基于促BMSCs成骨诱导分化作用的鹿茸药材品质评价研究

Review on quality of Lurong( cornu cervi Pantotrichum ) based on osteoinductive differentiatione effect of bone mesenchymal stem cells

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 influences of medicated serum of Lurong （cornu cervi Pantotrichum） in various specifications on proliferation and osteogenic differentiation of bone mesenchymal stem cells （BMSCs）, and conduct a cluster analysis on Lurong in 8 specifications. Methods SD rats （ n = 72） were randomly divided into 9 groups. The medical materials of Lurong in 8 specifications were screened according to the growth characteristics and processing methods of antlers. The medicated serum samples contained Lurong in various specifications were prepared respectively in rats, and BMSCs were cultured in these serum samples. The proliferation of BMSCs was detected by using methyl thiazolyl tetrazolium test （MTF）, and secretion volume of alkaline phosphatase （ALP）, bone morphogenetic protein-2 （BMP-2） and bone gla-protein （BGP） were detected by using enzymelinked immunosorbent assay （ELISA）. The test results were statistically analyzed by applying SPSS17.0 software and grey relational analysis respectively. Results The medicated serum of Lurong had an accelerative effect on proliferation of BMSCs, which was superior in Lurong wax-scale groups to that in inducing liquid group. The serum also improved the secretion of ALP, BGP and BMP-2 during the course of osteogenic differentiation of BMSCs. The ranking of Lurong in different qualities, according to the effects of tonifying the kidney and invigorating bone, showed that the results of quality reviews on samples of Lurong in 8 specifications conformed to the traditional quality partition of Lurong medical materials. Conclusion This method and model can be applied in quality review of medical materials of Lurong.