Résumé
OBJECTIVE:To investigate the inhibitory effect of siRNA expression vector inhibiting human insulin-like growth factor 2(IGF2)gene on the proliferation of hepatoma cell line Huh-7. METHODS:siRNA expression vector pGL3-hAFP-hTERT-siRNA3(“siRNA3”)which inhibited IGF2 gene by dual promoter regulation of recombinant human alpha-foetoprotein(hAFP)and human telomerase reverse transcriptase(hTERT)was transfected into the Huh-7 cell and normal hepatocyte L-02,and then a nega-tive control group(vector pGL3-hAFP-hTERT)and a blank control group were set up. IGF2 mRNA expression was detected by re-al-time fluorescent quantitative polymerase chain reaction 48 h after transfection into the cells in all groups;the activity of the cells by the microplate reader 0,24,48 and 72 h thereafter;and the cell cycle and apoptosis by the flow cytometer 48 h thereafter,and the changes in the protein levels of IGF2,PCNA,Cyclin E2,Cyclin D2,Cdc2 and Bcl-2 in the cell were detected by Western blot. RESULTS:Compared with the negative control group and blank control group,IGF2 mRNA expression in the Huh-7 cell transfected with siRNA3 was obviously weaker;at 48 and 72 h after transfection,the activity of Huh-7 cell signigicantly reduced, Huh-7 cells at G1 phase obviously increased and those at S phase markedly decreased;the occurrence of early,late and total apopto-sis in Huh-7 cells apparently increased,and the protein expression of IGF2,PCNA,Cyclin E2,Cyclin D2,Cdc2 and Bcl-2 in cells significantly weakened,with statistically significance(P0.05). CONCLUSIONS:siRNA which inhibited IGF2 gene by dual promoter regulation of recombinant hAFP and hTERT can specially inhibit IGF2 gene expression and the prolifer-ation of Huh-7 cells,which may be involved with down-regulated protein expression of cell proliferation-associated gene PCNA, cell cycle control-associated genes Cyclin E2,Cyclin D2 and Cdc2 and apoptosis regulation-associated gene Bcl-2 as a result of down-regulated IGF2 mRNA expression and protein expres-sion.
Résumé
ObjectiveTo evaluate the efficacy and safety of the angiotensin Ⅱ receptor blockers (ARB) in reducing portal hypertension ( PHT) in patients with cirrhosis.Methods PubMed,EMBASE,Web of Science,The Cochrane Central Register of Controlled Trials,Chinese BiomedicalDatabase,ChineseJournals Full-text Database and WanFang Digital Journal Full-text database were searched.Statistical analysis was performed by meta-analysis using RevMan4.2 software.ResultsAmong 8 randomized controlled trials ( RCT) including 282 patients met the inclusion criteria,4 trials were analyzed to compare the ARB with the placebo or no treatment and the other 4 trials were analyzed to compare the ARB with propranolol.Meta-analysis results were as follows.(1) The ARB resulted in more significant hepatic venous pressure gradient ( HVPG) reduction as compared with the placebo or no treatment [ WMD =1.87 mm Hg (1 mm Hg =0.133 kPa),95％CI ( 0.86-2.87 )mmHg,P =0.00003 ].Andthe ARB were similar to propranolol in reducing HVPG [ WMD =0.92 mm Hg,95％ CI ( - 0.41-2.26)mm Hg,P =0.17 ].(2)The ARB led to more significant reduction in mean arterial pressure than the placebo or no treatment [ WMD =8.89 mm Hg,95％ CI( 7.16-10.62)mm Hg,P ＜ 0.00001 ],but they were similar to propranolol had no significant difference.And the ARB had no significant effect on the heart rate of the patients,which was similar to no treatment group ( P ＞ 0.05 ).Whereas,propranolol could greatly decrease heart rate of the patients ( WMD =- 21.25,95％ CI - 25.83-16.68,P ＜ 0.000 01 ).( 3 ) No significant differences were found in serum bilirubin and creatinine levels between the ARB and the placebo or no treatment groups ( P ＞0.05).The rate of nonspecific adverse events was higher in the ARB groups than in the placebo or no treatment groups ( P =0.03 ),but it showed there was no difference between the ARB and propranolol groups (P =0.72).ConclusionThe ARB is effective in reducing portal hypertension in patients with cirrhosis,which is similar to propranolol.Their effects on mean arterial pressure is similar to propranolol without significant effects on hear rate,liver functionand kidney function,and with less nonspecific adverse events.The ARB could become a new choice for the treatment of portal hypertension.