双氢青蒿素治疗狼疮肾炎与SIGIRR诱导免疫负调控的相关性

Therapeutic effect of DHA on lupus nephritis and its relationship with SIGIRR inducing immune negative regulation

目的:探究双氢青蒿素(DHA)对狼疮肾炎的治疗作用与SIGIRR诱导的TLR4/NF-κB信号通路的相关性,并观察DHA对HK-2细胞炎性损伤的影响。方法:将MRL/lpr狼疮小鼠分为模型组、DHA(25、50、100 m/kg)治疗组、阳性对照组(强的松,5 mg/kg),另取C57BL/6小鼠为正常对照组,共给药12周。HE染色观察肾脏病理学改变,Western blot法检测肾脏SIGIRR,IRAK1和TRAF6蛋白表达变化。在体外,以10μg/ml LPS刺激人肾小管上皮细胞HK-2细胞,分别加入0.67、2.00、6.00μg/ml DHA,以Western Blotting法检测药物分别作用6、12和24 h后SIGIRR蛋白表达情况,ELISA方法检测上清IL-6、CCL2的分泌水平。结果:体内实验中,肾脏病理结果显示模型组小鼠肾脏受损,DHA 100 mg/kg治疗组肾脏损伤有所减轻。DHA 100 mg/kg组相比模型组,SIGIRR蛋白表达有一定的升高,且随DHA剂量增加该蛋白表达有升高趋势。体外实验中,不同浓度DHA能抑制CCL2的分泌,0.67μg/ml DHA作用24 h能显著增加SIGIRR的表达,且SIGIRR表达量随着时间延长有逐渐升高趋势。结论:DHA对狼疮肾炎有一定的治疗作用,且与肾脏SIGIRR表达升高抑制TLR4/NF-κB信号通路的过度活化有关;DHA对LPS诱导的HK-2炎症损伤的抑制与抑制CCL2分泌及增加SIGIRR表达有关。

Objective： To inspect the relationship between the therapeutic effect of DHA on lupus nephritis and the negative immune regulation of TLR4 / NF-κB signal pathway which was induced by SIGIRR; in vitro,to observe the effect of DHA on damaged HK-2 cell. Methods： In vivo,MRL / lpr mice were divided in model group,DHA groups（ 25,50,100 mg / kg）,positive group（ prednisone,5mg / kg）,and C57 BL /6 mice were taken as control group. Administrate drugs daily for 12 weeks. Examine the changes in renal pathology; the expression of SIGIRR,IRAK1,TRAF6 in kidneys were determined by Western blot. In vitro,treat human renal tubular epithelial cell HK-2 cells with LPS,and co-culture cells with DHA at the concentration of 0. 67 μg / ml to 6. 00 μg / ml for 6 h,12 h and 24 h. Detect SIGIRR expression by Western blot and the level of IL-6 and CCL2 of HK-2 cells by ELISA. Results： In vivo,renal pathology revealed that kidneys of model group were damaged,while treatment with 100 mg / kg DHA alleviated renal injury. Compared to model group,SIGIRR expression of DHA 100 mg / kg group increased a little,and the expression of this protein had a tendency to increase with the augment of DHA dose. In vitro,DHA treatment reduced secretion of CCL2 in HK-2 cells,and treatment of 0. 67 μg / ml DHA for 24 h increased SIGIRR expression significantly,which also showed a growing expression with time. Conclusion： DHA could inhibit development of mouse lupus nephritis through increasing SIGIRR expression which inhibited TLR4 / NF-κB signal pathway; DHA inhibited CCL2 secretion of HK-2 cells which were irritated by LPS,and it may be associated with increased expression of SIGIRR.