Memandangkan dah meluat nk review journal..nak x nak kena buat jgk...so aku arini aku try gabungkan dengan minat berblogging ni dgn review...baru smngt sket mo wat revision..hahaha!
Publication :
Molecular and Cellular Biochemistry 262: 165-176, 2004.
Author:
Jingxiang Bai and Arthur I.Cederbaum
Deparment of Pharmacology and Biological Chemistry, Mount Sinai School of Medicine, New York, USA
(Normally klu publisher tu dari USA..journal tu mmg boleh dipercayai..memandangkan byk jg researcher kuat "kencing" psl research dorg...so berbaloi baca journal ni..pilihan journal sebagai rujukan amat penting!)
Title: Adenovirus mediated overexpression of CYP2E1 increases sensitivity of HepG2 cells to acetaminophen induced cytotoxicity
Idea:
Boleh dijadikan rujukan sbb aku pun buat pasal
dengue&chang liver cells&acetaminophen aka PANADOL. Persamaannya..kitorg sm2 guna virus dan guna drugs yg sama itu acetaminophen.
Q: Tahukah anda PANADOL itu boleh menyebabkan kerosakan hati (hepatotoxicity)??? jika diambil lebih dari dos yg disarankan...tp ada gak kes melibatkan hepatotoxicity wpun pd dos yg rendah...itu kerana hati tersebut telah berisiko seblm tu (mungkin disebabkan pengambilan alcohol)..kira Pan ni akan menambahkan lagi risiko tersebut..so watch out!!! Makan PAN ketika perlu sahaja ^__^
METHOD:
Cell culture & infection
1) HepG2 were cultured in 10% MEM in 5%C02 at 37C. (routine cell culture)
2) Cells were seeded onto plate and grown to 60% confluence (normally, researcher akan incubate for at least 16hours incubation...)
3) Cells were then infected with Ad-CYP2E1 100 m.o.i (adenoviurus containing CYP2E1) for 48hours.
(kes aku lak letak dengue virus for all serotypes).
4) After 48hrs, cells were collected and assayed for CYP2E1 expression or treated with PAN
(biasanya, cells akan diinfeksi selama 1 jam sahaja dan kemudian diikuti dgn treatment)
Trypan blue exclusion assay (salah satu assay plg simple utk determine cell viability)- murah dan basic
To determine the cell viability, HepG2 cells were treated with different amounts of PAN after infection with Ad-CYP2E1. (0, 2.5,5,10, and 15 mM).
2) After 36hours, all cells were collected by trpsinized followed by centrifugation.
(basic utk detach cells dari permukaan plate dengan meletakkan trypsin....centrifugation adalah utk mendapatkan cells pellet sahaja)
3) Cells were resuspended in PBS and stained with trypan blue solution.
(langkah perwarnaan cells..biasa guna nisbah 1:1 (10ul of cells&10ul of TB)...so dilution factor 2/1= 2.
4) The number of cells excluding/staining positively for uptake of TB were counted under light microscope.
(prin: cells hidup tidak akan menerima colour TB, cells yg berwrna biru akan adalah cells mati)
5) Cell viability was expressed as percentage of cells excluding tyrpan blue out of the total cells.
(Cells hidup/jumlah cells x 100= cell viability (%), jumlah cells (cell hdp+cell mati)
Calculation = (average calculated cells x 2 x 10^4). etc. living cells = 1.5x10^4
Main point sini:
Choose only 1 the best concentration of PCM (0,5,10,15,20) - selamat digunakan
Toink2x...*__*
Bersambung....
