Protective effect of Angelica sinensis water extract medicated serum on corticosterone-damaged PC12 cells through modulation of sphingolipid metabolic pathway
Objective To clarify the protective effect of Angelica sinensis water extract medicated serum(ASWEMS)on PC12 cells damaged by corticosterone(CORT)through the regulation of sphingolipid metabolism.Methods Preparation of Angelica sinensis water extract and identification of its components by LC-MS method.Preparation of Angelica sinensis water extract medicated serum(ASWEMS):Twelve healthy male SD rats were randomly divided into blank serum(received 0.9％sodium chloride solution)group and ASWEMS(crude drug dose of 7.5 g·kg-1)group,acclimated for one week,and then intragastrically administered ASWEMS for six consecutive days.Blood was collected from the abdominal aorta of the rats,and the serum was separated and mixed within the same group.The serum was then heated at 56 ℃ for 30 min and filtered through a 0.22 μm microfiltration membrane to remove bacteria.A cell injury model was constructed by treating PC12 cells with CORT,with the cells divided into control(5％control serum),model(400 μmol·L-1 CORT+5％control serum),ASWEMS(400 μmol·L-1 CORT+5％ASWEMS)and fumonisin(FB1,400 μmol·L-1 CORT+5％control serum+10 μmol L·L-1 FB1)groups,which subjected to the treatment for 24 h.The cell survival rate was detected by CCK-8 method.Annexin FITC/PI flow cytometry was used to detect apoptosis rate.Changes in lipid metabolism profile and the relative content of sphingolipid metabolites in PC12.cells were determined by ultra-high performance liquid chromatography with quadrupole time-of-flight mass spectrometry(UPLC-Q-TOF-MS).The contents of Ceramide(Cer)in PC12 cells was detected by ELISA.Results LC-MS method identified 17 chemical components in Angelica sinensis water extract.Compared with the model group,ASWEMS and FB1 could significantly reverse the decreased cell viability and increased apoptosis rate caused by CORT injury in PC12 cells(P＜0.001).Sphingolipids metabolites were the metabolites that contributed more to the difference between the control group and the model group.Compared with the control group,the relative content of Dhsph-type and PhytoSph-type metabolites in PC 12 cells was significantly lower after CORT intervention for 24 h(P＜0.001),while the relative content of SM-type metabolites was significantly higher(P＜0.001),and the relative content and absolute content of Cer were both significantly higher(P＜0.01).Compared with the model group,the contents of all metabolites were significantly adjusted after ASWEMS or FB1 intervention(P＜0.01,0.001).Conclusion ASWEMS has a protective effect against CORT-induced PC12 cell injury through regulating sphingolipid metabolism pathway and inhibiting apoptosis.
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