Effects of scutellarin on PKCgamma in PC12 cell injury induced by oxygen and glucose deprivation.
OBJECTIVE
To evaluate the neuroprotective effect and mechanisms of scutellarin (Scu) against PC12 cell injury after oxygen and glucose deprivation followed by reperfusion (OGD-Rep).


METHODS
Undifferentiated rat pheochromocytoma PC12 cells, exposed to oxygen and glucose deprivation followed by reperfusion (OGD-Rep), used as an in vitro model of ischemia/reperfusion. Cell survival was evaluated by diphenyltetrazolium bromide (MTT) assay and the amount of LDH release was determined using assay kits. [Ca2+](i) was monitored using a fluorescent Ca2+-sensitive dye Fura-2 acetoxymethyl ester. Cell apoptosis was detected by a DNA ladder and by flow cytometric detection. The expression of protein kinase C (PKC)gamma was determined using both RT-PCR and Western blotting. The translocation of PKCgamma was assayed by subcellular fractionation and Western blotting.


RESULTS
OGD-Rep injury significantly elevated the level of LDH release, [Ca2+](i), mRNA expression and the translocation of PKCgamma compared in the PC12 cells with those of the normal group. Scu (10-100 micromol/L) exerted a protective effect against OGD-Rep injury by reducing LDH release, [Ca2+](i), the percent of apoptosis, and the translocation of PKCgamma.


CONCLUSIONS
Scu inhibits the increase of [Ca2+](i) and the activation of PKCgamma, exerting protective effects against PC12 cell injury induced by OGD-Rep.
Xu
Wei
W
State Key Laboratory of Drug Research, Shanghai Institute of Materia Medica, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 201203, China.
Zha
Ruo-Peng
RP
Wang
Wen-Yi
WY
Wang
Yi-Ping
YP
D000818:Animals; D047310:Apigenin / Q000302:isolation & purification / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects*; D001692:Biological Transport / Q000187:drug effects; D015153:Blotting, Western; D002118:Calcium / Q000378:metabolism; D015687:Cell Hypoxia; D002462:Cell Membrane / Q000378:metabolism; D002467:Cell Nucleus / Q000378:metabolism; D003600:Cytosol / Q000187:drug effects / Q000378:metabolism; D049919:Erigeron / Q000737:chemistry; D005434:Flow Cytometry; D005947:Glucose / Q000378:metabolism / Q000494:pharmacology; D005965:Glucuronates / Q000302:isolation & purification / Q000494:pharmacology*; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D018696:Neuroprotective Agents / Q000302:isolation & purification / Q000494:pharmacology*; D010100:Oxygen / Q000378:metabolism / Q000494:pharmacology; D016716:PC12 Cells; D010946:Plants, Medicinal / Q000737:chemistry; D011493:Protein Kinase C / Q000096:biosynthesis* / Q000235:genetics; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D051381:Rats; D020133:Reverse Transcriptase Polymerase Chain Reaction
D005965:Glucuronates; D018696:Neuroprotective Agents; D012333:RNA, Messenger; C484876:scutellarin; D047310:Apigenin; D007770:L-Lactate Dehydrogenase; C087857:protein kinase C gamma; D011493:Protein Kinase C; D005947:Glucose; D010100:Oxygen; D002118:Calcium