摘要
目的 :探讨重组人类肝细胞增殖因子 (rh HGF)刺激 ECV30 4细胞株表达基质金属蛋白酶 - 9(MMP- 9)的作用及他克莫思 (Tacrolimus,FK5 0 6 )的抑制作用。方法 :绘制正常 ECV30 4细胞株生长曲线 ,明确最适生长浓度及对数生长期 ;明确 rh HGF及 FK5 0 6的作用浓度 ;流式细胞仪 (FCM)检测 ECV30 4细胞株的 MMP- 9水平。结果 :1ECV30 4细胞的最适生长浓度为 1.0 E+ 4 / ml,对数生长期为 3～ 5 d;rh HGF的作用浓度为 8ng/ ml,IC50 为 8.2 7ng/ ml;FK5 0 6的作用浓度为 5 0 ng/ ml,IC50 为 5 1.6 3ng/ ml。 2对照组 MMP- 9的表达量为 39.74 % ;培养基中加入rh HGF 8ng/ ml后 ,细胞存活率为 1.38816 9± 0 .10 2 0 33(P<0 .0 1) ,MMP- 9的表达量为 4 0 .32 % ;培养基中加入FK5 0 6 5 0 ng/ ml后 ,细胞存活率为 0 .39876 4± 0 .0 92 4 76 (P<0 .0 1) ,MMP- 9的表达量为 14 .6 1% ;培养基中加入FK5 0 6 5 0 ng/ ml15 min后加入 rh HGF8ng/ ml,细胞存活率为 0 .76 72 0 3± 0 .0 2 6 39(P<0 .0 1) ,MMP- 9的表达量为 35 .0 8%。结论 :rh HGF可刺激 ECV30 4细胞增殖 ,使 MMP- 9的表达量增加 ;FK5 0 6可抑制 ECV30 4细胞增殖 ,使 MMP- 9的表达量降低 ;FK5 0 6可拮抗 rh HGF所引起的 ECV30 4细胞增殖 ,从而使 MMP-
Objective: To investigate the stimulating effect of recombinant human hepatocyte growth factor (rhHGF) on the expression of matrix metalloproteinase 9 (MMP 9) in ECV304 cells and the suppression effect of FK506. Methods: The growth curve of ECV304 cells was drawn and the valid concentration of rhHGF and FK506 were determined. The expression levels of MMP 9 were analyzed by flow cytometry (FCM). Results: ①The optimal growth concentration of ECV304 cell was 1.0E+4/ml and logarithm growth time was 3～5 days. The valid concentration of rhHGF was 8 ng/ml and IC 50 was 8.27 ng/ml. The valid concentration of FK506 was 50 ng/ml and IC 50 was 51.63 ng/ml. ② The expression level of MMP 9 was 39.74% in controls. Cell livability was 1 388169±0.102033 ( P <0.01) when 8ng/ml rhHGF was added to culture medium, and the expression level of MMP 9 was 40.32%. Cell livability was 0.398764±0.092476( P <0.01) when 50 ng/ml FK506 was added to culture medium, and the expression level of MMP 9 was 14.61%. Cell livability was 0.767203±0.02639 ( P <0.01) when 8 ng/ml rhHGF was added 15 minutes after 50 ng/ml FK506 was added to culture medium, and the expression level of MMP 9 was 35.08%. Conclusion: rhHGF can stimulate the proliferation of ECV304 cells and increase the protein expression level of MMP 9. FK506 can suppress the proliferation of ECV304 cells and decrease the expression level of MMP 9. FK506 can inhibit the proliferation of ECV304 cells induced by rhHGF and decrease MMP 9 protein level. [
出处
《浙江大学学报（医学版）》
CAS
CSCD
2004年第6期540-545,共6页
Journal of Zhejiang University(Medical Sciences)
基金
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