[1]周雪娟,夏颖,赵岩岩,等.雌二醇显著上调骨质疏松大鼠成骨细胞内抗氧化酶的表达[J].南方医科大学学报,2018,(04):402.
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雌二醇显著上调骨质疏松大鼠成骨细胞内抗氧化酶的表达()
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《南方医科大学学报》[ISSN:/CN:]

卷:
期数:
2018年04期
页码:
402
栏目:
出版日期:
2018-04-30

文章信息/Info

Title:
Estradiol significantly increases the expression of antioxidant enzymes in osteoporotic rats and osteoblasts in vitro
作者:
周雪娟夏颖赵岩岩古文清肖潇白晓春刘俊李明
关键词:
雌二醇抗氧化酶骨质疏松
Keywords:
estradiol antioxidant enzymes osteoporosis
摘要:
目的探讨雌二醇对成骨细胞内抗氧化酶表达的影响及在绝经后骨质疏松中的作用。方法构建去卵巢骨质疏松大鼠模 型,随机分为SHAM组(假手术组),OVX组(去卵巢手术组),OVX+E2组(雌二醇+去卵巢手术组),3月后检测以下常规指标鉴 定骨质疏松模型是否成功:体质量、血清Ca2+、P、碱性磷酸酶(ALP)、酸性磷酯酶(ACP)以及骨密度。同时检测血清活性氧 (H2O2)及抗氧化酶γ-GCS、GSH-px和GSH-ST水平;PCR和Western blot检测雌二醇对成骨样细胞MC3T3-E1、MG63和原代成 骨细胞(OB)内关键的抗氧化酶γ-GCS mRNA和蛋白表达的影响;mRNA 芯片检测雌二醇对成骨细胞前体MC3T3-E1内84种 抗氧化酶表达的影响;利用PCR验证筛选出的有变化的抗氧化酶;利用CCK8检测雌二醇和抗氧化剂NAC对MC3T3-E1细胞 增殖的影响。结果去卵巢手术组大鼠体质量增加,血清P、ALP、ACP升高,骨密度显著下降,表明造模成功。同时血清活性氧 (H2O2)水平显著增加,而抗氧化酶γ-GCS水平显著下降。造模的同时给予雌二醇预防性治疗,可逆转骨质疏松表型,同时抑制 血清活性氧水平,并提高抗氧化酶γ-GCS水平;雌二醇上调MC3T3-E1、MG63及OB中关键的抗氧化酶γ-GCS mRNA和蛋白水 平;mRNA芯片筛查到雌激素上调MC3T3-E1细胞内6种抗氧化酶(Gpx6、Gstk1、Nos2、Prdx2、Ngb和Ccs)表达;PCR检测发现雌 二醇均上调成骨样细胞MC3T3-E1、MG63及OB内Ccs和Ngb mRNA水平;雌二醇和抗氧化剂NAC促进成骨细胞MC3T3-E1 增殖。结论雌二醇显著上调成骨样细胞内抗氧化酶γ-Gcs、Ccs和Ngb的表达。绝经后骨质疏松与体内活性氧增高,抗氧化水平 下降密切相关。雌激素缺乏可能通过增加成骨细胞内活性氧水平,降低抗氧化酶水平,启动氧化应激级联反应,进而抑制成骨细 胞增殖,加重骨质疏松的进程。
Abstract:
Objective To investigate the effect of estradiol on the expression of antioxidant enzymes in osteoblasts and its role in postmenopausal osteoporosis. Methods Rat models of osteoporosis established by ovariectomy were treated with estradiol for 3 months, and the changes in serum levels of reactive oxygen species (H2O2) and antioxidant enzymes (γ-GCS, GSH-ST and GSH-px) were detected. The effects of estradiol on the expression of γ-GCS mRNA and protein in osteoblast-like cells MC3T3- E1, MG63 and OB were examined with PCR and Western blotting. Using a mRNA microarray, we analyzed the changes in the expressions of 84 antioxidant enzymes in the osteoblast cell line MC3T3-E1 following estradiol treatment, and the enzymes with significant changes were verified by PCR. CCK-8 kit was used to evaluate the effect of estradiol and antioxidant NAC on the proliferation of MC3T3-E1 cells. Results Rat models of osteoporosis were successfully established with ovariectomy. The osteoporotic rats showed significantly increased serum level of reactive oxygen species (H2O2) and decreased levels of antioxidant enzymes. Estrogen treatment of the osteoporotic rats obviously reversed the phenotype of osteoporosis, lowered serum level of reactive oxygen species, and increased the level of γ-GCS. In MC3T3-E1, MG63 and OB cells, estradiol treatment significantly upregulated the expression levels of γ-GCS mRNA and protein. In MC3T3-E1 cells treated with estrogen, the mRNA chip identified 6 upregulated antioxidant enzymes (Gpx6, Gstk1, Nos2, Prdx2, Ngb and Ccs), and the results of PCR verified that estradiol upregulated Ccs and Ngb mRNAs in MC3T3-E1, MG63 and OB cells. Estradiol and antioxidant NAC obviously promoted the proliferation of MC3T3-E1 cells. Conclusion Estradiol significantly increases the expression of antioxidase γ-Gcs, Ccs and Ngb in osteoblasts in vitro. Postmenopausal osteoporosis is closely related with the increase of reactive oxygen species and the decrease of antioxidant levels. In osteoblasts, estrogen deficiency may increase the level of reactive oxygen species, decrease the level of antioxidant enzymes, activate the oxidative stress cascade, and consequently inhibit the proliferation of osteoblasts to aggravate the condition of osteoporosis.

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更新日期/Last Update: 1900-01-01