中药续断对跟腱愈合过程中TGF-β1表达的影响研究

Role of Radix Dipsaci in healing process of achilles tendon rupture of rabbit models

目的探讨中药续断对跟腱断裂愈合的影响及其机制。方法16只8周龄雄性新西兰白兔(体重2~2.5kg)制作成跟腱断裂模型。实验动物随机分为实验组(续断治疗组)和对照组,实验组予以续断灌胃,对照组予生理盐水灌胃。术后6周采集标本。观察跟腱吻合口标本Masson染色及天狼星红染色下细胞形态和胶原分布,并采用Western blot法分析跟腱吻合口处TGF-β1的表达。结果Masson染色下实验组标本无差异,天狼星红染色可见对照组内不规则胶原纤维形成;续断组胶原纤维致密,按一定方向规则排列。续断组胶原分布较对照组整齐。续断组Western blot灰度值为(0.905±0.23),对照组为(0.590±0.21),两组比较差异有统计学意义(t=2.785,P=0.015)。以GAPDH作为内参基因,续断组qPCR为(23.15±2.38),对照组为(21.66±2.08),差异具有统计学意义(t=2.808,P=0.018);续断组qPCR相对表达值较对照组增高2.16倍。续断组TGF-β1较对照组显著上调,差异有统计学意义(P<0.05)。结论续断可能通过增加TGF-β1的表达增加跟腱的愈合能力,有望为跟腱断裂的治疗提供新的中医药参考。

Objective The objective of this study was to evaluate the role of radix dipsaci on the healing process of achilles tendon rupture and its mechanism.Methods 16 rabbits(weighing 2-2.5 kg,age 8 months) were made achilles tendon rupture model and randomly divided into the experimental group (radix dipsaci) and the control group.The experimental group was given intermittent gastric lavage,and the control group was given normal saline gastric lavage.Specimens were collected 6 weeks after operation.The morphology and collagen distribution of Achilles tendon anastomosis specimens were observed by Masson staining and Sirius red staining,and the expression of TGF-beta 1 at Achilles tendon anastomosis was analyzed by Western blot.Results There was no difference in the experimental group under Masson staining.Irregular collagen fibers were observed in the control group by Sirius red staining,and the collagen fibers in the intermittent group were dense and arranged regularly in a certain direction.The distribution of collagen in the intermittent group was neater than that in the control group.The gray value of Western blot in the intermittent group was (0.905 + 0.23) and that in the control group was (0.590 + 0.21).There was a significant difference between the two groups (t = 2.785, P = 0.015).With GAPDH as the internal reference gene,the qPCR in the intermittent group was (23.15 +2.38) and that in the control group was (21.66 +2.08),with significant difference (t=2.808,P=0.018);the relative expression of qPCR in the intermittent group was 2.16 times higher than that in the control group.The level of TGF-beta 1 in the intermittent group was significantly higher than that in the control group (P < 0.05).Conclusion The radix dipsaci may enhance the TGF-β1 induced healing process,which is expected to provide a new idea for the achilles tendon repair.