壁虎粉提取物作用于乙型肝炎病毒侵袭的HepG2和HL-7702细胞的实验研究

目的探讨壁虎粉提取物对乙型肝炎病毒侵袭过的人肝癌细胞(HepG2)和人肝细胞(HL-7702)中乙型肝炎病毒的复制能力和细胞生长状况的影响。方法实验分壁虎粉干预组、干扰素干预组、乙肝对照组和正常对照组四组,除正常对照组,其他三组以终浓度为1×107IU/mL含HBV-DNA的患者血清侵袭2小时后,再以一定浓度壁虎粉提取物或干扰素分别作用于上述两种细胞,收集各时段的培养液上清和(或)细胞,检测培养液上清AST、ALT、r-GT和LDH含量;采用荧光定量PCR法检测HBV-DNA;AO/EB进行细胞凋亡染色。结果两种细胞不同组别(除正常对照组外)培养6天、9天较培养3天AST显著增高(P<0.01),以两个干预组增高更为明显;HepG2干预组各时间段LDH含量高于HL-7702干预组(P<0.01),以壁虎粉干预HepG2组增高最为明显,同时凋亡细胞增多也较明显;两干预组细胞HBV-DNA含量较乙肝对照组低约1个数量级。结论壁虎粉提取物和干扰素对HBV复制具有抑制作用,对HL-7702细胞作用更为明显;壁虎粉提取物可诱导肝癌HepG2细胞早期凋亡、坏死。

The effect of gecko powder extract on HepG2 and HL-7702 cells attacked by hepatitis B virus

Objective To investigate the effect of gecko powder extract on cell growth of HepG2 and HL-7702 attacked by hepatitis B virus and the replication capacity of the hepatitis B virus. Methods There were four groups, including gecko powder extract group, interferon group, HBV control group and normal control group. The former three groups were attacked for two hours by HBV patients' serum, in which the HBV patients' serum concentration of the HBV-DNA was 1×10 7 IU/mL. And the cells were treated by gecko powder extract and interferon respectively. The supernatant of culture medium and(or) cells were collected on day 0, 3, 6 and 9. The content of AST, ALT, r-GT and LDH of the supernatant of culture medium were detected. The HBV-DNA was tested by Real-time Quantitative PCR. The AO/EB was used for apoptosis fluorescence staining assay. Results The AST content significantly increased in the two cell lines in different groups (except the normal control group) on day 6 and 9 than those on day 3(P<0.01). The AST content of the two intervention groups were much higher. The LDH content of the HepG2 intervention groups in different times were significantly higher than those of the HL-7702 intervention groups(P<0.01). The LDH content of gecko powder extract intervention group was the highest. And the increase of the apoptotic cells was also highest in the gecko powder intervention group. The HBV-DNA of the two intervention groups were one order of magnitude lower than that of the HBV control group. Conclusion The gecko powder extract and interferon can inhibit the HBV replication. The inhibition on the HL-7702 cells line was more significant. The gecko powder extract could induce the early apoptosis of the HepG2.