Effect Of Matrine On ERK1/2 Signaling Pathway And The Receptor Expression Of Marc-145 Cells Infected With PRRSV

Objective:In order to investigate the possible antiviral mechanism of Matrine on Marc-145 cell infected with PRRSV, we analyzed the effect of Matrine on ERK1/2 signal pathway and the receptor expression of CD163, CD151, Heparan sulfate (HS) and Vimentin.Methods:We established the cell model of PRRSV infection. The experiment included cell only control group, PRRSV only control group (100TCID50 PRRSV), Matrine only control group (0.75 mg/mL Matrine), PRRSV+Matrine group (100TCID50 PRRSV+0.75 mg/mL Matrine) and PRRSV+U0126 virus group (100TCID50 PRRSV+30?M U0126). The cell samples were collected following the infection by PRRSV at 3 h,6 h,12 h and 24 h. Real-time quantitative PCR (qRT-PCR) was used to check the effect of Matrine on the expression of CD 163, CD151, HS, Vimentin and PRRSV N gene. Western blot analysis was used to check the effect of Matrine on the expression of pERKl/2, CD163, CD151 and Vimentin.Results:1. We analyzed the expression of N-gene at four time points following the virus infection. The expression of N gene was increased with the infection time and reached the highest point at 24 h (p<0.05). Compared with the virus only control group, the expression of N gene was reduced at all time points in virus+Matrine group (p<0.05). These results indicate that Matrine could inhibit the expression of N gene and effect the replication of PRRSV.2. When Marc-145 cell infected with PRRSV at 6 h, compared with cell only control group, the expression of pERK1/2 was significantly increased in virus only control group and Matrine only control group (p<0.05); compared with virus only control group, the expression of pERK1/2 was significantly increased in Matrine only control group and virus+Matrine group (p<0.05), the expression of pERKl/2 was decreased in U0126 group (p<0.05); compared with Matrine only control group, the expression of pERKl/2 in virus+matrine group was significantly decreased (p<0.05). These results suggest that in Marc-145 cell, ERKl/2 signaling pathway can be activated by both Matrine and PRRSV, the activation ability of Matrine is higher than that of PRRSV. On Marc-145 cell infected with PRRSV, the ability of Matrine to activate ERKl/2 is weak than that in Matrine only group.3. In the cell model of PRRSV infection, compared with the virus only control group, at 3 h the expression of CD151 and Vimentin were significantly decreased in PRRSV+Matrine group (p<0.05); at 6 h the expression of HS, Vimentin and CD 163 were significantly decreased in PRRSV+Matrine group (p<0.05); at 24 h the expression of HS and CD151 were significantly decreased in PRRSV+Matrine group (p<0.05). These results suggest that at different time points following the infection of Marc-145 with PRRSV, Matrine regulated the expression of different cell receptors.Conclusions:1. Marc-145 cells infected with PRRSV 24 h, Matrine can inhibit the expression of PRRSV N gene to continually inhibit the replication of PRRSV in Marc-145 cell.2. When Marc-145 cells infected with PRRSV, Matrine can't inhibit the activated of ERK1/2 signal pathway to inhibit the replication of PRRSV.3. At different time points following the infection of Marc-145 with PRRSV, Matrine inhibited the expression of different cell receptors and inhibited the adsorption and the entrance of PRRSV to block the ability of PRRSV to attack the cells and uncoating and releasing virus to inhibit the replication of PRRSV.