间充质干细胞联合白藜芦醇对1型糖尿病小鼠周围神经病变保护作用

发布时间：2018-03-15 22:31

本文选题：间充质干细胞　切入点：白藜芦醇　出处：《青岛大学》2017年硕士论文　论文类型：学位论文

【摘要】：目的:糖尿病周围神经病变(Diabetic peripheral neuropathy,DPN)在病理生理学上与神经营养因子的缺乏以及炎症因子的水平升高有关。间充质干细胞(Mesenchymal stem cells,MSC)具有降低炎症因子,对受损神经提供营养支持的作用。白藜芦醇具有促进组织再生、分泌生长因子等生物作用。因此,本实验通过局部移植MSC及应用白藜芦醇来探究其对DPN的保护作用及相关机制。方法:饲养健康雄性NOD小鼠,诊断为1型糖尿病后随机分为对照组、胰岛素组、干细胞组、白藜芦醇组、联合治疗组织。对照组在发病后未给予任何处理;胰岛素组在发病后根据血糖水平给以胰岛素治疗;干细胞组于发病后第3天尾静脉注射h UCMSC 1×10^6;白藜芦醇组于发病后第3天起给与白藜芦醇200mg/kg·d灌胃;联合治疗组于发病后第3天尾静脉注射h UCMSC 1×10^6,同时给与白藜芦醇200mg/kg·d灌胃;待饲养结束后未发病的NOD小鼠分为正常组。除正常组及对照组外,均给予甘精胰岛素作为基础治疗。于发病8周后提取各组小鼠坐骨神经,并进行HE染色、免疫组化NF-κB、NGF、MBP染色及Masson染色,了解坐骨神经病变情况,并进行统计学分析。结果:在发病第56天,联合治疗组小鼠体重与正常组相差不大,其他四组均出现显著下降;在发病第1天,其他五组血糖均较正常组明显升高,在发病第56天,胰岛素组、干细胞组、联合治疗组血糖水平与正常组无明显差异;HE染色发现,实验组与其他五组相比较,神经纤维变性明显,神经纤维排列紊乱、稀疏,可见大量髓鞘空泡变;免疫组化NF-κB染色显示,对照组NF-κB表达较其他五组明显增强,五组之间表达无显著差异;免疫组化NGF染色显示,正常组、干细胞组以及联合治疗组NGF表达明显增强,而胰岛素组、白藜芦醇组与对照组无显著差别;免疫组化MBP染色显示,对照组MBP表达显著减少,而胰岛素组与对照组表达相近;Masson染色显示,胰岛素组、干细胞组、白藜芦醇组、联合治疗组有髓神经纤维计数、轴突直径及髓鞘厚度均高于对照组,而且联合治疗组有髓神经纤维计数明显高于干细胞组和白芦藜醇组。结论:间充质干细胞及白藜芦醇均对糖尿病受损坐骨神经起到保护作用。间充质干细胞可以降低血糖水平,改善糖尿病受损神经的炎症状态,分泌神经营养因子,促进神经纤维再生,改善脱髓鞘程度。白藜芦醇对降低炎症因子、修复受损神经也起到显著作用。并且二者联合应用,可以更好地保护受损神经,改善神经脱髓鞘状态,尤其是在促进神经纤维再生方面,明显优于单一治疗方案。
[Abstract]:Objective: the pathophysiology of diabetic peripheral neuropathies is related to the deficiency of neurotrophic factor and the increase of inflammatory factor level. Mesenchymal stem cells (MSCs) can reduce the inflammatory factor. The role of resveratrol in providing nutritional support to damaged nerves. Resveratrol promotes tissue regeneration, secretes growth factors and other biological effects. In this study, MSC was transplanted locally and resveratrol was used to investigate the protective effect of resveratrol on DPN. Methods: healthy male NOD mice were bred and randomly divided into control group, insulin group and stem cell group after diagnosis of type 1 diabetes mellitus. Resveratrol group, combined treatment tissue. The control group was not given any treatment after onset, the insulin group was treated with insulin according to the level of blood sugar after the onset. The stem cell group was injected with h UCMSC 1 脳 10 ^ 6 at the end of the third day after onset, the resveratrol group was given resveratrol 200 mg / kg 路d from the third day after onset, the combined treatment group was given UCMSC 1 脳 10 ^ 6 by intravenous injection at the end of the third day after the onset of the disease, and the resveratrol 200 mg / kg 路d was given intragastric administration at the same time. The NOD mice without disease after feeding were divided into normal group, except normal group and control group, all of them were treated with insulin glargine as the basic therapy. Sciatic nerve was extracted from each group 8 weeks after the onset of the disease, and HE staining was performed on the sciatic nerve of each group. The results showed that on the 56th day, the weight of mice in the combined treatment group was not significantly different from that in the normal group, and the other four groups were significantly decreased. On the first day of onset, the blood glucose levels in the other five groups were significantly higher than those in the normal group. On the 56th day of onset, there was no significant difference in blood glucose levels between the insulin group, stem cell group and combined treatment group, and HE staining showed that there was no significant difference between the experimental group and the other five groups, and the experimental group was compared with the other five groups. The denaturation of nerve fibers was obvious, the arrangement of nerve fibers was disordered and sparse, a large number of myelin vacuoles were observed, the expression of NF- 魏 B in the control group was significantly higher than that in the other five groups, and there was no significant difference between the five groups in the expression of NF- 魏 B. Immunohistochemical NGF staining showed that the expression of NGF was significantly increased in normal group, stem cell group and combined treatment group, but there was no significant difference in insulin group, resveratrol group and control group, and the expression of MBP in control group was significantly decreased by immunohistochemistry MBP staining. The expression of myelinated nerve fibers, axon diameter and myelin sheath thickness in insulin group, stem cell group, resveratrol group and combination group were higher than those in control group. Conclusion: both mesenchymal stem cells and resveratrol can protect the injured sciatic nerve of diabetes mellitus. Mesenchymal stem cells can reduce blood glucose level. Improve the inflammatory state of injured nerve in diabetes mellitus, secrete neurotrophic factor, promote regeneration of nerve fiber, improve demyelinating degree. Resveratrol also plays a significant role in reducing inflammatory factor and repairing damaged nerve. It can better protect the injured nerve and improve the demyelinating state of the nerve, especially in promoting the regeneration of nerve fibers, which is obviously superior to the single treatment.
【学位授予单位】：青岛大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R587.2;R745

【参考文献】