CAJ | 학술논문

目的：分析临床不同疾病程度慢性乙型肝炎（CHB ）患者和正常对照者外周血辅助性 T 细胞17（Th17）的比例和其效应分子白细胞介素17（IL-17）水平的变化与疾病严重程度、乙型肝炎病毒（HBV）DNA载量、肝脏损伤标志物等的关系，以进一步证实Th17细胞在CHB患者疾病进程中的作用。方法应用流式细胞术、实时荧光定量聚合酶链反应（PCR）和酶联免疫吸附试验（ELISA）等检测93例CHB患者（其中轻度21例、中度37例、重度35例）和28名正常对照者外周血Th17细胞比例、血清IL-17水平，纯化CD4＋T细胞经功能性抗CD3、抗CD28抗体刺激后IL-17和维甲酸相关孤儿核受体C（RORC）mRNA表达水平，纯化CD4＋T细胞刺激后培养液中IL-17水平。比较和分析上述检测数据在各组间的统计学差异及与HBV DNA载量、肝脏损伤标志物等的关系。结果与正常对照组相比，CHB组外周血Th17细胞比例明显升高，且随着疾病的加重，Th17细胞的比例也随之上升。CHB重度组Th17细胞比例、IL-17和RORC mRNA表达水平显著高于中度组、轻度组和正常对照组（P＜0．05），各组间血清IL-17水平差异无统计学意义（P＞0．05）；CHB组纯化CD4＋T细胞刺激后培养液中IL-17水平显著高于正常对照组（P＜0．05）；CHB重度组培养液IL-17水平明显高于中度组和轻度组（P＜0．05）。CHB组外周血Th17细胞比例与血清丙氨酸氨基转移酶（ALT）水平呈明显正相关（P＜0．01），而与血清HBV DNA载量无相关性。结论 CHB患者体内增高的Th17细胞可能是造成肝脏炎症、肝细胞损伤的主要原因之一，并与CHB的重症化密切相关。
목적：분석림상불동질병정도만성을형간염（CHB ）환자화정상대조자외주혈보조성 T 세포17（Th17）적비례화기효응분자백세포개소17（IL-17）수평적변화여질병엄중정도、을형간염병독（HBV）DNA재량、간장손상표지물등적관계，이진일보증실Th17세포재CHB환자질병진정중적작용。방법응용류식세포술、실시형광정량취합매련반응（PCR）화매련면역흡부시험（ELISA）등검측93례CHB환자（기중경도21례、중도37례、중도35례）화28명정상대조자외주혈Th17세포비례、혈청IL-17수평，순화CD4＋T세포경공능성항CD3、항CD28항체자격후IL-17화유갑산상관고인핵수체C（RORC）mRNA표체수평，순화CD4＋T세포자격후배양액중IL-17수평。비교화분석상술검측수거재각조간적통계학차이급여HBV DNA재량、간장손상표지물등적관계。결과여정상대조조상비，CHB조외주혈Th17세포비례명현승고，차수착질병적가중，Th17세포적비례야수지상승。CHB중도조Th17세포비례、IL-17화RORC mRNA표체수평현저고우중도조、경도조화정상대조조（P＜0．05），각조간혈청IL-17수평차이무통계학의의（P＞0．05）；CHB조순화CD4＋T세포자격후배양액중IL-17수평현저고우정상대조조（P＜0．05）；CHB중도조배양액IL-17수평명현고우중도조화경도조（P＜0．05）。CHB조외주혈Th17세포비례여혈청병안산안기전이매（ALT）수평정명현정상관（P＜0．01），이여혈청HBV DNA재량무상관성。결론 CHB환자체내증고적Th17세포가능시조성간장염증、간세포손상적주요원인지일，병여CHB적중증화밀절상관。
Objective To analyze T helper 1 7 (Th1 7 ) cell proportion and interleukin 1 7 (IL-1 7 ) levels in peripheral blood of patients with chronic hepatitis B(CHB)and healthy controls,and to evaluate their correlations with disease severity,serum hepatitis B virus(HBV)DNA load and liver injury markers,in order to investigate Th1 7 cells′role in the pathogenesis of CHB.Methods The Th1 7 cell proportion and IL-1 7 levels in peripheral blood of 93 CHB patients (the mild,moderate and severe CHB patients had 21 ,37 and 35 cases,respectively)and 28 healthy controls were determined by flow cytometry and enzyme-linked immunosorbent assay(ELISA).The mRNA expression levels of IL-1 7 and RAR-related orphan nuclear receptor C (RORC)of purified CD4 +T cells after stimulating with anti-CD3 and anti-CD28 antibodies were analyzed by real-time fluorescence polymerase chain reaction(PCR).The IL-1 7 levels in the supernatant of purified CD4 +T cells after stimulating with anti-CD3 and anti-CD28 antibodies were evaluated by ELISA. The data were analyzed statistically,and their relations with serum HBV DNA load,liver injury markers,and so on were analyzed.Results Compared with those of healthy control group,the proportion of Th1 7 cells in peripheral blood of CHB group was significantly higher,and increased with disease severity.The Th1 7 cell proportion,IL-1 7 levels and RORC mRNA levels in severe CHB group were significantly higher than those in moderate and mild CHB groups and healthy control group (P<0.05 ).There was no statistical significance in serum IL-1 7 levels among the groups(P>0.05).The IL-1 7 levels in the supernatant of purified CD4 +T cells after stimulating with anti-CD3 and anti-CD28 antibodies from CHB group were significantly higher than those from healthy control group(P<0.05),and the IL-17 levels in the supernatant from severe CHB group were higher than those from moderate and mild groups(P<0.05).The Th1 7 cell proportion in peripheral blood of CHB group showed a positive correlation with serum alanine aminotransferase (ALT)level (P<0.01 ),and showed no correlation with serum HBV DNA load.Conclusions The increased Th1 7 cell proportion in CHB patients may be the one of the factors for liver inflammatory response and liver injury and cause the progression of CHB.