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目的考察8种不同规格鹿茸含药血清对骨髓间充质干细胞(BMSCs)细胞增殖及成骨分化的影响,并对8种不同规格鹿茸进行聚类分析。方法雌性SD大鼠72只,随机分为9组,按鹿角的生长特点,加工方式等筛选8个不同规格的鹿茸药材,分别制得不同规格鹿茸的大鼠含药血清,以上述血清对BMSCs细胞进行培养,MTT法检测成骨细胞的增殖情况,ELISA法检测碱性磷酸酶(ALP)、骨形成蛋白2(BMP-2)、骨钙素(BGP)分泌量,分别采用SPSS 17.0及灰色关联度分析方法对实验结果进行统计分析。结果鹿茸含药血清对体外培养BMSCs增殖有促进作用,且鹿茸蜡片组作用优于阳性对照组,鹿茸含药血清能促进BMSCs成骨分化过程中ALP、BGP、BMP-2蛋白酶分泌。以“补肾健骨”功效对不同品质鹿茸进行排序,结果表明8种不同规格鹿茸药材样品的品质评价结果与传统鹿茸药材品质划分相符合。结论本方法及模型可用于鹿茸药材质量评价。
Objective To investigate the effect of 8 kinds of antler serum on the proliferation and osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs), and to analyze the clustering analysis of 8 kinds of antler. Methods Seventy-two female Sprague-Dawley rats were randomly divided into 9 groups. According to the growth characteristics and processing methods of the antlers, 8 different kinds of pilose antlers were screened to obtain the drug-containing serums of rats with different velvet antlers. The proliferation of osteoblasts was detected by MTT assay. The levels of alkaline phosphatase (ALP), bone morphogenetic protein 2 (BMP-2) and osteocalcin (BGP) were measured by ELISA. SPSS 17.0 and gray Correlation analysis method for statistical analysis of experimental results. Results The antler serum could promote the proliferation of BMSCs in vitro. The effect of antler serum group was better than that of positive control group. The antler serum could promote the secretion of ALP, BGP and BMP-2 in BMSCs during osteogenic differentiation. The results showed that the quality evaluation of 8 samples with different specifications of pilose antler was in accordance with the classification of the quality of traditional pilose antler. Conclusion The method and model can be used to evaluate the quality of pilose antler.