基于微流控芯片的菟丝子黄酮对大鼠HSC-T6细胞凋亡影响研究

Study on the Pro-apoptosis Effect of Semen Cuscutae Flavonoids of Fufangmuji Granule in HSC-T6 Cell based on Microfluidic Technology

目的:构建一种集成有浓度梯度及微阀结构的微流控芯片,并利用该芯片探讨菟丝子黄酮对大鼠肝星状细胞株(HSC-T6)凋亡坏死的影响。方法:设计和构建一种集成有浓度梯度及微阀结构的微流控芯片,以大鼠HSC-T6细胞活力、形态等特征检验芯片的适用性;同时利用芯片探讨8种不同浓度菟丝子黄酮作用HSC-T6细胞24、48和72h后的凋亡坏死情况。结果:细胞在芯片中生长状态良好,增殖活力强,存活率达到96%;菟丝子黄酮具有明显促细胞凋亡坏死作用,其作用细胞24h、48h及72h后,细胞凋亡坏死率与空白组比较有显著差异(P<0.05),且在一定浓度下,随着给药浓度的增加及作用时间的延长,呈现量-效和时-效关系。结论:该芯片重复性及适用性良好,可极大简化药效筛选中操作的复杂性,为中药新药药效评价提供一种新方法。

Objective:To explore the pro-apoptosis influence of semen cuscutae flavonoids in HSC-T6cell,a microfluidic chip integrated with a concentration gradient and a microfluidic valve structure was established.Methods:A microfluidic chip integrated with a concentration gradient and a microfluidic valve was design and structured,and the HSC-6cell viability and morphology were taken to verify its adaptability.Meanwhile,it was applied to explore the apoptosis and necrosis of HSC-6cell treated with 8different concentrations of semen cuscutae flavonoids for 24,36and 72h.Results:The cells grew well in this chip,the proliferation activity was strong,and the survival rate was 96%.After treatment with semen cuscutae flavonoids for 24,48and 72h,the apoptosis and necrosis rate on HSC-6、cells in semen cuscutae flavonoids group、had significant difference(P<0.05)between that in control group.In addition,at a certain concentration,the action concentration and time increased,the apoptosis and necrosis rate increased,presented a relationship between the quantity-effect and time-effect.Conclusion:The repeatability and applicability of this chip was good,it can greatly simplify the drug screening in complicated traditional Chinese medicine and its compound,it also provide experimental basis for theapplication of microfluidic chip in the field of drug development.