The Influence Of Sappanwood Ethyl Acetate Extract On Vascular Endothelial Injury And JNK Pathway In Rats

OBJECTIVE:To observe the effects of sappanwood ethyl acetate e-xtract on aortic pathological morphology,inflammatory cytokines (IL-10 and MCP-1),and aorta JNK mRNA,JNK and p-JNK proteins of vascular endothelial injury rats,and to explore the mechanism of sappanwood eth-yl acetate extract against vascular endothelial injury.METHODS:40 Wistar rats (male,180-220g)were randomly divided i-nto 4 groups according to random number table,with 10 rats in each g-roup,namely blank control group,model control group,sappanwood group, and simvastatin group.Animals of the blank control group were fed with normal diet and the remaining groups were given dietary methionine for model establishment.Six weeks after modeling,2 rats were randomly se-lected from the model control group,the sappanwood group,and the sim-vastatin group,respectively,and morphological detections were performed by HE staining to confirm the success of modeling. The corresponding drug intervention was administered after the modeling,once a day for 2 weeks.Pathological changes of aortic endothelial morphology were obser-ved by HE staining;serum MCP-1 and IL-10 levels were detected by E-LISA method;expression of aortic JNK mRNA was detected by real-time fluorescence quantitative PCR;and expressions of channel proteins JNK and p-JNK were determined by Western-Blot.RESULTS:1.The aortic endothelial morphology results showed that for the ra-ts of blank control group,the aortic vascular endothelial cells swelled a-nd shed slightly and the surface was not smooth with a small amount of inflammatory cells attached. The medial smooth muscle was slightly disordered and the number of nuclei was increased.The pathological inj-ury of vascular endothelium was significantly reduced compared with t-hemodel group.2.The ELISA results showed that the serum IL-10 level of the mo-del control group was significantly decreased and that of MCP-1 was s-ignificantly increased compared with the blank group(P< 0.05);serum le-vels of MCP-1 of the sappanwood group and the simvastatin group wer--e both reduced compared with the model control group and the differe-nces were statistically significant(P< 0.05);The serum levels of MCP-1 of the sappanwood group and the simvastatin group were comparable,w-ith no statistically significant difference(P> 0.05);and the serum level of IL-10 in the sappanwood group was lower than that of the simvasta-tin group and the difference was statistically significant(P<0.05).3.The real time fluorescent quantitative PCR results indicated that the expression of JNK mRNA of the model group was markedly higher than that of the blank control group with a statistically significant diff-erence(P<0.05);compared with the model control group,aorta JNK mRN-A expression levels of the sappanwood group and the simvastatin group were both decreased significantly and the differences were statistically significant(P<0.05);the reduced levels of aorta JNK mRNA expression of the sappanwood group and the simvastatin group were comparable a-nd the difference was not statistically significant(P> 0.05).4.Western Blot results showed that there were no significant differ-ences in the expressions of JNK protein between groups(P> 0.05).Comp-ared with the blank control group,the phosphorylation level of JNK in the model control group was significantly increased with a statistically significant difference(P< 0.05).The JNK phosphorylation levels of the r-at aortic tissues of the sappanwood group and the simvastatin group w-ere both increased compared with the model control group and the diff-erences were statistically significant(P< 0.05).The aortic JNK phosphory-lation levels of the sappanwood group and the simvastatin group were comparable and the difference was not statistically significant(P> 0.05).CONCLUSION:1.Sappanwood ethyl acetate extract can reduce the serum MCP-1 1-evel and increase the IL-10 level of the rats with vascular endothelial injury,therefore to inhibit the inflammatory reaction of vascular endothe-lial injury and play a role in the protection of vascular endothelium.2.Sappanwood ethyl acetate extract can down-regulate the JNK mR-NA expression and JNK phosphorylation and inhibit the activation of t-he JNK pathways of rats with vascular endothelial injury.