Abstract:
Objective To explore the proliferation, expression and effect of TRPC on passively sensitized human airway smooth muscle cells (HASMC).
Methods HASMC cells treated with TRPC6 blocker SKF96365 in the experimental group were grouped as follows: TRPC6 siRNA transfected HASMC cells; Control group TRPC6 wild type HASMC cells. The proliferation, expression and function of tritium thymine nucleoside were detected by real-time fluorescence quantitative polymerase chain reaction, western blot, cell count and tritium thymine nucleoside (3H-TdR) addition test.
Results Western blot analysis of TRPC6 protein showed that TRPC6 blocker SKF96365 and TRPC6 siRNA transfection could significantly reduced the expression of TRPC6 in HASMC (
P<0.05). When 3H-TdR was added to HASMC, it was found that the number of 3H-TdR labeled cells decreased significantly after 10 mol/L SKF96365 treatment for 72 h or TRPC6 siRNA transfection for 48 h (
P<0.05). Cell count experiment found that TRPC6 blocker SKF96365 reduced TRPC6 gene expression, the number of cells with the rising of SKF96365 concentration reduced (
P<0.01), in a single concentrations under the action of 10 μmol/L, cell number as SKF96365 processing time slowed growth rate (
P<0.01), and at the same time through TRPC6 siRNA transfection cell proliferation significantly reduced after the TRPC6 knocked out. After TRPC6 blocking agents SKF96365 and TRPC6 siRNA were transfected, the expression of PI3K, Akt, p-Akt and mTOR in HASMC were significantly reduced.
Conclusions TRPC6 had a significant inhibitory effect on the proliferation and expression of downstream genes in passively sensitized HASMC, through PI3K/Akt/mTOR pathway. Therefore, this study provided a new reference for the treatment of asthma.