慢加急性乙型肝炎肝衰竭患者中CD163和sCD163的临床表达研究

Study on the expression of CD163 and sCD163 in patients with clinical acute on chronic hepatitis B liver failure

目的检测慢加急性乙型肝炎肝衰竭(ACHBLF)患者中外周血单个核细胞表面CD163及外周血血浆中s CD163的表达,明确其在急性炎性反应过程中扮演的角色及临床意义,为临床治疗ACHBLF患者寻找新的临床监测因子。方法 ELISA方法检测ACHBLF患者、慢性乙型肝炎(CHB)患者及正常对照组(HCs)血浆中s CD163的表达水平;流式细胞术检测ACHBLF患者、CHB患者及HCs外周血单个核细胞CD163的阳性率和CD163平均荧光强度(MH),应用反转录聚合酶链反应(RT-PCR)测量ACHBLF患者、CHB患者及HCs外周血单个核细胞中CD163 mRNA的表达。结果外周血中CD163单核细胞阳性率在ACHBLF组[(47.89±16.58)%]要明显高于CHB患者[(33.24±11.39)%]及HCs[(16.57±6.38)%],具有统计学差异;外周血中CD163荧光强度在ACHBLF患者中较其他两组升高,但三组无统计学差异;CD163 RNA在ACHBLF患者中的表达(0.014 40±0.011 20)要明显高于CHB(0.005 23±0.001 41)及HCs(0.003 89±0.001 52),具有统计学差异;血浆中s CD163表达在ACHBLF患者中明显升高,且与患者预后有明显相关,在生存组患者的血浆中s CD163浓度要明显低于死亡组患者SCD163浓度[(5 435.84±1 562.28)ng/ml vs.(3 657.29±1 369.69)ng/ml,P=0.03],具有统计学差异;血浆中s CD163表达在ACHBLF患者中与反映患者预后的MELDScore呈正相关,与HBV-DNA正相关,凝血酶原活动度负相关,与总胆红素水平无相关;外周血中CD163阳性的单个核细胞比率在CHB患者中要明显高于HCs,具有统计学差异;CD163 mRNA在CHB患者中的表达要高于HCs,但并不具有统计学差异,血浆SCD163表达在CHB患者中明显升高,高于HCs,但是升高幅度远远小于其在ACHBLF患者中升高幅度,具有统计学差异。结论作为在巨噬细胞表面特异表达的CD163在ACHBLF患者、CHB患者发病过程中起重要作用,CD163与SCD163都可以作为ACHBLF患者有用的标记物,其中SCD163检测更为简单直接,组间差异更为明显,与疾病的严重程度明显相关。CD163与SCDI63都可以作为肝衰竭患者预后的评价指标,与患者预后呈负相关。

Objective To study the expression and slgnlticance otCDl63 in ACHBLF patients. We detected the surface CD163 in peripheral blood mononuclear cells （PBMCs） and sCD163 in peripheral blood in ACHBLF patients. We want to know the relationship between the expression of CD163, sCD163 and the prognosis of patients with ACHBLF patients, and their relationship with clinical indexes and clinical significance. Methods We detected the frequency of CD163＋ PBMCs using flow cytometry （FCM）. We also evaluated the average mean fluorescence intensity （MFI） of CD 163＋ PBMCs using FCM. Real-time transcripfion-polymerase chain reaction （RT-PCR） was performed to assess relative CD163 mRNA levels in PBMCs. We evaluated the plasma levels of sCD163 by enzyme linked immunosorbent assay （ELISA）. Results CD163 positive in PBMCs in patients with ACHBLF was significantly higher than CHB patients and healthy controls （l-ICs）, there were significant differences within three groups. However, there were no significant differences in MFI in CD163~ PBMCs within three groups. CD163 mRNA expression in ACHBLF patients was significantly higher than CHB patients and HCs, there were significant differences within three groups. Plasma sCD163 expression was significantly elevated in patients with ACHBLF, and was significantly correlated with the prognosis of patients. There were significant differences within three groups. Plasma levels of sCD163 were positively correlated with MELD score, plasma HBV-DNA levels and negatively correlated with PTA, but had no correlation with TBIL. CD163 positive in PBMCs in CHB patients was significantly higher than that in HCs. CD163 mRNA expression in CI-IB patients was higher than that of HCs, there was no significant difference between two groups. Plasma sCD163 expression was significantly elevated in patients with CHB, there was significant difference between two groups. Conclusion As expressed in the macrophage surface specific, CD163 plays an important role in patients with ACI-1BLF and CHB in the pathogenesis. The research suggested that CD163 and sCD163 can be used as useful biomarkers for ACHBLF. SCD163 detection is more simple and direct, with more significant group differences. It is significantly correlated with the severity of the disease. CD163 and sCD163 can be used as index of prognosis in patients with liver failure, and negatively correlated with the prognosis of the patients.