摘要
目的：研究小檗碱（Ｂｅｒ）对原代培养的大鼠大脑皮质和小脑颗粒神经元的直接作用及Ｂｅｒ与间接胆红素的相互作用。方法：采用二乙酸荧光素（ＦＤＡ）和Ｈｏｅｃｈｓｔ３３２５８ＤＮＡ染色法，观察神经元的存活率及形态学特征；用琼脂糖凝胶电泳和流式细胞仪分析Ｂｅｒ诱导神经元死亡的生化学特征。结果：Ｂｅｒ呈剂量（０～２０μｍｏｌ／Ｌ）依赖性地诱导原代培养的大鼠大脑皮质和小脑颗粒神经元死亡。其死亡的机制呈现坏死的特征：包括神经元胞体肿大、无明显核染色质浓缩、ＤＮＡ随机断裂而在凝胶电泳图上呈弥散样改变、流式细胞仪分析未见凋亡峰。用蛋白质合成抑制剂预处理神经元，不能阻断Ｂｅｒ的神经毒性作用。无毒性剂量下的Ｂｅｒ（１μｍｏｌ／Ｌ）与无毒性剂量下的胆红素相加，可产生神经元的毒性。结论：Ｂｅｒ诱导原代培养的大鼠大脑皮质和小脑颗粒神经元坏死，并增强胆红素的神经毒性作用。
Objective: The present study was aimed to examine the effect of berberine, the major ingredient of the Chinese herb huanglian (Coptis chinensis Franch), on primary cultured cortical and cerebellar granule neurons of rat and the interaction between berberine and bilirubin on the neurons. Methods: Neuronal viability and morphology were tested by fluorescent staining with fluorescein diacetate and Hoechst 33258. The biochemical features of berberine induced neuronal death were characterized by agarose gel electrophoresis and flowcytometric analysis. Results: We found that berberine induces neuronal death in a concentration dependent fashion at the range of 0～20 μmol/L. The neuronal death displayed the characteristics of necrosis, including an enlargement of cell bodies, without nuclear condensation, a diffuse agarose gel electrophoresis panel and without apoptosis peak in flowcytometric analysis. In addition, pretreatment of the primary cultures of cortical and cerebellar granule neurons with protein synthesis inhibitors, cycloheximide, did not block neuronal death induced by berberine. Moreover, treatment of the cultures with subtoxic concentrations of berberine (1 mg/L) and bilirubin produces neurotoxicity. Conclusions:Taken together, these data suggest that berberine induces neuronal death of primary cultured cortical and cerebellar granule neurons of rat via a necrosis mechanism. Berberine potentiates neurotoxicity induce by bilirubin.
出处
《中山医科大学学报》
CSCD
1998年第4期250-254,共5页
Academic Journal of Sun Yat-sen University of Medical Sciences
基金
国家自然科学基金
广东省自然科学基金
关键词
小檗碱
神经元
毒性
死亡
berberine/pharmacology
bilirubin/drug effects
neurons/drug effects
