The Role Of TRPV4 Channels For The Expression Of Enos In Mechanical Stretch Of Brain Microvascular Endothelial Cells

Objective: To screen out the subtype of transient receptor potential(TRPs) channel in stretch-induced injury of human brain microvascular endothelial cells(HBMEC) and to study its mechanism of the expression of eNOS protein effected by the TRPs.Methods: HBMEC was cultured in vitro. TRPs was screened out by Western blot and immunocytofluoresence method in HBMEC.Based onTRPs and mechanical stretch by Mini-type multi-functional bio-impact machine, cells were divided into unstretched group, simple stretch, stretch and low molecular weight heparin(LMWH), stretch and TRPs-specific inhibitor, stretch and TRPs-specific inhibitor and low molecular weight heparin(LMWH), stretch and SKF-96365 and stretch and N-nitro-l-arginine methyl ester(L-NAME) groups. The concentration of free calcium ion in cells were examined by the flow cytometry. The expression of eNOS protein was detected by Western blot..Results: The mRNA and protein of TRPV4 was detected in HBMEC by qRT-PCR,Western blot and immunocytofluoresence method.Flow cytometry show the concentration of free calcium ion was increased in the stretch group and decreased in stretch+TRPV4-specific inhibitor group(P=0.000),and in the stretch+SKF-96365 group; the analysis ofWestern blot shows the expression of eNOS protein was increased in the stretch group,and decreased in the stretch+TRPV4-specific inhibitor group and in stretch+L-NAME group.Conclusion: The activation of TRPV4 maybe play an important role in HBMEC with mechanical stretch, which cause the influx of calcium and increase the expression of eNOS protein.