雷公藤内酯醇对诱发性红斑狼疮小鼠模型的实验研究

Experimental Study of Triptolide on Induced Lupus Erythematosus Model in Mice

目的观察雷公藤内酯醇对Pristane诱导的BALB/c小鼠诱发性红斑狼疮的影响,分析其作用机制。方法 SPF级雌性BALB/c小鼠48只,将其中36只雌性BALB/c小鼠采用单次腹腔注射Pristane(0.5 m L/鼠)的方法建立诱发性红斑狼疮模型,注射后随机均分为模型组、羟氯喹组和雷公藤组,另取12只单次腹腔注射0.9%氯化钠注射液并设为对照组,成模后分别给予对应药物治疗4周。于末次给药后采小鼠眶静脉血,采用流式细胞术检测滤泡性辅助T淋巴细胞(Tfh细胞)表面标记物CXCR5在静脉血CD19+B细胞中的表达。采用ELISA法检测血清中滤泡归巢趋化因子(CXCL13)、细胞因子白细胞介素-6(IL-6)、白细胞介素-21(IL-21)、白细胞介素-27(IL-27)、双链DNA抗体(ds-DNA)滴度及免疫球蛋白G(Ig G)。取小鼠腹腔内脂肪肉芽肿结节病理组织,匀浆后采用ELISA法检测血管内皮生长因子(VEGF)及其受体1(VEGFR1)表达。结果雷公藤组CXCR5呈高表达,CXCL13水平、抗ds-DNA抗体滴度、Ig G均显著降低,与模型组比较差异均有统计学意义(均P<0.05)。雷公藤组细胞因子IL-6、IL-21、IL-27、VEGF和VEGFR1均显著降低,与模型组和羟氯喹组比较差异均有统计学意义(均P<0.05)。结论雷公藤内酯醇主要通过抑制归巢趋化因子CXCL13来调控Tfh细胞下游系列的免疫反应,并通过降低病变组织中VEGF水平而发挥对红斑狼疮的治疗作用。

Objective： To observe the effect of triptolide on induced lupus erythematosus induced by Pristane in BALB/c mice and to analyze its mechanism. Methods： Among 48 SPF female BALB/c mice,36 ones were made into models induced systemic lupus erythematosus by intraperitoneal injection of Pristane（0.5 m L/rats）. After injection,they were randomly divided into the model group,hydroxychloroquine group and Tripterygium wilfordii group;another 12 received single intraperitoneal injection of 0.9% normal saline and served as the control group.The models were given corresponding drugs for 4 weeks. After the last administration,the orbital blood of mice was collected,and the expression of follicle auxiliary T lymphocyte（Tfh cell） surface marker CXCR5 in venous blood CD19 ＋B cells was detected by flow cytometry. ELISA method was used to detect follicular homing factor CXCL13,cytokine IL-6,IL-21,IL-27,double chain DNA antibody（ds-DNA） titer and immunoglobulin G（Ig G）in serum. The pathological tissue of abdominal fat granuloma and the expression of vascular endothelial growth factor（VEGF） and its receptor 1（VEGFR1） were detected by ELISA. Results： CXCR5 in the Tripterygium wilfordii group was highly expressed,and CXCL13 level,anti-ds-DNA antibody titer and Ig G were significantly lower than those in the model group;compared with the model group,there were statistically significant differences（P〈0.05）. The cytokine IL-6,IL-21,IL-27,VEGF and VEGFR1 in Tripterygium wilfordii group were significantly lower than those in the model group and hydroxychloroquine group,with statistically significant differences（P〈0.05）. Conclusion： Triptolide regulates the downstream immune response of Tfh cells mainly by inhibiting the homing chemokine CXCL13,and plays a role in the treatment of lupus erythematosus by decreasing VEGF in the lesions.