BACKGROUND: Serum containing Shudi and Biejia up-regulates the gene expression of osteocalcin in osteoblasts; the mesangial cells supernatant can also up-regulate the gene expression of osteocalcin in osteoblasts.OBJECTIVE: To detect if there are additive effects of the mesangial cells supernatant and serum containing Shudi and Biejia on the gene and protein expression of osteocalcin in osteoblasts.METHODS: Primary cultured osteoblasts of SD rats were divided into four groups after identification. Control group contained blank rat serum. Shudi Biejia group contained rat serum containing Shudi Biejia. Mesangial cell supernatant group contained blank rat serum and mesangial cell supernatant. Shudi Biejia plus mesangial cell supernatant group contained mesangial cell supernatant and rat serum containing Shudi Biejia. Culture medium was collected on day 6 after cultivation and was well mixed with the culture medium collected on day 3 after cultivation for detection.RESULTS AND CONCLUSION: Compared with the control group, the mRNA expression of osteocalcin in Shudi Biejia group, mesangial cell supernatant group and Shudi Biejia plus mesangial cell supernatant group was significantly up-regulated (P < 0.05); the expression rate of osteocalcin mRNA in the Shudi Biejia plus mesangial cell supernatant group was significantly higher than that in the Shudi Biejia group and mesangial cell supernatant group. The absorbance value of osteoblastic osteocalcin concentration in the Shudi Biejia group and mesangial cell supernatant group was significantly higher than that in the control group (P < 0.05). Compared with Shudi Biejia group, the absorbance value of osteoblastic osteocalcin concentration in the Shudi Biejia plus mesangial cell supernatant group was significantly increased (P < 0.05). These findings indicate that the mesangial cell supernatant and serum containing Shudi Biejia both can increase the gene expression of osteocalcin in osteoblast; the combined action of the two can further up-regulate the gene expression of osteocalcin; meanwhile the osteocalcin secretion also increases significantly.%背景:熟地鳖甲含药血清可上调成骨细胞骨钙素基因表达,系膜细胞上清液亦可促进成骨细胞该基因的表达.目的:验证熟地鳖甲含药血清与系膜细胞上清液对成骨细胞骨钙素基因及蛋白表达的影响是否有叠加作用.方法:原代培养经鉴定的SD大鼠成骨细胞分为4组培养:①空白对照组含空白血清培养液.②含药血清组含熟地鳖甲含药血清培养液.③无药系膜组含空白血清培养液+空白系膜细胞上清.④含药系膜组含熟地鳖甲血清培养液+含药系膜细胞上清液.6 d后收集培养液,并与3 d所收集的培养液混匀进行检测.结果与结论:与空白对照组比较,含药血清组、无药系膜组、含药系膜组骨钙素mRNA表达率明显上调(P < 0.05),含药系膜组骨钙素mRNA表达率明显高于含药血清组或无药系膜组.含药系膜组成骨细胞骨钙素浓度/MTT A值明显高于空白对照组(P < 0.05).与含药血清组比较,含药系膜组成骨细胞骨钙素浓度/MTT A值明显升高(P < 0.05).提示系膜细胞上清液及含药血清可上调成骨细胞骨钙素基因表达,二者共同作用进一步上调骨钙素基因的表达量,此时骨钙素分泌量亦明显增加.
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