目的 研究环磷酰胺(cyclophosphamide, CP)引起小鼠神经管畸形(neural tube defects, NTDs)的动物模型, 探讨牛磺酸的保护作用及其作用机理, 为预防NTDs的发生提供理论依据。方法 建立CP致小鼠NTDs模型, 分析比较各组鼠胚神经管发育情况和畸形情况;用免疫组织化学和图像分析技术检测各组鼠胚神经管上皮细胞中Cx26和Cx43蛋白的表达, 分析其与NTDs发生的关系。结果 1)用CP建立小鼠NTDs模型的最佳剂量为15 mg/kg体重, 牛磺酸能有效减少NTDs的发生;2)Cx26和Cx43蛋白定位于神经细胞的胞膜和胞质, CP组Cx26和Cx43蛋白的表达明显高于对照组(P＜0.05);中、高剂量牛磺酸组Cx26和Cx43蛋白的表达显著降低(P＜0.05)。结论 Cx26和Cx43的过度表达可能是CP致NTDs的途径之一;牛磺酸可能拮抗CP的毒性, 保护神经细胞, 减少NTDs的发生。
Abstract
Objective To study the animal model of neural tube defects (NTDs) induced by cyclophosphamide (CD) in mice, and to explore the protective effects of taurine and its mechanism, and provide a theoretical basis on the prevention of human NTDs. Methods NTDs model was set up in mouse embryo by CP.The mouse embryonic neural tube development situation and the abnormal situation in each group was analysised and compared.Immunohistochemistry and image analysis technology were used to detected the expression of Cx26 and Cx43 protein in each group of mouse embryonic neural tube epithelial cells.Its relationship was analyzed with NTDs happening. Results 1)The best dose was 15 mg/kg body weight to establish the model of NTDs in mice with CP.Taurine could effectively reducing the occurrence of NTDs.2)Cx26 and Cx43 proteins were localized in the cell membrane and cytoplasm in nerve cells.The expressions of Cx26 and Cx43 proteins in CP group were significantly higher than those in the control group (P＜0.05).The expressions of Cx26 and Cx43 proteins in middle and high dosage taurine group were decreased significantly (P＜0.05). Conclusions Excessive expressions of Cx26 and Cx43 might be one of the important causes of NTDs with cyclophosphamide.Taurine can antagone the toxicity of cyclophosphamide and protect nerve cells, witch may reduce the occurrence of NTDs.
关键词
环磷酰胺 /
神经管畸形 /
牛磺酸 /
Cx26 /
Cx43
Key words
cyclophosphamide /
neural tube defects /
taurine /
Connexin26 /
Connexin43
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参考文献
[1] 朱锦明, 杨欣.徐州市2004年～2009年出生缺陷资料分析[J].中国优生与遗传杂志, 2010, 18(12):110-111.
[2] Xiao R, Yu HL, Zhao HF, et al.Developemental neurotoxitcity role of cyclophosphamide on post-neural tube closure of rodents in vitro and in vivo[J].Int J Dev Neurosci, 2007, 25(8):531-537.
[3] Kibar Z, Capra V, Gros P.Toward understanding the genetic basis of neural tube defects[J].Clin Genet, 2007, 71(4):295-310.
[4] Copp AJ, Greene ND.Genetics and development of neural tube defects[J].J Pathol, 2010, 220(2):217-230.
[5] 金岩.小鼠发育生物学与胚胎实验方法[M].北京:人民卫生出版社, 2005:145-147.
[6] 赵海峰, 肖荣, 杨燕, 等.环磷酰胺致神经管畸形及可能机理研究[J].中国公共卫生, 2002, 18(11):1302-1304.
[7] 郑敏, 管英俊, 李如江.WIF-1、SFRP-1在环磷酰胺致大鼠神经管畸形中神经上皮细胞表达的变化[J].中国妇幼保健, 2012, 27(27):42649-4268.
[8] 张家魁, 王维林, 张锦华, 等.5氮杂胞苷对长春新碱及环磷酰胺诱导神经母细胞瘤细胞凋亡的调节作用[J].中华肿瘤防治杂志, 2010, 17(1):17-19.
[9] Gajda Z, Szupera Z, Quinine, et al.Blocker of neuronal Cx36 channels, suppresses seizure activity in rat neocortex in vivo[J].Epilepsia, 2005, 46:1581-1591.
[10] 王亚辰, 朴丰源, 马宁.牛磺酸和维生素 C 对砷致小鼠小脑神经毒性保护作用[J].中国公共卫生, 2012, 28(7):942-944.
[11] Sun M, Xu CH.Neuroprotective mechanism of taurine due to upregulating calpastatin and down-regulating calpain and Caspase-3 during focal cerebral ischemia cell[J].Mol Neurobiol, 2008, 28(4):593-611.
[12] Takatani T, Takahashi K, Uozumi Y, et al.Taurine prevents the ischemia-induced apoptosis in cultured neonatal rat cardiomyocytes through Akt/caspase-9 pathway[J].Biochemical and Biophysical Research Communications, 2004, 316(2):484-489.
[13] 何乐, 杨君杰, 彭玉立, 等.氯乙烯诱导小鼠胚胎神经管发育畸形的研究[J].第三军医大学学报, 2010, 32(2):159-163.
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