金雀异黄酮对去势大鼠分离培养的成骨细胞增殖及分化的影响  

Effect of genistein on proliferation and differentiation of osteoblasts isolated and cultured from ovariectomized rats

作　　者：谢艳芳 游丽娟 张菊林 胡秦妮[1] 张涛[1] 李娟[1] 高玉海[3] XIE Yanfang;YOU Lijuan;ZHANG Julin;HU Qinni;ZHANG Tao;LI Juan;GAO Yuhai(Department of Clinical Laboratory,No.940 Hospital of Joint Logistic Support Force of Chinese PLA,Lanzhou,Gansu Province,730050;Health Management Center,Gansu Provincial Hospital of Maternal and Child Health,Lanzhou,Gansu Province,730050,China;Laboratory of Basic Medical Scineces,No.940 Hospital of Joint Logistic Support Force of Chinese PLA,Lanzhou,Gansu Province,730050)

机构地区：[1]中国人民解放军联勤保障部队第940医院检验科,兰州730050 [2]甘肃省妇幼保健院健康管理中心,兰州730050 [3]中国人民解放军联勤保障部队第940医院基础医学实验室,兰州730050

出　　处：《第三军医大学学报》2021年第24期2671-2676,共6页Journal of Third Military Medical University

基　　金：联勤保障部队第940医院民用项目(2021yxky050);甘肃省青年科技基金计划项目(20JR5RA589)。

摘　　要：目的研究金雀异黄酮(Genistein,GEN)对去势大鼠分离培养的成骨细胞增殖及分化的影响。方法选取3月龄SD雌性大鼠30只,分为假手术组(Sham)、切除卵巢模型组(OVX)和GEN治疗组(OVX+GEN)。双能X射线骨密度仪检测大鼠全身骨密度。各组选取3只最接近其骨密度均值的大鼠麻醉后处死,无菌分离培养成骨细胞。CCK-8检测细胞增殖能力,化学法检测碱性磷酸酶(ALP)活性,RT-PCR检测各组成骨细胞中骨形成相关基因Runx-2、BMP-2的表达情况;钙化结节染色观察钙化结节阳性克隆数并计算钙化面积。结果全身骨密度检测结果显示,OVX组全身骨密度显著低于Sham组,而OVX+GEN组全身骨密度显著高于OVX组,差异均有统计学意义(P<0.01)。细胞增殖、ALP活性检测、RT-qPCR检测成骨性基因的表达以及钙化结节面积结果均表明,OVX组与Sham组相比均显著降低(P<0.01),而OVX+GEN组与OVX组相比均显著增加(P<0.01)。结论 GEN提高去卵巢大鼠骨密度可能与其促进成骨细胞增殖和成熟矿化能力有关。Objective To determine the effects of genistein(GEN) on the proliferation and differentiation of osteoblasts isolated and cultured from ovariectomized rats.Methods Thirty 3-month-old female SD rats were divided into sham operation group(Sham),ovariectomy model group(OVX) and Gen treatment group(OVX+GEN).Their bone mineral density of whole body of was measured by dual energy X-ray absorptiometry.Three rats out of each group with bone mineral density closest to the mean value were killed after anesthesia,and osteoblasts were isolated and cultured aseptically.Cell proliferation was detected by CCK-8 assay,alkaline phosphatase(ALP) activity was detected by chemical method,and the expression of bone formation related genes Runx-2 and BMP-2 in the obtained osteoblasts was detected by RT-PCR.The number of positive clones of calcified nodules was observed after staining and the calcified area was calculated.Results The results of whole-body bone mineral density showed that the whole-body bone mineral density was significantly lower in the OVX group than the sham group,while that in the OVX+GEN group was significantly higher than that in the OVX group(P<0.01).The OVX group had significantly lower cell proliferation,ALP activity,expression levels of Runx-2 and BMP-2 and calcified nodule area when compared with the sham group(P<0.01),while the OVX +GEN group had obviously higher results of above tests than the OVX group(P<0.01).Conclusion Gen can improve the bone mineral density of osteoporosis in OVX rats,which may be related to its ability to promote osteoblast proliferation,maturation and mineralization.

关 键 词：金雀异黄酮 骨密度 成骨细胞 骨形成 

分 类 号：R282.71[医药卫生—中药学] R322.71[医药卫生—中医学]