丁型肝炎病人肝组织HDV和HBV表达及关系

Study of the Role and the Relationship between HDV and HBV in the Liver Tissues of Patients with Hepatitis D

目的观察丁型肝炎病人肝组织HDAg、HBsAg、HBcAg及HBV DNA表达,了解HDV和HBV的作用和关系。方法应用免疫组化双标记及原位杂交技术,检测48例丁型肝炎病人肝组织HDAg、HBsAg、HBcAg和HBV DNA表达,以52例乙型肝炎作疾病对照。结果丁型肝炎HBsAg、HBcAg、HBV DNA检测率分别为83％、67％、21％,对照组为94％、92％、44％,两组HBcAg、HBV DNA表达有差别意义(P<0.05)。HDAg主以核浆型表达,阳性细胞在门脉周围、碎屑状坏死处较密集,坏死灶边缘和气球样变性的肝细胞浆内有HDAg呈浆型强阳性表达。HBsAg主表达于肝细胞浆,阳性细胞分布类似HDAg阳性细胞,HBsAg和HDAg表达及分布有相关性。HBcAg主以核浆型表达,阳性细胞明显少于HDAg阳性细胞,分布呈单个细胞、点状(3—5个细胞)较多。HBV DNA在肝细胞浆或核内呈稀疏分布、着色浅淡,HBV DNA复制与HDAg表达强度呈负相关(P<0.05)。结论 HDV感染的HBV抑制状况,主要在HDV急性感染或HDAg强表达阶段;HDV致病机制中既有HBV和HDV相加作用,可能更主要有HDV的直接细胞毒性作用。

Objective To explore the role and the relationship between HDV and HBV in the liver tissues of patients with hepatitis D (HD). Methods HDAg, HBsAg, HBcAg in Paraffinembedded liver tissues from 48 liver biopsy or autopsy samples of the patients with HD were studied with immunohistochemical double labelling techniques; in situ hybrization was performed by using digocxinlabeled HBV DNA probes to observe the location and morphology of HBV DNA, meanwhile, compared with the expression of HBsAg, HBcAg and HBV DNA in liver tissues of contrast group. Results In HD group, positive rates for HBsAg, HBcAg and HBV DNA were 83%(40/48), 67%(32/48) and 21 (10/48). In contrast group, positive rates for HBsAg, HBcAg and HBV DNA were 94% (49/52), 92%(48/52) and 44%(23/52). There were a different singificance between HBcAg, HBV DNA expression in HD group and HB group (P<0.05). The positive signal of HDAg was mainly expressed in unclei and cytoplasm of hepatocyts, much of positive cells were obviously aggregated in periportal region, especially at the advanceing edges of areas of piecemeal necrosis, moreover, strong expression of HDAg was shown in the bollooning degeneration hepatocyts and hepatocyts near piecemeal necrosis region. HBsAg was mainly expressed in cytoplasm of hepatocytes, and positive cells distribution were similar to HDAg positive cells'. The expression and positive cells distribution of HDAg and HBsAg had close relationship. HBcAg was mainly expressed in unclei or in cytoplasm of hepatocytes, positive cells distribution were alone or focal type (3—5 cells) and much less wide than HDAg positive cells. But, HDAg, HBsAg and HBcAg were closely correlated with the activity of inflammation and the severity of the liver damage. HBV DNA positive granules were showed scarce of stained lightly in cytoplasm or unclei, the expression of HBV DNA was negatively co-related with HDAg expression (P<0.05). Conclusion HDV infection inhibits replication of HBV, especially during the active stage of HDV infection or during the strong expression phase of HDAg. Becides of interactions beteween HDV and HBV, it is possible that direct viral cytotoxicity of HDV may play a major pathogenetic role in pathogenetic mechanisms of HDV.