Study On The Effects Of Procyanidins On NF-?B Signaling Pathway In The Treatment Of Inflammatory Injury Of Lung BEAS-2B Cells Induced By Arsenic

Objective:Procyanidins(PC)have strong anti-inflammatory effects.In order to investigate the mechanism of NF-?B signaling pathway role in grape seed proanthocyanidins(GSPE)against arsenic induced lung injury,and to provide reference for the prevention and treatment of arsenic poisoning.Methods:In this study,human bronchial epithelial cells(BEAS-2B)as the object,according to the As2O3(0,5,10 and 20 mol/L),GSPE(0,25 and 50mg/L)and BAY 11-7082(NF-?B inhibitor,inhibits stimulus induced phosphorylation of I?B-?,0 and 5 mol/L)doses were divided into 48 groups.The survival rate of the cells was measured by MTT assay,and the apoptosis rate was measured by flow cytometry;The expression levels of IL-1?,IL-6,IL-10,TNF-?,CRP,ROS and LPO in BEAS-2B cell homogenates were determined by ELISA method;mRNA determination of I?B-?,IKK?,IKK?,NF-?Bp65 and NF-?Bp50 in BEAS-2B cell homogenates by qRT-PCR;Western-Blot was used to detect the expression of I?B-?,P-I?B-?,IKKa/(3,NF-?Bp65 and NF-?Bp50 protein in BEAS-2B cell homogenates.Results:1.With the increase of the dose of As2O3 exposure to 24h,the survival rate of BEAS-2B was decreased(100%to 65.26%,P all<0.05),and the apoptosis rate was increased(from 1.7%to 30.4%,P all<0.05);a similar trend in 48h.After applying for BAY 11-7082,the cell survival rate and apoptosis rate decreased;when GSPE was used to antagonize the toxic effect of arsenic,the cell survival rate increased(65.26%to 83.38%,P all<0.05),and the apoptosis rate decreased(from 30.4%to 13%,P all<0.05).2.Compared with the blank control group,the expression levels of proinflammatory cytokines(IL-1?,IL-6,TNF-?,and CRP),ROS and LPO were increased(P all<0.05),and the expression level of anti-inflammatory factor(IL-10)was decreased(P<0.05)in BEAS-2B24 hours;in addition,with the increase of arsenic exposure dose,the expression levels of proinflammatory cytokines,ROS and LPO were increased(P all<0.05)and the inflammatory factors decreased(P<0.05).Compared with the arsenic exposure group,the expression of proinflammatory cytokines,ROS and LPO were significantly decreased(P all<0.05)after the administration of BAY 11-7082,and the expression of inflammatory factors increased(P<0.05);after GSPE intervention,the expression levels of proinflammatory cytokines,ROS and LPO were decreased(P all<0.05),while the expression of proinflammatory cytokines increased(P<0.05).3.qRT-PCR results showed that compared with the blank control group,the mRNA levels of IKK??IKK??NF-?Bp65?NF-?Bp50(P all<0.05)were increased,however the levels of I?B-?(P<0.05)was decreased by arsenic.Compared with the arsenic exposure group,the mRNA levels of KK??IKK??NF-?Bp65?NF-?Bp50(P all<0.05)were decreased and I?B-?(P<0.05)was increased(mRNA)after the application of BAY 11-7082 or GSPE.4.Western-Blot results showed that compared with the control group,the ratio of the P-I?B-?,IKK?/?,NF-?Bp65,NF-?Bp50 to beta-actin(P all<0.05)were increased,the ratio of I?B-?(P<0.05)was reduced.Compared with the arsenic exposure group,after applying BAY 11-7082 or GSPE,the ratio of the P-I?B-?,IKK?/?,NF-?Bp65,NF-?Bp50 to beta-actin(P all<0.05)were decreased,the ratio of I?B-?(P<0.05)was inreased.Conclusion:Arsenic exposure will increase the apoptosis rate of BEAS-2B,reduce the survival rate may be due to the activation of NF-?B signaling pathway,so the expression level of proinflammatory factor increases,reduce the quantity of anti inflammation cytokines,leading to inflammatory injury;GSPE can effectively inhibit the activation of NF-kappa B signaling pathway induced by arsenic,decrease the expression of pro-inflammatory cytokines and increase the expression of proinflammatory cytokines,the results indicated that GSPE could inhibit the activation of NF-kappa B signaling pathway induced by arsenic,which could regulate the expression of inflammatory factors in the downstream,and antagonize the inflammatory damage induced by arsenic.