siRNA沉默c-jun基因对肝硬化大鼠肝切除术后肝再生的影响及其机制
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摘要
目的探讨si RNA沉默c-jun基因对肝硬化大鼠肝切除术后肝再生的影响及其机制。方法建立肝硬化大鼠模型,采用随机数字表法将50只肝硬化大鼠随机分为治疗组和对照组,每组25只。治疗组行肝大部分切除术的同时门静脉内注射慢病毒包装的Pcmv6-AC-GFP/c-jun-si RNA质粒。对照组行肝大部分切除术的同时门静脉内注射PBS。两组大鼠于术后14 d检测门静脉压力、血清ALT及肝重量体重比,采用Western blot法检测肝脏c-jun和增殖细胞核抗原(PCNA)蛋白相对表达量。两组间的数据比较采用t检验。结果治疗组术后平均门静脉压力为(1.18±0.11)k Pa,明显小于对照组的(1.67±0.24)k Pa(t=-26.74,P<0.05)。治疗组术后ALT为(43±5)U/L,明显低于对照组的(257±14)U/L(t=-31.11,P<0.05)。治疗组术后肝重量体重比为(6.94±0.31)%,明显大于对照组的(2.76±0.14)%(t=23.57,P<0.05)。治疗组术后c-jun和PCNA蛋白相对表达量分别为0.143±0.014、0.195±0.027,对照组相应为0.742±0.057、0.029±0.003,差异有统计学意义(t=-37.17,14.86;P<0.05)。结论慢病毒负载的c-jun-si RNA可能通过下调肝细胞c-jun,上调PCNA蛋白表达来促进肝再生,改善肝硬化大鼠肝切除术后肝功能。
Objective To investigate the influence and mechanism of c-jun gene silenced by si RNA on liver regeneration in liver cirrhotic rats after hepatectomy. Methods Liver cirrhotic rat models were established and 50 liver cirrhotic rats were randomized into the treatment group and the control group by random number table method with 25 rats in each group. The rats in the treatment group underwent major hepatectomy and were administered with lentivirus carrying plasmid Pcmv6-AC-GFP/ c-jun-si RNA by intraportal injection, while the rats in the control group underwent major hepatectomy and were administered with phosphate buffer saline(PBS) by intraportal injection. The portal pressure, serum ALT and liver weight/body weight of both groups were tested 14 d after surgery. The relative expression of c-jun in the liver and proliferating cell nuclear antigen(PCNA) protein was detected by Western blot. The data between the treatment group and the control group were compared using t test. Results The average portal pressure of the treatment group after surgery was(1.18±0.11) k Pa, which was significantly lower than(1.67±0.24) k Pa of the control group(t=-26.74, P<0.05). ALT of the treatment group after surgery was(43±5) U/L, which was significantly lower than(257±14) U/L of the control group(t=-31.11, P<0.05). Liver weight/body weight of the treatment group after surgery was(6.94±0.31)%, which was significantly greater than(2.76±0.14)% of the control group(t=23.57, P<0.05). The relative expression of c-jun and PCNA protein of the treatment group after surgery was respectively 0.143±0.014 and 0.195±0.027, and those of the control group was respectively 0.742±0.057 and 0.029±0.003, where significant difference was observed(t=-37.17, 14.86; P<0.05). Conclusion Lentivirus carrying c-jun-si RNA may promote liver regeneration and improve live function in cirrhotic rats after hepatectomy by down-regulating the c-jun in hepatocyte and up-regulating the PCNA protein.
Objective To investigate the influence and mechanism of c-jun gene silenced by si RNA on liver regeneration in liver cirrhotic rats after hepatectomy. Methods Liver cirrhotic rat models were established and 50 liver cirrhotic rats were randomized into the treatment group and the control group by random number table method with 25 rats in each group. The rats in the treatment group underwent major hepatectomy and were administered with lentivirus carrying plasmid Pcmv6-AC-GFP/ c-jun-si RNA by intraportal injection, while the rats in the control group underwent major hepatectomy and were administered with phosphate buffer saline(PBS) by intraportal injection. The portal pressure, serum ALT and liver weight/body weight of both groups were tested 14 d after surgery. The relative expression of c-jun in the liver and proliferating cell nuclear antigen(PCNA) protein was detected by Western blot. The data between the treatment group and the control group were compared using t test. Results The average portal pressure of the treatment group after surgery was(1.18±0.11) k Pa, which was significantly lower than(1.67±0.24) k Pa of the control group(t=-26.74, P<0.05). ALT of the treatment group after surgery was(43±5) U/L, which was significantly lower than(257±14) U/L of the control group(t=-31.11, P<0.05). Liver weight/body weight of the treatment group after surgery was(6.94±0.31)%, which was significantly greater than(2.76±0.14)% of the control group(t=23.57, P<0.05). The relative expression of c-jun and PCNA protein of the treatment group after surgery was respectively 0.143±0.014 and 0.195±0.027, and those of the control group was respectively 0.742±0.057 and 0.029±0.003, where significant difference was observed(t=-37.17, 14.86; P<0.05). Conclusion Lentivirus carrying c-jun-si RNA may promote liver regeneration and improve live function in cirrhotic rats after hepatectomy by down-regulating the c-jun in hepatocyte and up-regulating the PCNA protein.
引文
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[11]乔建梁,刘补报,张俊晶,等.显微镜下大鼠70%肝切除模型的制备[J].内蒙古医科大学学报,2014,36(4):302-305.
[12]郑玉宝,高志良.干细胞移植在肝衰竭治疗的现况及前景[J].临床内科杂志,2014,31(8):523-525.
[13]Wang W,Du Z,Yan J,et al.Mesenchymal stem cells promote liver regeneration and prolong survival in small-for-size liver grafts:involvement of C-Jun N-terminal kinase,cyclin D1,and NF-κB[J].PLo S One,2014,9(12):e112532.
[14]Mcgill MR,Du K,Xie Y,et al.The role of the c-Jun N-terminal kinases 1/2 and receptor-interacting protein kinase 3 in furosemideinduced liver injury[J].Xenobiotica,2014,DOI:10.3109/00498254.2014.986250[Epub ahead of print].
[15]潘华,景海娟.乌司他丁和前列地尔在肝硬化并肾病综合征患者行肝移植手术中保护性研究[J].河南外科学杂志,2014,20(5):71-72.
[16]Li Y,Ma J,Fang Q,et al.C-fos and c-jun expression in the liver of silver carp and the effect of microcystins[J].J Biochem Mol Toxicol,2014,28(4):157-166.
[17]Ferreira DM,Afonso MB,Rodrigues PM,et al.C-Jun N-terminal kinase 1/c-Jun activation of the p53/micro RNA 34a/sirtuin 1 pathway contributes to apoptosis induced by deoxycholic acid in rat liver[J].Mol Cell Biol,2014,34(6):1100-1120.
[2]马心逸,刘巧云,喻智勇.血流力学信号与肝再生的研究进展[J].中华消化外科杂志,2014,13(3):237-240.
[3]Yang QH,Hu SP,Zhang YP,et al.Effects of different therapeutic methods and typical recipes of Chinese medicine on activation of c-Jun N-terminal kinase in kupffer cells of rats with fatty liver disease[J].Chin J Integr Med,2012,18(10):769-774.
[4]袁晟光,梁科伟,刘杰,等.不同引流方式对梗阻性黄疸大鼠部分肝切除术后肝再生的影响[J].中华消化外科杂志,2013,12(12):956-962.
[5]刘立宝,黄蕾,张军航,等.Caspase-3 siR NA减少骨髓间充质干细胞稳定株细胞凋亡的实验研究[J].器官移植,2014,5(5):283-288.
[6]Morise Z,Sugioka A,Kawabe N,et al.Pure laparoscopic hepatectomy for hepatocellular carcinoma patients with severe liver cirrhosis[J].Asian J Endosc Surg,2011,4(3):143-146.
[7]Xue XJ,Zhou S,Li RR,et al.Vascular exclusion by preserving tumor-contralateral branch of hepatic artery in hepatectomy in treatment liver cancer with cirrhosis[J].Hepatogastroenterology,2012,59(117):1366-1367.
[8]耿明凡,高方媛,谷莉莉,等.终末期肝病的模型与评价指标的研究进展[J/CD].中国肝脏病杂志:电子版,2014,6(3):98-100.
[9]Li C,Mi K,Wen TF,et al.Outcome comparison of right hepatectomy for living liver donation versus for hepatic patients without cirrhosis[J].J Gastrointest Surg,2011,15(6):982-987.
[10]Cao J,Luo SM,Liang L,et al.Effects of parenteral nutrition without and with growth hormone on growth hormone/insulin-like growth factor-1 axis after hepatectomy in hepatocellular carcinoma with liver cirrhosis[J].JPEN J Parenter Enteral Nutr,2007,31(6):496-501.
[11]乔建梁,刘补报,张俊晶,等.显微镜下大鼠70%肝切除模型的制备[J].内蒙古医科大学学报,2014,36(4):302-305.
[12]郑玉宝,高志良.干细胞移植在肝衰竭治疗的现况及前景[J].临床内科杂志,2014,31(8):523-525.
[13]Wang W,Du Z,Yan J,et al.Mesenchymal stem cells promote liver regeneration and prolong survival in small-for-size liver grafts:involvement of C-Jun N-terminal kinase,cyclin D1,and NF-κB[J].PLo S One,2014,9(12):e112532.
[14]Mcgill MR,Du K,Xie Y,et al.The role of the c-Jun N-terminal kinases 1/2 and receptor-interacting protein kinase 3 in furosemideinduced liver injury[J].Xenobiotica,2014,DOI:10.3109/00498254.2014.986250[Epub ahead of print].
[15]潘华,景海娟.乌司他丁和前列地尔在肝硬化并肾病综合征患者行肝移植手术中保护性研究[J].河南外科学杂志,2014,20(5):71-72.
[16]Li Y,Ma J,Fang Q,et al.C-fos and c-jun expression in the liver of silver carp and the effect of microcystins[J].J Biochem Mol Toxicol,2014,28(4):157-166.
[17]Ferreira DM,Afonso MB,Rodrigues PM,et al.C-Jun N-terminal kinase 1/c-Jun activation of the p53/micro RNA 34a/sirtuin 1 pathway contributes to apoptosis induced by deoxycholic acid in rat liver[J].Mol Cell Biol,2014,34(6):1100-1120.