三七皂苷通过AMPK/DRP1介导的线粒体裂变减轻过敏性鼻炎

Panax notoginseng saponin R1 attenuates allergic rhinitis through AMPK/DRP1 mediated mitochondrial fission

目的探讨三七皂苷(notoginsenoside R1,PNS-R1)是否通过AMP激活蛋白激酶(AMP-activated protein kinase,AMPK)/线粒体裂变关键蛋白(dynamin-related protein 1,DRP1)介导的线粒体裂变减轻过敏性鼻炎(allergic rhinitis,AR)。方法利用不同剂量PNS-R1治疗卵清蛋白(Ovalbumin,OVA)诱导的AR小鼠,通过观察擦鼻、打喷嚏的过敏症状和鼻组织的HE染色探索PNS-R1在AR中的抑制作用。通过酶联免疫吸附法(ELISA)检测血清IgE水平和鼻灌洗液(nasal lavage fluid,NLF)炎性细胞因子水平,Western blot检测凋亡相关蛋白。在体外,利用IL-13刺激人鼻黏膜上皮细胞(human nasal epithelial cells,HNEpC),观察细胞凋亡、线粒体膜电位、细胞活性氧(reactive oxygen species,ROS)和线粒体ROS(mtROS)生成、AMPK/DRP1,TXNIP/NLRP3炎症小体的表达水平以及DRP1易位情况。结果PNS-R1减轻了AR小鼠的过敏症状,HE染色炎性细胞减少,降低血清OVA特异性IgE水平以及NLF中IL-4、IL-6和IL-8的水平。在体外,PNS-R1上调IL-13刺激后的线粒体膜电位,降低ROS和mtROS生成、减少cleaved-caspase-3、Bax和上调Bcl-2表达,以AMPK依赖性方式下调DRP1磷酸化(Ser 616)和DRP1在线粒体膜上的易位,减少TXNIP/NLRP3表达。结论PNS-R1通过抑制AMPK/DRP1信号轴及随后的TXNIP/NLRP3信号轴保护线粒体的完整性,缓解AR。

Aim To investigate whether notoginsenoside R1(PNS-R1)alleviates allergic rhinitis(AR)through AMP-activated protein kinase(AMPK)/mitochondrial fission critical protein(DRP1)-mediated mitochondrial fission.Methods Different doses of PNS-R1 were used to treat ovalbumin(OVA)-induced AR model mice,and the inhibitory effect of PNS-R1 on AR was investigated by observing allergic symptoms such as nasal rubbing and sneezing,as well as HE staining of nasal tissues.Serum IgE levels and nasal lavage fluid(NLF)inflammatory cytokine levels were detected by enzyme-linked immunosorbent assay(ELISA)and apoptosis-related proteins were detected by Western blot.In vitro human nasal epithelial cells(HNEpC)were stimulated with IL-13 to observe apoptosis,mitochondrial membrane potential,cellular ROS and mitochondrial ROS production,as well as the expression levels of AMPK/DRP1,expression levels of the TXNIP/NLRP3 inflammasomes and the translocation of DRP1.Results PNS-R1 attenuated allergic symptoms in AR mice,HE staining reduced inflammatory cells and reduced the levels of OVA-specific IgE in serum,and the levels of IL-4,IL-6,and IL-8 in NLF.PNS-R1 attenuated the apoptosis and ROS production of nasal epithelial cells in AR.In vitro PNS-R1 could up-regulate mitochondrial membrane potential after IL-13 stimulation,reduce ROS and mtROS production,the proportion of apoptotic positive cells,and reduce cleaved caspase-3,Bax,and up-regulate Bcl-2 expression,down-regulate DRP1 phosphorylation(Ser 616)and DRP1 translocation at the mitochondrial membrane in an AMPK-dependent manner,reducing TXNIP/NLRP3 expression.Conclusions PNS-R1 can protect mitochondrial integrity by inhibiting the AMPK/DRP1 signaling axis and its subsequent TXNIP/NLRP3 signaling axis,thereby alleviating rhinitis in AR mice.