AIM: To investigate the protective effect of stronger neo-minophafen C (SNMC) on fulminant hepatic failure (FHF) and its underlying mechanism. METHODS: A mouse model of FHF was established by intraperitoneal injection...AIM: To investigate the protective effect of stronger neo-minophafen C (SNMC) on fulminant hepatic failure (FHF) and its underlying mechanism. METHODS: A mouse model of FHF was established by intraperitoneal injection of galactosamine (D-Gal N) and lipopolysaccharide (LPS). The survival rate, liver function, inflammatory factor and liver pathological change were obtained with and without SNMC treatment. Hepatocyte survival was estimated by observing the stained mitochondria structure with terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate fluorescence nick end labeling (TUNEL) method and antibodies against cytochrome C (Cyt-C) and caspase-3. RESULTS: The levels of plasma tumor necrosis factor alpha (TNF-α), nitric oxide (NO), ET-1, interleukin-6 (IL-6), and the degree of hepatic tissue injury were decreased in the SNMC-treated groups compared with those in the model group (P < 0.01). However, there were no differences after different dosages administered at different time points. There was a significant difference in survival rates between the SNMC-treated groups and the model group (P < 0.01). The apoptosis index was 32.3% at 6 h after a low dose of SNMC, which was considerably decreased from 32.3% ± 4.7% vs 5% ± 2.83% (P < 0.05) to 5% on d 7. The expression of Cyt-C and caspase-3 decreased with the prolongation of therapeutic time. Typical hepatocyte apoptosis was obviously ameliorated under electron microscope with the prolongation of therapeutic time. CONCLUSION: SNMC can effectively protect liver against FHF induced by LPS/D-Gal N. SNMC can prevent hepatocyte apoptosis by inhibiting inflammatory reactionand stabilizing mitochondria membrane to suppress the release of Cyt-C and sequent activation of caspase-3.展开更多
目的:建立暴发性肝功能衰竭(FLF)小鼠模型并探讨复方甘草酸苷(SNM C)的保护作用及可能的作用机制。方法:采用D-氨基半乳糖(D-G a ln)和脂多糖(LPS)一次性腹腔注射构建FLF小鼠模型。利用光镜、电镜观察小鼠肝损伤情况;应用末端脱氧核苷...目的:建立暴发性肝功能衰竭(FLF)小鼠模型并探讨复方甘草酸苷(SNM C)的保护作用及可能的作用机制。方法:采用D-氨基半乳糖(D-G a ln)和脂多糖(LPS)一次性腹腔注射构建FLF小鼠模型。利用光镜、电镜观察小鼠肝损伤情况;应用末端脱氧核苷酸转移酶介导的dUTP缺口末端标记法(TUNEL)检测肝细胞原位凋亡情况;应用免疫组化法分别检测肝组织中细胞色素C和天冬氨酸特异性半胱氨酸蛋白酶-3(caspase-3)的表达。结果:D-G a ln加LPS可成功构建FLF小鼠模型。光镜及电镜可见肝细胞大量凋亡,且凋亡情况随治疗时间的延长明显改善。TUNEL检测结果表明,随着SNM C治疗时间的延长,凋亡指数逐渐降低。细胞色素C和caspase-3在模型组细胞阳性表达明显增加,随SNM C治疗时间延长表达逐渐减少。结论:利用D-G a ln和LPS可以构建理想的FLF小鼠模型。SNM C能有效抑制小鼠FLF模型中的肝细胞凋亡。SNM C可能通过稳定线粒体膜抑制细胞色素C释放及其随后caspase-3活化,从而阻断肝细胞凋亡的进行。展开更多
基金Supported by the Key Program During the Tenth Five-Year Plan of HeilongJiang Province, No. 200101031-00
文摘AIM: To investigate the protective effect of stronger neo-minophafen C (SNMC) on fulminant hepatic failure (FHF) and its underlying mechanism. METHODS: A mouse model of FHF was established by intraperitoneal injection of galactosamine (D-Gal N) and lipopolysaccharide (LPS). The survival rate, liver function, inflammatory factor and liver pathological change were obtained with and without SNMC treatment. Hepatocyte survival was estimated by observing the stained mitochondria structure with terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate fluorescence nick end labeling (TUNEL) method and antibodies against cytochrome C (Cyt-C) and caspase-3. RESULTS: The levels of plasma tumor necrosis factor alpha (TNF-α), nitric oxide (NO), ET-1, interleukin-6 (IL-6), and the degree of hepatic tissue injury were decreased in the SNMC-treated groups compared with those in the model group (P < 0.01). However, there were no differences after different dosages administered at different time points. There was a significant difference in survival rates between the SNMC-treated groups and the model group (P < 0.01). The apoptosis index was 32.3% at 6 h after a low dose of SNMC, which was considerably decreased from 32.3% ± 4.7% vs 5% ± 2.83% (P < 0.05) to 5% on d 7. The expression of Cyt-C and caspase-3 decreased with the prolongation of therapeutic time. Typical hepatocyte apoptosis was obviously ameliorated under electron microscope with the prolongation of therapeutic time. CONCLUSION: SNMC can effectively protect liver against FHF induced by LPS/D-Gal N. SNMC can prevent hepatocyte apoptosis by inhibiting inflammatory reactionand stabilizing mitochondria membrane to suppress the release of Cyt-C and sequent activation of caspase-3.
文摘目的:建立暴发性肝功能衰竭(FLF)小鼠模型并探讨复方甘草酸苷(SNM C)的保护作用及可能的作用机制。方法:采用D-氨基半乳糖(D-G a ln)和脂多糖(LPS)一次性腹腔注射构建FLF小鼠模型。利用光镜、电镜观察小鼠肝损伤情况;应用末端脱氧核苷酸转移酶介导的dUTP缺口末端标记法(TUNEL)检测肝细胞原位凋亡情况;应用免疫组化法分别检测肝组织中细胞色素C和天冬氨酸特异性半胱氨酸蛋白酶-3(caspase-3)的表达。结果:D-G a ln加LPS可成功构建FLF小鼠模型。光镜及电镜可见肝细胞大量凋亡,且凋亡情况随治疗时间的延长明显改善。TUNEL检测结果表明,随着SNM C治疗时间的延长,凋亡指数逐渐降低。细胞色素C和caspase-3在模型组细胞阳性表达明显增加,随SNM C治疗时间延长表达逐渐减少。结论:利用D-G a ln和LPS可以构建理想的FLF小鼠模型。SNM C能有效抑制小鼠FLF模型中的肝细胞凋亡。SNM C可能通过稳定线粒体膜抑制细胞色素C释放及其随后caspase-3活化,从而阻断肝细胞凋亡的进行。
