比较不同大鼠来源骨髓基质细胞对雌激素的反应性

Comparison the effects of estrogen on bone marrow stromal cells from different rats

目的探讨增龄和去卵巢大鼠骨髓基质细胞(BMSCs)对雌激素干预的反应。方法取3月龄、9月龄与去卵巢(卵巢切除6月后的9月龄)SD雌性大鼠,全骨髓培养法分离培养BMSCs,传代后给予10 nME2干预不同时间(3、7、11、15 d)。用油红O染色鉴定脂肪细胞,MTT法检测细胞的增殖能力,采用磷酸苯二钠法检测碱性磷酸酶的活性。结果①细胞的增殖能力随增龄和去卵巢依次递减,去卵巢组原代培养有细胞自发分化为脂肪细胞。②干预后细胞的增殖能力随时间延长逐渐增强,3月龄组和去卵巢组与相应对照组比较有显著差异。E2增强各组碱性磷酸酶活性,对去卵巢组影响更显著。结论大鼠BMSCs的增殖和分化能力随增龄和去卵巢下降,雌激素促进大鼠BMSCs的增殖和成骨分化,3月龄组和去卵巢组细胞对雌激素更敏感。

Objective The study was to explore the effects of estrogen on proliferation and osteogenic differentiation of bone marrow stromal cells （BMSCs） which came from different age healthy rats and ovariectomy rats. Methods Three months old, nine months old and ovariectomized （OVX） （nine months old, six months after ovariectomy） female Sprague-Dawley （SD） rats were used for the isolation of BMSCs by the method of the whole bone marrow culture. The passage BMSCs were treated with 10nM estradiol for 3 d, 7 d, 11 d, 15 d. The adipocytes were identified using the Oil Red O staining method. The proliferation of BMSCs was measured by MTY method, and the alkaline phosphatase activity was determined by disodium phenyl orthophosphate method. Results 1. The proliferation of BMSCs was declining with age and ovariectomy in turn. It was seen that a small number of cells had differentiated spontaneously into adipocytes in primary culture in OVX group. 2. After estradiol treatment, the capability of BMSCs proliferation was increasing with time, and there were significant differences when comparing those treated with non-treated cells in 3-month-old group and OVX group. Alkaline phosphatase activity was increased, and the effect of estradiol on OVX group was more evident than on the other groups. Conclusion The capacity of proliferation and osteogenic differentiation of rat BMSCs declines with age or ovariectomy. Estrogen promotes BMSCs proliferation and osteogenic differentiation, and the cells＇ responses to estrogen of 3-month-old group and OVX group are more sensitive than those of 9-month-old group.