黄精多糖与积雪草苷对糖尿病斑马鱼尾鳍再生的药效学研究

Pharmacodynamic Study of Polygonatum Sibiricum Polysaccharide and Asiaticoside on Caudal Fin Regeneration of Zebrafish with Type 2 Diabetes

目的 通过建立和使用斑马鱼2型糖尿病模型,探讨黄精多糖与积雪草苷对2型糖尿病斑马鱼尾鳍再生的促进作用。方法 首先对药物进行毒性分析,得出最佳给药浓度范围;通过显微注射链脲佐菌素方法构建斑马鱼2型糖尿病模型,切断斑马鱼尾鳍,经不同给药浓度干预后,测定斑马鱼空腹血糖水平并观察尾鳍再生情况;通过实时定量PCR(qRT-PCR)检测黄精多糖、积雪草苷及联合用药对斑马鱼尾鳍再生相关基因mRNA的表达水平。结果 各给药组均可有效降低斑马鱼空腹血糖水平,且能上调骨再生相关基因的表达,其中积雪草苷高浓度组对成纤维细胞生长因子的表达上调具有显著差异(P<0.01)。结论 黄精多糖、积雪草苷及其联合给药的高浓度组可有效降低斑马鱼空腹血糖水平且具有促进尾鳍再生的效果,为治疗2型糖尿病和促进组织修复方面药物的开发提供理论依据。

OBJECTIVE To explore the effects of polygonatum sibiricum polysaccharide(PSP)and asiaticoside(AS)on the regeneration of caudal fin in type 2 diabetic zebrafish.METHODS First,the safe dosages of the drugs were analyzed,and then a zebrafish type 2 diabetes model was constructed by microinjection of 0.3%streptozotocin(STZ).After the tail of the type 2 diabetic zebrafish was docked,PSP,AS and combined drugs were given,and real time quantitative PCR(qRT-PCR)was used to detect the effects on the expression of bone morphogenetic protein(bmp2b),fibroblast growth factor(fgf20a)and matrix metalloproteinase(mmp13)related to zebrafish tail fin regeneration.RESULTS Three days after STZ injection,the fasting blood glucose of zebrafish was 2.7264 mg·mL^(-1),indicating that the modeling was successful.Compared with the model group,the length of caudal fin regeneration increased significantly in the AS 150μg·mL^(-1),PSP 75μg·mL^(-1),AS 75μg·mL^(-1)+PSP 37.5μg·mL^(-1) groups.The expression of bmp 2b in the AS 75μg·mL^(-1),AS 75μg·mL^(-1)+PSP 37.5μg·mL^(-1) group was significantly increased,surpassing that of the metformin group.The expression of fgf 20a in the AS 300μg·mL^(-1) group was up-regulated to a very significant level(P<0.001),and the expression of mmp13 gene was up-regulated in the administration group,but the effect was not as good as that in the metformin group.CONCLUSION AS,PSP and their combination had the effects of lowering blood sugar and promoting tail fin regeneration in type 2 diabetic zebrafish,and the latter effect may be produced by promoting the gene expression of bmp 2b,fgf 20a,and mmp13.Overall,AS more effectively increased the expression of genes related to zebrafish tail fin regeneration.