虎杖苷调控PINK1-Parkin介导的线粒体自噬改善变应性鼻炎

Polydatin alleviates allergic rhinitis by regulating PINK1-Parkin-mediated mitophagy

目的探讨虎杖苷(Polydatin,PD)通过PINK1-Parkin信号通路调控线粒体自噬对变应性鼻炎(Allergic rhinitis,AR)的保护作用及机制,为临床治疗AR提供新靶点。方法将32只BALB/c小鼠随机分为4组,即对照组,OVA组,PD低剂量(30 mg·kg^(-1))和高剂量(45 mg·kg^(-1))治疗组。造模结束后,记录小鼠挠鼻和打喷嚏的总次数。HE染色观察鼻黏膜上皮的形态和嗜酸性粒细胞浸润情况。Western blot检测PINK1、Parkin、TOM20和线粒体凋亡相关蛋白Bax、Bcl-2、caspase-3、cleaved-caspase-3和Cytochrome C的表达量。免疫荧光观察鼻黏膜上皮细胞(HNEpC)中PINK1和cleaved-caspase-3的表达。结果AR小鼠挠鼻和打喷嚏的频率显著增加,鼻黏膜上皮增厚,嗜酸性粒细胞聚集,予以PD治疗后逆转了上述变化。Western blot结果显示,OVA组PINK1、Parkin和促凋亡蛋白Bax、caspase-3、cleaved-caspase-3、Cytochrome C表达增加,TOM20、Bcl-2的表达降低,PD上调了PINK1、Parkin、Bcl-2的水平,抑制了TOM20和促凋亡蛋白的表达,而予以线粒体分裂抑制剂1(Mdivi-1)预处理后,PINK1、Parkin和Bcl-2表达减少,TOM20和促凋亡蛋白表达增加,PD治疗作用不明显。免疫荧光证实了PINK1和cleaved-caspase-3的Western blot结果。结论PD通过激活PINK1-Parkin信号通路,促进线粒体自噬从而改善AR。

Objective To investigate the protective effect and mechanism of polydatin(PD)on allergic rhinitis(AR)by regulating mitophagy through PINK1-Parkin signaling pathway,and to provide a new target for clinical treatment of AR.Methods Thirty-two BALB/c murine were randomly divided into 4 groups:control group,OVA group,PD low-dose(30 mg·kg^(-1))and high-dose(45 mg·kg^(-1))treatment groups.At the end of modeling,the total number of sneezing and nasal rubbing of murine was recorded.HE staining was used to observe the morphology of nasal mucosal epithelium and eosinophil infiltration.Western blot was used to detect the expression of PINK1,Parkin,TOM20 and mitochondrial apoptosis-related proteins Bax,Bcl-2,caspase-3,cleaved-caspase-3 and Cytochrome C.The expression of PINK1 and cleaved-caspase-3 in nasal epithelial cells(HNEpC)was observed by immunofluorescence.Results The frequency of sneezing and nasal rubbing movements was significantly increased,the nasal mucosa epithelium was thickened,and eosinophils were accumulated in AR murine,these results were reversed after PD treatment.Western blot results showed that signaling proteins PINK1/Parkin and pro-apoptotic proteins,including Bax,caspase-3,cleaved-caspase-3 and Cytochrome C were significantly overexpressed,the expression of TOM20 and Bcl-2 was decreased in OVA group,and PD up-regulated the levels of PINK1,Parkin,as well as Bcl-2 and inhibited the expression of TOM20 and pro-apoptotic proteins,while after pretreatment with mitochondrial division inhibitor 1(Mdivi-1),the expression of PINK1 and Parkin was reduced,the expression of TOM20 was increased,while PD treatment did not significantly affect this effect.From the immunofluorescence results,it can be seen that the level of PINK1 was increased after IL-13 stimulation of HNEpCs compared with the control group,and PD further up-regulated the expression of PINK1,which was suppressed after pretreatment with Mdivi-1,while PD did not change this phenomenon.Western blot results for PINK1 and cleaved-caspase-3 were confirmed by immunofluorescence.Conclusion PD may activate mitophagy through the PINK1-Parkin signaling pathway,thereby protecting against AR.