CAJ | 학술논문

目的评价HBV基因组核苷酸C1913 A/G变异对乙型肝炎患者疾病进程及体外病毒复制力的影响。方法385例研究对象包括116例慢性乙型肝炎轻中度（ CHB-M）患者，123例慢性乙型肝炎重度（ CHB-S）患者和146例慢加急性肝衰竭（ ACLF）患者。从患者血清中提取HBV DNA，PCR扩增HBV DNA全长，统计分析G1896A，A1762T＋G1764A和C1913A/G变异的发生率，C1913A/G变异引起核心蛋白第5位氨基酸残基由脯氨酸变为苏氨酸或丙氨酸。统计各组变异与不同疾病进程患者间的单因素与多因素分析。挑选代表性C1913A/G变异株通过定点突变回复为野生型对照。用BspQⅠ/ScaⅠ双酶切HBV基因组，转染HepG2细胞，检测病毒复制力和HBsAg表达水平。结果 CHB-M、CHB-S和ACLF三组患者G1896A，A1762T＋G1764 A和C1913 A/G 变异检出率分别为17.45％、27.43％和51.63％（ P ＜0.01），21.32％、32.64％和78.61％（ P＜0.01），15.52％、33.33％和35.62％（ P＜0.01）， CHB 与 ACLF 患者年龄校正以后两组比较C1913 A/G检出率分别为26.34％和31.13％（ P＜0.01）。 C1913 A变异株的复制力和HBsAg水平较野生株分别降低了32.6％和9.7％，C1913 G分别降低了50.7％和9.2％。结论 C1913 A/G变异发生率随疾病程度加重依次增高且有统计学意义；C1913 A/G变异降低HBV病毒复制力并进而降低HBsAg表达水平，可能与核心蛋白结构发生改变有关，结果提示该变异可能与慢性乙型肝炎重症化发生机制相关。
목적평개HBV기인조핵감산C1913 A/G변이대을형간염환자질병진정급체외병독복제력적영향。방법385례연구대상포괄116례만성을형간염경중도（ CHB-M）환자，123례만성을형간염중도（ CHB-S）환자화146례만가급성간쇠갈（ ACLF）환자。종환자혈청중제취HBV DNA，PCR확증HBV DNA전장，통계분석G1896A，A1762T＋G1764A화C1913A/G변이적발생솔，C1913A/G변이인기핵심단백제5위안기산잔기유포안산변위소안산혹병안산。통계각조변이여불동질병진정환자간적단인소여다인소분석。도선대표성C1913A/G변이주통과정점돌변회복위야생형대조。용BspQⅠ/ScaⅠ쌍매절HBV기인조，전염HepG2세포，검측병독복제력화HBsAg표체수평。결과 CHB-M、CHB-S화ACLF삼조환자G1896A，A1762T＋G1764 A화C1913 A/G 변이검출솔분별위17.45％、27.43％화51.63％（ P ＜0.01），21.32％、32.64％화78.61％（ P＜0.01），15.52％、33.33％화35.62％（ P＜0.01）， CHB 여 ACLF 환자년령교정이후량조비교C1913 A/G검출솔분별위26.34％화31.13％（ P＜0.01）。 C1913 A변이주적복제력화HBsAg수평교야생주분별강저료32.6％화9.7％，C1913 G분별강저료50.7％화9.2％。결론 C1913 A/G변이발생솔수질병정도가중의차증고차유통계학의의；C1913 A/G변이강저HBV병독복제력병진이강저HBsAg표체수평，가능여핵심단백결구발생개변유관，결과제시해변이가능여만성을형간염중증화발생궤제상관。
Objective To evaluate the impact of hepatitis B virus ( HBV) genome C1913A/G mutation on disease progression and the viral replication capacity in vitro.Methods 385 patients were categorized into mild chronic hepatitis B(CHB-M,116),severe hepatitis B(CHB-S,123),and acute on chronic liver failure (ACLF,146) sub-groups.Full-length HBV genome extracted from patient′s serum was amplified by PCR and the G1896A,A1762T+G1764 A and C1913 A/G variant incidences were calculated .C1913 A/G mutation can cause the core protein′s fifth residue proline replacement by threonine or alanine .The difference in incidences of C 1913 A/G among groups was analyzed.The representative C1913A/G mutant was picked out for site-directed mutagenesis converted to wild type as controls by .HBV 1.0 full length genome derived by BspQ Ⅰ/Sca Ⅰ double digestion was transfected into HepG 2 cells to compare viral replication capacity and HBsAg expression between C 1913 A/G mutant and wild type . Results G1896 A, A1762 T +G1764 A and C1913 A/G mutation detection rates were 17.45%, 27.43% and 51.63%(P<0.01);21.32%,32.64% and 78.61%(P <0.01),15.52%,33.33% and 35.62% for CHB-M, CHB-S and ACLF respectively ( P<0.01 ) .The C1913 A/G detection rates were 26.34% and 31.13%( P<0.01 ) after the age-revised in CHB and ACLF groups .Replication capacity and HBsAg expression of C 1913 A mutants reduced by 32.6%and 9.7%compared to that of wild type;C1913 G reduced by 50.7% and 9.2% respectively . Conclusions Incidence of C1913 A/G mutation increased along with disease severity , which was statistically significant.C1913A/G mutation reduced viral replication capacity and HBsAg expression;it was possibly due to changes in the core protein structure .The results suggested that C 1913 A/G mutation may be associated with disease progression of chronic hepatitis B .