三七总皂苷对小鼠Treg细胞分化的影响及其机制初探

Preliminarily explore the effect of panax notoginseng saponins on differentiation of mouse treg cells and its mechanism

目的:观察三七总皂苷(panax notoginseng saponins,PNS)对CD_(4)^(+) T细胞向调节性T细胞(Treg)分化的影响,并初步探讨其效应与调节细胞脂肪酸氧化代谢的相关性。方法:分离C57BL/6小鼠的脾细胞并磁珠分选CD_(4)^(+) T细胞进行体外培养,采用CCK-8法筛选PNS的最佳给药浓度并检测其对细胞活力的影响,之后分别设对照组、PNS干预组(5,10,20μg·mL^(-1)),相应药物干预后,流式细胞术检测Treg细胞的比例,ELISA方法检测上清液中β-羟基丁酸(BHBA)水平,q-PCR方法检测细胞肉毒碱棕榈酰转移酶1A(CPT-1A)mRNA表达水平,Western Blot法检测细胞中CPT-1A蛋白表达水平。结果:5,10,20μg·mL^(-1)的PNS可显著增加CD_(4)^(+) T细胞向Treg细胞分化的比例,明显增加细胞上清液中BHBA水平,升高细胞中CPT-1A mRNA和CPT-1A蛋白的表达水平。结论:一定浓度的PNS可显著促进小鼠CD_(4)^(+) T细胞向Treg细胞分化,其作用机制可能与提高CD_(4)^(+) T细胞脂肪酸氧化代谢水平有关。

Objective:To observe the effect of panax notoginseng saponins(PNS)on the differentiation of CD_(4)^(+) T cells into regulatory T(Treg)cells and preliminarily explore its relationship with regulating cellular fatty acid oxidation metabolism.Methods:The spleen cells of C57 BL/6 mice were isolated and CD_(4)^(+) T cells were cultured in vitro by magnetic beads sorting.The optimal concentration of PNS was screened by the CCK-8 method and its effect on cell viability was tested.Then the cells were divided into control group and PNS intervention groups(5,10,20μg·mL^(-1)).After corresponding drug intervention,the proportion of Treg cells was detected by using flow cytometry,the level ofβ-hydroxybutyric acid(BHBA)in the supernatant was measured by using ELISA,cellular carnitine palmitoyl transferase 1 A(CPT-1 A)mRNA expression was detected by using q-PCR,and the expression of CPT-1 A protein was detected by using Western Blot method.Results:The differentiation ratio of Treg cells and BHBA level in the supernatants significantly increased in the PNS intervention groups(5,10,20μg·mL^(-1)).The increasing expressions of CPT-1 A mRNA and protein in the cells were also observed in the PNS intervention groups.Conclusion:Certain concentrations of PNS can significantly promote the differentiation of mouse CD_(4)^(+) T cells into Treg cells,and its mechanism may be related with promoting the fatty acid oxidation of CD_(4)^(+) T cells.