四妙汤对RA模型大鼠关节VDR、CYP27B1、CYP24A1、Foxp3、RORγt的影响

Effect of Simiao Decoction on VDR, CYP27B1, CYP24A1, Foxp3 and RORγt in the Joints of RA Model Rats

摘要: 目的：探讨四妙汤改善类风湿关节炎(RA)模型大鼠的作用机制。方法：采用胶原诱导法(CIA)建立RA模型大鼠，灌胃四妙汤水煎液低、中、高剂量组(3.15 g/kg、6.3 g/kg、12.6 g/kg)，维生素D组(25 μg/kg)。给药周期为28天，记录大鼠一般情况。苏木精–伊红(HE)染色法观察踝关节组织病理改变；酶联免疫吸附剂检测法(ELISA)检测大鼠血清白细胞介素-17 (IL-17)、白细胞介素-10 (IL-10)、转化生长因子-β (TGF-β)含量；实时荧光定量PCR法(RT-PCR)检测大鼠关节滑膜组织中维生素D受体(VDR)、1-α羟化酶(CYP27B1)、24-羟化酶(CYP24A1)、叉头/翼状螺旋转录因子3 (Foxp3)、维甲酸相关孤儿核受体γt (RORγt) mRNA表达。结果：与空白组相比，模型组血清IL-17含量水平显著上升(P < 0.05)，血清IL-10、TGF-β含量水平显著下降(P < 0.05)；关节滑膜CYP24A1、RORγt mRNA表达水平显著上升(P < 0.05)，VDR、CYP27B1、Foxp3 mRNA表达水平显著下降(P < 0.05)。与模型组相比，维生素D组及四妙汤高剂量组血清IL-17水平显著下降(P < 0.05)，维生素D组及四妙汤中、高剂量组血清IL-10水平显著上升(P < 0.05)，维生素D组及四妙汤高剂量组血清TGF-β水平显著上升(P < 0.05)；维生素D组及四妙汤中、高剂量组关节滑膜VDR、CYP27B1、Foxp3 mRNA表达水平显著上升(P < 0.05)，维生素D组及四妙汤低、中、高剂量组关节滑膜CYP24A1 mRNA表达水平显著下降(P < 0.05)，维生素D组及四妙汤中、高剂量组关节滑膜RORγt mRNA表达水平显著下降(P < 0.05)。结论：四妙汤通过调节关节滑膜组织中VDR、CYP27B1、CYP24A1、Foxp3、RORγt mRNA表达情况，改善血清IL-17、IL-10、TGF-β水平，发挥其干预RA的作用。

Abstract: Objective: To explore the mechanism of action of Simiao decoction in improving the rat model of RA. Methods: Rat models of rheumatoid arthritis (RA) were established by collagen-induced arthritis (CIA). The rats were then administered with low, medium and high doses of Simiao decoction (3.15 g/kg, 6.3 g/kg, 12.6 g/kg) and vitamin D (25 ng/kg) by gavage. The administration period was 28 days, and the general conditions of the rats were recorded. Observation of pathological changes in ankle joint tissues by hematoxylin-eosin (HE) staining method. The contents of interleukin-17 (IL-17), interleukin-10 (IL-10), and transforming growth factor-β (TGF-β) in the serum of rats were detected by enzyme-linked immunosorbent assay (ELISA). The mRNA expressions of VDR, CYP27B1, CYP24A1, Foxp3, and RORγt in the synovial tissue of rat joints were detected by real-time fluorescence quantitative PCR (RT-PCR). Results: Compared with the control group, the serum IL-17 content level in the model group significantly increased (P < 0.05), while the serum IL-10 and TGF-β content levels significantly decreased (P < 0.05); the mRNA expression levels of CYP24A1 and RORγt in the synovial membrane significantly increased (P < 0.05), and the mRNA expression levels of VDR, CYP27B1, and Foxp3 significantly decreased (P < 0.05). Compared with the model group, the serum IL-17 level in the vitamin D group and the high-dose Simiao decoction group significantly decreased (P < 0.05), the serum IL-10 level in the vitamin D group and the medium and high-dose Simiao decoction groups significantly increased (P < 0.05), and the serum TGF-β level in the vitamin D group and the high-dose Simiao decoction group significantly increased (P < 0.05); the mRNA expression levels of VDR, CYP27B1, and Foxp3 in the synovial membrane in the vitamin D group and the medium and high-dose Simiao decoction groups significantly increased (P < 0.05), the mRNA expression level of CYP24A1 in the synovial membrane in the vitamin D group and the low, medium, and high-dose Simiao decoction groups significantly decreased (P < 0.05), and the mRNA expression level of RORγt in the synovial membrane in the vitamin D group and the medium- and high-dose Simiao decoction groups significantly decreased (P < 0.05). Conclusion: Simiao decoction exerts its intervention effect on RA by regulating the mRNA expression of VDR, CYP27B1, CYP24A1, Foxp3, and RORγt in the synovial membrane and improving the levels of serum IL-17, IL-10, and TGF-β.