Bilirubin toxicity and differentiation of cultured astrocytes.
Bilirubin toxicity and differentiation of cultured astrocytes.
复制标题
DOI：
10.1038/sj.jp.7200180
复制
发表时间：
1999-04-01
期刊：
影响因子：
--
通讯作者：
Stevenson, D K
中科院分区：
文献类型：
--
作者：
Rhine, W D;Schmitter, S P;Stevenson, D K
目的：为了研究胆红素的毒性，在原代培养的新生大鼠大脑皮质星形胶质细胞。研究设计：原代培养的新生大鼠星形胶质细胞孵育胆红素浓度为0，1，5，10，25，50，100，200，和2000 μ M，在胆红素：白蛋白摩尔比为1.7。胆红素毒性测定细胞形态的变化，台盼蓝染色，乳酸脱氢酶（LDH）释放到培养基中在不同的时间孵育。为了确定星形胶质细胞的分化是否影响胆红素的毒性，培养物进行了处理与dibutyryl环磷酸腺苷。在胆红素浓度>或= 100 μ M时，实验培养物中的LDH释放在24小时内显著高于对照培养物（p < 0.05）。从1.5到24小时，200和2000 μ M培养物之间LDH释放的绝对量显著不同，之后暴露的持续时间似乎接管并且所有培养物接近最大值。较低浓度的LDH释放均在120小时达到最大值，但1 μ M培养物除外，其在整个研究期间未显示出显著高于对照的升高。在100和200 μ M胆红素，LDH释放未处理的细胞是显着高于（P < 0.05）比释放处理cells 36 h.CONCLUSION：未分化的星形胶质细胞似乎是更敏感的胆红素毒性，这可能与更大的敏感性，新生儿的kernicteric损伤。原代星形胶质细胞培养的研究可以提供胆红素敏感性如何随着大脑发育而变化以及胆红素脑病的细胞和生化机制的见解。
OBJECTIVE: To study the toxicity of bilirubin in primary cultures of newborn rat cerebral cortical astrocytes.STUDY DESIGN: Primary cultures of newborn rat astrocytes were incubated at bilirubin concentrations of 0, 1, 5, 10, 25, 50, 100, 200, and 2000 microM, at a bilirubin:albumin molar ratio of 1.7. Bilirubin toxicity was determined by changes in cellular morphology, trypan blue staining, and lactate dehydrogenase (LDH) release into the culture medium at various times of incubation. To determine if differentiation of astrocytes affects bilirubin toxicity, cultures were treated with dibutyryl cyclic adenosine monophosphate.RESULTS: All three indices of toxicity showed a bilirubin concentration dependence. LDH release in experimental cultures was significantly elevated (p < 0.05) above that of control cultures by 24 hours at bilirubin concentrations of > or = 100 microM. The absolute amount of LDH release differed significantly between the 200 and 2000 microM cultures from 1.5 to 24 hours, after which duration of exposure appeared to take over and all cultures approached maximum. LDH release for the lower concentrations all reached maximum by 120 hours, except for the 1 microM cultures, which showed no significant elevation above control throughout the study period. At 100 and 200 microM bilirubin, LDH release by untreated cells was significantly higher (p < 0.05) than release by treated cells by 36 hours.CONCLUSION: Undifferentiated astrocytes appeared to be more sensitive to bilirubin toxicity, which may correlate with the greater susceptibility of newborns to kernicteric injury. Studies with primary astrocyte culture may provide insight into how bilirubin sensitivity changes with brain development as well as the cellular and biochemical mechanisms of bilirubin encephalopathy.