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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。
AIM: To investigate the direct effect of berberine on primary cortical and cerebellar granule neurons cultured in vitro and the interaction between Ber and indirect bilirubin. Methods: The survival rate and morphological characteristics of neurons were observed by using fluorescein diacetate (FDA) and Hoechst33258 DNA staining. The biochemical characteristics of neuronal death induced by Ber were analyzed by agarose gel electrophoresis and flow cytometry. Results: Ber inhibited the neuronal death of cerebral cortex and cerebellar granule cells in primary culture in a dose-dependent manner (0 ~ 20μmol / L). The mechanism of its death showed the characteristics of necrosis: including neuronal cell body enlargement, no obvious nuclear chromatin condensation, DNA random rupture in the gel electrophoresis showed diffuse samples change, no apoptotic peak analysis by flow cytometry. Pretreatment of neurons with protein synthesis inhibitors failed to block the neurotoxic effect of Ber. The addition of Ber (1 μmol / L) at non-toxic doses to bilirubin at non-toxic doses can produce neuronal toxicity. Conclusion: Ber induced primary cultured rat cerebral cortex and cerebellar granule neurons necrosis, and enhance the neurotoxic effect of bilirubin.