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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:5×106~1×107/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。
OBJECTIVE: To observe the effect of right kidney drink combined with stem cell intervention on vascular repair and therapeutic effect of frozen and necrotic femoral head, and to explore the mechanism of this method in the treatment of frozen and necrotic femoral head. Methods: 24 Beagle dogs were randomly divided into 4 groups: group A (model group), group B (youguiyin group), group C (stem cell intervention group), and group D (stem cell intervention plus Youguiyin group). Liquid nitrogen freezing method was used to isolate, culture, and propagate bone marrow stromal cells (MSCs). After the model was completed, C and D groups were perfused with MSCs 1 mL (cell density: 5×10 6 to 1×10 7 /mL); Group B and D received 100 mL of right-depleted drink each day; they were killed in batches after 4 weeks and 8 weeks of treatment. DSA, pathology, immunohistochemistry and RT-PCR were used to observe the repair of femoral head necrosis, vascular repair, and the expression of VEGF and TGF-β. Results: DSA observation: The number and diameter of blood vessels in group D were significantly improved compared with group C (P<0.01). Histopathology and immunohistochemistry: The structure of each treatment group was significantly improved compared with group A, and group D was more than that of other groups. The positive expression rates of VEGF and TGF-β were significantly higher (P<0.01). RT-PCR: The expression of VEGF and TGF-β in D group was higher than that in other groups (P<0.01, P<0.05). The expression of each treatment group All were higher than group A (P<0.01). Discussion: Youguiyin combined with stem cell intervention can promote the expression of VEGF and TGF-β, which can effectively improve the blood supply of necrotic femoral head and promote the repair of necrosis.