楮实子含药血清对成骨细胞功能的影响  

Effect of Fructus Broussonetiae-containing serum on the function of osteoblasts

作　　者：颜志鹏 杜小康 杨鸿遒 温志刚 余腾烨 刘云 陈珺 YAN Zhipeng;DU Xiaokang;YANG Hongqiu;WEN Zhigang;YU Tengye;LIU Yun;CHEN Jun(School of Biosciences and Biopharmaceutics,Guangdong Pharmaceutical University,Guangzhou 5110006,China;Center of Biological Resources and Innovative Medicine Research,Guangdong Pharmaceutical University,Guangzhou 5110006,China)

机构地区：[1]广东药科大学生命科学与生物制药学院,广东广州510006 [2]广东药科大学生物资源与创新药物研究中心,广东广州510006

出　　处：《广东药科大学学报》2024年第1期8-13,共6页Journal of Guangdong Pharmaceutical University

基　　金：2021年第七届中国国际“互联网+”大学生创新创业大赛(202110571016)。

摘　　要：目的 研究楮实子对体外培养的成骨细胞增殖、分化及其功能的影响,为楮实子在骨代谢疾病中的应用提供实验依据。方法 采用酶消化法培养大鼠成骨细胞,MTT比色法检测不同浓度(5%、10%和20%)楮实子含药血清对细胞增殖的影响,并据此选择最佳浓度进行后续实验;以AKP试剂盒检测成骨细胞AKP活性、以茜素红染色检测成骨细胞形成矿化结节数量;Western blot法检测成骨细胞功能指标基因如成骨细胞特异性转录因子(osterix,OSX)、I型胶原(collagen type I, Coll-I)、碱性磷酸酶(alkaline phosphatase, ALP)、骨桥蛋白(osteopontin, OPN)、骨保护素(osteoprotegerin, OPG)和骨钙素(osteocalcin, OCN)等蛋白的表达水平。结果 楮实子含药血清可显著促进成骨细胞增殖(P<0.01)、提高AKP分泌水平(P<0.01)、促进矿化结节形成(P <0.01),并显著上调OSX、Coll-I、ALP、OPN和OPG等成骨功能指标相关基因的蛋白表达(P<0.05)。结论 楮实子含药血清能提高成骨细胞功能,具有较好的促成骨活性。Objective To study the effect of Fructus Broussonetiae on the proliferation,differentiation and function of osteoblasts,and to provide experimental basis for the application of Fructus Broussonetiae in bone metabolic diseases.Methods Osteoblasts were primarily cultured by enzymatic digestion.The effect of different concentrations(5%,10%,and 20%)of Fructus Broussonetiae-containing serum on cell proliferation was detected by methyl thiazolyl tetrazolium(MTT)assay,and the optimal concentration was selected for subsequent experiments.The AKP activity of osteoblasts was assessed by AKP kit,and the number of calcified nodules was assessed by Alizarin red staining.The expression levels of osteogenic marker genes of osteoblasts,including osterix(OSX),collagen type I(Coll-I),alkaline phosphatase(ALP),osteopontin(OPN),osteoprotegerin(OPG)and osteocalcin(OCN)were determined by western blot.Results Fructus Broussonetiae-containing serum significantly enhanced the proliferation of osteoblasts(P<0.01),promoted AKP activity(P<0.01)and promoted mineralized nodule formation in osteoblasts(P<0.01),as well as upregulated the expression of osteogenic proteins such as OSX,Coll-I,ALP,OPN and OPG(P<0.05).Conclusion Fructus Broussonetiae-containing serum could improve osteogenic function of osteoblasts,showing the beneficial effects on osteogenic activity.

关 键 词：楮实子 血清药理学 成骨细胞 成骨功能指标基因 

分 类 号：R285.5[医药卫生—中药学]