雌激素受体β介导植物雌激素骨代谢效应的流行病学与细胞学研究
发布时间：2019-06-11 04:36
【摘要】：背景:绝经后骨质疏松症(PMOP)是由更年期后女性机体雌激素水平下降引起,现阶段临床防治手段多使用外源性补充雌激素,但是该替代疗法可能产生诸如乳腺癌,子宫内膜癌的等恶性肿瘤毒副作用。而植物雌激素作为安全的雌激素替代物防治PMOP正逐渐受到关注。我国膳食摄取植物雌激素量虽然显著高于西方欧美人口,但是PMOP发病率并未明显降低。我们认为这可能涉及遗传因素。本课题组前期研究已经发现绝境后妇女ERα基因多态性与膳食中植物雌激素摄入的骨密度效应有关,且细胞研究发现植物雌激素对ERβ亲和力显著高于ERα,但是ERβ基因多态性是否与膳食植物雌激素摄入的骨密度效应有关并未见关报道。目的:本研究拟从流行病学水平探讨ERβ基因多态性与膳食植物雌激素骨密度调节效应的关系。方法:对选取符合调查要求的300名南昌市绝经后妇女,用PCR-RFLP技术检测ERβ基因Rsa I和Alu I限制性酶切位点基因多态性;根据调查数据比较ERβ不同基因型时骨密度差异,并分析不同基因型时植物雌激素摄入量与不同部位BMD骨密度关系。结果:在比较各基因型样本间一般情况与各部位骨密度发现,在校正体重指数、年龄、绝经时间后,RR基因型L4的BMD高于Rr、rr组(P0.05),L2,L3髋骨总体及股骨颈、大转子、ward三角在各摄入量组内无统计学差异。将各基因型样本依据膳食植物雌激素摄入量三分位数分组后进一步分析发现,在校正体重指数、年龄、绝经时间后,Alu I,Rsa I基因型样本植物雌激素摄入量与低中高三组不同骨组织的骨密度比较,差别均无统计学意义。结论:1.ERβ基因Rsa I位点的RR基因型样本L4 BMD高于Rr,rr组,其余部位BMD在个基因型组间无显著性差异。2.ERβ基因Alu I位点多态性与BMD无关。3.在ERβ基因Rsa I,Alu I位点的各基因型样本中,各部位与膳食植物雌激素摄入量无关。目的:本课题组流行病学研究显示ERα而非ERβ基因多态性影响膳食植物雌激素与BMD关联性,现有报道认为植物雌激素的生物学效应主要通过作用于ERβ,这与我们的结果相矛盾。本研究拟从细胞水平观察植物雌激素代表药金雀异黄酮(genestein,GEN)对成骨细胞和破骨细胞效应是否由ERα、ERβ所介导,从而为流行病学研究结果提供依据。。方法:体外诱导培养小鼠前成骨细胞MC3T3-E1分别给予各组细胞10-9-10-7mol/l的GEN,以及ERα阻断剂MPP,ERβ阻断剂PHTPP处理。成骨细胞骨形成效应,通过碱性磷酸酶(ALP)染色法进行观察;并通过矿化结节茜素红染色再次验证同时体外诱导培养小鼠单核/巨噬细胞系RAW264.7细胞分化为破骨细胞,分别给予10-9-10-7mol/l GEN,以及ERα阻断剂MPP,ERβ阻断剂PHTPP处理。通过抗酒石酸酸性磷酸酶(TRAP)染色实验与骨吸收陷窝实验观察破骨细胞的骨吸收效应。结果:ALP染色结果显示,与对照组相比,不同浓度的GEN可增加成功细胞的生成,差异有统计学意义(p0.01)。在加入MPP,PHTPP后,可取消GEN的这种效应(p0.05)。矿化结节染色实验结果与ALP染色实验类似。破骨细胞实验方面,TRAP染色试验中,加入不同浓度的GEN均能减少成熟破骨细胞的生成(p0.01),且呈浓度依赖性,加入MPP后可逆转GEN的破骨细胞抑制效应。骨吸收陷窝实验结果显示与对照组相比,加入不同浓度的GEN均可减少骨吸收陷窝面积,且呈浓度依赖性。而这一效应可以被MPP而非PHTPP所逆转。结论:1.金雀异黄酮(GEN)的促MC3T3-E1骨形成效应是由ERα、ERβ所共同介导。2.金雀异黄酮(GEN)的抗RAW264.7分化为破骨细胞是主要由ERα介导,其对RAW264.7分化破骨细胞的骨吸收陷窝抑制效应由ERα而非ERβ所介导。
[Abstract]:BACKGROUND: The post-menopausal osteoporosis (PMOP) is caused by the decrease of the estrogen level of the female body after the menopause. At present, the external complementary estrogen is used in the present clinical prevention means, but the replacement therapy can produce the malignant and toxic side effects such as breast cancer and endometrial cancer. The treatment of PMOP with phytoestrogen as a safe estrogen replacement is becoming more and more concerned. Although the amount of estrogen in the dietary intake of China was significantly higher than that of the Western European and American population, the incidence of PMOP was not significantly reduced. We think this may involve genetic factors. in that early study of the research group, the polymorphism of ER gene in the postmenopausal women has been found to be related to the bone mineral density effect of the estrogen intake of the plant in the diet, and the cell study found that the affinity of the phytoestrogen to ER is significantly higher than that of ER, However, whether the ER gene polymorphism is related to the bone mineral density effect of the dietary phytoestrogen intake is not reported. Objective: To study the relationship between the polymorphism of ER gene and the regulation of the bone mineral density in dietary plant from the level of epidemiology. Methods: The polymorphisms of the ER gene Rsa I and Alu I restriction enzyme sites were detected by PCR-RFLP in 300 women with post-menopausal women in Nanchang, which were in accordance with the survey requirements. The relationship between the intake of phytoestrogen and the BMD of different parts in different genotypes was also analyzed. Results: The BMD of RR genotype L4 was higher than that of Rr, rr group (P0.05), L2, L3 hip bone and femoral neck and large rotor after comparing the general conditions between the genotype samples and the bone mineral density of each site. There was no statistical difference in the intake group. according to the three-digit number of the estrogen intake of the dietary plant, each genotype sample is further analyzed and found, after the body weight index, the age and the menopause time are corrected, the estrogen intake of the Alu I and the Rsa I genotype sample is compared with the bone density of the three different bone tissues in the low middle and high three groups, The difference was not statistically significant. Conclusion:1. The BMD of the Rsa I site in the ER gene is higher than that of the Rr, rr group, and the BMD of the rest is no significant difference between the genotypes. In each genotype sample of the ER gene Rsa I, the Alu I site, each site was independent of the dietary phytoestrogen intake. Objective: To study the relationship between ER and non-ER gene polymorphism on the relationship between the estrogen and BMD of dietary plant, and the present report is that the biological effect of phytoestrogen is mainly applied to ER, which is in contradiction with our results. In this study, it is proposed to observe whether the effect of phytoestrogen on osteoblast and osteoclast effect is mediated by ER antigen and ER antigen from the cell level, so as to provide the basis for epidemiological study. Methods: The pre-cultured mouse osteoblast MC3T3-E1 in vitro was treated with 10-9-10-7 mol/ l of GEN and ER-blocking agent MPP, ER-blocking agent, PHTPP, respectively. Osteoblastic bone formation effect was observed by alkaline phosphatase (ALP) staining method, and the mouse mononuclear/ macrophage-based RAW264.7 cells were re-validated by the mineralization of nodular and red staining. The cells were differentiated into osteoclasts,10-9-10-7 mol/ l GEN and ER-type blocking agent MPP, respectively. The ER antagonist PHTPP was treated. The bone resorption of osteoclasts was observed by anti-tartrate-acid phosphatase (TRAP) staining and bone resorption. Results: The results showed that, compared with the control group, the different concentrations of GEN could increase the production of successful cells and the difference was of statistical significance (p0.01). This effect of GEN can be eliminated after the addition of MPP, PHTPP (p0.05). The results of the staining of the mineralized nodules were similar to those of the ALP staining. In the experiment of osteoclast, in the TRAP staining test, the generation of mature osteoclasts (p0.01) can be reduced by the addition of a different concentration of GEN, and the inhibitory effect of the osteoclast can be reversed after the addition of MPP. The results of bone resorption and depression showed that, compared with the control group, the addition of the different concentrations of GEN could decrease the area of the bone resorption pit and be in a concentration-dependent manner. And this effect can be reversed by the mpp rather than the pHTPP. Conclusion:1. The MC3T3-E1 bone formation effect of the genistein (GEN) is mediated by ER antigen and ER antigen. The anti-RAW264.7 differentiation of the genistein (GEN) is mainly mediated by ER, and its inhibitory effect on the bone resorption pit of the differentiation osteoclasts of RAW264.7 is mediated by ER antigen rather than ER antigen.
【学位授予单位】：南昌大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R580
本文编号：2497019
[Abstract]:BACKGROUND: The post-menopausal osteoporosis (PMOP) is caused by the decrease of the estrogen level of the female body after the menopause. At present, the external complementary estrogen is used in the present clinical prevention means, but the replacement therapy can produce the malignant and toxic side effects such as breast cancer and endometrial cancer. The treatment of PMOP with phytoestrogen as a safe estrogen replacement is becoming more and more concerned. Although the amount of estrogen in the dietary intake of China was significantly higher than that of the Western European and American population, the incidence of PMOP was not significantly reduced. We think this may involve genetic factors. in that early study of the research group, the polymorphism of ER gene in the postmenopausal women has been found to be related to the bone mineral density effect of the estrogen intake of the plant in the diet, and the cell study found that the affinity of the phytoestrogen to ER is significantly higher than that of ER, However, whether the ER gene polymorphism is related to the bone mineral density effect of the dietary phytoestrogen intake is not reported. Objective: To study the relationship between the polymorphism of ER gene and the regulation of the bone mineral density in dietary plant from the level of epidemiology. Methods: The polymorphisms of the ER gene Rsa I and Alu I restriction enzyme sites were detected by PCR-RFLP in 300 women with post-menopausal women in Nanchang, which were in accordance with the survey requirements. The relationship between the intake of phytoestrogen and the BMD of different parts in different genotypes was also analyzed. Results: The BMD of RR genotype L4 was higher than that of Rr, rr group (P0.05), L2, L3 hip bone and femoral neck and large rotor after comparing the general conditions between the genotype samples and the bone mineral density of each site. There was no statistical difference in the intake group. according to the three-digit number of the estrogen intake of the dietary plant, each genotype sample is further analyzed and found, after the body weight index, the age and the menopause time are corrected, the estrogen intake of the Alu I and the Rsa I genotype sample is compared with the bone density of the three different bone tissues in the low middle and high three groups, The difference was not statistically significant. Conclusion:1. The BMD of the Rsa I site in the ER gene is higher than that of the Rr, rr group, and the BMD of the rest is no significant difference between the genotypes. In each genotype sample of the ER gene Rsa I, the Alu I site, each site was independent of the dietary phytoestrogen intake. Objective: To study the relationship between ER and non-ER gene polymorphism on the relationship between the estrogen and BMD of dietary plant, and the present report is that the biological effect of phytoestrogen is mainly applied to ER, which is in contradiction with our results. In this study, it is proposed to observe whether the effect of phytoestrogen on osteoblast and osteoclast effect is mediated by ER antigen and ER antigen from the cell level, so as to provide the basis for epidemiological study. Methods: The pre-cultured mouse osteoblast MC3T3-E1 in vitro was treated with 10-9-10-7 mol/ l of GEN and ER-blocking agent MPP, ER-blocking agent, PHTPP, respectively. Osteoblastic bone formation effect was observed by alkaline phosphatase (ALP) staining method, and the mouse mononuclear/ macrophage-based RAW264.7 cells were re-validated by the mineralization of nodular and red staining. The cells were differentiated into osteoclasts,10-9-10-7 mol/ l GEN and ER-type blocking agent MPP, respectively. The ER antagonist PHTPP was treated. The bone resorption of osteoclasts was observed by anti-tartrate-acid phosphatase (TRAP) staining and bone resorption. Results: The results showed that, compared with the control group, the different concentrations of GEN could increase the production of successful cells and the difference was of statistical significance (p0.01). This effect of GEN can be eliminated after the addition of MPP, PHTPP (p0.05). The results of the staining of the mineralized nodules were similar to those of the ALP staining. In the experiment of osteoclast, in the TRAP staining test, the generation of mature osteoclasts (p0.01) can be reduced by the addition of a different concentration of GEN, and the inhibitory effect of the osteoclast can be reversed after the addition of MPP. The results of bone resorption and depression showed that, compared with the control group, the addition of the different concentrations of GEN could decrease the area of the bone resorption pit and be in a concentration-dependent manner. And this effect can be reversed by the mpp rather than the pHTPP. Conclusion:1. The MC3T3-E1 bone formation effect of the genistein (GEN) is mediated by ER antigen and ER antigen. The anti-RAW264.7 differentiation of the genistein (GEN) is mainly mediated by ER, and its inhibitory effect on the bone resorption pit of the differentiation osteoclasts of RAW264.7 is mediated by ER antigen rather than ER antigen.
【学位授予单位】：南昌大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R580
【参考文献】
相关期刊论文 前4条
1 王佳茵;张海珍;关云谦;谢峰;;汉族人群雌激素受体β基因多态性与子宫肌瘤的关系[J];首都医科大学学报;2010年05期
2 冯丽华;叶琳;刘艳广;齐英;金花;崔建林;李娜;胡伟军;周丽婷;;雌激素受体β基因多态性与子宫肌瘤相关性研究[J];中国妇幼保健;2009年20期
3 徐霞,贡沁燕,鲁映青,姚明辉;大豆异黄酮对去卵巢大鼠骨密度和雌激素活性的影响(英文)[J];中国新药与临床杂志;2002年06期
4 金慰芳,朱文菁,王洪复,骆宏丰,胡昌奇;补肾中药HU-ECS对培养成骨细胞增殖、分化及矿化功能的影响[J];中国骨质疏松杂志;2001年01期
相关博士学位论文 前1条
1 乐爱文;子宫内膜雌激素受体β基因多态性及表达与原因不明月经过少关系的探讨[D];重庆医科大学;2008年
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