胰岛素样生长因子1对酒精干预体外培养成骨细胞增殖和功能抑制的影响

目的探讨体外培养成骨细胞在不同血清浓度条件下酒精干预后增殖和功能的改变，以及胰岛素样生长因子1（insulin—likegrowth factor1，IGF-1）的保护作用。方法体外分离培养新生SD大鼠颅骨成骨细胞，分6组在不同条件下进行培养，分别为正常血清浓度组（F15组，含15％新生牛血清）、正常血清浓度加酒精组（F15／EtOH组，酒精浓度为100mmol／L）、血清饥饿组（F2组，含2％新生牛血清）、血清饥饿加酒精组（F2／EtOH组）、血清饥饿加IGF-1组（F2／IGF-1组，IGF-1浓度为25ng／m1）和血清饥饿、IGF-1加酒精组（F2／IGF-1／EtOH组）。于培养24、48、72、96h检测细胞增殖、细胞内碱性磷酸酶（alkalinephosphatase，ALP）活性和骨钙素（boneglaprotein，BGP）mRNA表达。结果各时间点F15／EtOH组成骨细胞吸光度（A）值及ALP活性、BGPmRNA表达较F。。组均下降，差异有统计学意义（P〈0．05）。F2组较F15组A值及ALP、BGPmRNA降低（P〈0．05），F2／EtOH组较F2组降低（P〈0．05），F2／IGF-1组较F2组增加（P〈0．05）；F2／IGF-1／EtOH组较F2／IGF-1组除24h外A值无明显降低（P〉0．05），ALP、BGPmRNA均降低（P〈0．05），A值及ALP、BGPmRNA较F2／EtOH组增加（P〈0．05）。结论酒精能引起成骨细胞增殖和功能抑制，加重血清饥饿对成骨细胞的抑制作用，可能是酒精性骨损害的机制之一。IGF-1能改善血清饥饿引起的成骨抑制，并抵抗酒精抑制细胞增殖的作用，可能成为探索治疗酒精性骨损害的途径之一。

EFFECTS OF INSULIN-LIKE GROWTH FACTOR 1 ON INHIBITION OF OSTEOBLASTIC PROLIFERATION AND FUNCTION BY ETHANOL

Objective To investigate the effects of insulin-like growth factor 1(IGF-1) and ethanol(EtOH) on the changes in the osteoblast proliferation and the osteoblast function under the normal serum concentration and serum starvation.Methods The osteoblasts harvested from the SD rat calvaria were incubated in the following six conditions according to the supplements in DMEM:the F15 group:15% newborn calf serum(NCS);the F15/EtOH group:100 mmol/L of EtOH added to 15% NCS;the F2 group:2% NCS;the F2/EtOH group:100 mmol/L of EtOH added to 2% NCS;the F2/IGF-1 group:25 ng/ml of IGF-1 added to 2% NCS;the F2/IGF-1/EtOH group:100 mmol/L EtOH added to 25 ng/ml IGF-1 and 2% NCS.The osteoblasts were analyzed by the MTT assay,alkaline phosphatase(ALP) activity,and RT-PCR at 24,48,72 and 96 hours after the culture.Results The absorbance(A),the ALP activity,and the expression of BGP mRNA(the proliferation and function indicators of the osteoblasts) were significantly decreased in the F15/EtOH group at all the time points when compared with those in the F15 the group(P<0.05);the above 3 indicators were significantly decreased in the F2 group when compared with those in the F15 group(P<0.05);they were significantly decreased in the F2/EtOH group when compared with those in the F2 group(P<0.05);however,the indicators in the F2/IGF-1 group were significantly increased when compared with those in the F2 group(P<0.05);the A value in the F2/IGF-1/EtOH group was not significantly decreased when compared with that in the F2/IGF-1 group,with an exception of the A value at 24 hours(P>0.05);however,ALP and BGP mRNA were significantly decreased(P<0.05).All the indicators were significantly increased when compared with those in the F2/EtOH group(P<0.05).Conclusion Ethanol can inhibit the osteoblast proliferation and the osteoblast function,and can increase the inhibition when the osteoblasts were cultured under the serum starvation.This may be one of the mechanisms for alcoholic bone disease.IGF-1 can prevent the inhibition of the osteoblasts under the serum starvation and counteract the ethanol-induced proliferation inhibition;therefore,IGF-1 is an alternative therapeutic intervention for alcoholic bone disease.