目的 探討羥氯喹對系統性紅斑狼瘡小鼠腎損傷的影響,并基于Toll樣受體9/髓樣分化因子88/核因子-κB(TLR9/MyD88/NF-κB)信號通路探索其潛在機制。方法 采用ip降植烷的方法構建系統性紅斑狼瘡雌性小鼠模型,設模型組、羥氯喹(ig 80 mg/kg)組、TLR9抑制劑(ODN2088,ip 4 mg/kg)組、羥氯喹+ODN2088組,另設對照組,每組8只。分別1次/d連續(xù)給藥5周后采集標本,稱體質量、腎臟質量并計算腎臟指數(RI),檢測24 h尿蛋白量(24 h-UTP)和血清肌酐(Scr)、尿素氮(BUN)水平;蘇木精-伊紅染色法(HE)和Masson染色觀察腎組織病變和纖維化程度,進行病理評分和膠原容積分數(CVF);檢測腎組織干擾素-γ(IFN-γ)、腫瘤壞死因子-α(TNF-α)、白細胞介素-2(IL-2)、白細胞介素-4(IL-4)水平;RT-PCR法和Western blotting法檢測腎組織TLR9、MyD88、NF-κB p65 mRNA和蛋白表達。結果 與模型組比較,羥氯喹組、ODN2088組和HCQ+ODN2088組小鼠體質量顯著升高,RI、24 h-UTP、血清Scr、BUN水平均顯著降低(P<0.05);腎組織病變和纖維化狀況均明顯改善,腎小球、腎小管病理評分和CVF均顯著降低(P<0.05);腎組織IFN-γ、TNF-α、IL-4水平顯著降低,IL-2水平顯著升高(P<0.05);腎組織TLR9、MyD88、NF-κB p65 mRNA和蛋白表達量均顯著降低(P<0.05)。羥氯喹+ODN2088組對系統性紅斑狼瘡小鼠各檢測指標的影響顯著優(yōu)于羥氯喹組和ODN2088組(P<0.05)。結論 羥氯喹對系統性紅斑狼瘡小鼠腎損傷具有保護作用,其機制可能與抑制TLR9/MyD88/NF-κB信號通路,減輕炎癥反應有關。

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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石家莊市科學技術研究與發(fā)展計劃項目(201200773)