Influence Of Sheshang Capsules Over Trimeresurus Stejnegeri Snakebite On Changing Expressions Of AMPK, NF-?B, ICAM-1 And MCP-1 In Vascular Endothelial Cells

Objective To observe the influence of Sheshang capsule on vascular endothelial cells AMPK,NF-?B,ICAM-1 and MCP-1 expression in Trimeresurus stejnegeri snakebite in order to base AMPK/NF-?B pathway to discuss the mechanism of vascular endothelial cell inflammatory reaction induced by the venom of Trimeresurus stejnegeri and the regulation of Sheshang capsule on the process.To provide the experimental basis for the clinical application and popularization of Sheshang capsule.Methods Rabbit serum containing Sheshang capsules and the cell model after Trimeresurus stejnegeri snakebite were prepared into five groups:the blank group(KB),the model group(MX)and the three groups with serum containing Sheshang capsules of low,middle and high dosage respectively(DY,ZY,GY),each with 10 samples.Specifically,10%of normal rabbit serum was added in the KB group;apart from the 10%serum,another 5ug/ml of snake venom was added into the MX group;DY,ZY and GY were cultured with the same composition as MX's in the first 6 hours,and 5%,10%and 15%of the rabbit serum containing Sheshang capsules were added to them respectively.After 72 hours' culture,the media of all the groups were collected to measure their level of AMPK,ICAM-1 and MCP-1 by means of enzyme linked immunosorbent assay(ELISA);cells were then collected and their total protein was extracted to measure the protein expressions of AMPK and NF-?B p65 by means of the Western Blot analysis.Results 1.Expression of ICAM-1 and MCP-1 by ELISA:ICAM-1 and MCP-1 of the MX group was significantly higher than that in the KB group and the difference was statistically significant(P<0.01);compared with the MX group,ICAM-1 and MCP-1 in the ZY group were significantly lower after the intervention treatment with the Sheshang capsules,and the difference was statistically significant(P<0.01).2.Expression of AMPK by ELISA:expression of AMPK in the MX group was significantly lower than that in KB group,and the difference was statistically significant(P<0.01);compared with the MX group,after the intervention treatment with the Sheshang capsules,AMPK in the treatment groups was raised at varied levels,among which that of the ZY and GY groups was statistically significant(P<0.05 and P<0.01 respectively).3.Expression of AMPK and NF-?B p65 protein by the Western Blot analysis:compared with the KB group,the expression of AMPK in the MX group was down-regulated and the expression of NF-?Bp65 in the MX group was up-regulated,and the difference was statistically significant(P<0.01 and P<0.05 respectively);compared with the MX group,after the intervention treatment with the Sheshang capsules,expression of AMPK in the treatment groups was up-regulated at varied levels,among which that of the GY group was statistically significant(P<0.01),and expression NF-?B p65 in the treatment groups was down-regulated,among which that of the ZY group was statistically significant(P<0.05).Conclusion Trimeresurus stejnegeri venom can induce inflammation in human umbilical vein endothelial cells;one of the mechanisms of Sheshang capsule in the treatment of inflammatory reaction of Trimeresurus stejnegeri vascular endothelial cells may be through the regulation of the AMPK,NF-?B and its downstream media,i.e.ICAM-1 and MCP-1.