第一部分：龍葵水萃物抑制乳癌細胞凋亡；第二部分：蘋果多酚對STZ 誘導糖尿病大白鼠腎病變之影響

PART Ⅰ:Aqueous Extract of Solanum Nigrum Caused　Apoptosis in Human Breast Cancer Cell.PART Ⅱ:The Ameliorative Effect of Apple Polyphenol on Streptozotocin Induced Diabetic Nephropathy

王旨吟(Chih-Ying Wang)

指導教授 ： 李彗禎

中山醫學大學/醫學院/生化暨生物科技研究所/碩士(2010年)

https://doi.org/10.6834/CSMU.2010.00074

摘要

第一部分：乳癌在台灣的死亡統計中排名第五名。龍葵，學名Solanum nigrum L.，始載於《唐本草》，在台灣常為草藥被使用。龍葵水萃物具有抗氧化、鎮痛解熱及抗發炎作用、保肝作用和抗癌作用。在最近的研究指出，龍葵水萃物具有抑制肝癌細胞的生長及肝硬化，但對於龍葵使是否影響乳癌尚未清楚。本篇研究探討龍葵水萃物在乳癌之增生和細胞凋亡調節的作用。在細胞增生的試驗中發現，龍葵水萃物會抑制人類乳癌細胞 (MDA-MB-231 cells) 的生長，其IC50值約為 3 mg/mL。在流式細胞儀分析DNA含量的試驗中，給予1、3、5 mg/mL 的龍葵水萃物培養24小時後，結果發現龍葵水萃物會引起細胞死亡的濃度為5 mg/mL的濃度，此濃度會使得sub G1增加90%。在 DAPI stain 以及流式細胞儀分析的結果顯示，龍葵水萃物會造成細胞凋亡而不影響其細胞週期。在西方點墨的結果顯示，龍葵水萃物能有效的影響 pro-apoptotic 蛋白 Bax 和 Bak 及 anti-apoptotic 蛋白 Bcl-xL 和 Bcl-2 的表現，顯示龍葵水萃物是藉由調節 Bcl-2 family 蛋白的表現來引發細胞凋亡。綜合以上實驗的結果顯示，龍葵水萃物能有效誘導乳癌細胞的凋亡，但對於龍葵水萃物是否可當作抗癌藥物需要再作更深入的評估其抗癌的功效。
第二部分：糖尿病為國內十大死因之一。糖尿病所引發的腎衰竭，又稱為糖尿病腎病變，已經成為全球嚴重的健康為題。近來研究指出糖尿病之併發症和氧化壓力有關係，而糖尿病腎病變會造成腎絲球肥大、腎間質增生及腎絲球基底膜肥厚。因此，本研究想要探討抗氧化劑是否能減緩氧化壓力所造成的糖尿病腎病變。先前文獻指出，蘋果多酚具有抗過敏、抗氧化、抗癌、預防心血管疾病以及美白功效。因此本實驗利用蘋果多酚降低糖尿病誘導腎病變之功效。以 65mg/kg 鏈佐菌素 (streptozotocin，STZ)c 注射老鼠，誘發成為第一型糖尿病老鼠。誘發糖尿病後，每日依飲食量給予 0.5%、1% 及 2% 蘋果多酚餵食10週。實驗結果顯示，蘋果多酚可以改善早期腎病變所引發的高微白蛋白及肌肝酸的濃度，在四週及十週分別降低 78% 及 69%。蘋果多酚也可降低三酸甘油脂，在四週可以降低 51%。給予蘋果多酚亦可降低血中尿素氮和血清肌酸酐比值，在第十週時可降低 45%。此外，在腎臟組織中亦發現蘋果多酚能降低氧化壓力的活性還可以減少糖尿病老鼠血液中的丙二醛。在腎臟組織切片發現，蘋果多酚可以減緩糖尿病腎病變所造成的腎絲球基底膜肥厚。綜合以上實驗的結果顯示，蘋果多酚可以改善糖尿病所造成的腎功能損傷。

關鍵字

乳癌 ； 凋亡 ； 糖尿病 ； 腎病變

並列摘要

PARTⅠ:Breast cancer is the 5th position of death causes in Taiwan. The water extract of Solanum nigrum (SNWE) possess the abilities in antioxidation, anti-inflammation, antipyretic effects, and anti-cancer. Recent studies show that SNWE inhibit hepatoma and liver cirrhosis, but there is no available information to address the effect of SNWE in breast cancer. In this study, SNWE is explored to inhibit growth of MDA-MB-231, a human breast cancer cell line, with an IC50 value of 3 mg/mL. Flow cytometric analysis for the DNA content in SNWE-treated cell indicated that following SNWE (1, 3, 5 mg/mL) treatment for 24 hr, the result showed that SNWE causes sub G1 increase to 90 %in the concentrations of 5 mg/mL but not regulates cell cycle. SNWE can significantly affect the levels of pro-apoptotic, Bax and Bak, and anti-apoptotic proteins, Bcl-xL and Bcl-2. Taken together, SNWE can induce apoptosis in breast cancer cells via regulating Bcl-2 family proteins. After clarifying the detail mechanism, SNWE might has the potential to apply as a cancer chemopreventive agent.
PART Ⅱ:Diabetes mellitus (DM) is one of the top ten lethal diseases in Taiwan. Diabetic nephropathy is a major complication to cause death in DM patients. Recent studies showed that high glucose caused reactive oxygen species (ROS) possessed important relationship between DM and its complications. The microscopic finding of diabetic nephropathy includes glomerular hypertrophy, mesangial matrix expansion, and glomerular basement membrane thickening. Therefore, the usage of antioxidative materials might elevate antioxidation status to decelerate the development of diabetic nephropathy. Previous studies showed apple polyphenol (AP) possesses anti-allergic, anti-oxidation, anti-cancer, prevent cardiovascular disease and bleach effect. In this study, we further examined the effect of AP on anti-nephropathy. In Sprague-Dawley (SD) Rat experiment, 65mg/kg of streptozotocin (STZ) induce type I DM. After completing the induction, 0.5%, 1% or 2% of AP was given with water for further 10 weeks.。The results showed that AP improved urine albumin/creatinine ratio (ACR, an early stage of diabetic nephropathy), which was decrease to 78% at 4 weeks and 69% at 10 weeks in diabetic rats. AP significantly decrease TG to 51% in diabetic rats at 4 weeks and BUN/Creatinine ratio to 45% at 10 weeks. AP also has the ability to improve the antioxidative condition and decreased reactive oxygen species activity significantly in kidney specimen and showed that AP decreased MDA level in serum. In histological examination, AP reduces the thickness of glomerular basement membrane in diabetic rats. These results implied that AP can ameliorate diabetic nephropathy progression.

並列關鍵字

Breast cancer ； Apoptosis ； Diabetic ； Nephropathy

參考文獻

行政院衛生署97年死亡統計上卷

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