摘要
为建立鸡胚胎尿绒膜微量血采集方法和检测胚胎血中同型半胱氨酸 (HCY)、叶酸、HCY - 2抗体及亚甲基四氢叶酸还原酶 (MTHFR)基因型变化情况 ,采用HPLC法测血浆总HCY ,PCR -RFLP法测纸血片MTHFR ,SLISA法测血清HCY - 2抗体和微生物法测纸血片血红蛋白叶酸盐。结果发现连续 7d给D .L -HCY后 ,鸡胚血浆总HCY浓度 ( 10 6 92 μmol/L)显著高于正常对照组 ( 14 16μmol/L) ;血清HCY - 2抗体水平( 0 4 2 μg/ml)亦明显高于对照组 ( 0 33μg/ml) ;未观察到MTHFR基因 667C→T突变。D .L -HCY对鸡血红蛋白叶酸盐含量影响不明显 (P >0 0 5) ;联合给药发现 ,D .L -HCY能显著降低叶酸所致的鸡胚血红蛋白叶酸盐水平 (P <0 0 1)。鸡胚尿绒膜微量采血方法具有实用、经济、简单易行等优点 ;高HCY血症可能是神经管畸形等疾病发生的一个重要危险因素。
The levels of homocysteine (HCY),hemoglobin folate and HCY-2 antibody and methylenetetrahydrofolate reductase(MTHFR) genotype in microblood specimens from chick embryonic choricallantoic blood.Were determined by HPLC,PCR-RFLP,ELISA and microbiological assay respectively.8 μmol of D.L-HCY and 5μg of folic acid were dropped on inner shell membrane,and the embryos were continuedly incubated to collection days.(100±20)μl blood sample was drawn from chorioallantoic blood vessels of embryos aged 10.5～11 days after continually dropping 7 days.The results of assay were as follows:the mean level of plasma total HCY was 106.92μmol/L in the treated embryos,which was obviously higher than control (14.16μmol/L),and the mean level of HCY-2 antibodies (0.42μg/ml) in sera was higher than the mean level of control (0.33μg/ml).The 677(C→T) mutation in the MTHFR gene was not observed in the treated and untreated embryonic samples.When 5μg of folic acid was placed on the inner shell membrane of day 10.5 embryos,there was a rapid rise in microtitre dried blood spot hemoglobin folate with a peak in 1 h at 22.3μg/g.During the total transient time course,D.L-HCY did not result in the obvious change of hemoglobin folate concentration(P>0 05).However,at the group the of combinative treatment of folic acid with D.L-HCY could significantly suppress the elevation of hemoglobin folate level by using 5μg folic acid(P<0 01).The established method was an effective,economic,simple and rapid.The exogenous D.L-HCY and folic acid could permeate the chick embryonic membrane and into the blood circulation of embryo.These findings support the hypothesis that hyperhomocysteinemia is an important risk factor for neural tube defects.
出处
《中国医师杂志》
CAS
2000年第5期262-264,共3页
Journal of Chinese Physician
