脱氢表雄酮在体外对成骨细胞代谢和IL-1β的影响

Effects of Dehydroepiandrosterone on Metabolism of Rat Osteoblasts and IL-1β in Vitro

目的探讨脱氢表雄酮(dehydroepiandrosterone,DHEA)对离体大鼠成骨细胞和IL-1β的影响。方法体外分离培养大鼠成骨细胞,取传一代细胞,分别给予10-5mmol/L、10-7mmol/L、10-9mmol/L DHEA培养72h,以10-8mmol/L雌二醇(estradiol,E2)为阳性对照,另设空白对照组。碱性磷酸酶(alkaline phosphatase,ALP)染色鉴定成骨细胞,MTT法检测成骨细胞的增殖能力,PNPP法测定ALP活性。ELISA法测定不同浓度DHEA培养液中IL-1β的水平。结果不同浓度DHEA组成骨细胞增殖能力和ALP的活性均有上升,其中以10-7mmol/L DHEA组最显著(P<0.01),其作用和E2相似(P>0.05);10-9mmol/L DHEA组单位细胞数的ALP活性高于空白对照组(P<0.05)。不同浓度的DHEA组IL-1β呈下降趋势,成骨细胞增殖能力及ALP活性和IL-1β成负相关。结论DHEA在体外促进大鼠成骨细胞生长和增殖,提高ALP活性,其作用可能和抑制IL-1β有关。

Objective To investigate the effects of dehydroepiandrosterone （DHEA） on rat osteoblasts and IL-1β in vitro. Methods Rat＇s osteoblasts were separated and cultured in vitro. Osteoblasts of first generation were treated with DHEA in different concentration （10^-5, 10^-7, 10^-9mmol/L） for 72 hours, E2 in 10^-8mmol/1 was adopted as positive control group, and blank control group was also set up. The biological characteristics of osteoblasts were evaluated by phase-contrast microscopy and alkaline phosphatase （ALP） histochemistry. Proliferation ability of osteoblasts was examined by MTY, activity of ALP was determined by PNPP. Level of IL-1 β in DHEA culture solution was determined by ELISA simultaneously. Results Proliferation and ALP activity of osteoblasts treated with DHEA in different concentration were raised, especially treated with DHEA in 10^-7 mmol/1 （P 〈 0.01 ）, which was similar to E2 （ P 〉 0.05 ）. ALP activity of unitary cell population treated with DHEA in 10^-9mmol/1 was higher than blank control group （P 〈 0.05 ）. Downtrend of IL-1 β could be seen in each DHEA group, and negative correlation between proliferation ability or ALP activity of OBs and IL-1β could also be found. Conclusion DHEA could stimulate proliferation and raise ALP activity of rat＇ s osteoblasts in vitro, whose mechanism might inhabit the expression of IL-1β.