中华跌打丸对斑马鱼骨折愈合作用研究

Study on Effect of Zhonghua Dieda Pill on Fracture Healing of Zebrafish

关键词:  中华跌打丸|斑马鱼|骨折愈合|抗炎|促血管形成|软骨损伤修复

Key words:  Zhonghua Dieda pill|zebrafish|fracture healing|anti-inflammatory|angiogenesis

摘要:

目的 应用斑马鱼研究中华跌打丸促骨折愈合、抗炎、促血管形成及软骨损伤修复作用。方法 采用顿性棱刀挤压斑马鱼尾鳍建立骨折模型，评价其骨折愈合及抗炎作用；用转基因血管荧光斑马鱼评价其促血管形成作用；地塞米松建立斑马鱼软骨损伤模型，评价其软骨损伤修复作用；通过Q-PCR检测中华跌打丸对TNF-α、IL-1β、BMP-2和BMP-7基因表达的影响。结果 与模型对照组比较，中华跌打丸在浓度为每尾2 mg时，骨折愈合促进作用为20.8%（P<0.05）；浓度在15.6，31.3和62.5 mg·mL^-1时炎症消退作用分别为44.6%，50.6%和52.6%（P<0.05），且伴随TNF-α和IL-1β基因的表达有不同程度下调（P<0.05或P<0.01）；浓度为1 000，2 000 mg·L^-1时，促血管形成作用分别为19.7%和49.9%（P<0.01或P<0.001）；中华跌打丸通过上调BMP-2和BMP-7基因的表达起到软骨修复作用（P<0.01或P<0.001），浓度为500 mg·L^-1时，软骨损伤修复作用为42.0%（P<0.05）。结论 中华跌打丸对斑马鱼有促进骨折愈合、抗炎、促血管及软骨损伤修复作用，且通过下调TNF-α和IL-1β的表达起到消炎作用，上调BMP-2和BMP-7基因的表达起到软骨修复作用。

Abstract:

OBJECTIVE To study the antifracture healing, anti-inflammatory, angiogenesis and repair of cartilage damage effects of Zhonghua Dieda pill by using zebrafish. METHODS The fracture model was established by squeezing the tail fin of zebrafish by a blunt edge knife, and the effect of Zhonghua Dieda pill on fracture healing and anti-inflammatory was then evaluated; transgenic vascular fluorescence zebrafish was used to evaluate the angiogenic effect; cartilage damage model was established by treating zebrafish with dexamethasone, and then the effect of cartilage damage reparing was evaluated; the effects of Zhonghua Dieda pill on the gene expressions of TNF-α, IL-1β, BMP-2 and BMP-7 were measured by Q-PCR. RESULTS Compared with the model control group, the fracture healing effect of the Zhonghua Dieda pill was 20.8%(P<0.05) when concentration was 2 mg per tail. With down-regulating the expression of TNF-α and IL-1β genes(P<0.05 or P<0.01), the anti-inflammatory effect of Zhonghua Dieda pill was 44.6%, 50.6% and 52.6% at the 15.6, 31.3 and 62.5 mg·mL^-1 concentration(P<0.05); the angiogenesis effect of Zhonghua Dieda pill was 19.7% and 49.9% at the 1 000, 2 000 mg·L^-1 concentration groups(P<0.01 or P<0.001); Zhonghua Dieda pill showed cartilage damage repairing effect by up-regulating the expression of BMP-2 and BMP-7 genes and result in 42.0% effect of cartilage damage repairing on zebrafish model at 500 mg·mL^-1 concentration(P<0.05). CONCLUSION Zhonghua Dieda pill can promote fracture healing, anti-inflammatory, angiogenesis and cartilage damage repairing in zebrafish, and play an anti-inflammatory role by down-regulating the expression of TNF-α and IL-1β and play a cartilage repairing by up-regulating the expression of BMP-2 and BMP-7 genes.