應用多光子自發螢光與二倍頻顯微影像技術於量化組織糖化現象

Imaging and Quantification of Tissue Glycation by Multiphoton Auto-fluorescence and Second Harmonic Generation Microscopy

曾若雅(Jo-Ya Tseng)

指導教授 ： 董成 淵

國立臺灣大學/理學院/物理研究所/碩士(2010年)

https://doi.org/10.6342/NTU.2010.00730

摘要

糖尿病是一種的內分泌紊亂的新陳代謝疾病，導致腎功能衰竭、失明、神經損傷和心血管疾病等慢性並發症。 而糖尿病併發症與還原糖和蛋白質之胺基所發生的梅納反應有密切的關係。梅納反應所產生不可逆性的糖化終產物，使蛋白質產生褐色、交聯和螢光。最近的研究發表中，非侵入式方法測量螢光，藉由螢光的強度來判讀組織中糖化終產物(AGEs)的累積程度。
本實驗利用多光子顯微鏡的優勢來提高診斷的可靠性，我們用不同部位的牛組織：眼角膜、皮膚的真皮層和大動脈，事先浸泡在0.5M的糖水中，來模擬糖尿病病人的組織。隨時間的變化，利用雙光子影像我們觀察到糖化組織有螢光增強及二倍頻訊號減弱的趨勢，並可以比較組織中膠原蛋白和彈性纖維的糖化速度。同時使用不同的激發波長來收雙光子螢光光譜，觀察到越短的激發波長能激發較多的螢光物質，組織糖化前後的光譜形狀也有所不同。
結語，雙光子顯微術在臨床上有診斷糖尿病的浅力和價值，希望未來藉由雙光子顯微術之優勢和AGEs之螢光作為非侵入性的光學檢測儀器。

關鍵字

多光子顯微術 ； 糖化 ； 膠原蛋白 ； 彈性蛋白 ； 糖尿病

並列摘要

Diabetes is a common endocrine disorder leading to chronic complications such as renal failure, blindness, nerve damage and vascular disease. The Maillard reaction is a process in which reducing sugars react spontaneously with amine-containing molecules and results in the irreversible production of advanced glycation end-products (AGEs) which play an important role in diabetic complication. Recent studies have shown that noninvasive spectroscopy offers a potential way of detection of diabetes by measuring the autofluorescence intensity of AGEs in the tissue.
In the present work the advantages of two-photon microscopy were used to improve the reliability of diagnosis. The two-photon autofluorescence (TPAF) and second harmonic generation (SHG) images of samples were obtained from bovine cornea, skin and aorta samples treated with 0.5 M ribose solution. Spectrally-resolved TPAF images were obtained for a range of excitation wavelengths.
Our results show that glycation resulted in increase of TPAF and decrease of SHG. The rates of glycation of collagen and elastin-rich tissues were derived from the changes of TPAF intensity and we found that the rate of glycation of collagen rich tissues exceeds that of elastin rich tissues. The maxima of spectra showed red shift depending on glycation level and excitation wavelength.
In conclusion, two-photon microscopy has the potential clinical implications for the diagnosis of diabetes and aging. The reported spectral features can be used in conjunction to conventional methods of non-invasive optical detection of AGEs.

並列關鍵字

multiphoton microscopy ； glycation ； collagen ； elastin ； diabetes

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