The Experimental Study Of The Effect Of Dan-shen Root Monome On NF-κB And Its Correlative Factors Of Cultural Human Vascular Endothelial Cell In Atherosclerosis

Objective:In order to illustrate the role of Salvianolic acid B (Sal B)and Tanshinone IIA(TanllA) which are extracted from dan-shen root in atherosclerosis and their mechanism and the different target, we use TNF- a -induced cultured ECV-304 to research for the effect of the two monomes on NF-KB and its correlative factors. Methods:By using of cultured ECV-304 ,We established the TNF- a -induced cell injury model. At the same time, the immunocytochemistry -, cell ELISA > RT-PCR x radioimmunoassay methods were used to analyze the expression of cells which were cultured with different denses of Salvianolic acid B and Tanshinone IIA of nuclear factor-kappa B , I-Kappa B a , ICAM-1 , VCAM-1 of ECV-304 and to detact the contents in the cultural medium of NO,ET-l,PGl2and TXAj. Results:1 .By establishing the injury model of Ecv-304 ,the effects of TNF- a on this injury were detected by current medical method which was significant when the density of TNF- a was 10 ng/ml.This effect could be inhibited by PDTC and Sal B or TanllA. Sal B or TanllA can suppress the TNF- a -induced cell injury in different extents.2.The results of indirect cell ELISA and immunocytochemical method showed that NF-KB was increased when the ECV-304 exposed to TNF- a ,but the I-Kappa B u was decreased This effect could be controled by PDTC. Sal B can also inhibit the expression of NF-icB and enhanced the expression of I-Kappa B a . TanllA had the same effect on this experiment by the higher density of it.3.The results of chemical and the radioimmunoassay methods showed that the systolic factors such as ET and TXA2 were increased when exposed to TNF- a ,but the diastolic factors such as NO and PGIjwere decreased PDTC, Sal B or TanllA can improve the disorder of the vascular activity substances ,but TanllA can not affect the content of PGI2.4.The results of RT-PCR method showed that the expression of the m RNA of ICAM-1 was significantly enhanced by 48 folds and the expression of the m RNA ofVCAM-1 was induced significantly .Comparing with the control groups,the expression of the m RNA of 1C AM -1 with the treatment of PDTC, Sal B or TanllA in two doses were decreased by about 1.4 or 4, 2.5 or 3.2 and 1 or 1.5 folds, and the expression of m RNA of VCAM-1 with the treatment of PDTC, Sal B or TanllA in two doses were decreased by about 3.2 or 4.5, 3, and 1 or 2 folds. Conclusion:1. ECV304 cell was injured by TNF-a through activating its NF-KB and adhesion molecules related to NF-KB .This effect could accelerate the genesis and progression of AS.2. PDTC could inhibit the effect of TNF a -induced cell injury by inhibiting the activation of NF-KB and its correlative factors.lt had the role of suppression on AS.3. Sal B has a significant effect on the inhibiting of AS by suppressing the activation of NF- K B and its correlative factors.4. TanllA has the same effect on the inhibiting of AS in some extent by suppressing the activation of NF-KB and its correlative factors, but it was some dependent on the dose.