摘要
ｐ２７蛋白是细胞周期素依赖性激酶（Ｃｄｋ）抑制蛋白家族中的一种，主要对外部促进或抑制细胞增殖的信号起反应。本研究应用流式细胞仪（ＦＣＭ）双标记的方法观察血管紧张素Ⅱ（ＡｎｇⅡ）、血管加压素（ＡＶＰ）和血小板源生长因子（ＰＤＧＦ）对血管平滑肌细胞（ＶＳＭＣｓ）细胞周期百分比和ｐ２７蛋白表达量的影响。静止状态培养的ＶＳＭＣｓ加入ＡｎｇⅡ，ＡＶＰ，ＰＤＧＦＢＢ后，在不同时间收集细胞，用碘化丙啶（ＰＩ）标记细胞ＤＮＡ，以确定细胞所处的周期。用ｐ２７蛋白的单抗和标记了ＦＩＴＣ的二抗标记细胞，通过流式细胞仪测定被激发出的荧光量来确定细胞ｐ２７蛋白表达的相对量。结果显示，ＡｎｇⅡ刺激ＶＳＭＣｓ增生，其蛋白含量增加了４３６％（Ｐ＜００１），但不抑制ｐ２７蛋白的表达；ＡＶＰ可轻度抑制ｐ２７的表达，有轻度促进ＶＳＭＣｓ增殖和增生的作用（Ｐ＜００５）；ＰＤＧＦ明显抑制ｐ２７的表达，引起细胞增殖。本研究结果提示，ｐ２７蛋白抑制ＶＳＭＣｓ通过Ｇ１期进入Ｓ期，是抑制ＶＳＭＣｓ增殖的重要调节因子。
This study is to investigate cell cycle distribution of the vascular smooth muscle cell(VSMCs) and negative regulator of cell proliferation p27 expression caused by platelet derived growth factor BB(PDGF BB), angiotensin Ⅱ(AngⅡ) and arginine vasopressin(AVP). Cultured VSMCss were deprived of fetal calf serum for 48 h to make quiescent. VSMCss were collected at different time after stimulation of AngⅡ, AVP and PDGF BB. Cell cycle distribution and p27 expression were determined with a flow cytometer. The results showed that the protein content of VSMCs was significantly increased (43 6%) by AngⅡ as a result of hypertrophy, but AngⅡ did not lead to downregulation of p27. AVP could downregulate p27 slightly. PDGF could inhibit p27 expression significantly and cause VSMCs hyperplasia. These results suggest that the progression of VSMCs through G 1 to S phase might be brought out by the inhibition of p27 during proliferation.
出处
《生理学报》
CAS
CSCD
北大核心
1999年第3期285-290,共6页
Acta Physiologica Sinica
基金
国家自然科学基金
