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中医不同治法诱导肌源性干细胞向成骨细胞诱导分化中SDF-1蛋白表达的比较研究
Comparative study of SDF-1 protein expression in differentiation of myogenic stem cells into osteoblasts induced by different methods of traditional Chinese medicine
  
DOI：10.3969/j.issn.1006-7108.2022.06.001
中文关键词:  中医中药  基质细胞衍生因子-1  补肾  健脾  肌源性干细胞  成骨细胞
英文关键词:traditional Chinese medicine  stromal cell-derived factor 1  tonifying the kidney  invigorating the spleen  myogenic stem cells  osteoblasts
基金项目:国家自然科学基金(81973719，82104709)；辽宁省科技厅项目（2019-ZD-0958）；沈阳市中青年科技创新人才计划（RC200160）
作者单位
付夜平 杨芳* 孙鑫 邸贵鑫 安勇 辽宁中医药大学，辽宁 沈阳 110032 
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中文摘要:
      目的 探究在中医补肾、健脾、补肾健脾的不同方法下，人骨形态发生蛋白2（BMP-2）诱导大鼠肌源性干细胞（MDSCs）成骨分化的过程中基质细胞衍生因子1（SDF-1）蛋白含量变化。方法 购置体外分离培养大鼠肌源性干细胞(MDSCs)。用随机数字表法将60只SPF级SD大鼠分为6组，依次为正常组、诱导液组、诱导液+补肾阴组（左归丸组）、诱导液+补肾阳组（右归丸组）、诱导液+健脾组（补中益气汤组）、诱导液+补肾健脾组。对大鼠进行7 d灌胃，大鼠胃容积为 1 mL /100 g，以此为大鼠配药及灌胃，制备含药血清。通过茜素红染色观察其细胞形态，使用qRT-PCR法、ELISA检测上述各组细胞中SDF-1mRNA及蛋白表达。结果 ①qRT-PCR检测结果显示：与正常组相比，诱导组、补肾阴组、补肾阳组、健脾组、补肾健脾组显著升高；与诱导组比较，除健脾组外，其余各组组中SDF-1均显著升高，且具有统计学意义(P<0.05)；②ELISA检测结果表明：与正常组相比，诱导组、补肾阴组、补肾阳组、健脾组、补肾健脾组中SDF-1mRNA相对表达量均高于正常组，除补肾阴组与补肾健脾组外均具有统计学意义(P<0.05)；与诱导组相比，除补肾健脾组外，补肾阴组、补肾阳组、健脾组均具有统计学意义（P<0.05）。结论 ①大鼠肌源性干细胞(MDSCs)向成骨细胞的转化与SDF-1的水平升高有关；②补肾阴、补肾阳、健脾和补肾健脾法大鼠肌源性干细胞(MDSCs)向成骨细胞诱导分化的过程中存在正向促进作用，可能是通过提高SDF-1水平来促进成骨细胞的转化。
英文摘要:
      Objective To explore the different methods of tonifying kidney, spleen, kidney and spleen, changes of Stromal cell-derived factor-1(SDF-1) protein during the osteogenic differentiation of rat muscle-derived stem cells (MDSCs) induced by human bone morphogenetic protein-2(BMP-2). Methods DSCs were isolated and cultured in vitro. Sixty SPF SD rats were divided into six groups by random number table method, the results were as follows: Normal Group, Induction Group, induction + Kidney Yin Group (Zuoguiwan group) , induction + Kidney Yang Group (youguiwan group) , induction + Spleen Strengthening Group (Bu Zhong Yi Qi Wan Group) , induction + Kidney Spleen strengthening group. The rats were given intragastric administration for 7 days with a gastric volume of 1 mL/100 g to prepare drug-containing serum. The Cell Morphology was observed by Alizarin red staining, and the expression of sdf-1 mRNA and protein was detected by qRT-PCR and Elisa. Results ①The results of qRT-PCR showed that Compared with the normal group, the levels of SDF-1 in the induction group, the kidney-yin-tonifying Group, the kidney-yang-tonifying Group, the spleen-tonifying Group and the spleen-tonifying group were significantly higher than those in the control group (P<0.05); ②ELISA test results showed that the expression of sdf-1 mRNA in Induction Group, kidney-yin Group, kidney-yang Group, spleen-strengthening Group and kidney-spleen-strengthening group was higher than that in normal group by Elisa, in comparison with the induction group, the kidney-yin-tonifying Group, the kidney-yang-tonifying Group and the spleen-tonifying group all had statistical significance (P<0.05) , except the kidney-yin-tonifying group and the spleen-tonifying Group. Conclusion ①The level of SDF-1 is related to the transformation of MDSCs into osteoblasts; ②The differentiation of MDSCs into osteoblasts in rats with the methods of tonifying Kidney Yin, tonifying kidney Yang, tonifying spleen and tonifying kidney spleen may be promoted by increasing the level of SDF-1.
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