中国人兽共患病学报
中國人獸共患病學報
중국인수공환병학보
CHINESE JOURNAL OF ZOONOSES
2009年
12期
1170-1173
,共4页
郭春霞%贺永文%彭程%李文庭%翁志宏
郭春霞%賀永文%彭程%李文庭%翁誌宏
곽춘하%하영문%팽정%리문정%옹지굉
乙型肝炎病毒%动物模型%液压法
乙型肝炎病毒%動物模型%液壓法
을형간염병독%동물모형%액압법
hepatitis B virus%hydro-dynamics based transfection%animal model
目的 采用尾静脉液压法建立小鼠急性HBV感染的动物模型.方法 以液压法将具有复制能力的HBV质粒pAAV-HBV1.2通过尾静脉注射到免疫功能正常的BALB/c小鼠体内,注射后第1、2、4、6、8d,分别采用改良赖氏法、时间分辨免疫荧光法、实时荧光定量PCR检测小鼠血清中谷丙转氨酶(ALT)、HBsAg、HBeAg、抗HBs、抗HBe、HBV DNA的水平,免疫组化检测肝组织HBsAg、HBcAg的表达.结果 16只小鼠注射pAAV-HBV1.2后,有14只(85.7%)小鼠在注射后第1d血清中可检测到HBsAg,小鼠血清中HBsAg和HBeAg水平在第1d达高峰,之后逐渐下降,第8d均未能检测到.小鼠血清中HBV DNA在第2d达高峰,之后仍维持在较高水平,至第8d时为1.9×10~4 copies/mL.至第8d肝组织中可见约5%的HBcAg阳性肝细胞和2%的HBsAg阳性肝细胞.结论 采用尾静脉液压法成功的建立了小鼠急性HBV感染的动物模型.
目的 採用尾靜脈液壓法建立小鼠急性HBV感染的動物模型.方法 以液壓法將具有複製能力的HBV質粒pAAV-HBV1.2通過尾靜脈註射到免疫功能正常的BALB/c小鼠體內,註射後第1、2、4、6、8d,分彆採用改良賴氏法、時間分辨免疫熒光法、實時熒光定量PCR檢測小鼠血清中穀丙轉氨酶(ALT)、HBsAg、HBeAg、抗HBs、抗HBe、HBV DNA的水平,免疫組化檢測肝組織HBsAg、HBcAg的錶達.結果 16隻小鼠註射pAAV-HBV1.2後,有14隻(85.7%)小鼠在註射後第1d血清中可檢測到HBsAg,小鼠血清中HBsAg和HBeAg水平在第1d達高峰,之後逐漸下降,第8d均未能檢測到.小鼠血清中HBV DNA在第2d達高峰,之後仍維持在較高水平,至第8d時為1.9×10~4 copies/mL.至第8d肝組織中可見約5%的HBcAg暘性肝細胞和2%的HBsAg暘性肝細胞.結論 採用尾靜脈液壓法成功的建立瞭小鼠急性HBV感染的動物模型.
목적 채용미정맥액압법건립소서급성HBV감염적동물모형.방법 이액압법장구유복제능력적HBV질립pAAV-HBV1.2통과미정맥주사도면역공능정상적BALB/c소서체내,주사후제1、2、4、6、8d,분별채용개량뢰씨법、시간분변면역형광법、실시형광정량PCR검측소서혈청중곡병전안매(ALT)、HBsAg、HBeAg、항HBs、항HBe、HBV DNA적수평,면역조화검측간조직HBsAg、HBcAg적표체.결과 16지소서주사pAAV-HBV1.2후,유14지(85.7%)소서재주사후제1d혈청중가검측도HBsAg,소서혈청중HBsAg화HBeAg수평재제1d체고봉,지후축점하강,제8d균미능검측도.소서혈청중HBV DNA재제2d체고봉,지후잉유지재교고수평,지제8d시위1.9×10~4 copies/mL.지제8d간조직중가견약5%적HBcAg양성간세포화2%적HBsAg양성간세포.결론 채용미정맥액압법성공적건립료소서급성HBV감염적동물모형.
A mouse model for acute hepatitis B virus (HBV) infection was established by using the hydrodynamical injection of mouse tail vein, in which the immunocompetent BALB/c mice were hydrodynamically injected with a competent replication plasmid pAAV-HBV1.2 having 1.2 fold over-length of HBV DNA. On day 1, 2, 4, 6 and 8 after injection, the levels of HBsAg, HBeAg and HBV DNA in blood serum were detected by using ELISA and fluorogenic quantitative PCR assay (FQ-PCR). And on day 8. HBsAg and HBeAg in liver tissue were assayed by immunohistochemical staining. It was found that HBsAg in blood serum could be detected on day 1 after infection in 14 of 16 mice (85.7%) injected with pAVV-HBV1.2 by using ELISA assay and the peak levels of HBsAg and HBeAg were attained during the first day after injection and then it dropped down gradually up to day 8 following injection. The titer of HBV DNA in blood serum attained its peak on day 2 and maintained a high level later on. On day 8 after injection, its titer was 1.9×10~4 copies/mL. The percentage of HBcAg-positive hepatocytes and HBsAg-positive hepatocytes in liver tissues were 5% and 2% respectively. Thus, by using the hydrodynamic injection with the competent replication plasmid, a mouse model for acute HBV infection is successively developed.
