Regulatory Effect of Buyang Hhuanwu Decoction on Rat Myocardial H9C2 Cell Activity Inhibition Induced by Trastuzumab / 广州中医药大学学报

ABSTRACT

Objective To observe the influence of Buyang Huanwu Decoction ( BYHWD) on the inhibition of rat myocardial H9C2 cell activity and SH2-containing tyrosine phosphatase-1 ( SHP-1) activity induced by trastuzumab, and to explore the possible regulatory mechanism after observing the intervention of BYHWD on rat myocardial H9C2 cell transfected with SHP-1 or SHPC/S-1 gene. Methods The eukaryotic expression vectors pcDNA3.1 (+)- SHP-1 and pcDNA3.1 (+) -SHPC/S–1 were constructed and then were transfected to rat myocardial H9C2 cells using the method of liposome transfection. The cells with positive clones were screened out with G418, and then were cultured with trastuzumab for maintaining growth. Using quantitative RT-PCR, we detected the expression of SHP-1 gene and SHPC/S - 1 gene in rat myocardial H9C2 cells. The phosphatase activity analysis was used for observing the regulatory effect of BYHWD on SHP-1 in myocardial cells. Furthermore, we observed the apoptosis of rat myocardial H9C2 cells by methyl thiazolyl tetrazolium (MTT) assay after treatment with BYHWD. Results Sequencing results indicated the successful construction of eukaryotic expression vectors, which had stable expression in myocardial H9C2 cells even under the intervention of trastuzumab. The results of phosphatase analysis showed that H9C2-SHP-1 had the highest activities of phosphatase, but the activities were decreased after the intervention with BYHWD ( P<0.05) . The results of MTT assay also showed the apoptotic rate of H9C2-SHP-1 cells was decreased after treatment with BYHWD ( P <0.05) . Conclusion BYHWD can promote the proliferation of myocardial H9C2 cells inhibited by trastuzumab, and can regulate the expression of SHP-1 in myocardial cells, which will supply reference to the further study of treatment of trastuzumab-induced cardiac toxicity.