补肾配伍化痰活血药对低血清培养SH-SY5Y神经细胞轴突生长的影响  

Effects of Chinese Medicine of Tonifying Kidney Combined with Dissolving Phlegm and Activating Blood Circulation on Axonal Outgrowth in Human Neuroblastoma SH- SY5Y Cells with Low Serum Medium

作　　者：刘蕾[1] 郑琦[1] 安辰[1] 赵晖[1] 张秋霞[1] 樊永平[2] 王蕾[1] 

机构地区：[1]首都医科大学中医药学院,北京100069 [2]首都医科大学附属北京天坛医院,北京100050

出　　处：《中医药信息》2014年第5期5-9,共5页Information on Traditional Chinese Medicine

基　　金：国家自然科学基金项目(No.81273742);北京市属高等学校高层次人才引进与培养计划项目-长城学者(No.CIT&TCD20140329)

摘　　要：目的:观察补肾益髓(BSYS)方(即补肾化痰活血方药)及其拆方补肾方(BS)、化痰活血(HTHX)方对低血清培养SH-SY5Y细胞致神经轴突生长抑制的作用。方法:将低血清培养基与SHSY5Y细胞培养24h,建立神经轴突生长抑制模型,再将BSYS方、BS方和HTHX方含药血清与损伤的SH-SY5Y细胞培养24h,CCK-8法测定细胞存活率,βtubulin-Ⅲ免疫荧光法观察细胞形态,计算神经突起细胞率、测量细胞轴突长度及积分光密度(IOD)值。结果:和空白对照组(NC)细胞比较,模型组(MO)细胞存活率显著下降(P<0.05),轴突变短(P<0.001),IOD值明显降低(P<0.001)。和MO细胞比较,BSYS方和BS方含药血清均能明显升高细胞存活率(P<0.001);BSYS方明显提高神经突起细胞率(P<0.05)、促进轴突生长(P<0.05)和升高IOD值(P<0.001);补肾方只在升高IOD值方面效果明显(P<0.01);而化痰活血方对以上指标的作用均不明显。结论:补肾配伍化痰活血方药和补肾方药对低血清致SH-SY5Y细胞轴突生长抑制的改善作用明显,其作用机制有待进一步探讨。Objective： To investigate effects of BSYS formula （Chinese medicine of tonifying kidney combined with dissolving phlegm and activating blood circulation）, formulas of BS （tonifying kidney） and HTHX （dis- solving phlegm and activating blood circulation） on inhibition of axonal outgrowth of SH -SY5Y cells with low serum medium. Methods： The cells were maintained in low serum medium for 24h to establish the model of axonal outgrowth inhibition, and then the cells were treated with serum containing medicine of BSYS, BS and HTHX formula for additional 24h. The cell viability was measured by the method of cell counting Kit - 8 （CCK- 8 ） ; the morphology of the cells was observed by 13tubulin - m immunofluorescence method; the via- bility of neurite bearing cells was calculated ; the neurite extension length and integral optical density （IOD） were measured. Results： Compared with the normal control （NC） group, the cell viability of model （MO） group was significantly decreased （ P 〈 0.05 ） ; the neurite length was shortened （ P 〈 0. 001 ） and IOD was decreased distinctly （P 〈 0. 001 ）. Compared with the cells in MO group, the serum containing medicine ofBSYS and BS significantly increased the cell viability （P 〈0. 001 ）. BSYS formula significantly increased the viability of neurite bearing cells （P 〈 0.05 ）, the neurite extension length （ P 〈 0.05 ） and IOD value （ P 〈 0. 001 ）. BS formula only showed its evidently rising effect on IOD （P 〈0.01 ） ,while HTHX formula had no significant effects. Conclusion： Chinese medicines of tonifying kidney combined with dissolving phlegm and activating blood circulation and tonifying kidney show significant effects on inhibition of axonal outgrowth of SH -SY5Y cells with low serum medium, and the mechanism needs to be further explored.

关 键 词：补肾益髓方 血清药理学 SH-SY5Y细胞 轴突生长 

分 类 号：R285.5[医药卫生—中药学]