中国骨伤2018，Vol.31Issue(3)：248-253,6.DOI:10.3969/j.issn.1003-0034.2018.03.011
骨碎补总黄酮通过激活mTOR信号通路促进大鼠腱骨愈合的实验研究
Osteopractic total flavone promoting rat extra-articular tendon-bone healing through mTOR pathway
摘要
Abstract
Objective:To explore function and related molecular mechanism of osteopractic total flavone (OTF) on tendon healing in rats.Methods:Ten male rats aged for 8 weeks were collected and weighted from 180 to 220 g.Tendon stem cells were cultivated,the third tendon stem cells were used for experiment.OTP treated with 0,0.1,1,10 ng/ml were added into tendon stem cells,and expression change of ALP,Runx2,OCN,VEGF,P-S6,P-4E/BP1 were detected after 14 days.Forty male rats aged for 8 weeks (weighted 180 to 220 g) were established extra-articular tendon-bone transplanting healing model,and divided into experimental group and control group.Experimental group were treated with OTF (100 mg·kg-1· d-1),while control group was treated by normal saline with the same volume.Tendon-bone healing degree were detected by biomechanical testing at 3 and 6 weeks after surgery,histological detection were applied to detect tendon-bone healing and number of new vessles.Results:After treated by OTP,ALP staining and active index detection showed there were statistical differences among 0,0.1,1,10 ng/ml group.After 14 days' cultivation,western blotting results showed mTOR downstream marker protein P-S6 protein expression were gradually increased with increase of density of OTP,expression of P-4E/BP1 was reduced,while expression of Runx2,OCN,VEGF were increased.Biological detection results showed that there was no significant difference in mechanical strength between experimental group (0.78±0.05) N/mm and control group (0.51 ±0.02) N/mm at 3 weeks after surgery,while mechanical strength in experimental group (1.36±0.09) N/mm was higher than control group (1.01±0.08) N/mm at 6 weeks after surgery.Histological results showed maturity of tendon-bone surface cell were higher at 3 and 6 weeks in experimental group,sharpey fiber growth more density,calcification extent of mesenchyme was high,and new bone,vessels were increased.Conclusion:OTF could promote osteogenic differentiation of tendon stem cells through mTOR signaling in vitro,and stimulate tendon-bone healing in bone tunnel and enhance connection quality between tendon and bone.关键词
骨碎补/黄酮/大鼠/跟腱Key words
Drynaria fortunei/Flavone/Rats/Achilles tendon引用本文复制引用
张新涛,江华基,梁祖儒,贺飞林,廖小卿,任玉香,张文涛..骨碎补总黄酮通过激活mTOR信号通路促进大鼠腱骨愈合的实验研究[J].中国骨伤,2018,31(3):248-253,6.基金项目
广东省中医药局科研课题(编号:20131027)Provided by Administration of Traditional Chinese Medicine of Guangdong Province(No.20131027) （编号:20131027）
