Effect of nuclear factor-κB on proliferation of vascular smooth muscle cells promoted by thrombin in spontaneously hypertensive rats

Abstract

Abstract: AIM: To study the dose-effect and time-effect on proliferation of vascular smooth muscle cells (VSMCs) in spontaneously hypertensive rats (SHR) promoted by hrombin and to explore the relationship between VSMCs proliferation and NF-κB path as well as their possible mechanism.METHODS: VSMCs were incubated with thrombin at the following concentrations of 0.01, 0.1, 0.2, 0.5, 1.0, 2.0 U/mL for 24 h and with 0.5 U/mL thrombin for 1, 2, 6, 12, 24, 48 h respectively;The proliferation rate of VSMCs was determined with both WST-1 metabolic activity and ³H-TdR incorporation efficiency;The expression of NF-κB in VSMCs was tested with both immumofluorescence and western blot in basic condition and with 0.5 U/mL thrombin intervention.Ang Ⅱ and PDGF on proliferation of VSMCs were used as positive control group.In order to research the relationship of NF-κB path and VSMCs proliferation induced by thrombin, after pretreatment with 100 μmol/L PDTC (NF-κB special inhibitor), VSMCs proliferation induced by thrombin was also investigated.RESULTS: Thrombin concentrations of 0.2-2.0 U/mL significantly promoted VSMCs proliferation respectively(P<0.05 or P<0.01).But VSMCs proliferation was not different in the thrombin at the concentrations of 0.5-2.0 U/mL at 24 h (P>0.05).VSMCs proliferation with 0.5 U/mL thrombin exhibited double hump when compared with the Control Group.The peak values at 1 h and 24 h showed that thrombin promotion VSMCs proliferation was not only rapid and brief but also delayed and retarded.The expression of NF-κB of VSMCs in basic condition was mainly distributed in cytoplasmwhile that of VSMCs interfered by thrombin was in nucleus.The PDTC pre-treatment could obviously inhibit VSMCs proliferation induced by thrombin (P<0.01).CONCLUSION: Thrombin promotes VSMCs proliferation in the manner of dose dependent at the concentrations of 0.2-0.5 U/mL.The proliferation curve induced by thrombin exhibits double hump which suggests that thrombin promotion VSMCs proliferation is delayed. Thrombin may activate NF-κB of VSMCs and make it transfer from cytoplasm to nucleus.VSMCs proliferation induced by thrombin is connected with NF-κB path.NF-κB may be the common path for the signal conduction inside the VSMCs proliferation.

Key words: NF-κB, thrombin, vascular smooth muscle cell, proliferation

CLC Number:

R965.2

HU Rong,HONG Hua-shan, XU Chang-sheng, WU Ke-gui. Effect of nuclear factor-κB on proliferation of vascular smooth muscle cells promoted by thrombin in spontaneously hypertensive rats[J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2008, 13(6): 627-633.

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