STZ誘發糖尿病小鼠之香瓜茄水萃物/間質幹細胞療法評估

Evaluation of treatment on streptozotocin induced mice diabetes by ppe/rats mesenchymal stem cells

馬金足(Chin-Tsu Ma)

指導教授 ： 郭士民

共同指導教授 ： 莊景文(Chi-Wen Chuang)

義守大學/電機資訊學院/電機工程學系/博士(2017年)

摘要

在本論文研究中嘗試利用兩種不同的方式來減輕糖尿病症狀或治療第2型糖尿病。首先，研究探討pepino polyphenolic extract（PPE）多酚提取物對糖尿病性神經病變的作用。PPE是一種多酚，其中含有香豆酰基和咖啡酰基衍生物。研究結果顯示PPE可以顯著改善糖尿病小鼠的血糖。經PPE治療後可以保留穀胱甘肽（GSH）水平、並減少硫代巴比妥酸反應物質（TBARS）水平、降低ROS、降低白細胞介素（IL）-6、腫瘤壞死因子（TNF）-α，以及果糖和糖化在糖尿病小鼠的坐骨神經中的晚期糖基化終末產物（AGE）的中間體和前體，例如甲基乙二醛（MG）和N-（羧甲基）賴氨酸（CML）。 PPE還具有改善糖尿病小鼠神經的作用-顯示促進許多小髓纖維束的分裂。 此外，PPE可以保持穀胱甘肽過氧化物酶（GPX）的蛋白和mRNA水平，並且減少坐骨神經中晚期糖基化終產物（RAGE）的醛糖還原酶（AR）和受體的表達。上述結果顯示PPE可以減輕糖尿病周圍神經中的氧化，炎症和糖原壓力。
由於間質幹細胞（MSCs）能夠分化成不同具功能性的細胞，因此，以此方式的細胞治療已成為組織修復與再生的應用潛力。本論文研究的第二部分即是探討結合第1型膠原蛋白奈米分子與菸鹼醯胺（NCT）和exendin-4（EX4）對MSCs分化成胰島素分泌細胞的影響，並評估此分化的胰島素分泌細胞對大鼠第2型糖尿病（T2DM）的治療效果。初步結果顯示，在結合第一型膠原蛋白奈米分與NCT，MSCs表達增加的Nkx6.1和早期PDX-1 mRNA表現，顯示MSCs能有效分化成具胰島素分泌的細胞。此外，添加第一型膠原蛋白奈米纖維可增加具胰島團簇之分化細胞數量。T2DM動物治療植入實驗中顯示：在植入NCT /第一型膠原蛋白奈米分子/ EX4所分化的細胞後，T2DM大鼠可恢復其正常的HbA1值，並顯示修復的胰島結構。簡言之，植入的細胞治療配方顯示可修復T2DM大鼠中被破壞的胰島，並能恢復葡萄糖的調節作用。

關鍵字

香瓜茄 ； 糖尿病 ； 間質幹細胞 ； 菸鹼醯胺

並列摘要

In this present study, we have utilized two different approaches tried to alleviate diabetes mellitus (DM) related stress or advanced to treat the DM. The effect of pepino polyphenolic extract (PPE) on diabetic neuropathy was examined first. PPE was demonstrated to contain coumaroyl and caffeoyl derivatives among polyphenols. The results demonstrated that PPE significantly improved glycaemic control in diabetic mice. These PPE treatments reserved the glutathione (GSH) level, and decreased the thiobarbituric acid reactive substances (TBARS) level, reactive oxygen species (ROS), interleukin (IL)-6, tumour necrosis factor (TNF)-alpha, fructose, and glycation intermediates and precursors of advanced glycation end products (AGEs), such as methylglyoxal (MG) and N-(carboxymethyl)lysine (CML), in the sciatic nerves of diabetic mice. PPE also had the effects in improving the nerves of diabetic mice, showing disorganization of the fascicle with numerous small myelinated fibers. The PPE can retain the protein and mRNA levels of glutathione peroxidase (GPX), and decrease the expressions of aldose reductase (AR) and the receptor for the advanced glycation end product (RAGE) in sciatic nerves. These findings support that pepino polyphenolic extract could attenuate oxidative, inflammatory and glycative stress in diabetic peripheral nerves.
Advances in mesenchymal stem cells (MSCs) and cell therapies have been promising approaches to treat (DM) since their potent capacities for differentiation into various functional cells match the demands of tissue repair and regeneration. The aim of this study is secondly to examine the effects of nano-sized type I collagen molecules in combination with nicotinamide (NCT) and exendin-4 (EX4) on MSCs differentiation into insulin-secreting cells in vitro and to evaluate their reparative effects against type 2 diabetes mellitus (T2DM) rat in vivo. The obtained results revealed that MSCs showed increased differentiation into insulin-secreting cells with higher mRNA expression for Nkx6.1 and early PDX-1 in the presence of NCT and nano-sized type I collagen molecules. Addition of nano-sized type I collagen fibrils increased morphologically islet-like clusters in differentiated cells. T2DM rats reverted to their normal HbA1 values and exhibited structurally repaired islets in the pancreas implanted with NCT/nano-sized collagen I molecule/EX4-incubated differentiated cells. In short, the combined recipe showed reparative actions on the destructive islet of Langerhans in the pancreas coupled with glucoregulatory effects in T2DM rats in vivo.

並列關鍵字

pepino ； diabetes mellitus ； mesenchymal stem cells ； nicotinamide ； Exendin-4

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