| 摘要註: |
大腦內產生過多的ROS (reactive oxygen species)已被證實是造成許多神經退化疾病的主要因素。由於腦內過多的ROS會損害蛋白質、核酸和膜上的多元不飽和脂肪酸(polyunsaturated fatty acid)，導致脂肪酸會被過氧化(peroxidation)，造成細胞膜完整性的喪失，甚至細胞凋亡。SNP (Sodium nitroprusside dehydrate)為一種可產生過多ROS的化學藥劑。將SNP加入Neuro-2a細胞處理12小時後造成neurite長度變短變細、分支數變少，增生(proliferation)降低，而p-Akt、Bcl-xL、SOD1、catalase蛋白表現量下降，cleaved caspase 3、p-P53、P38 MAPK表現量卻增加，這些情形都證明SNP對Neuro-2a有傷害性。白藜蘆醇 (Resveratrol，簡稱Res)具有抗癌、抗氧化、抗發炎的效力。將白藜蘆醇加入細胞處理12小時後造成neurite長度變長、neurite數變多，提高細胞增生，Bcl-2蛋白表現量等都有顯著變佳的狀態；p-Akt，p-P53，SOD1蛋白表現量也有大幅增加；cleaved caspase 3、Bcl-xL、catalase、p38 MAPK蛋白表現量些微增加。這些結果都證明Res對Neuro-2a細胞有益處。將SNP與白藜蘆醇共同處理12小時後，發現加入白藜蘆醇的組別neurite長度、neurite數都恢復成正常，增生也恢復正常，而p-Akt蛋白表現量顯著性的上升；catalase及p-P53蛋白表現量有增加；而且cleaved caspase 3蛋白表現量也顯著性的減少；Bcl-xL及SOD2些微增加；SOD1、Bcl-2及p38 MAPK蛋白表現量亦呈現些微減少，也都證明白藜蘆醇對SNP所造成的傷害有復原的效用。根據以上結果，一個in vitro的在Neuro 2a細胞中以SNP造成ROS傷害及可被Res回復的系統被建立。此系統將可提供一個簡單快速去檢測抑制ROS生成或加速代謝的有效藥物、中草藥或天然物的方法。 The main factors which cause neurodegenerative diseases have been proved having lots of ROS (reactive oxygen species) in brain. ROS in brain will damage protein, nucleic acids and polyunsaturated fatty acid. Then, damaged unpolysaturate would be peroxidated, lose cell membrane integrity, and even induce appotosis. SNP (Sodium nitroprusside dehydrate) is a chemical agent of which producing accumulation of ROS in cells. Neuro-2a cells, treating with SNP for 12 hours,appear retracting & diminishing of the neurite length, lessening of neurite number, reducing of cell proliferation, decreasing of protein expression, such as p-Akt, Bcl-xL, SOD1, catalase dramatically. Therefore, this treatments also increase cleaved caspase 3, p-P53 and p38 MAPK significantly. Those results demonstrate that SNP is harmful to Neuro-2a cell. Resveratrol (abbreviated to Res) possesses abilities of anti-cancer, anti-oxidant and anti-inflammatory. Neuro-2a cells, treating with Res for 12 hours, show elongation of neurite length, rising of neurite number and cell proliferation and increasing Bcl-2 obviously. Then, p-Akt, p-P53 and SOD1, also increase a lot. Expression of cleaved caspase 3, Bcl-xL, catalase and p38 MAPK improve slightly. The data shows that Resveratrol benefits on Neuro-2a cell. Neuro-2a cells, treating with both for 12 hours,results in restoring the neurite length and branch number, making proliferation back to normal, boosting p-Akt significantly and increasing expression of p-P53 and catalase largerly. Nevertheless, cleaved caspase 3 descends greatly. Bcl-xL, SOD2 raise lightly. SOD1, Bcl-2 and p38 MAPK decrease a little bit only. Those results prove that Resveratrol will recover Neuro-2a cells from SNP damage. According to all of results, system in vitro about SNP causing ROS damage and restoring from Res was established. This system will offer an easy and quick assy for drugs, herb, or natural products which might inhibit ROS production or accelerate ROS metabolism. |