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Background To confirm the proliferation of vascular smooth muscle cell (VSMC) lead by advanced glycation end products (AGEs) and investigate weather the mechanism is work through MAPK pathway. To investigate weather the prolification of VSMC lead by AGEs can be inhibited by reduced glutathione(GSH) and what the mechanisam is. Methods VSMC of rats were isolated and cultivated, separated in 8 groups, each group contained 12 samples. Density of cell was 1×105 /mL in each sample, cultivated with AGEs at different concentrations and intervened with GSH at different concentrations. In order to determine the mechanism and interventional factors of VSMCs, sandwich ELISA method was used to test the concentration of P-P38 and MTT colorimetry was adopted to evaluate the amount of VSMC. Results 1.Effect of AGEs to the OD value of MTT in VSMC: with stimulation of AGEs, OD valued of P-P38 in VSMC increased simultaneously (P<0.01), their value were 0.43±0.15, 0.49±0.16, 0.48±0.19 [L/(g·cm)]. With the increase of the dose of AGEs, there were no difference between groups B, C of MTT OD value(P<0.05). 2.Effect of GSH to the OD value of MTT in VSMC stimulated by AGEs: OD value of MTT decreased with the increase of GSH concentration, their value were 0.347±0.102, 0.333±0.108, 0.285±0.080 [L/(g·cm)] respectively, decreased by 45%, 56%, 60%(P<0.01)compared with value of AGEs control group. With the increasing of the dose of GSH, the MTT OD value had no difference between groups F, G and H (P>0.05). 3.Effect of AGEs to the OD value of P-P38 in VSMC: with stimulation of AGEs, OD valued of P-P38 in VSMC increased obviously (P<0.01), their value were 0.65±0.17, 0.85±0.26, 0.94±0.17 [L/(g·cm)]. With the increasing of the dose of AGEs, the P-P38 OD value increase simultaneously(P<0.05). 4.Effect of GSH on the OD value of P-P38 in VSMC stimulated by AGEs: OD value of P-P38 decreased with the increasing of GSH concentration, their value were 0.356±0.090, 0.281±0.070, 0.256±0.072 [L/(g·cm)] respectively, decreased by 45%, 56%, 60%(P<0.01)compared with the value of control group. With the increasing of the dose of GSH, the P-P38 OD value between groups F, G and H were decreased gradually (P<0.01). Conclusions 1.AGEs has the function of inducing the proliferation of vascular SMC, the activation of the P-P38 MAPK signal pathway may be the mechanism of the proliferation of VSMC. 2.GSH can inhibit the proliferation of VSMC lead by AGEs, The P-P38-MAPK pathway is being blocked by GSH, which is the mechanism of inhibiting the proliferation of VSMC lead by AGEs.
Background To confirm the proliferation of vascular smooth muscle cell (VSMC) lead by advanced glycation end products (AGEs) and investigate the mechanisms of the mechanism is through MAPK pathway. To investigate weather the prolification of VSMC lead by AGEs can be inhibited by reduced glutathione ( GSH) and what the mechanisam is. Methods VSMC of rats were isolated and cultivated, separated in 8 groups, each group contained 12 samples. Density of cell was 1 x 105 / mL in each sample, cultivated with AGEs at different concentrations and intervened with GSH at different concentrations. In order to determine the mechanism and interventional factors of VSMCs, sandwich ELISA method was used to test the concentration of P-P38 and MTT colorimetry was evaluated to amount of VSMC. Results 1. Effect of AGEs to the OD value of MTT in VSMC: with stimulation of AGEs, OD valued of P-P38 in VSMC increased simultaneously (P <0.01), their values were 0.43 ± 0.15, 0.49 ± 0.16, 0.48 ± 0.19 [L / (g · cm) ]. Wit h the increase of the dose of AGEs, there were no difference between groups B, C of MTT OD value (P <0.05). 2.Effect of GSH to the OD value of MTT in VSMC stimulated by AGEs: OD value of MTT with the increase of GSH concentration, their values were 0.347 ± 0.102, 0.333 ± 0.108, 0.285 ± 0.080 [L / (g · cm)] respectively, decreased by 45%, 56%, 60% (P <0.01) compared with value of AGEs control group. With the increasing of the dose of GSH, the MTT OD value had no difference between groups F, G and H (P> 0.05). 3.Effect of AGEs to OD value of P-P38 in VSMC: with stimulation of AGEs, OD valued of P-P38 in VSMC increased obviously (P <0.01), their values were 0.65 ± 0.17, 0.85 ± 0.26, 0.94 ± 0.17 [L / (g · cm) dose of AGEs, the P-P38 OD value increase simultaneously (P <0.05). 4.Effect of GSH on the OD value of P-P38 in VSMC stimulated by AGEs: OD value of P-P38 decreased with the increasing of GSH concentration , their value were 0.356 ± 0.090, 0.281 ± 0.070, 0.256 ± 0.072 [L / (g · cm)] respectively, decreased by 45%, 56%, 60% (P <0.01) compared with the value of control group. With the increasing of the dose of GSH, the P-P38 OD value between groups F (P <0.01). Conclusions 1.AGEs have the function of inducing the proliferation of vascular SMC, the activation of the P-P38 MAPK signal pathway may be the mechanism of the proliferation of VSMC. 2. GSH can inhibit the proliferation of VSMC lead by AGEs, The P-P38-MAPK pathway is being blocked by GSH, which is the mechanism of inhibiting the proliferation of VSMC lead by AGEs.