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【目的】观察壮骨强筋片含药血清对老年大鼠骨髓基质干细胞(bone marrow stromal stem cells,BMSCs)成脂分化的影响。【方法】取18月龄雄性SD大鼠,采用全骨髓贴壁法培养BMSCs,传代3次后用于实验。采用流式细胞仪检测细胞表面蛋白标志CD34、CD31、CD29。将细胞分为成脂诱导组、成脂诱导中药组、中药组、空白组。14 d后进行油红O染色,计算脂肪细胞转化率,并进行油红O染色定量分析;采用逆转录—聚合酶链反应(RT-PCR)法检测脂蛋白脂酶(lipoprotein lipase,LPL)和过氧化物酶增殖活化受体(peroxisome proliferator activated receptor,PPARγ2)mRNA表达。【结果】CD29表达为阳性,CD31和CD34表达均为阴性。油红O染色结果显示:成脂诱导组、成脂诱导中药组镜下观察可见有红色脂滴,成脂诱导中药组脂肪细胞转化率显著低于成脂诱导组(P<0.05)。油红O染色定量分析吸光度(D)值检测结果显示:成脂诱导中药组油红O含量显著低于成脂诱导组(P<0.01)。成脂诱导组LPL基因相对D值显著高于空白组(P<0.01)。成脂诱导中药组LPL基因相对D值较成脂诱导组显著降低(P<0.05),成脂诱导组PPARγ2基因相对D值显著高于空白组(P<0.01),中药组PPARγ2基因相对D值显著低于空白组(P<0.05),成脂诱导中药组PPARγ2基因相对D值显著低于成脂诱导组(P<0.01)。【结论】壮骨强筋片含药血清可抑制老年大鼠骨髓基质干细胞的成脂分化,可能是其防治老年性骨质疏松症的机制之一。
【Objective】 To observe the effect of Zhuanggugiangjin Tablet containing serum on adipogenic differentiation of bone marrow stromal stem cells (BMSCs) in aged rats. 【Methods】 18-month-old male Sprague-Dawley rats were used to culture BMSCs with whole bone marrow adherent method. Cell surface markers CD34, CD31 and CD29 were detected by flow cytometry. The cells were divided into adipogenic induction group, adipogenic induction group, traditional Chinese medicine group and blank group. Oil red O staining was performed 14 days later, and the adipocyte transformation rate was calculated. Oil red O staining was used for quantification. Reverse transcription-polymerase chain reaction (RT-PCR) was used to detect the expression of lipoprotein lipase (LPL) Peroxisome proliferator activated receptor (PPARγ2) mRNA expression. 【Results】 The expression of CD29 was positive, while the expressions of CD31 and CD34 were negative. The result of oil red O staining showed that there were red lipid droplets in the adipogenic and adipogenic groups, while the adipogenic rate in the adipogenic group was significantly lower than that in the adipogenic group (P <0.05). Oil Red O staining quantitative analysis of absorbance (D) value of the test results showed that: adipogenic induction of Chinese medicine group oil red O content was significantly lower than the adipogenic induction group (P <0.01). The relative D value of LPL gene in adipogenic induction group was significantly higher than that in blank group (P <0.01). The relative D value of LPL gene in Chinese herbal medicine group was significantly lower than that in adipogenic group (P <0.05), and the relative D value of PPARγ2 gene in adipogenic induction group was significantly higher than that in blank group (P <0.01) (P <0.05). The relative D value of PPARγ2 gene in the Chinese traditional medicine group was significantly lower than that in the adipogenic group (P <0.01). 【Conclusion】 Zhuanggu Qiangjin Tablet-containing serum can inhibit the adipogenic differentiation of bone marrow stromal cells in aged rats, which may be one of the mechanisms of prevention and treatment of senile osteoporosis.