Effects of Ginkgolide B on 6-OHDA-induced apoptosis and calcium over load in cultured PC12.
Ginkgolide B, one of the major components of Ginkgo biloba extracts, is a potent platelet-activating factor (PAF) receptor antagonist, which is also regarded as having neuroprotective effects on the CNS. The aim of this research is to observe the effects of Ginkgolide B on the PC12 apoptosis induced by 6-hydroxydopamine (6-OHDA) and to explore whether these effects are related to the changes of intracellular Ca(2+) and Calbindin D28K mRNA in PC12 cells. In the present work, the damage of PC12 cells was induced by 100 microM 6-OHDA. The cells survival rate was examined by MTT assays. The intracellular free calcium concentration in PC12 cells was measured by using the fluorescent Ca(2+) indicator fluo-3/AM. Semi-quantitative reverse transcriptase-polymerase chain reaction (RT-PCR) was employed to determine the expression of Calbindin D28K mRNA in PC12. The data show that the Ginkgolide B inhibited PC12 cells apoptosis induced by 6-OHDA in a dose-dependent manner, and decreased the activity of caspase-3. In addition, Ginkgolide B increased the expression of Calbindin D28K mRNA and inhibited 6-OHDA-induced elevation in the intracellular calcium concentration. Our results showed that the Ginkgolide B inhibited the apoptosis of PC12 induced by 6-OHDA, and the protective effects of Ginkgolide B on PC12 cells are mediated, at least in part, by up-regulating the Calbindin D28K mRNA and by decreasing the intracellular calcium concentration.
Meng
Haiwei
H
Department of Anatomy, School of Medicine of Shandong University, Jinan, China.
Li
Chuangang
C
Feng
Lei
L
Cheng
Baohua
B
Wu
Fengxia
F
Wang
Xiaohong
X
Li
Zhenping
Z
Liu
Shuwei
S
D000165:Acridine Orange; D000818:Animals; D017209:Apoptosis / Q000187:drug effects*; D064092:Calbindin 1; D064026:Calbindins; D002118:Calcium / Q000378:metabolism / Q000633:toxicity*; D053148:Caspase 3 / Q000378:metabolism; D061945:Caspase Inhibitors; D002470:Cell Survival / Q000187:drug effects; D005456:Fluorescent Dyes; D046934:Ginkgolides / Q000494:pharmacology*; D007783:Lactones / Q000494:pharmacology*; D018696:Neuroprotective Agents / Q000494:pharmacology*; D016627:Oxidopamine / Q000037:antagonists & inhibitors* / Q000633:toxicity*; D016716:PC12 Cells; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D051381:Rats; D020133:Reverse Transcriptase Polymerase Chain Reaction; D064030:S100 Calcium Binding Protein G / Q000096:biosynthesis / Q000378:metabolism; D013565:Sympatholytics / Q000633:toxicity*; D015854:Up-Regulation / Q000187:drug effects
C576500:Calb1 protein, rat; D064092:Calbindin 1; D064026:Calbindins; D061945:Caspase Inhibitors; D005456:Fluorescent Dyes; D046934:Ginkgolides; D007783:Lactones; D018696:Neuroprotective Agents; D012333:RNA, Messenger; D064030:S100 Calcium Binding Protein G; D013565:Sympatholytics; D016627:Oxidopamine; C045856:ginkgolide B; D053148:Caspase 3; D000165:Acridine Orange; D002118:Calcium