摘要
以重组的ＧＭＣＳＦ和ＩＬ３作为造血细胞存活因子，正常小鼠骨髓细胞与再障小鼠淋巴细胞共培养，同时设立正常骨髓细胞组、再障小鼠脾淋巴细胞培养组及正常－骨髓细胞和正常脾淋巴细胞共培养组等三个平行对照组，分别于培养第０、２、４、８、１２、１８、２４、３６ｈ收取标本，结果表明实验组第８ｈ后各时点骨髓造血细胞发生凋亡：（１）流式细胞仪所测凋亡细胞百分率，实验组在第８、１８、２４、３６ｈ分别为３８．７、４０．３、５４．５、７３．８、６７．９，明显高于各对照组。（２）电镜观察发现第８ｈ后实验组骨髓细胞发生皱缩，电子密度增高，质膜出现芽状突起，染色质分布于核的周边·（３）核ＤＮＡ裂解分析表明，第８ｈ后实验组的琼脂糖凝胶电泳呈现“梯状”条带，呈１８０ｂｐ的整数倍，ＤＮＡ裂解百分率也明显高于各对照组，对照组未出现“梯状”电泳条带·结果表明再障淋巴细胞诱导骨髓造血细胞发生凋亡。提示设法阻断或抑制造血细胞凋亡过程可望成为治疗再障的新靶点。
We reported previously that apastic anemia(AA)occured in BALB/c mice by transfusion of lymphocytes from H2 match but Mls unmatch strainDBA/2,and that the lymphocytes of AA mice suppressed the hematopoietic stem cells and progenitor cells of normal murine bone marrow.It is,however,still unclear how the hematopoietic cells are injured in the model.It is known that hematopoietic cells can survive in vitro in the presence of some survival factors such as recombinant GMCSF and IL3.Here we cocultured the hematopoeitic cells from bone marrow of normal mice with an equal number of the lymphocytes of AA mice in the presence of those survival factors.In the mean time,three parallel controls,hematopoetic cells of normal mice,splenic lymphocytes of AA mice,and combination of hematopoietic cells and lymphocytes from normal mice,were designed.At different time points,we detected the samples by flow cytometry(FACS),transmission electron microscopy(TEM)and gel electrophoretic analysis of DNA.The results in experimental mice 8 hours after coculture showed:(i)The percentages of apoptotic death cells increased.(ii)Hematopoietic cells appeared nuclear condensation,margination and blebbing under TEM.(iii)Agarose gel electrophoresis of DNA in the 13 00×g supernatant showed ladder pattern,which suggest the choromatin cleavage is at internucleosome sites.The percentage of DNA fragmentation measured by diphenylamine reaction also increased.Our experimental results indicate that apoptosis occurs in hematopoietic cells when induced by splenic lymphocytes of mice with aplastic anemia mediated immunologically,and suggest this phenomenon might contribute to the pathogenesis of hamatopoietic failure in AA,and that attempt to block the process of apoptosis could be a new clue of treating AA.
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
1997年第5期465-469,共5页
Chinese Journal of Pathophysiology
基金
国家自然科学基金
广东省科委青年科学基金
