(精选)microRNA 作用教学课件.pptVIP

Using anti-atherosclerosis microRNA for gene therapy 卓夙航 醫學遺傳學科 臨床醫學研究部 Atherosclerosis Atherosclerosis causes a restricted blood flow to vital organs. The cause of atherosclerosis: ?lack of physical activity ?smoking ?unhealthy eating ?age ?family history of heart disease U.S. Department of health human Services. National Institutes health 95年台灣十大死因統計 Drug discovery for atherosclerosis Notoginseng or sanchi, has a long history as a remedy in Oriental traditional medicine. ●treating trauma and bleeding from internal or external injury ●promoting blood circulation ●reducing blood clotting ●alleviating pain, especially cancer pain Therapy for Atherosclerosis Screening model for atherosclerosis(1) First screen Drug: Notoginseng crude extract Positive drug: Simvastatin Stimulator: Oxidized-LDL Cell model: Human aortic smooth muscle cell (HASMC) Method: WST-1 test for anti-proliferative activities of drug 40μg/ml of ox-LDL treatment (24hr) Control 20 40 60 80 100 ox-LDL (μg/ml) ? ? ? ? ? Screening model for atherosclerosis(1) Cell proliferation assay (MTT method) Screening model for atherosclerosis (2) Notoginseng Second screen Drug: Three different extracts from notoginseng crude extract Searching for new mechanisms for 三七 anti-atherosclerosis effect microRNA Array for screening ●High-throughput screening of microRNA array (667 microRNA) ●Performing gene quantitation (real-time RT-PCR) Differential expression of miR from array Ox-LDL treatment Ox-LDL plus Notoginseng treatment Performing Reverse Transcription (cDNA generation) Running the miR array (PCR) Real time data analysis Interpreting results RQ or 1.5 fold change 570 miR-2xx miR-1xxb miR-2xx miR-4xx miR-4xx miR-4xx miR-5xx miR-4xx miR-1xx miR-2xx 2.18 1.81 1.76 1.64 1.46 1.41 1.39 1.23 1.14 1.10 Fold change Top 10 Up regulation MicroRNA expression profile of HASMC after Notoginseng treatment miR-1xx miR-3xx miR-2xa miR-1xb