摘要
目的:探讨染料木黄酮对成骨细胞活性的影响及其可能机制,为染料木黄酮防治骨质疏松提供理论依据。方法:实验于2001-05/2003-01在卫生学环境医学研究所完成。无菌条件下用胰蛋白酶和Ⅱ型胶原酶分步消化乳鼠颅盖骨获得成骨细胞,传代后细胞用于实验。染料木黄酮组分别加入10-5～10-7mol/L染料木黄酮,对照组以吐温20为对照。分别用噻唑蓝比色法和3H-胸腺嘧啶掺入法测定成骨细胞增殖和DNA合成,用反转录-聚合酶链式反应和Westernblot测定雌激素受体mRNA和蛋白表达。结果:①成骨细胞培养基中加入(10-5～10-6mol/L)染料木黄酮,培养48h和72h后噻唑蓝的吸光度值与对照组相比均明显升高[培养48h:(0.39±0.03),(0.45±0.03),(0.46±0.02),(0.19±0.05);培养72h:(0.29±0.04),(0.32±0.02),(0.37±0.02),(0.15±0.04)]。②染料木黄酮组3H-胸腺嘧啶掺入量均显著增加[染料木黄酮组为(101.20±10.06),(844.60±366.90),(512.20±197.6);对照组为(68.67±10.39)],未发现成骨细胞雌激素受体基因和蛋白表达。结论:染料木黄酮不是通过促进雌激素受体基因和蛋白的表达来促进成骨细胞增殖。
AIM： To probe into the effect of genisten on the activity of osteoblasts and its possible mechanism, so as to provide theoretical basis for preventing and treating osteoporosis with genisten.
MRTHODS： This experiment was conducted at the Institute of Health and Environment Medicine from May 2001 to January 2003. Osteoblasts were isolated from neonatal rat calvaria through the digestion of trypsin and type Ⅱ collagenase. Cells were used for the experiment after passage. Different doses of genistein（10^-5 mol/L, 10^-6 mol/L and 10^-7 mol/L） was added in the genisten group and tween 20 was used in the control group. Proliferation and DNA synthesis of osteoblast were measured with thiazole blue and ^3H-TdR, respectively. Estrogen receptor mRNA and protein expression were measured with reverse transcription-polymerase chain reaction and Western blot respectively.
RESULTS： ①10^-5 - 10^-6 mol/L genistein was added into the culture medium of osteoblasts, The absorbance of thiazole blue was significantly increased as compared with control group 48 and 72 hours after culture [Culture for 48 hours： （0.39±0.03）, （0.45±0.03）, （0.46±0.02）, （0.19±0.05） ; Culture for 72 hours： （0.29±0.04）, （0.32±0.02）, （0.37±0.02）, （0.15±0.04）], ② Incorporation of 3H-TdR in the genistein group was significantly increased as compared with control group [Genistein group：（101,20±10,06）, （844.60±366,90）, （512.20±197.6）;Control group： （68,67±10,39）],We did not find estrogen receptorctandβmRNA and protein.
CONCLUTION： Genistein promotes the proliferation of osteoblast not through promoting the expression of estrogen receptor gene and protein.
出处
《中国临床康复》
CSCD
北大核心
2006年第13期70-72,i0004,共4页
Chinese Journal of Clinical Rehabilitation
