Therapeutic vasculogenesis using bone marrow mononuclear cells for painful diabetic neuropathy

糖尿病神经病变的特征是感觉兴奋性改变。阳性症状，包括痛觉过敏，通常出现在糖尿病的早期阶段。由于骨髓不仅是血管前体细胞的丰富来源，而且也是细胞因子的来源，我们研究了将骨髓来源的MNCs移植到后肢肌肉中是否改善了STZ诱导的糖尿病大鼠早期观察到的痛敏反应。2周后，将从骨髓或生理盐水中分离的单核细胞注入糖尿病大鼠一侧后肢骨骼肌。使用von Frey Hair每隔2或3天测量一次行为伤害性反应。注射MNCs 2周后，测定坐骨神经传导速度(NCV)和坐骨神经血流量(SNBF)。用实时定量聚合酶链式反应检测骨髓来源的单核细胞中多种细胞因子的基因表达。频率…STZ给药后，更多的足爪缩短率开始增加。与生理盐水注射侧相比，单核细胞移植可显著减少糖尿病大鼠注射MNCs侧的缩足次数。观察MNCs移植后3d至观察期对痛觉过敏的影响。电生理研究表明，移植单核细胞可明显改善糖尿病大鼠单核细胞注射侧受损的NCV和SNBF。MNC中碱性成纤维细胞生长因子和血管内皮细胞生长因子的mRNA表达增强，提示骨髓MNCs不仅具有治疗血管生成的作用，还可作为细胞因子的来源，提示骨髓MNCs移植可能是治疗糖尿病早期痛觉过敏的一种有效方法。较少

Diabetic neuropathy is characterized by altered sensory excitability. The positive symptoms, including hyperalgesia, are often present in the early stage of diabetes. Since bone marrow is a rich source of not only vascular progenitor cells but also cytokines, we have examined whether the transplantation of bone marrow-derived MNCs into hindlimb muscles improves hyperalgesia observed in the early stage of STZ-induced diabetic rats.Diabetes was induced by an intraperitoneal injection of STZ (60mg/Kg) into 6 week-old Sprague-Dawley rats. 2 weeks later, MNCs isolated from bone marrow or saline were injected into unilateral hindlimb skeletal muscles of diabetic rats. Behavioral nociceptive responses were measured every 2 or 3 days using von Frey hair. Two weeks after the injection of MNCs, sciatic nerve conduction velocity (NCV), and sciatic nerve blood flow (SNBF) were measured. mRNA expression of several cytokines in bone marrow-derived MNCs were examined using real time PCR. The frequenc … More y of paw withdrawal started to increase after STZ administration. The transplantation of MNCs significantly decreased the frequency of paw withdrawal in MNCs-injected side of diabetic rats compared with those in saline-injected side of diabetic rats. The effects of MNCs transplantation on hyperalgesia was observed from 3 days after MNCs transplantation until observation periods. Electrophysiological studies revealed that the trasnplantation of MNCs significantly ameliorated the impaired-NCV and SNBF in MNCs-injected side of diabetic rats compared with those in saline-injected side of diabetic rats. The mRNA expressions of the basic fibroblast growth factor and vascular endothelial growth factor were observed in MNCs.These results indicate that bone marrow-derived MNCs is useful for not only therapeutic vasculogenesis but also a source of cytokines, suggesting that the transplantation of bone marrow-derived MNCs may be an effective therapy for hyperalgesia in the early stage of diabetes. Less