甲状旁腺激素通过非依赖PLC的PKC途径抑制成骨细胞的凋亡  

Parathyroid hormone inhibits the apoptosis of osteoblast MC-3T3E1 cells through a non-PLC-dependent protein kinase C pathway

作　　者：胡少宇 童国军[1] 孟越[1] 郝松[1] 李威[1] 许伏龙 何友华[1] 陈建庭[1] 杨德鸿[1] 

机构地区：[1]南方医科大学南方医院脊柱骨科,广东广州510515

出　　处：《南方医科大学学报》2016年第6期785-789,共5页Journal of Southern Medical University

基　　金：国家自然科学基金(30973061;81272043)~~

摘　　要：目的观察甲状旁腺激素的非依赖PLC的PKC转导通路(PTH/non PLC/PKC)是否会对成骨细胞(MC3T3-E1)的凋亡以及细胞数量具有影响。方法培养MC3T3-E1细胞,以1.5×104密度接种到96孔板,然后置于培养箱培养3 d直到细胞达到汇合状态,随机分为5组:按100 nmol/L[Gly1,Arg19]h PTH(1-28);100 nmol/L[Gly1,Arg19]h PTH(1-34);100 nmol/L[Gly1,Arg19]h PTH(1-34)+1μmol/L Go6983,1μmol/L Go6983;空白对照组加入等体积的去离子水,分别刺激细胞1、24、48 h,然后使用细胞计数试剂盒(CCK-8)和Caspase-Glo&#174;3/7试剂盒(caspase-3)检测细胞的细胞数量与凋亡。结果 CCK-8检测结果显示,[Gly1,Arg19]h PTH(1-34)组与[Gly1,Arg19]h PTH(1-34)+Go6983组相比,在1 h和24 h具有提高细胞数量的趋势,但结果并没有统计学差异,48 h[Gly1,Arg19]h PTH(1-34)组与[Gly1,Arg19]h PTH(1-28)组相比,可以明显提高细胞的数量(P<0.05)。进一步的研究发现在[Gly1,Arg19]h PTH(1-34)组添加抑制剂Go6983后,细胞数量增多的效应消失(P<0.05)。Caspase-3检测凋亡结果显示,[Gly1,Arg19]h PTH(1-34)组与[Gly1,Arg19]h PTH(1-34)+Go6983组相比,在1 h和24 h具有抑制细胞凋亡(细胞凋亡受到抑制),但是差异无统计学意义。48 h检测细胞凋亡显示,[Gly1,Arg19]h PTH(1-34)组与[Gly1,Arg19]h PTH(1-28)组相比,前者可以明显抑制细胞凋亡(P<0.05),给予PKC抑制剂Go6983后,其抑制凋亡现象消失。结论 PTH的non PLC/PKC信号转导通路可能在长时间(48 h)作用于MC3T3-E1细胞时,可抑制细胞凋亡,提高细胞的数量。Objective To investigate the effect of the non- PLC- dependent protein kinase C（PKC） pathway of parathyroid hormone（PTH） on the apoptosis and proliferation of osteoblast MC-3T3E1 cells. Methods MC-3T3E1 cells were seeded in 96-well plates at the density of 1.5×10^4cells/m L and incubated for 3 day. The cells were then exposed to 100 nmol/L of [Gly^1, Arg^19]h PTH（1-28）, 100 nmol/L of [Gly^1, Arg^19]h PTH（1-34）, 100 nmol/L of [Gly^1, Arg^19]h PTH（1-34）＋1 μmol/L Go6983, 1 μmol/L Go6983,or deionized water（control） for 1, 24 or 48 h. After the treatments, cell counting kit- 8（CCK- 8） and Caspase- Glo#174;3/7 Assay（Caspase-3） were used to examine the proliferation and apoptosis of MC3T3-E1 cells. Results CCK-8 results showed that h PTH（1-34） increased the number of MC3T3-E1 cells compared with h PTH（1-34）＋Go6983 at 1 h and 24 h, but this difference was not statistically different. At 48 h, treatment with h PTH（1-34）, as compared with h PTH（1-28）, significantly increased the number of MC3T3- E1 cells（P〈0.05）, and this effect was blocked by the PKC inhibitor Go6983（P〈0.05）. h PTH（1- 34） did not result in significant inhibition of MC3T3- E1 cell apoptosis at 1 h and 24 h as compared with h PTH（1- 34） ＋ Go6983, but significantly inhibited the cell apoptosis as compared with h PTH（1- 28）（P〈0.05）; this inhibitory effect was blocked by Go6983（P〈0.05）.Conclusions A relatively long time（for 48 h） of exposure to PTH can inhibit apoptosis and promote the proliferation of MC3T3-E1 cells through a non-PLC-dependent PKC pathway.

关 键 词：甲状旁腺激素 凋亡 信号转导通路 

分 类 号：R96[医药卫生—药理学]