用血清药理学方法观察中药脑血通颗粒对凝血酶致C6细胞损伤的保护作用

【作者】 骆守真；

【导师】 姜亚军；

【作者基本信息】 南京中医药大学 ， 中西医结合临床， 2005， 硕士

【摘要】 目的： 本实验观察了中药脑血通颗粒含药血清对凝血酶致C6胶质瘤细胞损伤的保护作用，以探讨脑出血后凝血酶损伤机理及中药干预治疗作用机制。 方法： ①在含10％小牛血清的1640培养液中培养C6细胞株。取SD大鼠，灌胃给药(脑血通)，制成含药血清。在C6细胞培养系中，分别加入100u／ml凝血酶以及不同浓度的含药血清。 ②观察细胞形态，MTT法检测细胞存活力，LDH试剂盒测定LDH值，流式细胞仪检测细胞凋亡及钙离子浓度。 结果： 凝血酶浓度为100u／ml时对C6胶质瘤细胞产生一定的毒性作用，表现为细胞乳酸脱氢酶释放增加，细胞内钙离子浓度增高，并诱导细胞凋亡。脑血通能抑制这些现象的发生，阻止细胞过度凋亡。 结论： 研究提示，凝血酶对体外培养的C6胶质瘤细胞具有毒性损伤作用，脑血通含药血清可以抑制细胞凋亡，从而发挥保护C6胶质瘤细胞的作用。更多还原

【Abstract】 Background and purpose:The appropriate therapy for intracerebral hemorrhage （ICH） to reduce disability rate and mortality rate remains a subject of debate. Many researches have discovered that HuoxueHuayu Method of TCM has broad prospects to be used in the treatment of ICH. However, the specific mechanism remains to be further explored. So, the purpose of this work is to study the NaoXueTong protective mechanism on the C6 Glioma Cells injured by thrombin on molecular level, to clarify injured mechanism by thrombin and intervention mechanism of herbal medicine. Methods:We applied the experimental method of "serological pharmacology", which was combined with the theory and laboratory technique of modern cytobiology and molecular biology.（1）The C6 Glioma Cells were cultured by 1640 culture medium containing 10% calf serum. SD rats were administrated NaoXueTong by intragastric method to make the serum of drug, and later adding 100u/ml thrombin and the drug serum of different concentration in the C6 Glioma Cells.（2）MTT was used to measure the survival rate. The level of LDH in culture system was detected by the LDH kit. C6 Glioma Cells’ apoptosis and concentration of Ca²⁺ were detected by the flow cytometry （FCM）. Results:The results have shown that NaoXueTong could increase MTT, lower LDH, inhibit apoptosis and Ca²⁺ delivery in the culture system. Conclusions:In summary, the results of current study demonstrate that thrombin could induce apoptosis of C6 Glioma Cells. NaoXueTong may inhibite this apoptosis, protect the C6 Glioma Cells from being injured by the thrombin.更多还原