雷公藤多苷联合人参皂苷对胶原诱导关节炎大鼠外周血CD4+CD25+Foxp3+调节性T细胞和白介素-17的影响

Effect of Tripterygium Glycosides Combined with Ginsenoside on CD4+CD25+Foxp3+Regulatory T Cells and Interleukin-17in Peripheral Blood of Collagen Induced Arthritis in Rats

目的观察植物中药有效成分配伍——雷公藤多苷(TG)与人参皂苷(GS)对胶原诱导关节炎(CIA)大鼠外周血CD4+CD25+Foxp3+调节性T细胞(Treg)和白介素-17(IL-17)的影响。方法选择体质量150～180 g的Wistar大鼠24只,皮内注射鸡Ⅱ型胶原建立CIA模型,随机分为模型组、TG+GS组、甲氨蝶呤(MTX)组,每组8只。另设8只正常大鼠为正常对照组。正常对照组和模型组予生理盐水灌胃,TG+GS组、MTX组分别予TG+GS、MTX灌胃,共给药21 d。给药结束后测量各组大鼠足爪厚度,与造模前比较并计算关节肿胀率;分离外周血单个核细胞,采用流式细胞术检测CD4+CD25+Foxp3+Treg;采用酶联免疫吸附试验(ELISA)检测血清IL-17水平。结果给药结束后与模型组相比,TG+GS组和MTX组大鼠足爪肿胀率明显降低(P<0.01)、外周血CD4+CD25+Treg百分率和CD4+CD25+Foxp3+Treg百分率均显著增高(P<0.01)、血清IL-17水平降低(P<0.05);TG+GS组和MTX组上述指标比较差异均无统计学意义(P>0.05)。CD4+CD25+Foxp3+Tregs和血清IL-17水平呈显著负相关(r=0.423,P<0.05)。结论 TG联合GS对大鼠CIA具有明显治疗效应,其作用机制可能与调节Treg/Th17平衡有关。

Objective To observe effect of plant effective components of Chinese Herbal Medicines - tripterygi- um glycosides （TG） and ginsenosides （GS） on peripheral blood CD4 ＋ CD25 ＋ Foxp3 ＋ regulatory T cells （Treg） and IL-17 of collagen induced arthritis （ C IA ） in rats. Methods A total of 24 Wistar rats （weight 150-180 g） were given intradermal injection of chicken II collagen to build the CIA models, and were randomly divided into model group （ group A, n = 8） , tripteryginm glycosides combined with ginsenoside group （ group B, n = 8 ） and Methotrexate （MTX） group （ group C, n = 8 ） , and normal rats were as control group （ n = 8 ）. Rats in control group and group A were given intragas- tric administration with normal saline, group B were given intragastric administration with TG ＋ GS, and group C were given MTX gavage. Treatment period for the four groups was 21d. Rats＇ claws thicknesses of the four groups were meas- ured after treatment, and were compared with those before treatment. Joint swelling rates were also calculated; mononu- clear cells separated from peripheral blood, CD4 ＋ CD25 ＋ Foxp3 ＋ Treg were detected by flow cytometry; the level of se- rum IL-17 was detected by enzyme linked immunosorbent assay （ELISA）. Results After treatment rats＇ tumescence rates in group B and C were significantly reduced compared with that in group A （P 〈 0.01 ） , while the percentages of CD4 ＋ CD25 ＋ Tregs and CD4 ＋ CD25 ＋ Foxp3 ＋ Treg of peripheral blood in the two groups were significantly increased compared with those in group A （P 〈 0.01 ） , and serum levels of IL-17 in the two groups were lower than that in group A （ P 〈 0.05 ）. There was no statistical significance of above indexes in group B and C （P 〉 0. 05 ）. There was significantnegative correlation between CD4 ＋ CD25 ＋ Foxp3 ＋ Tregs and serum levels of IL-17 （ r = - 0. 423 ,P 〈 0.05 ）. Conclu- sion TG combined with GS in treatment of CIA in rats has significantly therapeutic effect, its mechanism may be related to regulating balance of Treg/Th17 ceils.