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目的:观察赤雹根总皂苷(total saponins of Thladiantha dubia root,TSTR)治疗牛II型胶原诱导性关节炎(CIA)大鼠,血液及膝关节滑膜中破骨细胞(OC)正反馈因子白细胞介素-15(IL-15),IL-34的水平变化,探讨TSTR治疗大鼠CIA的作用机制。方法:雄性Wistar大鼠80只,随机取10只作为正常组,其余大鼠均于尾根部及背部皮内多点注射牛II型胶原与完全弗氏佐剂混匀的乳剂。7 d后同法操作,以首次注射后14 d,大鼠关节炎指数(AI值)≥4分为造模成功,将模型大鼠随机分为模型组,TSTR低、中、高剂量组(20,40,80 mg·kg~(-1))和雷公藤多苷(TG)阳性药组(12 mg·kg~(-1)),各药物组均灌胃给予相应药物35 d,每天1次。观察大鼠足爪肿胀度,评价AI值;采用苏木精-伊红染色法(HE)观察滑膜病理变化;酶联免疫吸附测定(ELISA)检测大鼠血清中IL-15,IL-34含量;逆转录-聚合酶链式反应(RT-PCR)检测大鼠滑膜中IL-15,IL-34 mRNA表达水平;蛋白质印迹法(Western blot)检测滑膜中IL-15,IL-34蛋白表达水平。结果:与正常组比较,模型组大鼠AI值显著增高(P<0.01),滑膜细胞中炎性细胞的数量增多,在血清和滑膜组织中,IL-15,IL-34的含量、基因和蛋白表达均显著增高(P<0.01);与模型组比较,TSTR各剂量组和TG组AI值显著降低(P<0.01);且TSTR低剂量组和TG组滑膜中炎性细胞数量及血管数量明显减少;TSTR低剂量组和TG组大鼠血清中IL-15,IL-34表达和滑膜中IL-15,IL-34蛋白表达,IL-15,IL-34 mRNA的表达均显著降低(P<0.05,P<0.01)。结论:TSTR对CIA大鼠的治疗作用,可能与抑制血液和滑膜组织中IL-15,IL-34的表达有关。
OBJECTIVE: To observe the effects of total saponins of Thladiantha dubia root (TSTR) on osteoporotic (OC) positive feedback factor leukocytes in blood and knee synovium of rats with bovine type II collagen induced arthritis (CIA) Interleukin-15 (IL-15), IL-34 level changes in rats to explore the mechanism of TSTR treatment of CIA rats. Methods: Eighty male Wistar rats were randomly selected as the normal group. The rest of the rats were injected with emulsion of bovine type II collagen and complete Freund’s adjuvant in the root of the tail and the back. After 7 days, the model rats were randomly divided into model group and model group with low, middle and high dosage of TSTR (control group) 20, 40 and 80 mg · kg -1) and TG-positive group (12 mg · kg -1) respectively. Each drug group was given gavage for 35 days, 1 time. The degree of swelling of the paws was observed and the AI value was evaluated. The synovial pathological changes were observed by hematoxylin-eosin staining (HE staining). The levels of IL-15 and IL-34 in sera were detected by enzyme linked immunosorbent assay (ELISA) The expression of IL-15 and IL-34 mRNA in synovium of rats was detected by reverse transcription-polymerase chain reaction (RT-PCR). The levels of IL-15 and IL-34 Protein expression levels. Results: Compared with the normal group, the AI in the model group was significantly increased (P <0.01), and the number of inflammatory cells in the synovial cells increased. The levels of IL-15 and IL-34 in serum and synovial tissue, (P <0.01). Compared with the model group, the AI of TSTR and TG groups decreased significantly (P <0.01). The number of inflammatory cells in synovium of TSTR low dose group and TG group 15 and IL-34, IL-15 and IL-34 mRNA in synovium of TSTR low dose group and TG group were significantly decreased Significantly decreased (P <0.05, P <0.01). Conclusion: The therapeutic effect of TSTR on CIA rats may be related to the inhibition of the expression of IL-15 and IL-34 in blood and synovial tissue.