窒息新生猪淋巴细胞凋亡与[Ca^(2+)]i内流的实验研究  

Study On Apoptosis And [Ca^(2+) ]i Infulx In Lymphocytes Of The Neonatal Piglets Following Hypoxia Ischemia

作　　者：王晓蕾[1] 刘晓平[1] 刘凯[1] 唐方[1] 张俊梅[1] 余世禄[1] 

机构地区：[1]成都市第三人民医院,610031

出　　处：《中华围产医学杂志》1999年第4期214-217,共4页Chinese Journal of Perinatal Medicine

基　　金：四川省卫生厅及成都市卫生局科研基金

摘　　要：目的　探讨窒息新生儿Ｔ淋巴细胞亚群的变化是否由于淋巴细胞凋亡异常启动所致，以及缺氧缺血（ＨＩ）后［Ｃａ２＋ ］ｉ内流对淋巴细胞凋亡的影响。　方法　用琼脂电泳及流式细胞术（ＦＣＭ）对ＨＩ新生猪淋巴细胞凋亡及淋巴细胞内游离钙（ＩＦＣａ）水平作动态定量、定性分析。　结果　ＨＩ新生猪淋巴细胞经体外培养２４～７２小时后，其最大凋亡率（ＭＡＲ）较对照组及尼莫地平治疗组明显增加（ｎ＝ １５， １２， １２）； （３１．０９±１７．０３）％ ， （５．５２±６．１７）％ ， （１６．９５±１１．３）％ ； Ｆ＝ １３．４８０， （Ｐ＜ ０．００１）；日平均凋亡率（ＤＡＲ）较其它两组显著增高（２４．０８±１０．４４）％ ， （４．０５±３．５７）％ ， （１３．２０±６．７９）％ ，Ｆ＝ ２２．４０６，Ｐ＜ ０．００ １）； ＨＩ后４小时，７２小时活体内淋巴细胞ＩＦＣａ 水平明显增加，（２９３±７２，１７６±４３，１００±４３， Ｆ＝ ３３．５１４ ９， Ｐ＜ ０．０００ １； ２４０±１１３， １７３±５５， １３７±５０， Ｆ＝ ５．９０９， Ｐ＜０．００５ ９）； ＭＡＲ 及ＤＡＲ 与ＨＩ后４小时活体内ＩＦＣａ 呈显著正相关， （ｒ＝ ０．３６１，Ｐ＝ ０．０２?Objective To determine whether the reason for the changes of T lymphocyte subsets in the neonates following prenatal asphyxia is probably the accelerated apoptosis of lymphocytes and whether the apoptosis is correlative to the intracellular free Calcium(IFCa) level. Methods The 'DNA Ladder' was observed by gel electrophoresis and dynamic changes of the apoptosis ratio and IFCa level in lymphocytes were investigated via Flow Cytometry(FCM) in the neonatal piglets following hypoxia ischemia. Results The max apoptosis rate (MAR) in lymphocytes (incubated for 24～72 hr in vitro) post HI was higher than control group and treatment group ( n=15, 12,12, ±s =31.09% ±17.03%,5.52%±6.17, 16.95±11.39%, F=13.48, P <0.000 1); the daily apoptosis ratio(DAR) was also more than the others( ±s =24.08±10.44%, 4.05%±3.57%, 13.20% ±6.79%, F=22.406, P <0.00 1); there was a significant increasing IFCa fluorescent intensity on the neonatal piglets' lymphocytes in viro 4hr later and 72 hr later post HI ( ±s =293±72, 176±70, 100±43, F=33.5149, P<0.000 1; ±s =240±113, 173±55, 137±50, F=5.908 8, P = 0.005 9) ; MAR and DAR were positively correlative to IFCa fluorescent intensity in vivo 4 hr later post HI( r=0.3614, P=0.024; r=0.3946, P =0.013); the appearance of ‘DNA Ladder’ was corresponded to hypodiploid peak observed by FCM. Conclusion The results suggest that there is accelerated apoptosis in lymphocytes of the neonates following prenatal asphyxia and the apoptosis is correlative to activation of Calcium channel; and that proper usage of Calcium channel blockers Nimodipin can partially inhibit this apoptosis.

关 键 词：新生儿窒息 新生猪 细胞凋亡 钙离子 T细胞亚群 

分 类 号：R722.120.2[医药卫生—儿科]