雨來菇藻多醣萃取物通過調節粒線體動態增加幹細胞功能來改善經由D-(+)-galactose誘導的心臟細胞老化

Abstract

背景：隨著年紀的上升，許多生理的功能衰退都與老化息息相關，如衰老會造成心臟和心血管系統功能衰退，故衰老是與心功能衰竭相關的病理過程之一。幹細胞具有很好的再生、分化、自我更新能力，因此幹細胞移植是治療老化心臟的治療策略之一。許多研究證明，雨來菇含有多種有益成分。從雨來菇中萃取出的多醣無毒，並具有調節生理功能，可以增強免疫功能、促進細胞增殖和抗氧化作用。目的：本研究旨在探討經雨來菇多醣萃取物(NCPE)預處理的間質幹細胞是否比未經雨來菇多醣萃取物預處理的幹細胞對老化心肌細胞是否具有更好的保護作用。方法：使用半乳糖(D-(+)-galactose)作為誘導心肌細胞老化模型，利用幹細胞移植(使用人類臍帶血間質幹細胞Human Wharton's jelly stem cells, WJSCs)，來當作心臟老化治療的方法，並使用NCPE強化WJSCs。 結果：經由細胞存活率測試(MTT assay)以及西方墨點法(Western blot)研究發現在10 g/L D-(+)-galactose誘導48小時會使心肌細胞老化(大量表現p53老化蛋白質)並且不會造成凋亡(細胞凋亡蛋白質Caspase 3不活化)。另研究顯示0.0003 μg/ml NCPE通過活化膜蛋白胰島素樣 C-X-C 趨化因子受體 4 (CXCR-4) 來增加WJSCs活性。更進一步將老化的心肌細胞與幹細胞共培養後發現WJSCs能有效治療老化的心肌細胞，並且經由NCPE預處理過的WJSCs其治療效果更好。 結論: 以0.0003 μg/ml NCPE通過活化膜蛋白CXCR-4增加幹細胞活性，提高幹細胞治療老化心臟的效果。未來可以利用NCPE預處理的幹細胞進行動物實驗，印證在細胞實驗所發現的機制。As age increases, many physiological functional declines are closely related to aging, especially the influence on the heart and cardiovascular system. Aging is one of the many pathologies associated with heart failure. Stem cells have good regeneration, differentiation, and self-renewal abilities. Therefore, stem cell transplantation is one of the therapeutic strategies for treatment of heart aging. Several studies have indicated that Nostoc commune contains various beneficial components. The polysaccharides extracted from Nostoc commune are non-toxic and contains physiological functions which can increase immune function, promote cell proliferation, and activate antioxidant effects.Aim: The aim of this study is to investigate whether mesenchymal stem cells pretreated with Nostoc commune extract have a better cardio-protection effect on aging heart than stem cells without the Nostoc commune extract pretreatment.Methods: D-(+)-galactose was used to induce aging of cardiomyocytes. Human Wharton’s jelly stem cells (WJSCs) were used for heart aging treatment, and Nostoc commune polysaccharide extract (NCPE) was used to enhance functions of WJSCs.Results: The results from MTT assay and Western blot analysis showed that cardiomyocytes induced with 10 g/L of D-(+)-galactose for 48 hours did increase aging marker p53, however, apoptotic marker caspase 3 did not show significance. These findings indicated that cardiomyocytes did age and did not undergo apoptosis. NCPE at the concentration of 0.0003 μg/ml can increase stem cell viability through the expression of membrane protein insulin-like C-X-C chemokine receptor type 4 (CXCR-4). After co-culture of aged cardiomyocytes and stem cells, p53 expression did decrease in MTT assay and Western blot, which indicated that WJSCs can cure aged cardiomyocyte. Furthermore, the effects are enhanced in WJSCs pretreated with NCPE.Conclusion: The concentration of 0.0003 μg/ml of NCPE can increase stem cell viability through CXCR-4 and enhance the effects of stem cells in aging heart. In the future, NCPE pretreated stem cells can be used in animal experiments and its mechanisms can be further investigated.