自噬调节白介素-33的表达与子痫前期发病机制的关系

Relationship between expression of autophagy regulation interleukin-33 and the pathogenesis of preeclampsia

目的:探究白介素-33(interleukin-33,IL-33)在子痫前期(preeclampsia,PE)胎盘中的表达以及自噬通过调节IL-33的表达参与PE发病的分子机制。方法:应用免疫组化法及Western blot检测PE患者与正常妊娠胎盘中IL-33,微管相关蛋白LC3(LC3)的表达,应用Western blot检测PE患者与正常妊娠胎盘中IL-33受体ST2和核转录因子-κB(nuclear factor,NF-κB)的表达。使用3-Methyladenine(3-MA)、巴佛洛霉素A1(bafilomycin A1,BAF)处理人类绒毛膜外滋养细胞株HTR8/SVneo,检测IL-33、ST2和NF-κB蛋白表达情况以及自噬水平的高低。结果:(1)Western blot检测结果显示IL-33平均吸光度(absorbance,A)值PE组低于对照组(t=2.830,P=0.010),LC3B/A平均A值PE组高于对照组(t=3.470,P=0.005),ST2蛋白表达PE组高于对照组(t=2.650,P=0.026),NF-κB表达水平PE组高于对照组(t=3.180,P=0.005);(2)BAF处理HTR8/SVneo细胞株,IL-33蛋白水平较正常对照组明显降低(ta=3.740,Pa=0.006),LC3B/A比值水平较二甲基甲砜(dimethyl sulphoxide,DMSO)组明显升高(td=2.500,Pd=0.032)。3-MA对IL-33、NF-κB以及自噬水平的表达情况影响不明显。结论:过度自噬可能降低IL-33水平,导致IL-33抗炎作用减弱,进而出现过度炎症反应,并且这一过程与PE的发病存在一定关系。

Objective：To characterize the deregulation of interleukin-33 （IL-33） expression which regulated by autophagy in the pathological process of preeclampsia（PE）. Methods：The expressions of IL-33 protein and autophagy were detected in placentas of normal term and pregnancies complicated with PE by immunohistochemistry and Western blot. The levels of ST2, specific receptor of IL-33 ,and nuclear factor（NF-κB） were tested in placentas of normal term and pregnancies complicated with PE by Western blot. The autophagy pharmacological cell model was buih in vitro by using HTR8/SVneo cell line which treated with 3-methyladenine（the inhibition of autophagy） and bafilomycin A1 （the inhibition of autophagy body material degradation）;then the expressions of IL-33, ST2 and NF-κB were detected by Western blots. Results：The expressions of IL-33 protein were reduced,expressions of LC3B/A were increased, expressions of ST2 protein and NF-κB were increased in placentas of pregnancies complicated with PE than in placentas of normal term （t=2.830, P=0.010; t=3.470, P=O.O05 ; t=2.650, P=0.026; t=3.180, P=0.005 ）. In addition, BAF treatment exerted negative influence on the level of IL-33 in HTR8/SVneo cells （ta=3.740, Pa=0.006）, however, 3-MA treatment had no significant function. Conclusion： Autophagy plays a role in the pathophysiology mechanism of PE through the down-regulating the levels and anti-inflam- matory effects of IL-33.