周期性应力下磷脂酶C-γ1及其抑制剂对大鼠髓核细胞增殖能力的影响

Effect of the phospholipase C-γ1 protein and its inhibitor on rat nucleus pulposus cells proliferation under periodic mechanical stress

目的 观察周期性应力下磷脂酶C-γ1（PLCγ1）及其抑制剂对大鼠髓核细胞增殖的影响.方法 将细胞玻片随机分成4组：对照组、加压0.5h组、加压1h组、加压2h组.Western blot法检测各组PLCγ1、磷酸化PLCγ1表达.另取细胞玻片,随机分为3组：对照组、加压6h组、U73122＋加压6h组.使用荧光实时定量聚合酶链反应（FQ-PCR）检测各组细胞外基质的表达.结果 PLCγ1磷酸化水平在加压0.5h组（0.109 ±0.010）、加压1h组（0.138±0.001）、加压2h组（0.201 ±0.014）较对照组（0.086 ±0.005）增高.加入U73122后,细胞外基质的基因表达,以对照组最低,加压6h组最高,U73122＋加压6h组居中,差异有统计学意义（P＜0.05）.结论 周期性压力能促进髓核细胞的增殖,PLCγ1在压力传导中起信号传递作用.

Objective To evaluate the effect of the phospholipase C-γ1 （PLCγ1） protein and its inhibitor on rat nucleus pulposus cells proliferation under periodic mechanical stress.Methods Some cell slides were randomized into four groups：the control group,0.5 h stress group,1 h stress group and 2 h stress group.The expression of PLCγ1 and pPLCγ1 was detected by using Western blotting.The other slides were randomized into three groups：the control group,6 h stress group and U73122 ＋ 6 h stress group.Type Ⅱ collagen and aggrecan gene expression levels were evaluated through real-time fluorescent quantitative polymerase-chain reaction.Results The pPLCγ1/PLCγ1 ratio in 0.5 h stress group （0.109 ± 0.010）,1 h stress group （0.138 ±0.001） and 2 h stress group （0.201 ±0.014） was significantly higher than in control group （0.086 ± 0.005）.The gene expression levels of type Ⅱ collagen and aggrecan in U73122 ＋ 6 h stress group were significantly higher than those in control group,but lower than those in 6 h stress group （P 〈 0.05）.Conclusion Periodic mechanical stress can promote the proliferation of rat nucleus pulposus cells.PLCγ1 plays important roles in the mechanical transductions.