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| 左归丸和右归丸对大鼠脂肪干细胞增殖及成骨分化影响的比较 |
| Comparison of the effect of Zuogui pill and Yougui pill on the proliferation and osteogenic differentiation of rat adipose-derived stem cells |
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| DOI：10.3969/j.issn.1006.7108.2013.11.008 |
| 中文关键词: 左归丸 右归丸 脂肪干细胞 成骨分化 Runx2 |
| 英文关键词:Zuogui pill Yougui pill Adipose-derived Stem Cells Osteogenic differentiation Runx2 |
| 基金项目:“中医骨伤科学”国家重点学科(100508)，国家自然基金项目(81073114 , 81001527 , 81102603 , 81173621)，上海领军人才项目(041)，上海高校“中医脊柱病损研究”创新团队建设项目(2009-26)、上海市科委重点项目(09dZ1973800,08411950300,11DZ1973400,11XD 1404600) |
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| 中文摘要: |
| 目的 比较左归丸、右归丸对大鼠脂肪干细胞( ASCs)增殖和成骨性分化的影响。方法 成年大鼠15只，随机分为3组，分别以左归丸(ZGW组)、右归丸(YGW组)和生理盐水(CON组)连续灌胃后制备含药血清。胶原酶消化法分离大鼠脂肪干细胞，成骨诱导21 d后，茜素红染色对细胞进行鉴定。分别采用CCK-8法、PNPP法测定加药诱导后ASCs的增殖能力及碱性磷酸酶表达差异；比色法检测细胞外钙离子浓度。Western Blot法测定加药诱导7d后的Runx2蛋白表达量。结果ASCs细胞呈长梭形生长，增殖旺盛；茜素红染色显示为阳性。CCK-8法检测结果显示，加药24 h及48 h时，YGW组OD值显著高于ZGW组及CON组(P<0.05);加药72 h时，ZGW组、YGW组OD值均显著高于CON组(P<0.05)，而两个加药组间并无显著性差异(P > 0. 05)。PNPP检测结果显示，ZGW组、YGW组OD值均显著高于CON组(P<0.05);其中YGW组OD值显著高于ZGW组(P < 0. 05)。YGW组细胞外钙离子浓度显著升高(P < 0. 05 )。 Western Blot结果显示，YGW组Runx2蛋白表达显著上调(P<0.05)。结论 与滋肾阴法相比，温肾阳法在促进ASCs增殖及成骨分化方面均表现出明显优势。 |
| 英文摘要: |
| Objective To compare the effect of Zuogui pill and Yougui pill on the proliferation and osteogenic differentiation of rat adipose-derived stem calls(ASCs).Methods Fifteen adult rats were randomly divided into 3 groups:Zuogui pill group(ZGW group )，Yougui pill group(YGW group )，and normal saline group(CON group).Drug containing-serum of rats was acquired after continuous lavage with decoction of Zuogui pill and Yougui pill，and physiological saline，respectively. Rat ASCs were isolated using collagenase digestion method .After 21-day ostegenic induction，the cells were identified using Alizarin red staining .The proliferation of ASCs and the expression of alkaline phosphatasc afterdrug intervention were detected using CCK-8 and PNPP methods. Extraccllular Ca2+ concentration was detected using colorimetric method. The expression of Runx2 protein after 7-day drug induction was detected using Western blotting. Results The ASCs were mainly spindle-shaped and proliferated quickly .The result of Alizarin red staining was positive. The results of CCK-8 method showed that the OD value was significantly higher in YGW group than that in both ZGW group and CON group after 24-hour and 48-hour culture ( P < 0.05).And after 72-hour culture，the OD value in both YGW group and ZGW group was higher than that in CON group(P < 0. 05)，while no significant difference between the two drug-intervened groups was observed ( P > 0.05).The result of PNPP detection showed that the OD value in both YGW group and ZGW group was significantly higher than that in CON group ( P < 0. 05).Compared with that in ZGW group，the OD value in YGW group was higher(P<0.05). The extracellular Ca2+ concentration was higher in YGW group (P <0.05).The result of Western blotting showed that the expression of Runx2 protein increased significantly in YGW group(P<0. 05).Conclusion Compared with the method of nourishing the kidney-yin, the method of warming the kidney-yang has obvious advantage in promoting both the proliferation and the osteogenic differentiation of ASCs. |
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