紅光與近紅外光光照對大鼠骨髓間葉幹細胞增生影響之比較及機制探討

Comparative Study of the Effects on the Proliferation and Its Mechanism of Mesenchymal Stem Cells Derived from Rat Bone Marrow by Red Light and Near Infrared Light Irradiation

吳家隆(Jia-Lung Wu)

指導教授 ： 李文婷

中原大學/工學院/醫學工程研究所/碩士(2010年)

https://doi.org/10.6840/CYCU.2010.00601

摘要

低能量光照刺激 (Low level light irradiation, LLLI) 能引發生物效應，進行生物調節作用，以達到修復組織及減緩發炎反應的效果，然而不同組織間最能發揮效用之光照參數以及光對生物組織之間的作用機制仍不甚了解，因此有必要針對最佳刺激參數下之作用機制深入探討。本研究採用先前本實驗室所獲得的最佳光照參數-紅光630 nm (15 mW/cm2, 4 J/cm2) 及近紅外光850 nm (10 mW/cm2, 4 J/cm2)，比較紅光及近紅外光刺激對體外培養之大鼠骨髓間葉幹細胞增生及其促進增生之機制。結果發現兩組刺激參數均能促進幹細胞增生，第 7 天時近紅外光實驗組的細胞數比控制組增加 43.48 %，紅光實驗組則增加 20.95 %；在細胞群落數目方面，近紅外光組的群落數目比紅光多 17.38 %。在活性氧分子 (ROS) 方面則有迥然不同的結果，近紅外光組使 ROS 產量提高 36.9 %，但紅光組則抑制 27.5 %。在基因表現方面，紅光與近紅外光皆不明顯活化熱休克蛋白 70；而原致癌基因 c-fos 在紅光與近紅外光刺激過後 0.5 小時達到最大表現量，並隨即在刺激後約 1 小時回到原來的水平；在 cyclin D1 表現方面，紅光光照在刺激後 1 小時表現最高，而近紅外光光照則可在刺激後立即提升 cyclin D1 的表現。紅光在光照後 6 小時會明顯提升 SOD1 與 SOD2 的表現，而近紅外光在光照後 1 小時可顯著增加 SOD1 表現，SOD2 則在刺激結束後隨即明顯表現。ALP 表現則不受紅光與近紅外光影響。粒線體膜電位在受光照後，皆有明顯提升；而細胞週期分析顯示兩光源皆可促進幹細胞進入 S 期。本研究目前只針對各種可能之增生路徑做初步偵測，因此仍需更深入探討其完整作用機制，期望能釐清低能量光照刺激對大鼠間葉幹細胞增生之影響及其作用機制，以提升光照刺激應用於幹細胞療法之潛能。

關鍵字

發光二極體 ； 間葉幹細胞 ； 紅光 ； 近紅外光 ； 低能量光照

並列摘要

Low level light irradiation (LLLI) has been shown to elicit biostimulatory and biomodulatory effects, which can enhance tissue repair and decrease inflammation. However, it remains unclear regarding the optimal irradiation parameters as well as the mechanism related photomodulation for different tissues. It is important to perform the mechanistic study under the optimal LLLI parameters. In this study, the optimal LLLI parameters obtained previously, 630 nm irradiation at 15 mW/cm2 and 4 J/cm2 as well as 850 nm irradiation at 10 mW/cm2 and 4 J/cm2, had been used to stimulate mesenchymal stem cells (MSCs) derived from rat bone marrow. The effect on the proliferation and related mechanism were studied after MSCs received red light and near-infrared (NIR) irradiation. The result showed that both red light and NIR irradiation could stimulate MSCs proliferation. The number of MSCs with NIR illumination increased 43.48 % while the one with red light stimulation increased 20.95 % compared to non-irradiated control group at the seventh day post-irradiation. The potential of colony-forming unit-fibroblast (CFU-F) was also promoted by LLLI. The number of CFU-F in NIR group was 17.38 % higher than that in red light group. The level of reactive oxygen species (ROS) in NIR group increased by 36.9 % while red light reduced by 27.5 %. The results from gene expression study demonstrated that neither red light nor NIR irradiation led to hsp70 expression. As to the proto-oncogene c-fos expression, maximal expression was observed at 30 minutes while the expression leveled off at 1 hour post-irradiation. Red light group exhibited maximal cyclin D1 expression level at 1 hour. Unlike red light groups, NIR irradiation could raise the cyclin D1 expression immediately after illumination. Six hours after the red light irradiation, SOD1 and SOD2 expression were significantly enhanced. The SOD1 expression was significantly increased at 1 hour, while SOD2 expression was elevated immediately after near-infrared light stimulation. ALP expression was not significantly affected by both red light and near-infrared irradiation. Mitochondrial membrane potential was improved significantly, while the cell cycle of MSCs was promted to enter the S phase after light exposure. The study provides preliminary data regarding the possible pathway leading to MSCs proliferation. Further investigation is required to shine a light on the mechanism related to MSCs proliferation before LLLI can be applied as an alternative to enhance the efficacy of stem cell therapy.

並列關鍵字

red light ； light-emitting diode ； low level light irradiation ； mesenchymal stem cell ； near-infrared

參考文獻

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