高糖条件下牛视网膜毛细血管周细胞增殖与凋亡的研究

Study on proliferation and apoptosis in retinal capillary pericytes(PC) with high glucose condition

目的观察高糖对牛视网膜毛细血管周细胞(pericyte,PC)增殖和凋亡的影响。方法建立牛视网膜毛细血管周细胞体外培养模型,将传至第三代的周细胞用常规培养液制作细胞悬液、计数并接种于96孔培养板(或培养瓶)中,待细胞贴壁后,分组加入含不同葡萄糖浓度(5.5、10、20、30、40mmol/L)的DMEM培养液,分别培养2～8天后收集细胞。用MTT法、流式细胞仪(FCM)检测,研究高糖对周细胞的影响。结果①高糖10mmol/L培养4、6、8天组和高糖20、30L、40(mmol/L)培养2、4、6、8天组周细胞增殖均有明显抑制作用,其抑制率以40mmol/L、8天组最高(P<0.01)。②高糖10、20、30、40mmol/L培养2～8天均能促进周细胞的凋亡,其凋亡细胞数以40mmol/L、8天组最多(P<0.01)。结论①高糖对周细胞的增殖有明显抑制作用,高糖以剂量依赖性方式使周细胞减少,随着葡萄糖浓度升高和时间延长,周细胞形态学改变更明显,高糖对周细胞增殖的抑制率也增大。②高糖能促进周细胞凋亡,其凋亡百分率随糖浓度的增高和培养时间的延长而增加。

OBJECTIVE To investigate the effect on proliferation and apoptosis of retinal capillary pericytes（PC） to high glucose.METHODS Retinas were dissected from eyes of fresh killed catde,separated and cultured retinal capillary pericytes.Using eyes of cattle,we build cells model of PC cultured in vitro. Using low glucose culturing liquid made the third pericytes cells into suspension cell, counted cells and planted them in 96 hole culture- bottles, then made DMEM culturing liquid of different glucose（5.5,10,20,30,40 mmol/L） .Cultured after 2 - 8 days in vitro,we collected cells and then studied the effect of high glucose for them. RESULTS 1. Retinal pericytes proliferation have been restricted in various high glucose（ 10, 20, 30, 40 mmol/L）, and the proportion of restriction in 40mmol/L for 8 days was high . 2. High glucose（10,20,30,40 mmol/L） for 2- 8 days could accelerate the apoptosis of PC.And the apoptosis in 40 mmol/L for 8 days was the most in all（ P 〈 0.01 ）. CONCLUSIONS 1. High glucose could restrain retinal capillary pericytes proliferation,and reduce the amount of PC by dose dependence.With the concentration rising and time lasting, cells have distinct changes in morphology and the proportion of restriction increase. 2. High glucose could accelerate the apoptosis of retinal capillary pericytes. The more glucose proportion and long culturing time, the more the sum of apoptosis of PC is.