Ox-LDL Upregulates PHF2 Expression Through The PKA/PKC Pathway To Promote Inflammation In THP-1 Macrophages

Objective: This experiment was to explore the role of protein kinase(PK)/PHF2 pathway in ox-LDL-induced inflammatory reaction in THP-1 macrophages,so as to further clarify the mechanism of atherosclerosis.Methods: We used ox-LDL to induce inflammatory reaction in THP-1 macrophages,using RNA interference and specific inhibitors,Western blotting technique and PCR was used to explore PKA?PKC?PHF2?TNF-??CXCL4?CXCL10 expression levels and the change of activity.Results: 1.Western Blotting showed that ox-LDL can increase the protein levels of PKA and PKC in THP1 macrophages in dose-dependent manner(p < 0.05),the difference is statistically significant.2.Western Blotting results showed the protein levels of PKA and PKC in the ox-LDL group were significantly higher than that in PBS group(p<0.05).The increased levels of PKA and PKC protein in the ox-LDL group were significantly reduced by PKA inhibitors or PKC inhibitors(p < 0.05).Western Blotting and RT-PCR showed that the protein and mRNA level of TNF-alpha in THP1 macrophages were elevated by ox-LDL treatment,which could be inhibited by PKA inhibitors or PKC inhibitors(p < 0.05).3.Western Blotting results showed that PHF2 protein expression level in ox-LDL group was significnalt increased compared to in PBS group and nLDL group(p < 0.05),while the increased PHF2 level can be down-regulated by PHF2 siRNA(p < 0.05),the difference is statistically significant.4.Western Blotting and PCR resdults showed that elevated levels of PHF2 protein and mRNA induced by ox-LDL could be reduced by PKA inhibitors or PKC inhibitors(p < 0.05),the difference is statistically significant.5.The PCR results showed that mRNA levels of TNF-alpha,CXCL4 and CXCL10 were increased in ox-LDL group compared to in PBS and nLDL group(p < 0.05),the difference is statistically significant.PHF2 siRNA could down-regulated ox-LDL-induced elevations in TNF-alpha,CXCL4 and CXCL10 mRNA levels(p < 0.05),the difference is statistically significant.Conclusion: ox-LDL upregulates PHF2 expression through the PKA/PKC pathway to promote inflammation in THP-1 macrophages.