Kim Hye Jin, Yoo Hwa Seung, Kim Jin Chul, Park Chan Su, Choi Mi Sun, Kim Mijee, Choi Hyangsoon, Min Jung Sun, Kim Yong Soo, Yoon Seong Woo, Ahn Jeong Keun
Department of Microbiology, Chungnam National University, Daejeon 305-764, South Korea.
J Ethnopharmacol. 2009 Jul 15;124(2):189-96. doi: 10.1016/j.jep.2009.04.046. Epub 2009 May 3.
A medicinal herb Curcuma longa Linn has been used for treating various liver diseases caused by hepatitis B virus (HBV) in Asia.
The study was performed in order to investigate the antiviral activity of Curcuma longa Linn against HBV replication in liver cells.
Aqueous extract of Curcuma longa Linn (CLL) was prepared and used to analyze its antiviral activity against HBV replication in HepG 2.2.15 cells, which contain HBV genomes. The inhibitory effect of CLL on HBV replication was examined by testing the levels of secreted HBV surface antigens (HBsAg), HBV DNAs, and HBV RNAs in HepG 2.2.15 cells using ELISA, Southern blot, and Northern blot analyses. Cytotoxic activities of CLL extract on various liver cells were analyzed by MTT assay. To dissect the inhibitory mechanism of CLL extract on HBV replication, the levels of p53 protein and p53 mRNAs were analyzed by Western blot and RT-PCR in HepG 2.2.15 cells. The repression of CLL extract on HBV transcription was analyzed by RT-PCR and CAT assay.
CLL extract repressed the secretion of HBsAg from HepG 2.2.15 cells. CLL extract also suppressed the production of HBV particles and the level of intracellular HBV RNAs in HepG 2.2.15 cells, suggesting that CLL extract inhibits HBV replication. We found that the anti-HBV activity of CLL extract is mediated through enhancing the cellular accumulation of p53 protein by transactivating the transcription of p53 gene as well as increasing the stability of p53 protein. It turned out that CLL extract repressed the transcription of HBx gene by suppressing HBV enhancer I and X promoter through p53 protein. In addition, CLL extract did not have any cytotoxic effects on liver cells.
These data showed that CLL extract represses HBV replication through enhancing the level of p53 protein. CLL extract can be used as a safe and specific drug for patients with liver diseases caused by HBV infection.
在亚洲，药用植物姜黄已被用于治疗由乙型肝炎病毒（HBV）引起的各种肝脏疾病。
进行本研究以调查姜黄对肝细胞中HBV复制的抗病毒活性。
制备姜黄的水提取物（CLL），并用于分析其对含有HBV基因组的HepG 2.2.15细胞中HBV复制的抗病毒活性。通过使用ELISA、Southern印迹和Northern印迹分析检测HepG 2.2.15细胞中分泌的HBV表面抗原（HBsAg）、HBV DNA和HBV RNA水平，来检查CLL对HBV复制的抑制作用。通过MTT法分析CLL提取物对各种肝细胞的细胞毒性活性。为了剖析CLL提取物对HBV复制的抑制机制，通过蛋白质印迹法和RT-PCR分析HepG 2.2.15细胞中p53蛋白和p53 mRNA的水平。通过RT-PCR和CAT测定分析CLL提取物对HBV转录的抑制作用。
CLL提取物抑制了HepG 2.2.15细胞中HBsAg的分泌。CLL提取物还抑制了HepG 2.2.15细胞中HBV颗粒的产生和细胞内HBV RNA的水平，表明CLL提取物抑制HBV复制。我们发现CLL提取物的抗HBV活性是通过激活p53基因的转录以及增加p53蛋白的稳定性来增强p53蛋白的细胞积累来介导的。结果表明，CLL提取物通过p53蛋白抑制HBV增强子I和X启动子，从而抑制HBx基因的转录。此外，CLL提取物对肝细胞没有任何细胞毒性作用。
这些数据表明，CLL提取物通过提高p53蛋白水平来抑制HBV复制。CLL提取物可作为治疗HBV感染所致肝病患者的安全且特异性药物。
