| ObjectiveObjective to investigate the protein expression of Notch1/NFk B signaling pathway and the m RNA expression of TNF-α、IL-1β in BV-2 microglia under the condition of oxygen glucose deprivation / reoxygenation(4h / 3h),and the effect of rhynchophylline(RIN)on its expression.MethodsBV-2 microglia cell line was cultured in vitro.The concentration range of RIN that did not affect the normal growth activity of BV-2 microglia cells was screened by CCK-8 method.BV2 microglia cells were divided into normal control group,OGD/R(4h/3h)group,OGD/R(4h/3h)+ RIN(30um)group.The m RNA expression of TNF-α and IL-1β in microglia was detected by real-time PCR.The protein expression of Notch-1 and its downstream intracellular Notch receptor domain(NICD),RBP-JK,Hes-1,NFKB p65 and NFKB p-p65 was used to detect by Western blot.and the effect of RIN intervention.Results1.CCK-8 method was used to detect the cytotoxicity of RIN on BV-2 microglia.The results showed that RIN(1,3,10,30um)had no toxic effect on BV-2 microglia,while RIN(100um)significantly decreased the activity of BV-2 microglia.2.Real time PCR detection showed that the m RNA expression levels of inflammatory factors TNF-α and IL-1β in BV2 microglia of OGD/R group were significantly higher than those of normal control group(P<0.05),while the m RNA expression levels of inflammatory factors TNF-α and IL-1β in BV2 microglia of OGD/R + RIN group were significantly lower than those of OGD/R group(P<0.05).3.Western blot results showed that the protein expression levels of Notch1,NICD,RBP-JK,HES-1 and NFKB p-p65 in BV2 microglia of OGD/R group were significantly higher than those of normal control group(P<0.05);the protein expression levels of Notch1 Signaling Pathway and NFk B p-p65 in BV2 microglia of OGD/R + RIN group were significantly lower than those of OGD/R group(P<0.05);the protein expression levels of NFk B p-p65 in BV2 microglia of each group were significantly lower than those of OGD /R group(P<0.05)There was no significant difference in p65 protein expression(P>0.05)Conclusions1.Rhynchophylline can down regulate the m RNA expression of TNF-α、IL-1β in BV-2 microglia under the condition of oxygen glucose deprivation / reoxygenation,It is suggested that rhynchophylline may inhibit the inflammatory response of microglia after oxygen glucose deprivation / reoxygenation.2.Rhynchophylline can significantly inhibit the proteins expression of Notch1 /NFk B signaling pathway in BV-2 microglia induced by oxygen glucose deprivation/ reoxygenation. |
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