多花黄精多糖对斑马鱼2型糖尿病合并骨质疏松症模型的药效学研究

Pharmacodynamics study of polysaccharidefromPolygonatum cyrtonemaon zebrafish model with type2diabetic and osteoporosis

目的研究多花黄精多糖对斑马鱼幼鱼2型糖尿病合并骨质疏松症的作用。方法用链脲佐菌素和泼尼松龙建立斑马鱼幼鱼2型糖尿病合并骨质疏松症模型,设置对照组、模型组、二甲双胍+依替膦酸二钠组和多花黄精多糖(60、120、240μg/mL)组,考察药物对斑马鱼早期发育毒性的影响;采用试剂盒测定各组幼鱼组织的糖含量;采用茜素红染色法考察各组斑马鱼头骨矿化面积和骨密度;采用qRT-PCR法检测各组斑马鱼骨质疏松相关基因Runx家族转录因子2b(Runx family transcription factor 2b,Runx2b)、Sp7转录因子(Sp7 transcription factor,Sp7)、I型胶原蛋白α2(collagen type Iα2,col1a2)、富含半胱氨酸的酸性分泌蛋白质因子(cysteine-rich acidic secreted protein,sparc)和维生素D受体b(vitamin D receptor b,vdrb)mRNA表达情况。结果显微注射链脲佐菌素和浸泡泼尼松龙能够提高幼鱼组织糖浓度,降低头骨矿化面积和骨密度。与模型组比较,多花黄精多糖显著上调斑马鱼幼鱼2型糖尿病合并骨质疏松症模型Runx2b、Sp7、vdrb、sparc、col1a2mRNA表达水平(P<0.05、0.01、0.001)。结论多花黄精多糖对斑马鱼幼鱼2型糖尿病合并骨质疏松症模型有较好的降血糖及抗骨质疏松症作用。

ObjectiveTo study the effect of polysaccharidefromPolygonatum cyrtonema(PSP)on type 2diabetes and osteoporosis in juvenile zebrafish.MethodsA zebrafish juvenile type 2 diabetes model with osteoporosis was established with streptozotocin and prednisolone,which were divided into control group,model group,metformin+etidronate disodium group and PSP(60,120,240μg/mL)groups,the effect of drugs on early developmental toxicity of zebrafish was investigated;Kit was used to determine the sugar content of juvenile fish tissues in each group;Alizarin red staining method was used to investigate the mineralized area and bone mineral density in bones of zebrafish in each group;qRT-PCR was used to detect osteoporosis-related genes Runxfamily transcription factor 2b(Runx2b),Sp7 transcription factor(Sp7),type I collagenα2(col1a2),cysteine-rich acidic secreted protein(sparc)and vitamin D receptor b(vdrb)mRNA expressions inzebrafish.Results Microinjection of streptozotocin and immersion of prednisolone increasedthe sugar concentration of juvenile fish tissues,reducedthe mineralized area and bone density of skull.Compared with model group,PSPsignificantly up-regulated mRNA expression levels of Runx2b,Sp7,vdrb,sparcand col1a2in zebrafish juvenile type 2 diabetes mellitus with osteoporosis model(P<0.05,0.01,0.001).Conclusion PSPhas a good hypoglycemic and anti-osteoporosis effect on zebrafish juvenilewithtype 2 diabetes and osteoporosis.