安徽中医药大学教学课件.ppt

Protective effect of ligustilide against glutamate-induced apoptosis in PC12 cells Contents 1、Introduction 2、Methods and Results 3、 Conclusion Necessity ：investigation of the pathogenesis, prevention and treatment of cerebrovascular disease. Conception： Blockages Dysmetabolism Characteristic：high morbidity、high mortality、high disability、high relapse rates CBF Active constituent： TQHXD in vivo： Lig can get through the BBB Ligustilide TQHXD i.g HPLC 3-nButylphthalide (NBP) Z-Ligustilide (LIG) PC12 Glutamate Control Glutamate group LIG（1、5、15μM） Ca2+ ROS Apoptosis MMP Bcl-2 Bax Caspase-3 MTT Western blotting Flow Cytometry Inverted microscope Survival rate DCFH-DA and FLUO-3 AM Histological morphology # #P＜0.01 vs. control group；**P＜0.01 vs. group treated with glutamate alone. Effect of LIG on the viability of damaged PC12 cells elicited by glutamate Effect of LIG on the release of LDH from PC12 cells damaged by glutamate # #P＜0.01 vs. control group；* P＜0.05，**P＜0.01 vs. group treated with glutamate alone. Cell morphology was examined by light microscope(×200) a b d c e f (a) PC12 cells were incubated with culture medium only; (b) 15mM glutamate alone; (c) 10μM NBP+15mM glutamate; (d) 1 μM LIG+15mM glutamate; (e) 5μM LIG+15mM glutamate; (f) 15μM LIG+15mM glutamate. Morphological changes in apoptotic cells assessed by Hoechst 33258 staining(magnification, ×40) (a) PC12 cells were incubated with culture medium only; (b) 15mM glutamate alone; (c) 10μM NBP+15mM glutamate; (d) 1μM LIG+15mM glutamate; (e) 5μM LIG+15mM glutamate; (f) 15μM LIG+15mM glutamate. b Effect of LIG on glutamate-induced PC12 cell apoptosis (a) PC12 cells were incubated with culture medium only; (b) 15mM glutamate alone; (c) 10μM NBP+15mM glutamate; (d) 1μM LIG+15mM glutamate; (e) 5μM LIG+15mM glutamate; (f) 15μM LIG+15mM glutamate. # #P＜0.01 vs. control group；**P＜0.01 vs. group treated with g