艾灸调控G6PD介导的磷酸戊糖途径对佐剂性关节炎大鼠CD4^(+)T细胞增殖的影响

Effects of moxibustion on CD4^(+)T-cell proliferation by G6PD-mediated pentose phosphate pathway in adjuvant arthritis rats

目的:观察艾灸对佐剂性关节炎模型大鼠血浆和脾脏中葡萄糖-6-磷酸脱氢酶(G6PD)及还原型烟酰胺腺嘌呤二核苷磷酸(NADPH)水平、脾脏中CD4^(+)T细胞数量影响,探讨艾灸通过G6PD介导的磷酸戊糖途径发挥调节CD4^(+)T细胞增殖治疗类风湿性关节炎(RA)的效应机制。方法:将27只Sprague-Dawley雄性大鼠根据随机数字表法分为空白组、模型组和艾灸组,每组9只。模型组和艾灸组大鼠采用不完全弗氏佐剂致炎。空白组和模型组不干预。艾灸组悬灸双侧足三里、关元和阿是穴30 min,每日1次,共24次。采用苏木素-伊红染色评价大鼠滑膜组织病理改变;通过测量足趾容积观察大鼠足趾肿胀程度;采用免疫印迹法和酶联免疫吸附法分别检测各组大鼠脾脏和血浆中G6PD和NADPH含量;运用流式细胞术检测脾脏CD4^(+)T细胞数量。结果:与空白组比较,模型组血浆和脾脏中G6PD和NADPH含量明显升高,脾脏中CD4^(+)T细胞数量显著增加(P<0.01或P<0.05)。与模型组比较,艾灸组脾脏和血浆中NADPH含量以及脾脏中CD4^(+)T细胞数量显著降低(P<0.05或P<0.01),G6PD含量在血浆中显著降低(P<0.05),在脾脏中差异无统计学意义(P>0.05)。结论:艾灸可调节免疫,改善RA的关节滑膜炎症。其机制可能是通过调控CD4^(+)T细胞内G6PD介导的磷酸戊糖途径减少代谢产物NAPDH的产生,进而抑制初始CD4^(+)T细胞的增殖。

Objective:To observe the effects of moxibustion on the levels of glucose-6-phosphate dehydrogenase(G6PD)and reduced nicotinamide adenine dinucleotide phosphate(NADPH)in the plasma and spleen and the CD4^(+)T-cell number in the spleen of rats with adjuvant arthritis,thus to explore the mechanism in rheumatoid arthritis(RA)treatment with moxibustion by regulating the CD4^(+)T-cell proliferation through G6PD-mediated pentose phosphate pathway.Methods:Twenty-seven male Sprague-Dawley rats were randomly divided into a blank group,a model group,and a moxibustion group,with 9 rats in each group.Incomplete Freund’s adjuvant was used to induce inflammation in the model group and the moxibustion group.The blank group and the model group were not intervened.In the moxibustion group,suspended moxibustion was performed at bilateral Zusanli(ST36),Guanyuan(CV4),and Ashi points for 30 min,once a day for 24 times in total.Hematoxylin-eosin staining was used to evaluate the histopathological changes of rat synovial tissue;the swelling degree of the rat toes was observed by measuring the toe volume;G6PD and NADPH in the spleen and plasma were detected by Western blotting and enzyme-linked immunosorbent assay.Flow cytometry was used to detect the CD4^(+)T-cell number in the spleen.Results:Compared with the blank group,the levels of G6PD and NADPH in the plasma and spleen and the CD4^(+)T-cell number in the spleen were significantly increased in the model group(P<0.01 or P<0.05).Compared with the model group,the NADPH level in the spleen and plasma and the CD4^(+)T-cell number in the spleen in the moxibustion group decreased significantly(P<0.05 or P<0.01),and the G6PD level in the plasma decreased significantly(P<0.05),but there was no significant difference in the G6PD level in the spleen(P>0.05).Conclusion:Moxibustion can regulate immunity and improve joint synovial inflammation in RA.The mechanism may be that the G6PD-mediated pentose phosphate pathway reduces the production of metabolite NAPDH in CD4^(+)T cells,thereby inhibiting the proliferation of naive CD4^(+)T cells.