Effect of Zuogui Jiangtang Shuxin Formula Medicated Plasma on Foaming and Apoptosis of Macrophage Induced by ox-LDL Through PERK/ eIF2α/ATF4 Signaling Pathway

OBJECTIVE To investigate the effect of Zuogui Jiangtang Shuxin Formula medicated plasma on ox-LDL-induced PERK/eIF2α/ATF4 integrated stress response and activation of apoptosis key signaling molecules CHOP and Caspase-12. METHEOD J774A.1 cells were randomly divided into 5 groups: normal group， model group， TUDCA group， Zuogui Jiangtang Shuxin Formula medicated plasma group and western medicine combination (metformin+atorvastatin calcium) medicated plasma group. All the drug intervention groups were pretreated with ox-LDL to establish an experimental model for macrophage foam cell formation. The cells of drug intervention groups were cultured in 10% different drug containing plasma， respectively. Cells of normal and model groups were cultured in normal culture medium. The growth inhibition of J774A.1 were tested by CCK-8 essay after 24 h culture. Oil Red O Dyeing experiment was used to show the cellular lipid droplets. The expression of p-PERK， p-eIF2α， ATF4， CHOP and Caspase-12 were detected by Western blot. RESULTS 100 mg/L ox-LDL suppressed the growth of J774A.1 cells with an inhibitory rate of (50.06±0.09)%. Oil Red O staining showed that the model group cells had more lipid droplets than normal group (P<0.01). Drug intervention groups effectively prevented the pervasion of ox-LDL to reduce lipid accumulation. Oil red O staining showed that lipid aggregated in Zuogui Jiangtang Shuxin Formula medicated plasma， compared with western medicine combination medicated plasma group (P<0.01)， and the effect was equivalent to TUDCA group (P＞0.05). ox-LDL up-regulated the expression of p-PERK， p-eIF2α， ATF4， CHOP and Caspase-12. Zuogui Jiangtang Shuxin Formula medicated plasma significantly restrained the expression of p-PERK， p-eIF2α and ATF4， and apparently decreased endoplasmic reticulum associated apoptosis protein CHOP and Caspase-12 expression (P<0.01). There was no significant difference in Zuogui Jiangtang Shuxin Formula medicated plasma group and TUDCA group. And Zuogui Jiangtang Shuxin Formula medicated plasma group was superior to the western medicine combination medicated plasma group (P<0.01). CONCLUSION Zuogui Jiangtang Shuxin Formula medicated plasma can suppress foam cell formation and apoptosis in J774A.1 cells， which may be related to the regulation of PERK/eIF2α/ATF4 signaling pathway and the expression of CHOP and Caspase-12.