摘要
目的
通过对慢性乙型肝炎(CHB)患者和慢加急性肝衰竭患者血清IL-10含量水平的测定及其启动子区甲基化状态的研究,探讨IL-10在肝病患者的炎症反应中的作用,比较IL-10基因启动子区甲基化水平与其含量的关系,进一步揭示肝病患者血清中IL-10水平变化的机制。
方法
肝衰竭患者组25例,其中男22例,女3例；慢乙肝患者组25例,其中男19例,女6例；正常对照组10例,均为身体健康者,其中男女各5例。通过酶联免疫吸附法(ELISA)测定血清中IL-10的含量,分析三组之间含量的差异。并将肝衰竭患者组的IL-10含量与其ALT,HBV-DNA,TBIL,PTA,MELD进行相关性分析,探讨IL-10水平与病情程度的关系。运用甲基化特异性聚合酶链反应(MSP)检测三组中IL-10启动子甲基化状态,与其血清中IL-10水平进行比较,揭示肝病患者血清中IL-10水平变化的可能机制。
结果
1、肝衰竭组与正常对照组之间比较IL-10含量明显升高,(P<0.05)有统计学意义；慢性乙型肝炎组较正常对照组血清IL-10含量明显升高,(P<0.05)有统计学意义；肝衰竭者与慢乙肝组之间比较有升高,但(P>0.05),无统计学意义。
2、在肝衰竭组中,IL-10含量与ALT水平无明显相关性(r=-0.022,P>0.05)；IL-10含量与HBV-DNA水平无明显相关性(r=0.033,P>0.05)；IL-10含量与TBIL水平呈明显正相关(r=0.566,P<0.05)；IL-10含量与PTA水平呈明显负相关(r=-0.581,P<0.05)；IL-10含量与MELD水平呈明显正相关(r=0.443,P<0.05)。
3、肝衰竭患者组甲基化分布状态与慢性乙型肝炎患者组和正常对照组相比都具有显著性差异(P<0.05),而慢性乙型肝炎患者组与正常对照组相比没有显著性差异(P>0.05)
4、IL-10血清含量与启动子甲基化状态呈负相关(r=-0.878,P<0.05)。
结论
1、在两患者组血清中,IL-10水平较正常对照组明显升高,并且肝衰竭患者较慢性乙型肝炎患者血清中的IL-10含量有一定程度的升高。
2、在肝衰竭患者中,IL-10水平与TBIL,PTA,MELD水平有明显的相关性,提示IL-10水平随肝衰竭程度的加重而升高。
3、IL-10血清含量与启动子甲基化状态呈明显的负相关,说明IL-10启动子区CpG岛甲基化可能是基因失活的重要机制。
Objective:
Detect the serum level and relativity of IL-10 and the methylation of its promoter, to investigate the effect and machanism of IL-10 the inflammatory reaction of chronic hepatitis B (CHB) and acute on chroinc hepatic failure patients.
Methods:
There are three groups of patients:the hepatic failure group, the CHB group, the healthy control group, with total of a 25 patients (22 men and 3 women),25 patients (19 men and 6 women),10 patients (5 men and 5 women), respectively.
1、detect the serum level of IL-10 via ELISA in all patients, to analyze the difference among the three groups;
2、detect the relativity of serum level of IL-10 in hepatic group with other related parameters as ALT, HBV-DNA, TBIL, PTA, MELD, etc, via correlation analysis, to investigate the relationship between level of IL-10 and degree of patient conditions;
3、detect the methylation of IL-10 promoter via MSP in three groups, compared with the serum level of IL-10 to investigate the mechanism of IL-10 in hepatitis patients.
Results:
1、Both the hepatic failure group and the CHB group have significant increase in serum level of IL-10 compared with the control group (p<0.05); the serum level of IL-10 in CHB group is higher than the hepatic failure group, however without statistical significance (p>0.05);
2、the serum level of IL-10 in hepatic failure group has no significant relativity with level of ALT and HBV-DNA (r=-0.022, r=0.033, respectively; P>0.05); a positive correlation with level of TBIL (r=0.566, P<0.05); and negative correlations with level of PTA and MELD (r=-0.581, r=0.443, respectively; P<0.05);
3、the distribution of the methylation of IL-10 promoter in hepatic failure group is significantly different from the other two; while no significant difference between the CHB group and the control group;
4、the serum level of IL-10 has a significantly negetive correlation with its methylation (r=-0.878, P<0.05)
conclusions:
1、the serum level of IL-10 in hepatitis patients is significantly higher; the hepatic failure patients have a certain increase in IL-10 compared with CHB;
2、The serum level of IL-10 in hepatic failure patients has a signifiant relativity with levels of TBIL, PTA and MELD, suggesting that the serum level of IL-10 is increasing with the degree of hepatic failure;
3、The negetive correlation between level of IL-10 and methylation of IL-10 promoter suggests that the CpG methylation in promoter may be important in the machanism of gene inactivation.
引文
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