Effect Of Nicotine On Growth、Function And Cellular Signal Transduction Pathway Of HUVEC And The Protection Of Astragalus Against HUVEC Injured By Nicotine

Objective:Whether or not studies of nicotine in HUVEC injury and there is the intervention of Astragalus.Methods:（1） MTT method was used to research nicotine and Astragalus on the growth’s effect of HUVEC.（2） ELISA was used to detect content of ICAM-1、vWf、ET-1、P38MAPK、JNK、NF-κB in supernate fluid of cultured cell.（3） Nitric acid depoxidize method was used to detect content of the total NO in supernate fluid of cultured cell.（4） Chemistrical colorimetry was used to detect content of eNOS in supernate fluid of cultured cell.（5） RT-PCR was used to detect the mRNA ofP38MAPK、NF-κB and JNK.Results:（1） The concentration of the nicotine from 10^-3mol/L to 10^-5mol/L injure HUVEC, but the concentration of the nicotine as 10^-4mol/L is the most significant injury（P＜0.01）.（2） The concentration of Astragalus from 50μg/ml to 200μg/ml proliferate HUVEC,the concentration of Astragalus as 200μg/ml is the best effect（P＜0.01）.（3） Compared with the blank group,the cell proliferation of the nicotine treatment group is significantly declined（P＜0.01）;the cell proliferation of Astragalus treatment group is significantly promoted（P＜0.01）;compared with Astragalus treatment with the nicotine group,the cell proliferation of the nicotine treatment group is declined（P＜0.05）.（4） Compared with the blank group,the content of P38MAPK、JNK、NF-κB、ET-1、ICAM-1、vWf is significantly promoted in supernate fluid of the nicotine treatment group（P＜0.01）;compared with the blank group,the content of ET-1、vWf、P38MAPK、JNK、NF-κB、ICAM-1 is significantly declined in supernate fluid of the Astragalus treatment group（P＜0.01）;compared with Astragalus treatment with the nicotine group,the content of ET-1、vWf is promoted in supernate fluid of the nicotine treatment group（P＜0.05）.（5） The content of the total NO is significantly declined in supernate fluid of the nicotine treatment group（P＜0.01）;the content of the total NO is promoted in supernate fluid of the Astragalus treatment group（P＜0.05）;compared with Astragalus treatment with the nicotine group,the content of the total NO is declined in supernate fluid of the nicotine treatment group（P＜0.05）.（6） The content of eNOS is significantly declined in supernate fluid of the nicotine treatment group（P＜0.01）;the content of eNOS is significantly promoted in supernate fluid of the Astragalus treatment group（P＜0.01）;compared with Astragalus treatment with the nicotine group,the content of eNOS is declined in supernate fluid of the nicotine treatment group（P＜0.05）.（7）Compared with the blank group,the mRNA expression of P38MAPK、JNK、NF-κB is significantly promoted the nicotine treatment group（P＜0.01）.Conclusion:（1） Nicotine is capable of injuring HUVEC.（2） Astragalus is capable of protecting HUVEC from nicotine injured.（3） The nicotine injures HUVEC by combining with nicotine receptor and activates MAPK and NF-κB transduction pathway.