PLSCR1与乙肝病毒LS相互作用结构域的筛选
摘要
乙型肝炎病毒(hepatitis B virus,HBV)属嗜肝DNA病毒科,是已知感染人类的最小DNA病毒。HBV感染不仅可以引起急、慢性肝炎,而且还与肝硬化、肝癌的发生、发展密切相关。
根据文献报道,乙肝病毒表面抗原所含LS蛋白是导致肝细胞损伤、死亡和纤维化病变的主要原因。一些宿主蛋白与LS有相互作用,能够抑制乙肝病毒的感染、复制。
PLSCR1(磷脂爬行酶1,phospholipid scramblase 1)属于Ca2+结合的棕榈化Ⅱ型膜蛋白,非棕榈化的PLSCR1蛋白可定位于细胞核内,并与基因组DNA序列结合。PLSCR1蛋白广泛分布于细胞膜与细胞核中,参与了细胞的分化增殖,并且与信号的转导有关联。最初文献报道该蛋白有抗白血病作用[25]。在本实验前期工作中,通过应用Far-westen-2D结合质谱鉴定技术,从正常人肝细胞中筛选乙肝表面抗原相互作用蛋白,共获得了15个候选蛋白分子。磷脂爬行酶1(Phospholipid scramblase 1,PLSCR1)为上述实验筛选出的一个蛋白分子,与乙型肝炎病毒表面抗原(surface HBV antigens, HBsAg)有相互作用,继而通过酵母双杂交技术验证其有体外抑制HBV的作用。
本论文通过构建PLSCR1的分段真核表达载体,并和带有myc抗体标签的乙肝病毒打蛋白(LS蛋白)共表达于细胞中,采用免疫共沉淀实验筛选PLSCR1和HBsAg有效结构域。另外,通过酶联免疫吸附实验(enzyme linked immuno-sorbent assay,ELISA)验证了PLSCR1各片段对HBsAg的抑制作用。结果显示,部分PLSCR1片段对HBV的抑制作用比PLSCR1全长更显著,部分片段表现为促进宿主细胞中HBsAg的表达。其中1-118aa、81-118aa、61-118aa、31-118aa位置片段有抑制乙肝病毒HBsAg的作用;1-80aa、1-60aa、1-30aa片段有促进HBsAg分泌的作用。
综上所述,本研究选取人体组织中和HBsAg具有相互作用的蛋白PLSCR1,并筛选了PLSCR1与HBsAg的有效结构域,为抗乙肝的靶标药物提供候选靶标;同时,筛选出的对HBV中HBsAg的表达表现为促进作用的片段,可以为建立新的研究乙肝的细胞模型提供基础。
Hepatitis B virus (HBV), the prototype for the family Hepadnaviridae, is known as the smallest DNA virus that infects human beings. HBV infection causes acute and chronic necroinflammatory liver disease. Infected individuals are at high risk of developing liver cirrhosis and eventually, hepatocellular carcinoma.
LS was the main reason on damage ,die and dissolve of hepatocyte. The HBV infection and replication can be inhibited by some host Protein-LS interaction .
Phospholipid scramblase 1 (PLSCR1) is a calcium-binding, multiply palmitoylated type II endofacial plasma membrane protein, while unpalmitoylated PLSCR1 protein can import into the nucleus, where it binds to genomic DNA. Although the original work showed that PLSCR1 contributes to the transbilayer movement of phospholipids.
In summary, we reached the interaction between PLSCR1 and LS, and studied its biological significance in the process. Preliminary findings of this study, exogenous PLSCR1 inhibits secretion of HBsAg, after sub-expression, we found that the inhibition of some fragments of PLSCR1 on HBsAg was more significant than the full-length. The results may provide further evidence for potential roles of PLSCR1 ,and for the possibility of PLSCR1 as a new potential target for anti-HBV drugs.
根据文献报道,乙肝病毒表面抗原所含LS蛋白是导致肝细胞损伤、死亡和纤维化病变的主要原因。一些宿主蛋白与LS有相互作用,能够抑制乙肝病毒的感染、复制。
PLSCR1(磷脂爬行酶1,phospholipid scramblase 1)属于Ca2+结合的棕榈化Ⅱ型膜蛋白,非棕榈化的PLSCR1蛋白可定位于细胞核内,并与基因组DNA序列结合。PLSCR1蛋白广泛分布于细胞膜与细胞核中,参与了细胞的分化增殖,并且与信号的转导有关联。最初文献报道该蛋白有抗白血病作用[25]。在本实验前期工作中,通过应用Far-westen-2D结合质谱鉴定技术,从正常人肝细胞中筛选乙肝表面抗原相互作用蛋白,共获得了15个候选蛋白分子。磷脂爬行酶1(Phospholipid scramblase 1,PLSCR1)为上述实验筛选出的一个蛋白分子,与乙型肝炎病毒表面抗原(surface HBV antigens, HBsAg)有相互作用,继而通过酵母双杂交技术验证其有体外抑制HBV的作用。
本论文通过构建PLSCR1的分段真核表达载体,并和带有myc抗体标签的乙肝病毒打蛋白(LS蛋白)共表达于细胞中,采用免疫共沉淀实验筛选PLSCR1和HBsAg有效结构域。另外,通过酶联免疫吸附实验(enzyme linked immuno-sorbent assay,ELISA)验证了PLSCR1各片段对HBsAg的抑制作用。结果显示,部分PLSCR1片段对HBV的抑制作用比PLSCR1全长更显著,部分片段表现为促进宿主细胞中HBsAg的表达。其中1-118aa、81-118aa、61-118aa、31-118aa位置片段有抑制乙肝病毒HBsAg的作用;1-80aa、1-60aa、1-30aa片段有促进HBsAg分泌的作用。
综上所述,本研究选取人体组织中和HBsAg具有相互作用的蛋白PLSCR1,并筛选了PLSCR1与HBsAg的有效结构域,为抗乙肝的靶标药物提供候选靶标;同时,筛选出的对HBV中HBsAg的表达表现为促进作用的片段,可以为建立新的研究乙肝的细胞模型提供基础。
Hepatitis B virus (HBV), the prototype for the family Hepadnaviridae, is known as the smallest DNA virus that infects human beings. HBV infection causes acute and chronic necroinflammatory liver disease. Infected individuals are at high risk of developing liver cirrhosis and eventually, hepatocellular carcinoma.
LS was the main reason on damage ,die and dissolve of hepatocyte. The HBV infection and replication can be inhibited by some host Protein-LS interaction .
Phospholipid scramblase 1 (PLSCR1) is a calcium-binding, multiply palmitoylated type II endofacial plasma membrane protein, while unpalmitoylated PLSCR1 protein can import into the nucleus, where it binds to genomic DNA. Although the original work showed that PLSCR1 contributes to the transbilayer movement of phospholipids.
In summary, we reached the interaction between PLSCR1 and LS, and studied its biological significance in the process. Preliminary findings of this study, exogenous PLSCR1 inhibits secretion of HBsAg, after sub-expression, we found that the inhibition of some fragments of PLSCR1 on HBsAg was more significant than the full-length. The results may provide further evidence for potential roles of PLSCR1 ,and for the possibility of PLSCR1 as a new potential target for anti-HBV drugs.
引文
1.吴琳琳黄正明杨新波抗乙肝病毒治疗的研究进展[J].解放军药学学报2006 (22)373-376
2. Ganem D, Prince A M. Hepatitis B virus infection--natural history and clinicalconsequences[J]. N Engl J Med, 2004, 350 (11): 1118-29.
3. Seeger C, Mason WS. Hepatitis B Virus Biology Microbiology and molecularbiology reviews[J]. 2000, 64(1): 51-68.
4. Gripon P, Cannie I, Urban S. Efficient inhibition of hepatitis B virus infection by acylated peptides derived from the large viral surface protein[J]. J Virol. 2005, 79(3): 1613-22.
5. Chouteau P.Le Seyec J.Cannie I,et at. A short N-proximal region in the envelope harbors a determinant that contributes to the species specificity of human hepatitis B virus.J Virol,2001,75(23):11565-11572
6. Sieczkarski S B, Whittaker G R. Viral entry. Curr Top Microbiol Immunol, 2005, 285: 1-23.
7.丁晓然,杨静,伯晓晨等,靶向ASGPR的反义核酸与抗HBV药物联合用药的体外抗HBV作用[J].2006,22,(1)
8. .HUANG Ying,ZHAOQian,CHENGuo-Qiang et al.Phospholipid scramblase 1.Acta physiologica Sinica,December 25,2006,58(6):501-510
9.王升启,杨静,朱向前等,磷脂爬行酶1在制备抗乙型肝炎病毒感染药物中的用途[P].中国
10.成军,杨守纯.现代肝炎病毒分子生物学[M].北京:人民军医出社, 1997.
11.黄越前,符业阳。拉米夫定治疗慢性乙型肝炎临床疗效分析[J].河北医学,2007,13(3)272-274.
12. Xiong W, Wang X, Liu X Y, et al. Analysis of Gene Expression in Hepatitis B Virus Transfected Cell Line Induced by Interferon[J].Acta Biochimica et Biophysica Sinica, 2003, 35(12): 1053-1060.
13. Petit M A, Capel F, Dubanchet S. PreS1-specific binding proteins as potential receptors for hepatitis B virus in human hepatocytes[J]. Virology, 1992, 187(1): 211-222.14.刘庄,傅希贤,张乃临等,中草药叶下珠不同品种和制剂体外抗乙型肝炎病毒的实验研究[J]中华实验和临床病毒学杂志,1997,11(3):282.-385
14. Petit M A, Capel F, Dubanchet S. PreS1-specific binding proteins as potential receptors for hepatitis B virus in human hepatocytes[J]. Virology, 1992, 187(1): 211-222.
15.张士猛,王升启乙型肝炎病毒与宿主细胞蛋白相互作用的研究进展[J].军事医学科学院院刊2006,30(3).16.林汝仙,王升启反义核酸抗肝炎病毒研究进展[J].1999,26,(2)
16. Budkowska A, Quan C, Groh F, et al. Hepatitis B virus ( HBV)binding factor in human serum candidate for a soluble formal hepatocyte HBV receptor[J]. J Virol, 1993, 67(7): 4316-4322.
17. Xiong W, Wang X, Liu X Y, et al. Analysis of Gene Expression in Hepatitis B Virus Transfected Cell Line Induced by Interferon[J].Acta Biochimica et Biophysica Sinica, 2003, 35(12): 1053-1060.
18..邹向阳抗乙型肝炎病毒药物研究进展[J]抗感染药学,2005,9,2,(3)
19. Cooper A, Paran N, Shaul Y. The earliest steps in hepatitis B virus infection[J]. Biochim Biophys Acta. 2003, 1614(1): 89-96.
20.丁晓然,杨静,伯晓晨等,靶向ASGPR的反义核酸与抗HBV药物联合用药的体外抗HBV作用[J].2006,22,(1)21王升启,杨静,朱向前等,磷脂爬行酶1在制备抗乙型肝炎病毒感染药物中的用途[P].中国
21.张士猛,王升启乙型肝炎病毒与宿主细胞蛋白相互作用的研究进展[J].军事医学科学院院刊2006,30(3).
22.金奇.医学分子病毒学[M].北京:科学出版社, 2001.
23.Treichel U, Buschenfelde K H. The asialoglycoprotein recertor mediates hepatic binding and uptake of natural hepatitis B virus particles derived from viraemic carriers[J]. Journal of general virology, 1994, 75(1): 3021-3029.
24. Treichel U, Buschenfelde K H. The asialoglycoprotein recertor mediates hepatic binding and uptake of natural hepatitis B virus particles derived from viraemic carriers[J]. Journal of general virology, 1994, 75(1): 3021-3029.
25. Xiong W, Wang X, Liu X Y, et al. Analysis of Gene Expression in Hepatitis B Virus Transfected Cell Line Induced by Interferon[J].Acta Biochimica et Biophysica Sinica, 2003, 35(12): 1053-1060.
26.苏婧,杨静,张松等,用Far-western印迹技术筛选筛选人肝组织中与乙肝病毒表面抗原PreS1相互作用的蛋白,2010,21,(3),355-358
27.王升启,杨静,朱向前等,磷脂爬行酶1在制备抗乙型肝炎病毒感染药物中的用途[P].
28. Gripon P, Cannie I, Urban S. Efficient inhibition of hepatitis B virus infection by acylated peptides derived from the large viral surface protein[J]. J Virol. 2005, 79(3): 1613-22.
1..张明华抗乙肝病毒药物疗效[J].内蒙古医学杂志2004,4.
2.黄越前,符业阳,曾辉等拉米夫定治疗慢性乙肝[J].中国医疗导报,2002,11
3. Tompkins WA. Immunomodulmechanism and the rapeutic effects of oral use of interferon alpha:mechanism of action [J].Interferon Cytokine Res,1999,19(8):817-820
4.刘树人,李灼亮大剂量胸腺肽对慢性乙型肝炎的疗效与免疫调节的关系[J]中国新药杂志2002,11(4) 308-310
5.黄越前,符业阳。拉米夫定治疗慢性乙型肝炎临床疗效分析[J].河北医学,2007,13(3)272-274.
6.毛日成,尹有宽,张继明.乙型肝炎病毒对阿德福韦耐药的研究进展.中华肝脏病杂志,2007,15 (4):318-320.
7.徐文胜,王国俊,廖晓辉等。氧化苦参碱对HeG2.2.15细胞中乙型肝炎病毒DNA表达量的影响。第二军医大学学报2002,,23(1)72-75
8.刘庄,傅希贤,张乃临等,中草药叶下珠不同品种和制剂体外抗乙型肝炎病毒的实验研究[J]中华实验和临床病毒学杂志,1997,11(3):282.-385
9.唐霓,黄爱龙,张秉强,等。应用RNA干扰技术抑制乙型肝炎病毒抗原表达的实验研究[J]中华医学杂志,2003,83(15)1309
10.林汝仙,王升启反义核酸抗肝炎病毒研究进展[J].1999,26,(2)
11. Wagner R W, Flanagan W M. Antisense technology and prospects for therapy of viral infections and cancer[J]. Mol Med Today, 1997, 3(1): 31-38.
12.邹向阳抗乙型肝炎病毒药物研究进展[J]抗感染药学,2005,9,2,(3)
13.Gripon P, Cannie I, Urban S. Efficient inhibition of hepatitis B virus infection by acylated peptides derived from the large viral surface protein[J]. J Virol. 2005, 79(3): 1613-22.
14.Chouteau P.Le Seyec J.Cannie I,et at. A short N-proximal region in the envelope harbors a determinant that contributes to the species specificity of human hepatitis B virus.J Virol,2001,75(23):11565-11572
15.Sieczkarski S B, Whittaker G R. Viral entry. Curr Top Microbiol Immunol, 2005, 285: 1-23.
16.Petit M A, Capel F, Dubanchet S. PreS1-specific binding proteins as potentialreceptors for hepatitis B virus in human hepatocytes[J]. Virology, 1992, 187(1): 211-222.
17.Treichel U, Buschenfelde K H. The asialoglycoprotein recertor mediates hepatic binding and uptake of natural hepatitis B virus particles derived from viraemic carriers[J]. Journal of general virology, 1994, 75(1): 3021-3029.
18.丁晓然,杨静,伯晓晨等,靶向ASGPR的反义核酸与抗HBV药物联合用药的体外抗HBV作用[J].2006,22,(1)
19.张士猛,王升启乙型肝炎病毒与宿主细胞蛋白相互作用的研究进展[J].军事医学科学院院刊2006,30(3).
20.Yang J, Ding X R, Wang S Q, et al. Fibronectin is essential for hepatitis B virus infection and replication——Maybe a potential drug target?[J] Biochem. Biophys, 2006, 344(3): 757-764.
21. Budkowska A, Quan C, Groh F, et al. Hepatitis B virus ( HBV)binding factor in human serum candidate for a soluble formal hepatocyte HBV receptor[J]. J Virol, 1993, 67(7): 4316-4322.
22.Cooper A, Paran N, Shaul Y. The earliest steps in hepatitis B virus infection[J]. Biochim Biophys Acta. 2003, 1614(1): 89-96.
23.王峰,杨静等. Fibronectin的分段表达及乙肝表面抗原的相互作用研究[J].军事医学科学院院刊, 2007, 31(1): 24-27
24.HUANG Ying,ZHAOQian,CHENGuo-Qiang et al.Phospholipid scramblase 1.Acta physiologica Sinica,December 25,2006,58(6):501-510
25.王升启,杨静,朱向前等,磷脂爬行酶1在制备抗乙型肝炎病毒感染药物中的用途[P].中国
26.成军,杨守纯.现代肝炎病毒分子生物学[M].北京:人民军医出社, 1997.
27.金奇.医学分子病毒学[M].北京:科学出版社, 2001.
28.Xiong W, Wang X, Liu X Y, et al. Analysis of Gene Expression in Hepatitis B Virus Transfected Cell Line Induced by Interferon[J].Acta Biochimica et Biophysica Sinica, 2003, 35(12): 1053-1060.
2. Ganem D, Prince A M. Hepatitis B virus infection--natural history and clinicalconsequences[J]. N Engl J Med, 2004, 350 (11): 1118-29.
3. Seeger C, Mason WS. Hepatitis B Virus Biology Microbiology and molecularbiology reviews[J]. 2000, 64(1): 51-68.
4. Gripon P, Cannie I, Urban S. Efficient inhibition of hepatitis B virus infection by acylated peptides derived from the large viral surface protein[J]. J Virol. 2005, 79(3): 1613-22.
5. Chouteau P.Le Seyec J.Cannie I,et at. A short N-proximal region in the envelope harbors a determinant that contributes to the species specificity of human hepatitis B virus.J Virol,2001,75(23):11565-11572
6. Sieczkarski S B, Whittaker G R. Viral entry. Curr Top Microbiol Immunol, 2005, 285: 1-23.
7.丁晓然,杨静,伯晓晨等,靶向ASGPR的反义核酸与抗HBV药物联合用药的体外抗HBV作用[J].2006,22,(1)
8. .HUANG Ying,ZHAOQian,CHENGuo-Qiang et al.Phospholipid scramblase 1.Acta physiologica Sinica,December 25,2006,58(6):501-510
9.王升启,杨静,朱向前等,磷脂爬行酶1在制备抗乙型肝炎病毒感染药物中的用途[P].中国
10.成军,杨守纯.现代肝炎病毒分子生物学[M].北京:人民军医出社, 1997.
11.黄越前,符业阳。拉米夫定治疗慢性乙型肝炎临床疗效分析[J].河北医学,2007,13(3)272-274.
12. Xiong W, Wang X, Liu X Y, et al. Analysis of Gene Expression in Hepatitis B Virus Transfected Cell Line Induced by Interferon[J].Acta Biochimica et Biophysica Sinica, 2003, 35(12): 1053-1060.
13. Petit M A, Capel F, Dubanchet S. PreS1-specific binding proteins as potential receptors for hepatitis B virus in human hepatocytes[J]. Virology, 1992, 187(1): 211-222.14.刘庄,傅希贤,张乃临等,中草药叶下珠不同品种和制剂体外抗乙型肝炎病毒的实验研究[J]中华实验和临床病毒学杂志,1997,11(3):282.-385
14. Petit M A, Capel F, Dubanchet S. PreS1-specific binding proteins as potential receptors for hepatitis B virus in human hepatocytes[J]. Virology, 1992, 187(1): 211-222.
15.张士猛,王升启乙型肝炎病毒与宿主细胞蛋白相互作用的研究进展[J].军事医学科学院院刊2006,30(3).16.林汝仙,王升启反义核酸抗肝炎病毒研究进展[J].1999,26,(2)
16. Budkowska A, Quan C, Groh F, et al. Hepatitis B virus ( HBV)binding factor in human serum candidate for a soluble formal hepatocyte HBV receptor[J]. J Virol, 1993, 67(7): 4316-4322.
17. Xiong W, Wang X, Liu X Y, et al. Analysis of Gene Expression in Hepatitis B Virus Transfected Cell Line Induced by Interferon[J].Acta Biochimica et Biophysica Sinica, 2003, 35(12): 1053-1060.
18..邹向阳抗乙型肝炎病毒药物研究进展[J]抗感染药学,2005,9,2,(3)
19. Cooper A, Paran N, Shaul Y. The earliest steps in hepatitis B virus infection[J]. Biochim Biophys Acta. 2003, 1614(1): 89-96.
20.丁晓然,杨静,伯晓晨等,靶向ASGPR的反义核酸与抗HBV药物联合用药的体外抗HBV作用[J].2006,22,(1)21王升启,杨静,朱向前等,磷脂爬行酶1在制备抗乙型肝炎病毒感染药物中的用途[P].中国
21.张士猛,王升启乙型肝炎病毒与宿主细胞蛋白相互作用的研究进展[J].军事医学科学院院刊2006,30(3).
22.金奇.医学分子病毒学[M].北京:科学出版社, 2001.
23.Treichel U, Buschenfelde K H. The asialoglycoprotein recertor mediates hepatic binding and uptake of natural hepatitis B virus particles derived from viraemic carriers[J]. Journal of general virology, 1994, 75(1): 3021-3029.
24. Treichel U, Buschenfelde K H. The asialoglycoprotein recertor mediates hepatic binding and uptake of natural hepatitis B virus particles derived from viraemic carriers[J]. Journal of general virology, 1994, 75(1): 3021-3029.
25. Xiong W, Wang X, Liu X Y, et al. Analysis of Gene Expression in Hepatitis B Virus Transfected Cell Line Induced by Interferon[J].Acta Biochimica et Biophysica Sinica, 2003, 35(12): 1053-1060.
26.苏婧,杨静,张松等,用Far-western印迹技术筛选筛选人肝组织中与乙肝病毒表面抗原PreS1相互作用的蛋白,2010,21,(3),355-358
27.王升启,杨静,朱向前等,磷脂爬行酶1在制备抗乙型肝炎病毒感染药物中的用途[P].
28. Gripon P, Cannie I, Urban S. Efficient inhibition of hepatitis B virus infection by acylated peptides derived from the large viral surface protein[J]. J Virol. 2005, 79(3): 1613-22.
1..张明华抗乙肝病毒药物疗效[J].内蒙古医学杂志2004,4.
2.黄越前,符业阳,曾辉等拉米夫定治疗慢性乙肝[J].中国医疗导报,2002,11
3. Tompkins WA. Immunomodulmechanism and the rapeutic effects of oral use of interferon alpha:mechanism of action [J].Interferon Cytokine Res,1999,19(8):817-820
4.刘树人,李灼亮大剂量胸腺肽对慢性乙型肝炎的疗效与免疫调节的关系[J]中国新药杂志2002,11(4) 308-310
5.黄越前,符业阳。拉米夫定治疗慢性乙型肝炎临床疗效分析[J].河北医学,2007,13(3)272-274.
6.毛日成,尹有宽,张继明.乙型肝炎病毒对阿德福韦耐药的研究进展.中华肝脏病杂志,2007,15 (4):318-320.
7.徐文胜,王国俊,廖晓辉等。氧化苦参碱对HeG2.2.15细胞中乙型肝炎病毒DNA表达量的影响。第二军医大学学报2002,,23(1)72-75
8.刘庄,傅希贤,张乃临等,中草药叶下珠不同品种和制剂体外抗乙型肝炎病毒的实验研究[J]中华实验和临床病毒学杂志,1997,11(3):282.-385
9.唐霓,黄爱龙,张秉强,等。应用RNA干扰技术抑制乙型肝炎病毒抗原表达的实验研究[J]中华医学杂志,2003,83(15)1309
10.林汝仙,王升启反义核酸抗肝炎病毒研究进展[J].1999,26,(2)
11. Wagner R W, Flanagan W M. Antisense technology and prospects for therapy of viral infections and cancer[J]. Mol Med Today, 1997, 3(1): 31-38.
12.邹向阳抗乙型肝炎病毒药物研究进展[J]抗感染药学,2005,9,2,(3)
13.Gripon P, Cannie I, Urban S. Efficient inhibition of hepatitis B virus infection by acylated peptides derived from the large viral surface protein[J]. J Virol. 2005, 79(3): 1613-22.
14.Chouteau P.Le Seyec J.Cannie I,et at. A short N-proximal region in the envelope harbors a determinant that contributes to the species specificity of human hepatitis B virus.J Virol,2001,75(23):11565-11572
15.Sieczkarski S B, Whittaker G R. Viral entry. Curr Top Microbiol Immunol, 2005, 285: 1-23.
16.Petit M A, Capel F, Dubanchet S. PreS1-specific binding proteins as potentialreceptors for hepatitis B virus in human hepatocytes[J]. Virology, 1992, 187(1): 211-222.
17.Treichel U, Buschenfelde K H. The asialoglycoprotein recertor mediates hepatic binding and uptake of natural hepatitis B virus particles derived from viraemic carriers[J]. Journal of general virology, 1994, 75(1): 3021-3029.
18.丁晓然,杨静,伯晓晨等,靶向ASGPR的反义核酸与抗HBV药物联合用药的体外抗HBV作用[J].2006,22,(1)
19.张士猛,王升启乙型肝炎病毒与宿主细胞蛋白相互作用的研究进展[J].军事医学科学院院刊2006,30(3).
20.Yang J, Ding X R, Wang S Q, et al. Fibronectin is essential for hepatitis B virus infection and replication——Maybe a potential drug target?[J] Biochem. Biophys, 2006, 344(3): 757-764.
21. Budkowska A, Quan C, Groh F, et al. Hepatitis B virus ( HBV)binding factor in human serum candidate for a soluble formal hepatocyte HBV receptor[J]. J Virol, 1993, 67(7): 4316-4322.
22.Cooper A, Paran N, Shaul Y. The earliest steps in hepatitis B virus infection[J]. Biochim Biophys Acta. 2003, 1614(1): 89-96.
23.王峰,杨静等. Fibronectin的分段表达及乙肝表面抗原的相互作用研究[J].军事医学科学院院刊, 2007, 31(1): 24-27
24.HUANG Ying,ZHAOQian,CHENGuo-Qiang et al.Phospholipid scramblase 1.Acta physiologica Sinica,December 25,2006,58(6):501-510
25.王升启,杨静,朱向前等,磷脂爬行酶1在制备抗乙型肝炎病毒感染药物中的用途[P].中国
26.成军,杨守纯.现代肝炎病毒分子生物学[M].北京:人民军医出社, 1997.
27.金奇.医学分子病毒学[M].北京:科学出版社, 2001.
28.Xiong W, Wang X, Liu X Y, et al. Analysis of Gene Expression in Hepatitis B Virus Transfected Cell Line Induced by Interferon[J].Acta Biochimica et Biophysica Sinica, 2003, 35(12): 1053-1060.