多发性骨髓瘤、非霍奇金淋巴瘤及Castleman病例hepcidin mRNA的表达

【作者】 王凤丹；

【导师】 周道斌； 沈悌； 武永吉；

【作者基本信息】 北京协和医学院 ， 临床医学， 2009， 博士

【摘要】 研究背景：慢性病贫血(ACD),也叫炎性贫血(AOI)是临床上最常见的贫血类型,铁利用障碍是其发病机制之一,hepcidin为调节铁代谢的激素,在其中起着重要作用。国内外研究均提示多发性骨髓瘤贫血的原因之一是ACD,病人血清中IL-6水平往往升高,白介素6(IL-6)在体外和动物实验中证实能促进hepcidin表达。但多发性骨髓瘤患者体内hepcidin的表达不详。贫血也是非霍奇金淋巴瘤及Castleman病常见的临床表现,文献报道其血清IL-6升高,hepcidin是否在这两种疾病贫血中也有作用,目前尚不清楚。研究目的：1,多发性骨髓瘤血清对肝腺瘤细胞系Hep-3b细胞hepcidin表达的影响。如有变化,是否能被IL-6单抗或重组人促红细胞生成素(rhEPO)抑制。2,其它疾病如非霍奇金淋巴瘤及Castleman病hepcidin的表达。研究方法：1,样本收集：多发性骨髓瘤、非霍奇金淋巴瘤患者的临床资料及血清；一例Castleman病例的临床资料、血清、肝活检及受累淋巴结组织；2,肝腺瘤细胞系Hep-3b细胞的培养；3, Hep-3b细胞培养液中按10%浓度加入多发性骨髓瘤及非霍奇金淋巴瘤患者血清；提取Hep-3b细胞的mRNA及Castleman病例肝组织、受累淋巴结组织的mRNA,进行RT-PCR,产物电泳后荧光分析,以hepcidin与内参β-actin PCR产物的比值进行半定量分析。研究结果：1,未治疗的多发性骨髓瘤患者血清与Hep-3b细胞共同培养的hepcidin mRNA表达量高于正常对照组,这种升高作用能被IL-6单抗或rhEPO抑制。2,治疗后的多发性骨髓瘤患者血清与Hep-3b细胞共同培养的hepcidin mRNA表达量显著低于未治疗者。3,多发性骨髓瘤患者短期随访中,规律治疗能稳定血红蛋白,降低患者血清对Hep-3b细胞hepcidin mRNA的影响。4,非霍奇金淋巴瘤患者(未治疗及治疗后)血清与Hep-3b细胞共同培养的hepcidin mRNA表达量较正常对照组降低。5, Castleman病例肝脏hepcidin mRNA表达量升高,受累淋巴结检测到hepcidin mRNA表达。结论：1,未治疗的多发性骨髓瘤患者血清能促进肝腺瘤细胞系Hep-3b细胞hepcidin表达,这种促进作用可被IL-6单抗拮抗,提示IL-6可能导致Hep-3b细胞hepcidin表达量升高,从而造成ACD。这种促进作用也能被rhEPO抑制,rhEPO可能对ACD有治疗作用。治疗后的多发性骨髓瘤患者不能促进Hep-3b细胞hepcidin表达,多发性骨髓瘤患者短期随访中,血红蛋白稳定,患者血清对Hep-3b细胞hepcidin mRNA的影响减小,可能治疗使病情稳定,改善了ACD。2,非霍奇金淋巴瘤患者血清不能促进hepcidin表达,提示ACD可能不是该组非霍奇金淋巴瘤患者贫血的原因之一3,研究的Casleman病例的肝脏hepcidin表达量升高,淋巴结肿物也检测到hepcidin的表达,可能大量表达的hepcidin导致了该病例严重的ACD。更多还原

【Abstract】 Background:Anemia of chronic disease (ACD), also called anemia of inflammation (AOI) is the most common cause of anemia in clinic. A specific feature of ACD is iron disorder, which probably is caused by hepcidin, an iron-regulation hormone. Hepcidin can be elevated by interleukin6(IL-6) both in vivo and in vitro, the cytokine proved to elevate in multiple myeloma, non-Hodgkin’s lymphoma and Castleman disease. It’s unclear whether hepcidin is also elevated in these diseases.Purpose:1. The effect of multiple myeloma patients’serum on hepcidin expression of Hep-3b hepatoma cell line. If hepcidin expression is elevated, try to decrease it by adding IL-6antibody or recombinant human erythropoietin (rhEPO).2. Hepcidin expression in non-Hodgkin’s lymphoma patients and a Castleman disease patient.Methods:1. Sample collection:Collect the clinic information and serum of multiple myeloma and non-Hodgkin’s lymphoma. Collect the clinic information, serum, liver biopsy tissue and lymphnode mass tissue of a Castleman disease patient.2. Culture Hep-3b hepatoma cell line.3. Add multiple myeloma and non-Hodgkin’s lymphoma patients’serum the medium. Extract mRNA from Hep-3b cell and tissue, and detect hepcidin mRNA using RT-PCR.Results:1. The serum of untreated multiple myeloma patients elevated hepcidin expression of Hep-3b cell, compared to health control. This effect was neutralized by human IL-6antibody or rhEPO. 2. The serum of treated multiple myeloma patients did not elevate hepcidin expression of Hep-3b cell.3. The hemoglobin was stable during the follow up period of regularly treated multiple myeloma patients and the serum effect on Hep-3b cell hepcidin expression was reduced.4. The serum of non-Hodgkin’s lymphoma patients did not elevate hepcidin expression of Hep-3b cell.5. The Castleman disease patient’s Liver hepcidin expression was elevated, and there was hepcidin expression in the lymph node.Conclusion:Hepcidin could be increased by treated multiple myeloma patients’ serum and decreased by rhEPO, which indicates rhEPO is a potent treatment for ACD. IL-6/hepcidin pathway in Castleman disease patient’s liver and lymph node probably led to her severe anemia.更多还原