Study On The Mechanism Of Buyang Huanwu Decoction Fumigation In Treating Ischemic Stroke Based On VEGF,BDNF And MMP-9 Expression

Objective: To study on the mechanism of Buyang Huanwu Decoction Fumigation in treating ischemic stroke,at the molecular level,the expression of VEGF,BDNF and MMP-9was observed in the present research.Meantime,at the gene level,the correlation of the polymorphism of rs17577,rs3918242 in MMP-9 gene and ischemic stroke was further explored.The results of the experiment would provide the molecular biology and gene characteristics for the prevention and treatment of ischemic stroke in western Guangxi,and find a suitable point for future prevention and treatment of ischemic stroke.Methods: From the Department of Neurology and Rehabilitation Medicine of Affiliated Hospital of Youjiang National Medical University,41 patients with ischemic stroke and 41 healthy subjects were enrolled and the serum was collected.Then,the levels of VEGF,BDNF and MMP-9 in serum of ischemic stroke patients and normal control group were compared by ELISA.Further,41 patients with ischemic stroke were enrolled to treatment with Buyang Huanwu Decoction Fumigation in this study.The scores of ADL and the levels of serum VEGF,BDNF and MMP-9 were measured before and after treatment.Finally,152 patients with ischemic stroke and 152 healthy subjects were enrolled.The polymorphism of rs17577,rs3918242 in MMP-9 gene was analyzed by PCR-RFLP technique.The correlation of the polymorphism of rs17577,rs3918242 in MMP-9 gene and ischemic stroke was analyzed.Results: At the molecular level,ELISA assays showed that:(1)The levels of serum VEGF(78.65 ± 6.74)and BDNF(1567.66 ± 247.17)in ischemic stroke group were significantly lower than those in normal control group(161.61 ± 47.44;2030.94 ±300.97),and the difference was statistically significant(P < 0.05).The level of serum MMP-9 in ischemic stroke group was significantly higher than that in normal control group(7987.62 ± 4080.87),the difference was statistically significant(P < 0.05).(2)After the treatment with Buyang Huanwu Decoction Fumigation,the ADL score(61.92 ± 5.01)was significantly higher than that before treatment(43.26 ± 3.27),and the difference was statistically significant(P <0.05).The expression levels of serum VEGF(114.21 ± 30.72)and BDNF(2014.46 ± 301.96)were significantly higher than those before treatment(79.66± 7.69 and 1828.35 ± 287.39),and the difference was statistically significant(P <0.05).The expression levels of serum MMP-9(9083.54 ± 1616.99)was significantly lower than that before treatment(10356.30±2515.68),and the difference was statistically significant(P<0.05).At the gene level,gene polymorphism showed that:(1)The MMP-9 gene locus rs17577 and rs3918242 completely linked.(2)The genotype frequency and allele frequency of MMP-9 gene locus rs17577 and rs3918242 between ischemic stroke group and normal control group were compared as follows: the frequency of MMP-9 rs17577 genotype in ischemic stroke group was 75.0% in G / G type,24.3% in A / G type and 0.7% in A / A type;the allele frequency,G Was 87.2% and A was 12.8%.And the frequency of MMP-9 rs17577 genotype in normal control group was 76.3% in G / G type,21.7% in A / G type and 1.9% in A / A type;the allele frequency,G Was 87.2% and A was 12.8%.The frequency of MMP-9rs3918242 genotype in ischemic stroke group was 75.0% in C / C type,24.3% in T / C type and 0.7% in T / T type;the allele frequency,C Was 87.2% and T was 12.8%.And the frequency of MMP-9 rs3918242 genotype in normal control group was 76.3% in C / C type,21.7% in T / C type and 1.9% in T / T type;the allele frequency,C Was 87.2% and T was12.8%.(3)There was no significant difference in frequency and allele frequency of MMP-9gene rs17577 and rs3918242 between ischemic stroke group and control group(P> 0.05).Conclusion: At the molecular level,compared with the normal control group,the expression of MMP-9 in ischemic stroke patients was significantly increased and theexpression of VEGF and BDNF in ischemic stroke patients were significantly decreased.After the treatment with Buyang Huanwu Decoction Fumigation,the ADL score and the expression of serum VEGF,BDNF in patients was significantly improved,and the expression of serum MMP-9 in patients was significantly reduced.These results suggested that traditional chinese medicine Buyang Huanwu Decoction has a good therapeutic effect on ischemic stroke.At the level of gene,the MMP-9 gene locus rs17577 was completely linked to rs3918242,but the polymorphism of rs17577 and rs3918242 in the MMP-9 gene was not correlated with ischemic stroke.