CAJ | 학술논문

目的观察胡黄连总苷抑制HepG 2.2.15细胞内HBV共价闭合环状DNA(HBVeeeDNA)的作用效果及特点.方法分别用50 mg/L胡黄连总苷或5 mg/L阿德福韦酯的细胞培养液作用于HepG 2.2.15细胞,加药后2 d、5 d收集细胞及培养上清液,实时荧光定量PCR方法检测上清液和细胞内HBV DNA、细胞内HBV cccDNA和前基因组RNA(pgRNA)水平,分别计算抑制率.均值比较用t检验.结果胡黄连总苷作用2 d和5 d后,上清液HBV DNA抑制率为49.74%(t=4.723,P<0.05)和79.48%(t=7.512,P<0.05);细胞内HBV cccDNA抑制率为43.55%(t=5.216,P<0.05)和56.43%(t=7.262,P<0.05),总DNA抑制率为43.39%(t=4.137,P<0.05)和63.86%(t=7.861,P<0.05),pgRNA抑制率为54.72%(t=4.532,P<0.05)和56.08%(t=4.833,P<0.05).阿德福韦酯作用2 d和5 d后,上清液HBV DNA抑制率为25.56%(t=2.874,P<0.05)和92.44%(t=10.276,P<0.05);细胞内HBV cccDNA抑制率为18.54%(t=2.736,P<0.05)和47.19%(t=6.852,P<0.05),总DNA抑制率为21.20%(t=3.206,P<0.05)和71.47%(t=8.332,P<0.05),pgRNA抑制率为11.14%(t=1.761,P>0.05)和37.61%(t=3.632,P<0.05).结论胡黄连总苷能明显抑制HepG 2.2.15细胞内HBV复制,尤其对cccDNA具有抑制作用,在作用时相上早于阿德福韦酯,可能存在不同的作用机制.
목적 관찰호황련총감억제HepG 2.2.15세포내HBV공개폐합배상DNA(HBVeeeDNA)적작용효과급특점.방법 분별용50 mg/L호황련총감혹5 mg/L아덕복위지적세포배양액작용우HepG 2.2.15세포,가약후2 d、5 d수집세포급배양상청액,실시형광정량PCR방법검측상청액화세포내HBV DNA、세포내HBV cccDNA화전기인조RNA(pgRNA)수평,분별계산억제솔.균치비교용t검험.결과 호황련총감작용2 d화5 d후,상청액HBV DNA억제솔위49.74%(t=4.723,P<0.05)화79.48%(t=7.512,P<0.05);세포내HBV cccDNA억제솔위43.55%(t=5.216,P<0.05)화56.43%(t=7.262,P<0.05),총DNA억제솔위43.39%(t=4.137,P<0.05)화63.86%(t=7.861,P<0.05),pgRNA억제솔위54.72%(t=4.532,P<0.05)화56.08%(t=4.833,P<0.05).아덕복위지작용2 d화5 d후,상청액HBV DNA억제솔위25.56%(t=2.874,P<0.05)화92.44%(t=10.276,P<0.05);세포내HBV cccDNA억제솔위18.54%(t=2.736,P<0.05)화47.19%(t=6.852,P<0.05),총DNA억제솔위21.20%(t=3.206,P<0.05)화71.47%(t=8.332,P<0.05),pgRNA억제솔위11.14%(t=1.761,P>0.05)화37.61%(t=3.632,P<0.05).결론 호황련총감능명현억제HepG 2.2.15세포내HBV복제,우기대cccDNA구유억제작용,재작용시상상조우아덕복위지,가능존재불동적작용궤제.
Ohjective To observe the inhibition effect of total glucosides of Picrorhiza on hepatitis B virus covalently closed circular DNA (HBV cccDNA) in HepG 2.2.15 cell line. Methods HepG 2.2.15 cells were incubated with culture medium containing 50 mg/L of picrosides or 5 mg/L of adefovir dipivoxil for 2 or 5 days. HBV DNA in the supernatant, intracellular cccDNA, relaxed circular DNA (rcDNA) and pregenomic RNA (pgRNA) were quantified by specific real-time polymerase chain reaction (RT-PCR) and inhibition rates were calculated. The means were compared by t test. Results After treated with picrosides for 2 and 5 days, the inhibition rates of HBV DNA in thesupernatant were 49. 74% (t=4.723, P<0.05) and 79.48% (t = 7.512, P<0.05), respectively. The inhibition rates of intracellular cccDNA were 43.55% (t = 5.216, P<0.05) and 56.43% (t=7.262, P<0.05), respectively, while those of intracellular rcDNA were 43.39% (t=4.137, P<0.05) and 63.86% (t=7.861, P<0.05), respectively, and those of intracellular pgRNA were 54.72% (t=4.532, P<0.05) and 56.08% (t=4.833, P<0.05), respectively. Comparatively, after treatment with adefovir dipivoxil for 2 and 5 days, the inhibition rates of HBV DNA in the supernatant were 25.56% (t=2.874, P<0.05) and 92.44% (t =10.276, P<0.05), respectively. Those of cccDNA were 18.54% (t=2.736, P<0.05) and 47.19% (t=6.852, P<0.05), respectively. Those of rcDNA were 21. 20% (t=3.206, P<0.05) and 71.47% (t=8.332, P<0.05), respectively, pgRNA were 11.14% (t=1.761, P>0.05) and 37.61%(t=3.632, P<0.05) respectively in HepG2.2.15 cells. Conclusions Pierosides may inhibit the replication cycle of HBV, including the formation of cccDNA in HepG 2.2.15 cells. The mechanism of pierosides on cccDNA may differ from adefovir dipivoxil's due to its earlier inhibition time phase.