摘要
目的探讨肝活素(hepassocin,HPS)基因敲除对小鼠肝再生能力的影响。方法利用CRISPR/Cas9技术建立HPS基因敲除小鼠模型,分析小鼠体质量、肝质量及肝病理结构等。建立小鼠70%肝切除模型,收集肝切除后各个时间点肝组织,通过蛋白质免疫印迹(Western blotting,WB)检测p-HDAC3和PCNA的蛋白表达,利用免疫组化检测5-溴脱氧尿苷(Brd U)掺入率,及增殖细胞核抗原(PCNA)和Ki67的表达。采用1%CCl_4肝损伤模型,损伤后24 h,收集外周血及肝组织,检测ALT、AST和肝组织HE染色,利用不同浓度的CCl_4刺激小鼠肝实质细胞,检测ALT和LDH水平。结果 HPS^(-/-)小鼠与HPS^(+/+)小鼠相比体质量明显增加;HPS基因敲除会延缓70%肝切除手术后小鼠的肝再生并加重CCl_4诱导的急性肝损伤。结论 HPS基因敲除造成小鼠肝再生能力减弱,并加重CCl_4诱导的肝损伤,提示HPS在肝细胞增殖、抗损伤中发挥重要作用。
Objective To investigate the effects of hepassocin（ HPS） gene deficiency on liver development and liver regeneration after 70% liver hepatectomy. Methods An HPS knockout mouse model was established using CRISPR/Cas9 strategy. The body mass,liver mass,and the pathological structure of the liver of mice were analyzed. A 70% hepatectomy model was established in mice. Hepatic tissues at various time points after hepatectomy were collected. The protein levels of p-HDAC3 and proliferating cell nuclear antigen（ PCNA） were detected by Western blotting（ WB）. Brd U incorporation,PCNA and Ki67 expression were detected by immunohistochemistry. Using the 1% CCl_4 liver injury model,peripheral blood and liver tissue were collected 24 hours after injury. ALT, AST and hepatic HE staining were detected. Hepatic parenchymal cells were challenged with different concentrations of CCl_4 to detect ALT and LDH levels. Results HPS-/-mice had significant differences in body mass. HPS gene knockout resulted in a delayed liver regeneration in 70%hepatectomy. HPS knockout aggravated CCl_4-induced acute liver injury. Conclusion HPS gene knockout results in weakened liver regeneration in mice and aggravates CCl_4-induced hepatic injury,suggesting that HPS plays an important role hepatocyte proliferation and in the protection against liver injury.
作者
卜雯婧
陈慧
刘贤
王宪
孟晓竹
陈思聪
杜涵
杨晓明
于淼
BU Wen-jing;CHEN Htti;LIU Xian;WANG Xian;MENG Xiao-zhu;CHEN Si-cong;DU Han;YANG Xiao-ming;YU Miao(Graduate School, Anhui Medical University, Hefei 230032, China;National Center for Protein Seienees, State Key Lab of Proteomies, Institute of Radiation Medicine, Academy of Military Medical Sciences, Aeademy of Military Scienees, Beijing 100850, China;Beijing C＆N International Sei-Tech Co. , Ltd. , Beijing 102206, China;Air Force Military Medical University,Xi＇an 710032, China)
出处
《军事医学》
CAS
CSCD
北大核心
2018年第3期161-168,共8页
Military Medical Sciences
基金
北京市自然科学基金资助项目(7182124)
蛋白质组学国家重点实验室合作研究课题(SKLP-K201404)
