Apoptosis and its pathway in X gene-transfected HepG_2 cells

AIM: To investigate the effect of hepatitis B virus (HBV) X gene on apoptosis and expressions of apoptosis factors in X gene-transfected HepG2 cells.METHODS: The HBV X gene eukaryon expression vector pcDNVA3-Xwas transiently transfected into HepG2 cells by lipid-media transfection. Untransfected HepG2 and HepG2 transfected with pcDNA3 were used as controls. Expression of HBx in HepG2 was identified by PT-PCR. MTT and TUNEL were employed to measure proliferation and apoptosis of cells in.three groups. Semi-quantified RT-PCR was used to evaluate the expression levels of Fas/FasL, Bax/Bcl-xL,and c-myc in each group.RESULTS: HBV X gene was transfected into HepG2 cells successfully. RT-PCR showed that HBx was only expressed in HepG2/pcDNA3-X cells, but not expressed in HepG2 and HepG2/pcDNA3 cells. Analyzed by MTT, cell proliferation capacity was obviously lower in HepG2/pcDNA3-X cells (0.08910±0.003164) than in HepG2 (0.14410±0.004927)and HepG2/pcDNA3 cells (0.12150±0.007159) (P＜0.05and P＜0.01). Analyzed by TUNEL, cell apoptosis was much more in HepG2/pcDNA3-X cells (980/2 000) than HepG2 (420/2 000), HepG2/pcDNA3 cells (520/2 000) (P＜0.05 and P＜0.01). Evaluated by semi-quantified RT-PCR, the expression level of Fas/FasL was significantly higher in HepG2 cells transfected with HBx than in HepG2 and HepG2/pcDNA3 cells (P＜0.05 and P＜0.01). Bax/Bcl-xL expression level was also elevated in HepG2/pcDNA3-X cells (P＜0.05and P＜0.01). Expression of c-myc was markedly higher in HepG2/pcDNA3-X cells than in HepG2 and HepG2/pcDNA3 cells (P＜0.05 and P＜0.01).CONCLUSION: HBV X gene can impair cell proliferation capacity, improve cell apoptosis, and upregulate expression of apoptosis factors. The intervention of HBV X gene on the expression of apoptosis factors may be a possible mechanism responsible for the change in cell apoptosis and proliferation.

Meta-analysis on clinical efficacy of Simiao Yong'an decoction in treatment of peripheral arterial occlusive diseases

Objective:To systematically evaluate the clinical efficacy and safety of Simiao Yong'an Decoction in the treatment of peripheral occlusive disease(PAOD).Methods:Eight randomized controlled clinical trials(RCT)of Simiao Yong'an Decoction in the treatment of PAOD were searched and screened from domestic and foreign databases(all from database construction to March 2020).The quality of the retrieved original studies was evaluated according to the evaluation criteria of Cochrane Handbook 5.1.0,and the included studies were meta-analyzed by RevMan5.3 software.Results:A total of 350 articles were retrieved,among which 14 studies met the inclusion criteria,with a total sample size of 1254 cases.The results of meta-analysis showed that:compared with conventional western Med,combined with Simiao Yong'an Decoction on the basis of western Med treatment can significantly improve the total clinical response rate of patients[RR=1.20,95%CI(1.14,1.27),P<0.00001],improve ankle brachial index(ABI)level[MD=0.79,95%CI(0.66,0.92),P<0.00001]and toe brachial index(TBI)level[RR=0.13,95%CI(0.10,0.16),P<0.00001],decreased c-reactive protein levels[MD=-8.55,95%CI(-8.99,-8.11),P<0.00001]and LDL levels[MD=-0.41,95%CI(-0.62,-0.19),P=0.0002],and increased HDL levels[MD=0.32(0.22,0.43),P<0.00001].There was no statistically significant difference in the incidence of adverse reactions[RR=0.50,95%CI(1.15,1.64),P=0.25].Conclusion:Simiao Yong'an Decoction combined with conventional western Med is more effective than conventional western Med in the treatment of PAOD.However,in view of the limitations of the quality of the analyzed literature,the positive results obtained in this study still need to be further verified by a large sample and multi-center clinical trial with a reliable research program.