Background
To evaluate the protective effect and underlying mechanism of panax notoginseng saponins (PNS) on hydrocortisone (HC)-induced injury in human bone microvascular endothelial cells (HBMECs).HBMECs were isolated from femoral heads resected in total hip arthroplasty and identified by morphology and immunofluorescence.
Methods
HBMECs were injured by hydrocortisone (HC), treated with PNS in different concentrations, and transfected with APOA1 or siAPOA1 to explore the role of APOA1 in the process where PNS protected HC-injured HBMECs. The viability and apoptotic rate were determined by CCK-8 and flow cytometry assays, respectively. The migration and tube length of HBMECs were evaluated by wound healing and tube formation assays, respectively. The mRNA expression of APOA1 was determined by qRT-PCR. The proteins levels of APOA1, Bcl-2, Bax and Caspase-3 were determined by Western blot.
Results
HBMECs exhibited spindle-shaped morphology. CD31 and vWF served as positive control and CD133 served as negative control. The viability of HBMCS inhibited by HC was balanced with the lower mRNA expression of APOA1 in a dose-dependent manner. PNS reversed the inhibitory effect of HC on the mRNA expression of APOP1, cell viability, migration, tube length, and the stimulative effect of HC on apoptosis of HBMECs. APOA1 overexpression further promoted the protective effect of PNS on HC-inhibited cell viability, migration and tube length, and HC-promoted apoptosis, whereas APOA1 silencing inhibited the protective effect of PNS and further promoted the adverse effect of HC on HBMECs.
Conclusion
PNS can efficiently suppress HC-induced injury in HBMECs, which may be associated with over-expressed APOA1.
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