灯盏细辛单体成分对混合培养鼠视网膜神经节细胞的影响

Effect of 4-Pyrone-3-β-D-Glucopyraoside and Neo-breviscapine onrat retinal ganglion cells in vitro

目的 观察灯盏细辛主要单体成分4－吡喃酮-3-β-1)－吡喃葡萄糖甙(PCP)和新灯盏花素(NBC)对混合培养鼠视网膜神经节细胞(RGCs)的影响。方法 采用胰酶消化法将12只生后2－3天的SD乳鼠视网膜制成细胞悬液,接种于鼠尾胶原包被的96孔培养板培养。分为对照组和各种质量浓度梯度(1×10－3g/L－1×10-8g/L)的单体成分实验组,于1,3,5天,用MTT微量自动比色法测量存活细胞的吸光度A值。结果 培养在鼠尾胶原上的细胞生长良好,上述质量浓度的PGP和NBC对培养细胞形态无影响。各培养时期实验组吸光度A值与对照组比较无显著性差异(P＞0. 05)。结论 灯盏细辛单体成分PGP和NBC无直接促鼠RGCs在体外存活的作用。

Objective To study the effects of 4-Pyrone-3-β-D-Glucopyraoside (PGP) and Neo-hreviscapine (NBC) , the twoelements of main integrants of Erigeron Breviseapus(Vant.) Hand-Mazy,on the survival of rat retinal ganglion cells( RGCs) in vitro andto screen for their effective concentrations. Methods RGCs were isolated from postnatal(2~3d) Sprague-Dawley rat eyes. RGCs incell culture were identified by morphologic criteria and immunocytochemical staining. Various concentrations(1 × 10~8 g/L ~ 1 × 10^(-3) g/L) of PGP and NBG were added individually to the cultures. MTT was added to each culture well on 1 ,3 and 5 days,respectively ,thenthe survival RGGs were evaluated by the absorbance(A) values with ELISA-Reader. The A values in wells in which PGP and NBC hadbeen added were compared with those from control wells. Results The cells cultured in the wells by murine-tail collagen grew verywell. Most of them stained positively for NSE and Thy-1 and negatively for CFAP,indicating RGCs. There was no significant variance inthe A values between the wells added various concentrations of PGP and NBC of the control wells( P >0. 05). Conclusion PGP andNBC have no direct effect on the survival of rat RGCs in vitro.