右归饮联合干细胞介入法对冷冻坏死股骨头血管修复的实验研究

目的：观察右归饮联合干细胞介入法对冷冻坏死股骨头的血管修复及疗效,探讨该方法在治疗冷冻坏死股骨头的作用机理。方法：毕格犬24只,随机分为4组：A组（模型组）、B组（右归饮组）、C组（干细胞介入组）、D组（干细胞介入加右归饮组）。液氮冷冻法造模;分离、培养、增殖骨髓基质干细胞（MSCs）;造模完成,C、D组行动脉灌注MSCs 1mL（细胞密度：5×10^6-1×10^7/mL）;术后,B、D组每天予以右归饮100mL;治疗4周及8周后分批处死。采用DSA、病理学,免疫组织化学和RT-PCR观察股骨头坏死修复、血管修复及VEGF、TGF-β表达情况。结果：DSA观察：D组的血管数量及直径较C组明显改善（P〈0.01）;组织病理学及免疫组织化学：各治疗组较A组的结构有明显改善,D组较其他各组的VEGF、TGF-β阳性表达率都明显增高（P〈0.01）;RT-PCR：D组VEGF、TGF-β较其他各组表达量增多（P〈0.01,P〈0.05）,各治疗组的表达都高于A组（P〈0.01）。讨论：右归饮联合干细胞介入法能够促进VEGF、TGF-β的表达增强,能够有效地改善坏死股骨头的血供,促进坏死修复。

Experimental Study of Repairing Vessels with You Gui Yin and MSCs in Combination with Artery Perfusion for Osteonecrosis of Femoral Head

Objective：To observe the effect and the treatment of You Gui Yin and MSCs in Combination with Artery perfusion for ONFH.To explore the mechanism of this treatment.Methods：24 healthy male adult Beagle dogs divided into 4 groups： group A（model group）,group B（You Gui Yin recipe group）,group C（Artery perfusion with MSCs gtoup）,group D（You Gui Yin ＋MSCs in Combination with Artery perfusion group）.Use the liquid nitrogen to refrigerate the right femoral head to set up ONFH.At the same time,derive,culture and proliferate MSCs.After 3 weeks,gtoup C and group D had given MSCs with artery perfusion MSCs 1ml（5×106~1×107/mL）.After this operation,gave 100mL You Gui Yin to B group and D group.After 4 weeks or 8 weeks,put Beagle dogs to death.Observe the changes of main arteries of the femoral head,and the expression of VEGF、TGF-βby immunohistochemistry,and the expression of VEGF、TGF-βby RT-PCR.All data were analyzed by SPSS13.0.Results：DSA：The quantity and diameter of main arteries of femoral head in group D were improved than group C（P〈0.01）.Histology and immunohistochemistry：Each treatment group were improved compared with group A,especially group D（P〈0.01）.After 8 weeks treatment,the rate of VEGF、TGF-βwere significantly higher（P〈0.01）.RT-PCR：The expression of VEGF、TGF-β in group D significantly higher（P〈0.01,P〈0.05）.The expression of VEGF、TGF-βin each treatment group were higher compared with group A（P〈0.01）.And after 8 weeks treatment,the expression of VEGF were significantly higher than 4 weeks treatment（P〈0.01）.Conclusion：The effect of the You Gui Yin and MSCs in Combination with Artery perfusion for ONFH was significant.It can promote the expression of VEGF、TGF-β,and improve the circulation of femoral head and the repair of femoral head.