本文選題：間充質(zhì)干細(xì)胞　切入點(diǎn)：白藜蘆醇　出處：《青島大學(xué)》2017年碩士論文　論文類型：學(xué)位論文

【摘要】：目的:糖尿病周圍神經(jīng)病變(Diabetic peripheral neuropathy,DPN)在病理生理學(xué)上與神經(jīng)營養(yǎng)因子的缺乏以及炎癥因子的水平升高有關(guān)。間充質(zhì)干細(xì)胞(Mesenchymal stem cells,MSC)具有降低炎癥因子,對受損神經(jīng)提供營養(yǎng)支持的作用。白藜蘆醇具有促進(jìn)組織再生、分泌生長因子等生物作用。因此,本實(shí)驗(yàn)通過局部移植MSC及應(yīng)用白藜蘆醇來探究其對DPN的保護(hù)作用及相關(guān)機(jī)制。方法:飼養(yǎng)健康雄性NOD小鼠,診斷為1型糖尿病后隨機(jī)分為對照組、胰島素組、干細(xì)胞組、白藜蘆醇組、聯(lián)合治療組織。對照組在發(fā)病后未給予任何處理;胰島素組在發(fā)病后根據(jù)血糖水平給以胰島素治療;干細(xì)胞組于發(fā)病后第3天尾靜脈注射h UCMSC 1×10^6;白藜蘆醇組于發(fā)病后第3天起給與白藜蘆醇200mg/kg·d灌胃;聯(lián)合治療組于發(fā)病后第3天尾靜脈注射h UCMSC 1×10^6,同時給與白藜蘆醇200mg/kg·d灌胃;待飼養(yǎng)結(jié)束后未發(fā)病的NOD小鼠分為正常組。除正常組及對照組外,均給予甘精胰島素作為基礎(chǔ)治療。于發(fā)病8周后提取各組小鼠坐骨神經(jīng),并進(jìn)行HE染色、免疫組化NF-κB、NGF、MBP染色及Masson染色,了解坐骨神經(jīng)病變情況,并進(jìn)行統(tǒng)計學(xué)分析。結(jié)果:在發(fā)病第56天,聯(lián)合治療組小鼠體重與正常組相差不大,其他四組均出現(xiàn)顯著下降;在發(fā)病第1天,其他五組血糖均較正常組明顯升高,在發(fā)病第56天,胰島素組、干細(xì)胞組、聯(lián)合治療組血糖水平與正常組無明顯差異;HE染色發(fā)現(xiàn),實(shí)驗(yàn)組與其他五組相比較,神經(jīng)纖維變性明顯,神經(jīng)纖維排列紊亂、稀疏,可見大量髓鞘空泡變;免疫組化NF-κB染色顯示,對照組NF-κB表達(dá)較其他五組明顯增強(qiáng),五組之間表達(dá)無顯著差異;免疫組化NGF染色顯示,正常組、干細(xì)胞組以及聯(lián)合治療組NGF表達(dá)明顯增強(qiáng),而胰島素組、白藜蘆醇組與對照組無顯著差別;免疫組化MBP染色顯示,對照組MBP表達(dá)顯著減少,而胰島素組與對照組表達(dá)相近;Masson染色顯示,胰島素組、干細(xì)胞組、白藜蘆醇組、聯(lián)合治療組有髓神經(jīng)纖維計數(shù)、軸突直徑及髓鞘厚度均高于對照組,而且聯(lián)合治療組有髓神經(jīng)纖維計數(shù)明顯高于干細(xì)胞組和白蘆藜醇組。結(jié)論:間充質(zhì)干細(xì)胞及白藜蘆醇均對糖尿病受損坐骨神經(jīng)起到保護(hù)作用。間充質(zhì)干細(xì)胞可以降低血糖水平,改善糖尿病受損神經(jīng)的炎癥狀態(tài),分泌神經(jīng)營養(yǎng)因子,促進(jìn)神經(jīng)纖維再生,改善脫髓鞘程度。白藜蘆醇對降低炎癥因子、修復(fù)受損神經(jīng)也起到顯著作用。并且二者聯(lián)合應(yīng)用,可以更好地保護(hù)受損神經(jīng),改善神經(jīng)脫髓鞘狀態(tài),尤其是在促進(jìn)神經(jīng)纖維再生方面,明顯優(yōu)于單一治療方案。
[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.
【學(xué)位授予單位】：青島大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：R587.2;R745

【參考文獻(xiàn)】

相關(guān)期刊論文 前1條

1 ;Differentiation of bone marrow-derived mesenchymal stem cells from diabetic patients into insulin-producing cells in vitro[J];Chinese Medical Journal;2007年09期

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本文編號：1617144