- 學位論文
紫蘇醛對高糖及硫酸吲哚酚誘導EA.hy926人類血管內皮細胞內皮間質轉化、內質網壓力依賴性細胞凋亡及血管新生作用之影響
Effects of perillaldehyde on high glucose and indoxyl sulfate-induced endothelial to mesenchymal transition and endoplasmic reticulum stress-dependent apoptosis and angiogenesis in EA.hy926 human vascular endothelial cells
摘要
糖尿病腎病變(diabetic nephropathy, DN)是糖尿病併發症之一，也是造成末期腎病(end-stage renal disease, ESRD)最常見因素。臨床研究顯示，在慢性腎病(chronic kidney disease, CKD)患者中，心血管疾病(cardiovascular disease, CVD)之發生率相當高。硫酸吲哚酚(indoxyl sulfate, IS)為一種會和蛋白質結合之親蛋白性尿毒素(protein-bound uremic toxin, PBUT)主要成分。已知IS可促使血管內皮細胞纖維化(fibrosis)、內質網壓力(endoplasmic reticulum stress, ER stress)所介導之細胞凋亡(apoptosis)，造成內皮細胞功能受損。然而IS對血管內皮細胞內皮間質轉化(endothelial to mesenchymal transition, EndMT)之影響亦仍不清楚。本研究利用EA.hy926人類血管內皮細胞探討紫蘇醛(perillaldehyde, PAH)對高糖及IS所誘發的EndMT、纖維化、內質網壓力介導之細胞凋亡及血管新生作用之影響。結果顯示，以30 mM葡萄糖合併0.5 mM IS處理細胞後，可顯著降低細胞生存率、VE-cadherin蛋白表現，同時誘發EndMT及纖維化相關蛋白如α-SMA、Slug、FAP；內質網壓力調控蛋白如p-PERK、p-eIF2α、CHOP及細胞凋亡蛋白如cleaved-caspase 3之表現。此外，30 mM葡萄糖合併0.5 mM IS處理細胞後亦顯著誘發α-SMA、FAP、CHOP、Bim、Bax/Bcl-2 mRNA表現、細胞核內Snail1之累積及細胞凋亡情形，預處理PAH則可抑制上述之現象。而在30 mM葡萄糖下，PAH可逆轉IS所抑制之血管形成作用(tube formation)。以CHOP siRNA將CHOP 基因knockdown或處理內質網壓力抑制劑4-phenyl butyric acid (4-PBA)可顯著減少IS所誘發cleaved-caspase 3表現。綜合上述結果，PAH可抑制高糖及IS所誘發之EA.hy926細胞EndMT、纖維化、內質網壓力依賴性細胞凋亡，以及恢復血管新生作用，以保護血管內皮細胞功能。
並列摘要
Diabetic nephropathy (DN) is considered to be one of the complications of diabetes, and it is the most common cause of end-stage renal disease (ESRD). Clinical studies indicate that a high incidence of cardiovascular disease (CVD) is observed in patients with chronic kidney disease (CKD). Indoxyl sulfate (IS), one of the major protein bound uremic toxins (PBUT), has been found to promote fibrosis, endoplasmic reticulum stress (ER stress)-mediated apoptosis in vascular endothelial cells, leading to the decline of endothelial cells function. However, the effects of IS on endothelial to mesenchymal transition (EndMT) in vascular endothelial cells is still unclear as well. In the present study, EA.hy926 human vascular endothelial cells were used to investigate the effects of perillaldehyde (PAH) on IS-induced EndMT, fibrosis, ER stress-mediated apoptosis and angiogenesis in the presence of high concentration of glucose. The results showed that 30 mM glucose combined with 0.5 mM IS can significantly decrease cell viability and VE-cadherin protein expression, whereas elevate the level of EndMT and fibrosis-related proteins such as α-SMA, Slug, FAP, and ER stress-related proteins such as p-PERK, p-eIF2α, CHOP as well as apoptosis-related protein such as cleaved-caspase 3. Moreover, 30 mM of glucose combined with 0.5 mM IS significantly increases the mRNA level of α-SMA, FAP, CHOP, Bim, Bax/Bcl-2, and the nuclear accumulation of Snail1, as well as apoptotic cell death, whereas these induction were suppressed by PAH treatment. Interestingly, PAH is able to reverse the suppression of tube formation by IS in the presence of 30 mM glucose in EA.hy926 cells. Knockdown of CHOP with CHOP siRNA or treatment with ER stress inhibitor -4-phenyl butyric acid (4-PBA) can significantly reduce IS-induced the expression of cleaved-caspase 3. Taken together, these results suggest that PAH can inhibit high glucose and IS-induced EndMT, fibrosis, ER stress-dependent apoptotic cell death and restore angiogenesis in EA.hy926 cells to protect vascular endothelial function.
參考文獻
延伸閱讀
- 黃萾芬（2007）。紅甘藷葉介入對經中強度運動後體內氧化壓力、細胞激素及熱休克蛋白表現之影響〔碩士論文，臺北醫學大學〕。華藝線上圖書館。https://www.airitilibrary.com/Article/Detail?DocID=U0007-1707200713362500
- Huang, C. Y. (2004). 烏蘇酸對第一介白質-β和腫瘤壞死因子-α所誘發的大鼠腦纖維癌瘤母細胞株轉移之抑制作用;利用Ames Test篩選四種茶類萃取物與五種多酚類化合物對PhIP和2-AAF致突變反應之抑制作用探討 [master's thesis, National Taiwan University]. Airiti Library. https://doi.org/10.6342/NTU.2004.00276
- 劉安正（2014）。吳茱茰鹼於惡性神經膠質細胞瘤U87-MG細胞誘發細胞凋亡與存活性細胞自嗜之分子訊息探討〔博士論文，臺北醫學大學〕。華藝線上圖書館。https://www.airitilibrary.com/Article/Detail?DocID=U0007-0502201420172300
- Predescu, D. N., Neamu, R., Bardita, C., Wang, M., & Predescu, S. A. (2012). Impaired Caveolae Function and Upregulation of Alternative Endocytic Pathways Induced by Experimental Modulation of Intersectin-1s Expression in Mouse Lung Endothelium. Biochemistry Research International, 2012(), 416-429. https://doi.org/10.1155/2012/672705
- Marino, F., Guasti, L., Tozzi, M., Schembri, L., Castiglioni, L., Molteni, E., Piffaretti, G., Castelli, P., & Cosentino, M. (2013). Gene Expression of Adhesion Molecules in Endothelial Cells from Patients with Peripheral Arterial Disease Is Reduced after Surgical Revascularization and Pharmacological Treatment. International Journal of Vascular Medicine, 2013(), 33-40. https://doi.org/10.1155/2013/412761