The protective effect of essential oil of rhizome from Stahlianthus involucratus on endothelial cell damage based on Nrf2 target and mitochondria quality control system
Objective To study the mechanism of Nuclear E2-related factor 2(Nrf2)mitochondria quality control system regulating the protective effect of essential oil from Stahlianthus involucratus rhizomes(EOSIR)on human umbilical vein endothelial cells(HUVECs)injury.Methods HUVECs were induced by oxidized low-density lipoprotein(ox-LDL)to establish a cell injury model,the expression of Nrf2 factor was silenced by siRNA transfection.The cultured cells were divided into control group,model group,EOSIR group,transfection group and transfection negative control group.The protein and mRNA expression of Nrf2 and its downstream factors were detected by Western blot and qRT-PCR;Griess method was used to detect the content of nitric oxide(NO);ELISA was used to analyze the content of human endothelin(ET-1)and human prostacyclin(PGI2);The cells were divided into control group,ox-LDL-induced group,low dose EOSIR group,medium dose EOSIR group,high dose EOSIR group and aspirin positive control group in the mitochondrial quality test.Transmission electron microscopy was used to observe the mitochondrial morphology of HUVECs,the expression levels of mitofusin 1(Mfn1),mitofusin 1(Mfn2),dynamin-related protein 1(Drp1),Optic atrophy 1(Opa1),mitochondrial transcription factor A(TFAM),peroxisome proliferator activated receptorγcoactivator-1α(PGC-1α)and autophagy related proteins LC3A/B,P62 was detected by Western blot.Results Compared with the model group,the expression levels of Nrf2 and its downstream factors,and the contents of NO and PGI2 in the EOSIR group were significantly increased,and the levels of ET-1 were significantly decrease than those of model group.After silencing Nrf2,the effect of EOSIR on the above indicators was inhibited;transmission electron microscopy observing the mitochondrial morphology,the number of autophagosomes in the EOSIR group was significantly reduced compared with the model group,and the mitochondrial morphology returned to normal.At the same time,the EOSIR group significantly improved the changes in mitochondrial-related protein expression induced by ox-LDL(P<0.05).Conclusions EOSIR may play a protective role against ox-LDL-induced endothelial cell injury by activating the Keap1/Nrf2/ARE signaling pathway and regulating the mitochondrial quality control system.
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