A Comparative Study On The Effects Of Jiedu Huayu Recipe Containing Serum And Plasma On PC12 Cell Injury Model

Objective: Through thrombin combined hypoxia establishing the PC12 cell injury model,to observe what effects serum and plasma containing Jieduhuayu have on the vitality,PAR-1,ERK1/2mRNA and protein expression of the PC12 cell injury model, and compare the differences between serum and plasma containing Jieduhuayu.Methods: 1. Experimental study of thrombin combined hypoxia inducible PC12 cell injury: The PC12 cells were divided into 0 U / ml thrombin, 50 U / ml thrombin, 100 U / ml thrombin, 150 U / ml thrombin and 200 U / ml thrombin group, each group containing thrombin were added to a concentration of 0U / ml, 50 U / ml, 100 U / ml, 150 U / ml, 200 U / ml of serum-free medium sugar, placed in 37 ℃, 5 % CO2,1% O2,94% N2 incubator, were cultured 1h, 6h, 12 h, 24 h after the removal, MTT assay was used to detect cell viability of each group, Tunel method to detect the apoptotic rate. 2.Preparation of serum and plasma containing Jieduhuayu through gavage to rats. The combined use of thrombin and hypoxia is to establish the thrombin-induced hypoxic PC12 cell injury model. Grouping for the control group, model group, serum containing Jieduhuayu group, plasma containing Jieduhuayu group, cell viability of each group was detected by MTT assay; fluorescent quantitative real-time PCR to detect the expression levels of each group PAR-1,ERK1/2mRNA; immunofluorescence to detect the expression of PAR-1,ERK1/2 protein.Results: 1. 150 U / ml thrombin 12 h, 24 h, and 200 U / ml thrombin 6h, 12 h, 24 h after compared with the same period in the control group(0 U / ml) decreased cell viability(P <0.05 or P <0.01), apoptosis rate increased(P <0.05 or P <0.01). 2.Compared with the control group, the vitality of model group decreased(P <0.01),the expression of PAR-1,ERK1/2mRNA and protein in model group were enhanced(P <0.01);Compared with the model group,the vitality of serum containing Jieduhuayu and plasma containing Jieduhuayu representing an increase(P <0.05 or P <0.01),PAR-1, ERK1 / 2 mRNA and protein expression were decreased(P <0.05 or P <0.01);Among them, the vitality of plasma containing Jieduhuayu group compared with serum containing Jieduhuayu group increased significantly(P <0.05);the expression of PAR-1,ERK1/2mRNA and protein in plasma containing Jieduhuayu group compared with serum containing Jieduhuayu group decreased significantly(P<0.05).Conclusion: 1. 150 U / ml thrombin combined effect of 12 h after hypoxia for studying thrombin combined hypoxia on PC12 cell injury model of optimal conditions. 2.The combined use of thrombin and hypoxia decreased the viability of PC12 cell, enhanced ERK1 / 2 mRNA and protein expression, plasma containing Jieduhuayu prescription was superior to its containing serum against the injury of PC12 cell induced by hypoxic and thrombin. Therefore, prompting that plasma containing drug is more realistic response to drug efficacy, when experiments can using the drug-containing plasma and serum at the same time.