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目的:研究黄芪桂枝五物汤(HGW)对胶原诱导型关节炎(CIA)模型大鼠关节滑膜细胞凋亡的影响,并探讨其作用机制。方法:选用SD大鼠72只,随机分为正常组,模型组,HGW低、中、高剂量组,Tofacitinib(TFN)组,HGW低、中、高剂量组药物剂量分别为16,32,64 g·kg~(-1),相当于临床常人用量的3,6,12倍,TFN给药剂量为15 mg·kg~(-1),正常组、模型组每日给予10 m L·kg~(-1)蒸馏水。以牛Ⅱ型胶原加弗氏完全佐剂建立CIA大鼠模型,经HGW治疗4周后光镜下观察各组大鼠踝关节病理形态;通过流式细胞术检测白细胞介素(IL)-4,IL~(-1)0含量,免疫组化法检测B淋巴细胞瘤-2(B-cell lymphoma-2,Bcl-2),Bcl-2相关X蛋白(Bax)蛋白表达,DNA断裂的原位末端标记法(TUNEL)染色检测滑膜细胞的凋亡。结果:与正常组比较,模型组,HGW低、中、高剂量组,TFN组大鼠滑膜中Bcl-2蛋白表达增多(P<0.05),模型组Bax蛋白表达减少(P<0.05);与模型组比较,HGW高剂量组,TFN组大鼠滑膜中Bcl-2蛋白表达减少,Bax蛋白表达增多(P<0.05)。滑膜细胞凋亡可见,与正常组比较,模型组,HGW低、中、高剂量组,TFN组滑膜细胞增多,凋亡率下降(P<0.05);与模型组比较,HGW低、中、高剂量组,TFN组大鼠滑膜细胞凋亡率上升(P<0.05);与HGW低剂量组比较,HGW高剂量组滑膜细胞凋亡率显著上升(P<0.01);与HGW-M组比较,HGW高剂量组滑膜细胞凋亡率上升(P<0.05)。结论:黄芪桂枝五物汤可以促进RA大鼠异常亢进的滑膜细胞凋亡。
Objective: To study the effect of Huangqi Guizhi Wuwu Decoction (HGW) on the apoptosis of synovial cells in rats with collagen-induced arthritis (CIA) and to explore its mechanism. Methods: Seventy-two SD rats were randomly divided into normal group, model group, HGW low, medium and high dose group, Tofacitinib (TFN) group, HGW low dose, medium dose and high dose group dosage respectively 16,32,64 g · kg ~ (-1), equivalent to 3, 6 and 12 times of the clinical dosage, and the dosage of TFN was 15 mg · kg ~ (-1). In the normal group, the model group was given 10 m L · kg ~ (-1) distilled water. CIA rat models were established with Cat Ⅱ type Ⅱ collagen plus complete Freund’s adjuvant. The pathological changes of the ankle joint were observed under light microscope after 4 weeks of treatment with HGW. The levels of interleukin (IL) -4, IL The expression of Bcl-2, Bcl-2 and Bcl-2 protein was detected by immunohistochemistry. The apoptosis of synovial cells was detected by TUNEL staining. Results: Compared with normal group, the expression of Bcl-2 protein in synovium of model group, HGW low, middle and high dose group and TFN group increased (P <0.05), while the expression of Bax decreased in model group (P <0.05). Compared with model group, the expression of Bcl-2 protein and Bax protein in synovium of HGW high-dose group and TFN group were decreased (P <0.05). Compared with the normal group, synovial cells increased in the model group, HGW low, medium and high dose groups and TFN group, the apoptosis rate decreased (P <0.05). Compared with the model group, the HGW was lower (P <0.05). Compared with HGW low dose group, the apoptosis rate of synovial cells in HGW high dose group increased significantly (P <0.01) Compared with M group, the apoptosis rate of synovial cells in HGW high dose group increased (P <0.05). Conclusion: Huangqi Guizhi Wuwu Decoction can promote the apoptosis of synoviocytes in patients with abnormally hyperthyroidism.