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目的 :本研究观察大鼠肾小球系膜细胞 (MC)在不同环境条件培养下 ,其增殖、分泌t-PA及PAI -1的影响 ,寻找较合适的培养条件 ,为体外研究纤溶系统的调节机制提供模型和摸索合适的条件。方法 :在本实验中以MTT比色法观察了在不同的糖浓度下大鼠肾小球系膜细胞的增殖状况。其它的实验部份包括 :①以不同浓度的葡萄糖液 (5 ,1 0 ,2 0 ,30mmol/L)培养MC2 4 ,4 8,72及 1 6 8小时 (即一周 )。②以不同浓度的胎牛血清 (FBS) (5 ,1 0 ,2 0 ,2 5 % )培养MC2 4 ,4 8,72及 1 6 8小时。以上培养物的上清液内PAI - 1和t -PA的含量以ELISA法测定。结果 :①高糖对MC增殖有害 ,即有抗增殖作用。②高糖刺激MC分泌PAI - 1 ,其含量持续升高 ,维持一周 ,而t -PA量则在一周培养后有所下降。③增加FBS的浓度可促进PAI- 1的分泌 (r =0 81 3)至降低t-PA ,以 2 0 %的浓度量为合适。④如上②所述 :高糖条件下MC释放PAI - 1显示随时间依赖式增加 ,72小时达高峰 ,并至少在一周内维持较高的水平 ,而t-PA则在培养到一周时发生变化 (下降 )。结论 :在实验室中 ,高糖对MC增殖有抑制作用而增加了PAI - 1的表达 ,由此对细胞外基质 (ECM)的降解亦有抑制
Objective: This study was to observe the effects of proliferation and secretion of t-PA and PAI-1 in rat mesangial cells (MCs) under different environmental conditions, and to find more appropriate culture conditions for the study of fibrinolytic system The regulatory mechanism provides models and groping for the right conditions. Methods: In this experiment MTT colorimetric method was used to observe the proliferation of rat mesangial cells under different concentrations of glucose. Other experimental sections included: ① MC2, 48, 72, and 186 hours (ie one week) were cultured in different concentrations of glucose solution (5,1,0,2,0,30 mmol / L). ②MMC2 4, 48, 72 and 186 hours were cultured in different concentrations of fetal bovine serum (FBS) (5, 10, 20, 25%). The contents of PAI-1 and t-PA in the above culture supernatants were determined by ELISA. Results: ① high glucose harmful to MC proliferation, that is anti-proliferative effect. ② High glucose stimulated the secretion of PAI - 1 by MC, its content continued to increase for one week while the amount of t - PA decreased after a week of culture. ③ Increasing the concentration of FBS can promote the secretion of PAI-1 (r = 0 81 3) to reduce the t-PA, 20% of the concentration is appropriate. ④ As described in ② above, MCI release of PAI - 1 under high glucose showed a time - dependent increase, reaching a peak at 72 hours and maintained at a high level for at least one week, while t - PA changed during the first week of culture (decline). CONCLUSION: High glucose in the laboratory inhibited the proliferation of MCs and increased the expression of PAI - 1, thereby inhibiting the degradation of extracellular matrix (ECM)