恩替卡韦对肝移植受者血清及外周血单个核细胞内HBV DNA的抑制作用

Inhibition effect of Entecavir on hepatitis B virus DNA in serum and peripheral blood mononuclear cells of posttransplant patients

目的研究恩替卡韦对乙型肝炎相关性终末期肝病患者肝移植术后血清及外周血单个核细胞(PBMC)内HBV DNA的抑制作用。方法选取从2007年8～12月的20例HBsAg阳性肝移植受者作为研究对象(围手术期组),给予口服恩替卡韦每天0.5 mg联合肌注乙肝免疫球蛋白作为HBV再感染的预防方案,分别于术前1 d和术后1、4、12周采用实时荧光定量PCR法检测血清及PBMC内HBV DNA定量。同时回顾性分析2006年8月至2007年8月共34例行同种异体原位肝移植,术后长期使用恩替卡韦联合肌注乙肝免疫球蛋白的患者(随访组),随访时间4.0～22.5个月,同样的方法检测血清及PBMC内HBV DNA定量。结果20例受者血清HBV DNA在恩替卡韦治疗12周时全部转阴。术前1 d和术后1、4、12周PBMC内HBV DNA阳性率分别为85.0%(17/20)和45.0%(9/20)、45.0%(9/20)、40.0%(8/20),术后1周与术前比较差异有统计学意义(P=0.004),但1周以后阳性率无显著变化;围手术期PBMC内HBV DNA定量均值分别为104.07±2.07和101.69±1.96、101.51±1.72、101.30±1.63拷贝/106细胞,同样术后1周与术前比较差异有统计学意义(P=0.01),但随术后时间的延长,下降趋缓,术后4周后定量值的下降差异无统计学意义(P>0.05)。随访组平均随访13.6个月,均未发现HBV再感染(血清HBV DNA均阴性),PBMC内HBVDNA阳性率为32.4%(11/34),定量均值101.03±0.26拷贝/106细胞。结论恩替卡韦对肝移植术后血清及PBMC内HBVDNA均具有较好的抑制效果,但PBMC内HBVDNA下降趋势在术后4周后即维持在相对稳定状态,不能完全被清除。

Objective To explore the inhibition effect of Entecavir on hepatitis B virus （ HBV ） DNA in serum and peripheral blood mononuclear cells （PBMCs） of patients with HBV-related end-stage liver disease after transplantation. Methods From August to December 2007, 20 HBsAg positive liver graft recipients （perioperative group） were treated with Entecavir 0.5 mg/d and additional intramuscular hepatitis B immunoglobulin （HBIG） for prophylaxis of HBV recurrence. HBV DNA in serum and PBMCs was quantified by specific real-time fluorescence PCR 4 times（ on the day before operation and in 1, 4 and 12 weeks after transplantation）. Another 34 patients （retrospective group） who underwent liver transplantation from August 2006 to August 2007 and were treated with Entecavir were followed up from 4.0 to 22.5 months. HBV DNA copies in serum and PBMCs were quantified by the same method. Results In the perioperative group, HBV DNA in serum became negative in all 20 cases during Enteeavir treatment. The positive rates of HBV DNA in PBMCs on the day before operation and in 1,4 and 12 weeks after liver transplantation （LT） were 85.0% （17/20）, 45.0% （9/20）, 45.0% （9/20） and 40.0% （8/20）, respectively. Significant difference in the positive rates of HBV DNA in PBMCs was noted between the 1-week posttransplant group and the preoperative group （P = 0. 004, X^2 test）, but no significant difference between any posttransplant groups. The level of HBV DNA in the PBMCs of 4 times were 10^4.07±2.07 , 10^1.69±1.96, 10^1.51 ±1.72 and 10^1.30±1.63 copies/10^6 cells, respectively. The level of the 1-week posttransplant group was significantly lower than that of the preoperative group （P =0.01 ）, but the decrease became no significant difference 4 weeks after transplantation. The retrospective group had been traced no HBV recurrence on an average follow-up period of 13.6 months （the positive rate of HBV DNA in serum was 0）. The average positive rate in the PBMCs was 32.4% （11/34） and the level of HBV DNA in the PBMCs was 10^103±0.26 copies/10^6 cells. Conclusions Enteeavir has remarked inhibitory effect on HBV DNA both in serum and PBMCs of pesttransplant patients. However it cannot eliminate all the HBV in PBMCs, as the level of HBV DNA fluctuated in a very low range 4 weeks after transplantation.