Huo-Luo-Xiao-Ling-Dan Protects H9C2 Cardiomyocytes Against Hypoxia/Reoxygenation Injury

Objective:Huo-Luo-Xiao-Ling-Dan(HLXLD)is a classic prescription which was written by Zhang Xichun.The prescription is consist of frankincense,myrrh,salvia and angelica.It has the effect of promoting blood circulation and relieving pain.This study established a hypoxia/reoxygenation(H/R)injury model of rat myocardiocytes(H9C2s)in vitro to investigate the effect of HLXLD on myocardial ischemia and reperfusion injury(MIRI).We investigated the pharmacological effects of the prescription on myocardiocytes and related molecular mechanisms to elucidate the protective effect of HLXLD on H/R-injured H9C2 s.Methods:MTT assay detected the effect of different concentration gradients of HLXLD on the survival rate of normal H9C2 s to determine whether HLXLD had toxic reaction.We used physical method to establish the H/R model.MTT assay detected the effect of different concentrations of HLXLD on the survival rate of H/R injury H9C2 s.We investigated the pharmacological effect of different concentration gradients of HLXLD on H/R-injured H9C2 s was detected by the activity of NO,LDH and T-SOD.We used real-time PCR methods to investigate the effects of HLXLD on the expression of GSK-3? m RNA,Axin1 m RNA,?-catenin m RNA and C-myc m RNA in Wnt/?-catenin signaling pathway of H/R-injured H9C2 s.Results:HLXLD has no toxic effect on normal H9C2 s and it has the proliferative effect on H9C2 s.The choice of hypoxia for 24 h and reoxygenation for 2h is the best modeling condition.HLXLD can increase the survival rate of H/R injury H9C2 s.HLXLD could increase the activity of NO and T-SOD,and reduce the activity of LDH on the injured H9C2 s HLXLD could not only up-regulated the activity of GSK-3? m RNA and Axin1 m RNA expression in Wnt/?-catenin signaling pathway for 6h-hypoxic and 2h-reoxygenation but also inhibited the activity of ?-catenin m RNA and C-myc m RNA expression for 12h-reoxygenation and 2h-reoxygenation.Conclusions: 1.HLXLD has no toxic reaction on H9C2 s,and it has certain proliferation effect.HLXLD could increase the survival rate of H/R injury H9C2 s In addition,HLXLD could regulate the activity of NO,LDH and T-SOD on H/R-injured H9C2 s to alleviate the damage of H9C2 s in H/R injury process.2.The mechanism of the anti-H/R injury of HLXLD may be related to the regulation of Wnt/?-catenin signaling pathway.