Antifibrotic effects of ZK14, a novel nitric oxidedonating biphenyldicarboxylate derivative, on rat HSC-T6 cells and CCI4-induced hepatic fibrosis

• Published in: Acta pharmacologica Sinica no. 1; pp. 27 - 34
• Main Author: Li DAI Hui JI Xiang-wen KONG Yi-hua ZHANG
• Format: Journal Article
• Language: English
• Published: 2010
• Subjects: HSC; 四氯化碳; 联苯双酯; 肝纤维化; 衍生物
• ISSN: 1671-4083; 1745-7254

Aim： To study the pharmacologic effect of ZK14, a novel nitric oxide-donating biphenyldicarboxylate （DDB） derivative, on HSC-T6 cells and on CCI4-induced hepatic fibrosis. Methods： Inhibition of HSC-T6 cell growth by ZK14 was evaluated by MTT assay. The effect of ZK14 on the percentage of HSC-T6 cells undergoing apoptosis was measured using Annexin-V/PI double-staining and TUNEL assay. Mitochondrial membrane potential （MMP） and caspase activities were tested. Hepatic fibrosis was induced in Sprague-Dawley rats by intraperitoneal injection with 14% CCI4. Rats with hepatic fibrosis were randomly divided into four groups： model control, ZK14 （20 mg/kg）, ZK14 （10 mg/kg） and DDB （5 mg/kg）. Levels of aspartate aminotransferase （AST）, alanine aminotransferase （ALT）, hyaluronic acid （HA）, type Ⅲ collagen （PCⅢ）, and nitric oxide （NO） were assessed, and liver samples were stained with hematoxylin-eosin. The NO level in cells treated with ZK14 in vitro was also measured. Results： The effect of ZK14 on HSC-T6 cell apoptosis was concentration- and time-dependent, with up to 50% of cells becoming apoptotic when exposed to 100 μmol/L ZK14 for 18 h. ZK14 treatment resulted in mitochondrial membrane depolarization and activation of caspases 3 and 9. At a dose of 20 mg/kg, ZK14 significantly decreased serum transaminase （AST, ALT） activities and fibrotic index （HA, PCⅢ） levels and significantly inhibited fibrogenesis. Conclusion： These data indicate that ZK14, a novel NO-donating DDB derivative, promotes HSC-T6 apoptosis in vitro through a signaling mechanism involving mitochondria and caspase activation and it inhibits CCl4-induced hepatic fibrosis in vfvo. The results suggest that ZK14 has potential therapeutic value in the treatment of hepatic fibrosis.