Zheng Linfeng, Zhang Jin, Yuan Xiangning, Tang Juan, Qiu Sisi, Peng Zhangzhe, Yuan Qiongjing, Xie Yanyun, Mei Wenjuan, Tang Yiting, Meng Jie, Hu Gaoyun, Tao Lijian
Department of Nephrology Medicine, Xiangya Hospital, Central South University, Changsha, Hunan, China.
Department of Gastroenterology Medicine, Xiangya Hospital, Central South University, Changsha, Hunan, China.
Nephrology (Carlton). 2018 Jun;23(6):573-584. doi: 10.1111/nep.13062.
We explored whether Fluorofenidone reduced interleukin-1β (IL-1β) production by interacting with NLRP3 inflammasome in unilateral ureteral obstruction (UUO).
Ureteral obstruction rats were treated with Fluorofenidone (500 mg/kg per day) for 3, 7 days. Morphologic analysis and leukocytes infiltration were assessed in ligated kidneys. Furthermore, plasmids of NLRP3, ASC, pro-Caspase-1, pro-IL-1β were co-transfected into 293 T cells, and then treated with Fluorofenidone (2 mM). The expression of NLRP3, ASC, pro-caspase-1, cleavage caspase-1, pro-IL-1β and cleavage IL-1β were measured by Western blot or real-time PCR in vivo and in vitro. Moreover the interaction of NLRP3 inflammasome-assembly was detected by co-immunoprecipitation and confocal immunofluorescence.
Fluorofenidone treatment significantly attenuated renal fibrosis and leukocytes infiltration in UUO model. Fluorofenidone had no effect on the expression of pro-IL-1β. Interestingly, Fluorofenidone inhibited the activation of NLRP3 inflammasome, downregulated Caspase-1 levels and thereby decreased the cleavage of pro-IL-1β into IL-1β in vivo and in vitro. Fluorofenidone treatment distinctively weakened the interaction between NLRP3 and ASC, as well as ASC and pro-Caspase-1 in vivo. However, Fluorofenidone treatment only significantly weakened the interaction between ASC and pro-Caspase-1 in co-transfected 293 T cells.
Fluorofenidone serves as a novel anti-inflammatory agent that attenuates IL-1β production in UUO model by interacting with NLRP3 inflammasome.
我们探究了氟非尼酮是否通过与单侧输尿管梗阻（UUO）中的NLRP3炎性小体相互作用来减少白细胞介素-1β（IL-1β）的产生。
用氟非尼酮（每天500毫克/千克）对输尿管梗阻大鼠进行3天、7天的治疗。对结扎肾脏进行形态学分析和白细胞浸润评估。此外，将NLRP3、ASC、前半胱天冬酶-1、前白细胞介素-1β的质粒共转染到293T细胞中，然后用氟非尼酮（2毫摩尔）处理。通过蛋白质免疫印迹法或实时聚合酶链反应在体内和体外测量NLRP3、ASC、前半胱天冬酶-1、裂解的半胱天冬酶-1、前白细胞介素-1β和裂解的白细胞介素-1β的表达。此外，通过免疫共沉淀和共聚焦免疫荧光检测NLRP3炎性小体组装的相互作用。
氟非尼酮治疗显著减轻了UUO模型中的肾纤维化和白细胞浸润。氟非尼酮对前白细胞介素-1β的表达没有影响。有趣的是，氟非尼酮在体内和体外均抑制了NLRP3炎性小体的激活，下调了半胱天冬酶-1水平，从而减少了前白细胞介素-1β裂解为白细胞介素-1β。氟非尼酮治疗在体内显著减弱了NLRP3与ASC以及ASC与前半胱天冬酶-1之间的相互作用。然而，在共转染293T细胞中，氟非尼酮治疗仅显著减弱了ASC与前半胱天冬酶-1之间的相互作用。
氟非尼酮作为一种新型抗炎剂，通过与NLRP3炎性小体相互作用，在UUO模型中减轻IL-1β的产生。
