| Abstrakt: |
Hypotonic stimulation induces airway constriction in normal and asthmatic airways. However, the osmolarity sensor in the airway has not been characterized. TRPV4 (also known as VR-OAC, VRL-2, TRP12, OTRPC4), an osmotic-sensitive cation channel in the transient receptor potential (TRP) channel family, was recently cloned. In the present study, we show that TRPV4 mRNA was expressed in cultured human airway smooth muscle cells as analyzed by RT-PCR. Hypotonic stimulation induced Ca2+ influx in human airway smooth muscle cells in an osmolarity dependent manner, consistent with the reported biological activity of TRPV4 in transfected cells. In cultured muscle cells, 4α-phorbol 12,13-didecanoate (4-αPDD), a TRPV4 ligand, increased intracellular Ca2+ level only when Ca2+ was present in the extracellular solution. The 4-αPDD-induced Ca2+ response was inhibited by ruthenium red (1 µM), a known TRPV4 inhibitor, but not by capsazepine (1 µM), a TRPV1 antagonist, indicating that 4-αPDD-induced Ca2+ response is mediated by TRPV4. Verapamil (10 µM), an L-type voltage-gated Ca2+ channel inhibitor, had no effect on the 4-αPDD-induced Ca2+ response, excluding the involvement of L-type Ca2+ channels. Furthermore, hypotonic stimulation elicited smooth muscle contraction through a mechanism dependent on membrane Ca2+ channels in both isolated human and guinea pig airways. Hypotonicity-induced airway contraction was not inhibited by the L-type Ca2+ channel inhibitor nifedipine (1 µM) or by the TRPV1 inhibitor capsazepine (1 µM). We conclude that functional TRPV4 is expressed in human airway smooth muscle cells and may act as an osmolarity sensor in the airway. [ABSTRACT FROM AUTHOR] |
