便移植对PD小鼠有神经保护作用,其机制可能与调节Th17/Treg平衡、DA与5-HT含量有关。

Objective:To explore the effect of fecal flora transplantation on Th17/Treg balance and Parkinson’s disease(PD)mice neuroprotection.Methods:Eighteen mice were divided into normal control group,fecal flora transplantation control group and fecal flora transplantation group,among which the latter two groups would establish PD model.After establishing PD model,mice in fecal flora transplantation group were perfused with fecal flora suspension of mice in the normal control group and fecal flora transplantation control group was given normal saline;pole-climbing test was adopted to evaluate the action ability of mice,and traction test was to evaluate the muscle strength and balance ability of mice;the expression of RORγt,Foxp3 m RNA and protein in mouse substantia nigra compact part(SNpc)was detected by Real-time PCR and Western blot;the expression of Th17 cells(RORγt+CD4+)and Treg cells(Foxp3+CD4+)in SNpc was detected by immunofluorescence double staining;ELISA was to detect the expression of DA and 5-HT in mouse striatum and the expression of IL-17 A,IL-22,TGF-βand IL-10 in peripheral blood and SNpc.Results:Compared with fecal flora transplantation control group,the time of fecal flora transplantation group of returning to the pole bottom in the pole-climbing test shortened significantly(P<0.01);the score in the traction test increased significantly(P<0.01);the expression of RORγt m RNA and protein in SNpc decreased sharply(P<0.01);the expression of Foxp3 m RNA and protein increased significantly(P<0.01);the contents of DA and 5-HT in mouse striatum increased significantly(P<0.01);the contents of IL-17 A and IL-22 in peripheral blood and SNpc decreased sharply(P<0.01),and IL-10 and TGF-βincreased significantly(P<0.01);Th17 cells(RORγt^+CD4^+)in mouse SNpc decreased and Treg cells(Foxp3^+CD4^+)increased.Conclusion:Fecal flora transplantation has a neuroprotective effect on PD mice by regulating Th17/Treg balance.