中华微生物学和免疫学杂志
中華微生物學和免疫學雜誌
중화미생물학화면역학잡지
CHINESE JOURNAL OF MICROBIOLOGY AND IMMUNOLOGY
2014年
3期
186-193
,共8页
邢培祥%王传新%王立水%王洪春%邢全台
邢培祥%王傳新%王立水%王洪春%邢全檯
형배상%왕전신%왕립수%왕홍춘%형전태
丙型肝炎病毒%白细胞介素-1B%白细胞介素-1RN%基因多态性%基因芯片
丙型肝炎病毒%白細胞介素-1B%白細胞介素-1RN%基因多態性%基因芯片
병형간염병독%백세포개소-1B%백세포개소-1RN%기인다태성%기인심편
HCV%Interleukin-1β%Interleukin-1RN%Gene polymorphism%Gene chip
目的:探讨IL-1β、IL-1ra血清水平及白细胞介素-1B( IL-1B)和白细胞介素-1RN( IL-1RN)基因多态性与HCV相关肝病发生发展的关系。方法以酶联免疫吸附试验( ELISA)测定IL-1β、IL-1ra血清水平;基因芯片技术检测310例HCV肝病和324例不相关联的健康对照人群的IL-1B-31 C/T、IL-1B-511C/T单核苷酸多态性(SNP);琼脂糖凝胶电泳检测IL-1RN内含子2(intron 2)基因多态性( VNTR)。聚合酶链反应-限制性片段长度多态性( PCR-RFLP)随机分析部分样本验证基因芯片分型结果;HCV基因分型采用直接测序法;Lightcycler 特异性荧光探针PCR检测HCV RNA载量;血清丙氨酸氨基转移酶( alanine aminotransferase ,ALT)使用ROCHE cobas 8000全自动生化分析仪检测。结果(1)病例组IL-1β血清水平(22.6±7.3) pg/ml显著高于对照组(13.7±4.2) pg/ml( P<0.01);IL-1ra血清水平(286.30±55.10) pg/ml显著高于对照组(185.55±48.32) pg/ml(P<0.01)。(2)在病例组、轻中型肝炎、重型肝炎、肝硬化合并肝癌组、1b型组及2a型组,IL-1B-511T等位基因携带者血清IL-1β水平均显著高于其相应CC纯和子及对照组( P<0.05),且HCV感染各组-511 T携带者IL-1ra/IL-1β比值显著低于对照组(P<0.05);IL-1B-511T携带者在1b型组的IL-1β水平较2a型组显著增高(P<0.05);IL-1B-511T携带者IL-1ra血清水平显著高于相应对照组(P<0.05);IL-1B-511CC携带者血清IL-1β水平在各组别间差异无统计学意义(P>0.05)。(3)病例组IL-1B-511TT基因型频率显著高于对照组( P<0.05,OR=1.55,95%CI=1.10~2.18),两组间T等位基因频率差异有统计学意义(P<0.05,OR=1.31,95%CI=1.05~1.63)。 IL-1RN VNTR二组间差异无统计学意义(P>0.05)。(4)IL-1B-511C/T SNP基因型频率在不同转归类型中差异有统计学意义( P<0.005)。与相应的CT+CC基因型比较,不同转归类型各组IL-1B-511TT频率均显著高于对照组,其OR(95%CI)分别为:轻中型组2.17(1.48~3.19);重型组2.11(1.05~4.26);肝硬化组2.98(1.77~4.99);肝癌组4.33(2.16~8.67)。 IL-1B-511T频率组间差异有统计学意义(P<0.005)。与对照组比,IL-1 B-511 T 在HCV感染各组OR (95%CI )依次分别是:1.80(1.38~2.36);1.80(1.08~3.01);2.62(1.76~3.89);3.49(1.96~6.23)。(5)IL-1B SNP、IL-1RN VNTR与1b或2a基因型HCV感染差异无统计学意义;IL-1 B-511 C/T SNP与抗病毒治疗前HCV RNA复制、抗病毒治疗的反应性及血清ALT水平无关。结论 IL-1B-511T患者血清IL-1β、IL-1ra水平显著升高,而IL-1ra/IL-1β比值显著降低;IL-1B-511 SNP与HCV肝病慢性化及不同临床转归相关,IL-1B-511TT/T能增加HCV慢性感染及其临床转归的危险,IL-1B-511CC/C则是抗慢性感染和抗慢性化转归的保护因素;IL-1B-511T等位基因与1b型HCV具有协同升高IL-1β水平的作用。
目的:探討IL-1β、IL-1ra血清水平及白細胞介素-1B( IL-1B)和白細胞介素-1RN( IL-1RN)基因多態性與HCV相關肝病髮生髮展的關繫。方法以酶聯免疫吸附試驗( ELISA)測定IL-1β、IL-1ra血清水平;基因芯片技術檢測310例HCV肝病和324例不相關聯的健康對照人群的IL-1B-31 C/T、IL-1B-511C/T單覈苷痠多態性(SNP);瓊脂糖凝膠電泳檢測IL-1RN內含子2(intron 2)基因多態性( VNTR)。聚閤酶鏈反應-限製性片段長度多態性( PCR-RFLP)隨機分析部分樣本驗證基因芯片分型結果;HCV基因分型採用直接測序法;Lightcycler 特異性熒光探針PCR檢測HCV RNA載量;血清丙氨痠氨基轉移酶( alanine aminotransferase ,ALT)使用ROCHE cobas 8000全自動生化分析儀檢測。結果(1)病例組IL-1β血清水平(22.6±7.3) pg/ml顯著高于對照組(13.7±4.2) pg/ml( P<0.01);IL-1ra血清水平(286.30±55.10) pg/ml顯著高于對照組(185.55±48.32) pg/ml(P<0.01)。(2)在病例組、輕中型肝炎、重型肝炎、肝硬化閤併肝癌組、1b型組及2a型組,IL-1B-511T等位基因攜帶者血清IL-1β水平均顯著高于其相應CC純和子及對照組( P<0.05),且HCV感染各組-511 T攜帶者IL-1ra/IL-1β比值顯著低于對照組(P<0.05);IL-1B-511T攜帶者在1b型組的IL-1β水平較2a型組顯著增高(P<0.05);IL-1B-511T攜帶者IL-1ra血清水平顯著高于相應對照組(P<0.05);IL-1B-511CC攜帶者血清IL-1β水平在各組彆間差異無統計學意義(P>0.05)。(3)病例組IL-1B-511TT基因型頻率顯著高于對照組( P<0.05,OR=1.55,95%CI=1.10~2.18),兩組間T等位基因頻率差異有統計學意義(P<0.05,OR=1.31,95%CI=1.05~1.63)。 IL-1RN VNTR二組間差異無統計學意義(P>0.05)。(4)IL-1B-511C/T SNP基因型頻率在不同轉歸類型中差異有統計學意義( P<0.005)。與相應的CT+CC基因型比較,不同轉歸類型各組IL-1B-511TT頻率均顯著高于對照組,其OR(95%CI)分彆為:輕中型組2.17(1.48~3.19);重型組2.11(1.05~4.26);肝硬化組2.98(1.77~4.99);肝癌組4.33(2.16~8.67)。 IL-1B-511T頻率組間差異有統計學意義(P<0.005)。與對照組比,IL-1 B-511 T 在HCV感染各組OR (95%CI )依次分彆是:1.80(1.38~2.36);1.80(1.08~3.01);2.62(1.76~3.89);3.49(1.96~6.23)。(5)IL-1B SNP、IL-1RN VNTR與1b或2a基因型HCV感染差異無統計學意義;IL-1 B-511 C/T SNP與抗病毒治療前HCV RNA複製、抗病毒治療的反應性及血清ALT水平無關。結論 IL-1B-511T患者血清IL-1β、IL-1ra水平顯著升高,而IL-1ra/IL-1β比值顯著降低;IL-1B-511 SNP與HCV肝病慢性化及不同臨床轉歸相關,IL-1B-511TT/T能增加HCV慢性感染及其臨床轉歸的危險,IL-1B-511CC/C則是抗慢性感染和抗慢性化轉歸的保護因素;IL-1B-511T等位基因與1b型HCV具有協同升高IL-1β水平的作用。
목적:탐토IL-1β、IL-1ra혈청수평급백세포개소-1B( IL-1B)화백세포개소-1RN( IL-1RN)기인다태성여HCV상관간병발생발전적관계。방법이매련면역흡부시험( ELISA)측정IL-1β、IL-1ra혈청수평;기인심편기술검측310례HCV간병화324례불상관련적건강대조인군적IL-1B-31 C/T、IL-1B-511C/T단핵감산다태성(SNP);경지당응효전영검측IL-1RN내함자2(intron 2)기인다태성( VNTR)。취합매련반응-한제성편단장도다태성( PCR-RFLP)수궤분석부분양본험증기인심편분형결과;HCV기인분형채용직접측서법;Lightcycler 특이성형광탐침PCR검측HCV RNA재량;혈청병안산안기전이매( alanine aminotransferase ,ALT)사용ROCHE cobas 8000전자동생화분석의검측。결과(1)병례조IL-1β혈청수평(22.6±7.3) pg/ml현저고우대조조(13.7±4.2) pg/ml( P<0.01);IL-1ra혈청수평(286.30±55.10) pg/ml현저고우대조조(185.55±48.32) pg/ml(P<0.01)。(2)재병례조、경중형간염、중형간염、간경화합병간암조、1b형조급2a형조,IL-1B-511T등위기인휴대자혈청IL-1β수평균현저고우기상응CC순화자급대조조( P<0.05),차HCV감염각조-511 T휴대자IL-1ra/IL-1β비치현저저우대조조(P<0.05);IL-1B-511T휴대자재1b형조적IL-1β수평교2a형조현저증고(P<0.05);IL-1B-511T휴대자IL-1ra혈청수평현저고우상응대조조(P<0.05);IL-1B-511CC휴대자혈청IL-1β수평재각조별간차이무통계학의의(P>0.05)。(3)병례조IL-1B-511TT기인형빈솔현저고우대조조( P<0.05,OR=1.55,95%CI=1.10~2.18),량조간T등위기인빈솔차이유통계학의의(P<0.05,OR=1.31,95%CI=1.05~1.63)。 IL-1RN VNTR이조간차이무통계학의의(P>0.05)。(4)IL-1B-511C/T SNP기인형빈솔재불동전귀류형중차이유통계학의의( P<0.005)。여상응적CT+CC기인형비교,불동전귀류형각조IL-1B-511TT빈솔균현저고우대조조,기OR(95%CI)분별위:경중형조2.17(1.48~3.19);중형조2.11(1.05~4.26);간경화조2.98(1.77~4.99);간암조4.33(2.16~8.67)。 IL-1B-511T빈솔조간차이유통계학의의(P<0.005)。여대조조비,IL-1 B-511 T 재HCV감염각조OR (95%CI )의차분별시:1.80(1.38~2.36);1.80(1.08~3.01);2.62(1.76~3.89);3.49(1.96~6.23)。(5)IL-1B SNP、IL-1RN VNTR여1b혹2a기인형HCV감염차이무통계학의의;IL-1 B-511 C/T SNP여항병독치료전HCV RNA복제、항병독치료적반응성급혈청ALT수평무관。결론 IL-1B-511T환자혈청IL-1β、IL-1ra수평현저승고,이IL-1ra/IL-1β비치현저강저;IL-1B-511 SNP여HCV간병만성화급불동림상전귀상관,IL-1B-511TT/T능증가HCV만성감염급기림상전귀적위험,IL-1B-511CC/C칙시항만성감염화항만성화전귀적보호인소;IL-1B-511T등위기인여1b형HCV구유협동승고IL-1β수평적작용。
Objective To study the characteristics of IL-1B-31/-511 single nucleotide polymor-phisms (SNPs) and the variable number tandem repeat (VNTR) in intron 2 of the IL-1ra gene (IL-1RN) in patients with HCV-related liver diseases .Methods The concentration of IL-1βand IL-1ra in serum sam-ples was measured by ELISA assay .The SNPs of IL-1B gene (-31C/T,-511C/T) from 310 cases with HCV infection and 324 unrelated healthy controls were determined by using gene chip analysis , and the results for some randomly selected specimens were compared with those by using polymerase chain reaction -restriction fragment length polymorphism ( PCR-RFLP) assay.The VNTR polymorphism of IL-1RN intron 2 was ana-lyzed by PCR-RFLP assay.The serum level of alanine aminotransferase (ALT), an indicator of hepatocellu-lar injury, was detected by ROCHE cobas 8000 analyzer.HCV replication was measured by using specific fluorescence PCR .The genotypes of HCV were determined by direct nucleotide sequencing test .Results Compared with control group, the serum level of both IL-1β[(22.6 ±7.3) vs (13.7 ±4.2)] pg/ml and IL-1ra [(286.30 ±55.10) vs (185.55 ±48.32)] pg/ml were significantly increased in patients with HCV infection ( P<0.01 ) .There were two predominant genotypes identified among 310 patients including HCV 1b (75.5%) and HCV 2a (22.3%).The serum level of IL-1βand IL-1ra in IL-1B-511T carriers from four groups including case group , mild and moderate Hepatitis C group , severe Hepatitis C , and cirrhosis and hepatocellular carcinoma (HCC) group were significantly higher than those from IL-1B-511CC carriers and control group (P<0.05).The ratio of IL-1ra to IL-1βin all IL-1B-511T carriers with HCV infection were lower than those from healthy controls (P<0.05).IL-1B-511T carriers with HCV genotype 1b infec-tion showed a higher serum level of IL-1βas compared with those with HCV genotype 1a infection ( P<0.05).Compared with control group, they also showed an increase in IL-1ra level (P<0.05).There was no significant difference in the serum level of IL-1βamong IL-1B-511CC carriers from each group ( P>0.05).The frequency of IL-1B-511TT genotype (P<0.05, OR=1.55, 95% CI =1.10-2.18) and IL-1B-511T allele (P<0.05,OR=1.31,95% CI=1.05-1.63) in patients with HCV infection were signifi-cantly higher than those in healthy controls .IL-1B-511C/T SNP showed a significant association with the outcomes of HCV infection (P<0.005).Compared with IL-1B-511CC and IL-1B-511CT, IL-1B-511TT was a major risk factor for mild and moderate Hepatitis C [ OR=2.17 ( 1.48-3.19 ) ] , severe Hepatitis C [OR=2.11(1.05-4.26)], cirrhosis [OR=2.98(1.77-4.99)] and HCC [4.33(2.16-8.67)].IL-1B-511 T allele was significantly associated with mild and moderate Hepatitis C [ 1.80 ( 1.38-2.36 ) ] , severe Hepatitis C [1.80(1.08-3.01)], cirrhosis [2.62(1.76-3.89)] and HCC [3.49(1.96-6.23)].The fre-quency of IL-1B-511T allele showed significant difference among each group (P<0.005).No association was found between any of the other polymorphisms and HCV infection .Conclusion The serum level of IL-1βand IL-1ra were significantly associated with HCV infection .IL-1B-511T allele in patients with HCV in-fection up-regulated the serum level of IL-1β.IL-1B-511TT and IL-1B-511T allele were major risk factors for mild and moderate Hepatitis C, severe Hepatitis C, cirrhosis and HCC, but IL-1B-511CC/C had oppo-site effects.