目的 探讨羟氯喹对系统性红斑狼疮小鼠肾损伤的影响,并基于Toll样受体9/髓样分化因子88/核因子-κB(TLR9/MyD88/NF-κB)信号通路探索其潜在机制。方法 采用ip降植烷的方法构建系统性红斑狼疮雌性小鼠模型,设模型组、羟氯喹(ig 80 mg/kg)组、TLR9抑制剂(ODN2088,ip 4 mg/kg)组、羟氯喹+ODN2088组,另设对照组,每组8只。分别1次/d连续给药5周后采集标本,称体质量、肾脏质量并计算肾脏指数(RI),检测24 h尿蛋白量(24 h-UTP)和血清肌酐(Scr)、尿素氮(BUN)水平;苏木精-伊红染色法(HE)和Masson染色观察肾组织病变和纤维化程度,进行病理评分和胶原容积分数(CVF);检测肾组织干扰素-γ(IFN-γ)、肿瘤坏死因子-α(TNF-α)、白细胞介素-2(IL-2)、白细胞介素-4(IL-4)水平;RT-PCR法和Western blotting法检测肾组织TLR9、MyD88、NF-κB p65 mRNA和蛋白表达。结果 与模型组比较,羟氯喹组、ODN2088组和HCQ+ODN2088组小鼠体质量显著升高,RI、24 h-UTP、血清Scr、BUN水平均显著降低(P<0.05);肾组织病变和纤维化状况均明显改善,肾小球、肾小管病理评分和CVF均显著降低(P<0.05);肾组织IFN-γ、TNF-α、IL-4水平显著降低,IL-2水平显著升高(P<0.05);肾组织TLR9、MyD88、NF-κB p65 mRNA和蛋白表达量均显著降低(P<0.05)。羟氯喹+ODN2088组对系统性红斑狼疮小鼠各检测指标的影响显著优于羟氯喹组和ODN2088组(P<0.05)。结论 羟氯喹对系统性红斑狼疮小鼠肾损伤具有保护作用,其机制可能与抑制TLR9/MyD88/NF-κB信号通路,减轻炎症反应有关。

Objective To investigate the effect of hydroxychloroquine on renal injury in systemic lupus erythematosus mice, and explore the potential mechanism based on toll-like receptor 9/myeloid differentiation factor 88/nuclear factor-κB (TLR9/MyD88/NF-κB) pathway. Methods The female mice model with systemic lupus erythematosus was established by ip of nophytanes, and the model group, hydroxychloroquine (ig 80 mg/kg hydroxychloroquine) group, TLR9 inhibitor (ip 4 mg/kg ODN2088) group, hydroxychloroquine + ODN2088 group, and the normal control group was set up, with 8 mices in each group. The samples were collected after 5 weeks of continuous administration once daily. The body mass, renal mass was weighted and the RI was calculated. 24 h-UTP and the level of Scr, BUN in serum were measured. The renal tissue histopathological changes and fibrosis were observed through HE staining or Masson staining, and the pathological score and CVF were performed. The level of IFN-γ, TNF-α, IL-2, IL-4 in renal tissues were measured. The mRNA and protein expressions of TLR9, MyD88, NF-κB p65 in renal tissues were detected by RT-PCR or Western blotting. Results Compared with the model group, the body mass in hydroxychloroquine group, ODN2088 group, and hydroxychloroquine + ODN2088 group was significantly increased, while the RI and the level of 24 h-UTP, Scr, BUN were significantly decreased (P < 0.05). The pathological changes and fibrosis of renal tissue were significantly improved, and the pathological scores of glomeruli, renal tubules and the CVF were significantly decreased (P < 0.05). The level of IFN-γ, TNF-α, IL-4 in renal tissue were significantly decreased, while the IL-2 was significantly increased (P < 0.05). The mRNA and protein expressions of TLR9, MyD88, NF-κB p65 in renal tissue were significantly decreased (P < 0.05). The effects of hydroxychloroquine + ODN2088 group on the detection indexes of systemic lupus erythematosus mice were significantly better than those of hydroxychloroquine group and ODN2088 group (P < 0.05). Conclusion Hydroxychloroquine has protective effect on renal injury in systemic lupus erythematosus mice, which mechanism may be related to inhibiting TLR9/MyD88/NF-κB signaling pathway and reducing inflammatory response.

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石家庄市科学技术研究与发展计划项目(201200773)