1.Reactive Oxygen Species are Involved in Y-27632-induced Neurite Outgrowth in PC12 Cells.
So Yeong PARK ; Seong Ah MOON ; Jeong Mi AN ; Du sik KIM ; Jeong Taeg SEO
International Journal of Oral Biology 2016;41(4):231-236
Inhibition of Rho-associated coiled coil-containing kinase (ROCK) has been reported to promote differentiation of neuronal cells. Here, we examined the effect of Y-27632, a ROCK inhibitor, on the outgrowth of neurites in PC12 cells. Y-27632 caused a rapid induction of neurite outgrowth in PC12 cells in a time-dependent manner. The neurite outgrowth, triggered by Y-27632, was accompanied by Rac1 activation, and was attenuated by Rac1 inhibitor NSC23766, in a concentration-dependent manner. Y-27632 also induced an increase in the production of reactive oxygen species (ROS). Pretreatment with N-acetylcysteine, an ROS scavenger, inhibited the ROS generation and neurite outgrowth in response to Y-27632. These results indicate that the activation of Rac1 and the generation of ROS contribute to the neurite outgrowth triggered by Y-27632 in PC12 cells.
Acetylcysteine
;
Animals
;
Neurites*
;
Neurons
;
PC12 Cells*
;
Phosphotransferases
;
Reactive Oxygen Species*
2.Cysteinyl leukotriene receptor 1 is involved in rotenone-induced injury of PC12 cells.
Shu-ying YU ; Bing ZHAO ; Xia-yan ZHANG ; Xaio-yan ZHANG ; Yan-fang WANG ; Li-hui ZHANG ; Yun-bi LU ; Er-qing WEI
Journal of Zhejiang University. Medical sciences 2012;41(2):139-145
OBJECTIVETo investigate whether cysteinyl leukotriene receptor 1 (CysLT₁ receptor) is involved in rotenone-induced injury of PC12 cells.
METHODSAfter 24 h treatment with rotenone or with rotenone and the CysLT₁ receptor antagonist montelukast, PC12 cell viability was determined by the colorimetric MTT reduction assay. After PC12 cells were treated with various concentrations of rotenone for 24 h or with 3 μmol/L rotenone for various durations, the expression of CysLT(1) receptor was determined by Western blotting, and its intracellular distribution was detected by immunocytochemistry.
RESULTSRotenone (0.3-30 μmol/L) induced PC12 cell injury; this injury was significantly attenuated by montelukast at 1 and 5 μmol/L.The expression of CysLT(1) receptor increased after rotenone treatment at 1-10 μmol/L, or at 3 μmol/L for 3 and 24 h. Rotenone caused concentration-and time-dependent translocation of CysLT₁ receptor from the nucleus to the cytosol.
CONCLUSIONCysteinyl leukotriene receptor 1 is involved in rotenone-induced injury of PC12 cells.
Animals ; PC12 Cells ; Rats ; Receptors, Leukotriene ; metabolism ; physiology ; Rotenone ; toxicity
3.Effect of different sterilization methods on the surface morphology of PPDO-hybrid-PLGA nanofiber scaffold and attachments of PC12 cell.
Juhyon LEE ; Hyungi MIN ; Juyoung JUNG ; Nara KANG
Journal of the Korean Association of Oral and Maxillofacial Surgeons 2008;34(6):635-639
OBJECTIVES: the effect of different sterilization methods on the surface morphology of PPDO-hybrid-PLGA nanofiber scaffold and attachments of PC12 cell were investigated. METHODS: Poly (p-dioxone)-hybrid-Poly (lactide-glycolide) (PPDO-hybrid-PLGA) nanofiber scaffold, fabricated in a tube form with 1.5 mm internal diameter, 0.2 mm thickness and 5 mm length, was prepared using electrospinning method. To study the surface morphology using SEM, The study group and control group in respective were; Control:Non-sterilized scaffold, Group I:scaffold sterilized with 70% Alcohol, Group II: scaffold sterilized with Ethylene Oxide at 65 degrees C, and Group III: scaffold sterilized with Ethylene Oxide at 37 degrees C. To investigate viability of the PC12 cell on the scaffold, The study group and control group in respective were; Control: sterilized with 70% Alcohol, Group I: sterilized with Ethylene Oxide at 65 degrees C, and Group II: sterilized with Ethylene Oxide at 37 degrees C. RESULTS: 1. The surface morphology was slightly changed in Group I, II and GroupIII, compared with control. 2. The attachment of PC12 cells in Group I, II was not higher than in control DISCUSSION: The attachment of PC12 cell is not influenced by different sterilization methods.
Animals
;
Ethylene Oxide
;
Ethylenes
;
Nanofibers
;
PC12 Cells
;
Sterilization
5.Enhancement of ATP-induced Currents by Phospholipase D1 Overexpressed in PC12 Cells.
Jin Bong PARK ; Young Rae KIM ; Byeong Hwa JEON ; Seung Kiel PARK ; Sae Ock OH ; Young Geun KIM ; Sang Do LEE ; Kwang Jin KIM
The Korean Journal of Physiology and Pharmacology 2003;7(4):223-230
Using phospholipase D1 (PLD1) -overexpressing PC12 (PLD1-PC12) cells, the regulatory roles of PLD1 on ATP-induced currents were investigated. In control and PLD1-PC12 cells, ATP increased PLD activity in an external Ca2+ dependent manner. PLD activity stimulated by ATP was substantially larger in PLD1-PC12 cells than in control cells. In whole-cell voltage-clamp mode, ATP induced transient inward and outward currents. The outward currents inhibited by TEA or charybdotoxin were significantly larger in PLD1-PC12 cells than in control cells. The inward currents known as Ca2+ permeable nonselective cation currents were also larger in PLD1-PC12 cells than in control cells. However, the difference between the two groups of cells disappeared in Ca2+ -free external solution, where ATP did not activate PLD. Finally, ATP-induced 45Ca uptakes were also larger in PLD1-PC12 cells than in control cells. These results suggest that PLD enhances ATP-induced Ca2+ influx via Ca2+ permeable nonselective cation channels and increases subsequent Ca2+ -activated K+ currents in PC12 cells.
Adenosine Triphosphate
;
Animals
;
Charybdotoxin
;
PC12 Cells*
;
Phospholipases*
;
Tea
6.Protective effect of α-asarone and β-asarone on Aβ -induced inflammatory response in PC12 cells and its.
Jianhong SHI ; Ruizhi LI ; Yuanxiao YANG ; Liting JI ; Changyu LI
Journal of Zhejiang University. Medical sciences 2021;50(5):591-600
To investigate effects of α-asarone and β-asarone on induced PC12 cell injury and related mechanisms. Aβ toxic injury cell model was induced by Aβ in PC12 cells. PC12 cells were divided into blank control group, model control group, α-asarone group (0.5, 1.0, β-asarone group (6.3, 12.5, vasoactive intestinal peptide (VIP) group, and VIP antagonist control group. Cell survival rate was detected by CCK-8 kit; cell apoptosis rate was detected by flow cytometry. The levels of inflammatory cytokines interleukin (IL)-1, , tumor necrosis factor (TNF)-α, oxidation-related inducible nitric oxide synthase (iNOS), nitric oxide (NO), apoptosis factors caspase-3 and p53 were detected by ELISA method. The expressions of C-Jun N-terminal kinase (JNK) and p38 mitogen-activated protein kinase (p38MAPK) were detected by Western blotting. Compared with model control group, cell survival rates of group, β-asarone group and VIP group increased; the cell apoptosis rate decreased; levels of apoptosis-related factors caspase-3, p53, inflammatory factors IL-1, TNF-α decreased; IL-10 level increased; levels of oxidization-related factors iNOS and NO decreased; the expression of JNK and p38MAPK protein decreased (all <0.05). After VIP antagonist intervention, the survival rate of β-asarone group decreased; apoptosis rate increased; apoptosis related factors caspase-3, p53, inflammatory factors IL-1, TNF-α increased; IL-10 decreased; oxidation related factors iNOS and NO increased; the expression of JNK and p38MAPK protein increased (all <0.05); while there were no significant changes in these indicators of α-asarone group (all >0.05). α-asarone and β-asarone have protective effects on PC12 cell injury induced by Aβ. β-asarone may inhibit inflammatory factors and oxidation-related factors through promoting VIP secretion, regulating JNK/MAPK pathway, and reducing PC12 cell apoptosis; however, the effect of α-asarone may be not related to VIP secretion.
Allylbenzene Derivatives
;
Animals
;
Anisoles/pharmacology*
;
Apoptosis
;
PC12 Cells
;
Rats
7.Reserpine treatment activates AMP activated protein kinase (AMPK).
Rackhyun PARK ; Kang Il LEE ; Hyunju KIM ; Minsu JANG ; Thi Kim Quy HA ; Won Keun OH ; Junsoo PARK
Natural Product Sciences 2017;23(3):157-161
Reserpine is a well-known medicine for the treatment of hypertension, however the role of reserpine in cell signaling is not fully understood. Here, we report that reserpine treatment induces the phosphorylation of AMP activated protein kinase (AMPK) at threonine 172 (T172) in PC12 cells. Phosphorylation of AMPK T172 is regulated by upstream signaling molecules, and the increase of phospho-T172 indicates that AMPK is activated. When we examined the FOXO3a dependent transcription by using the FHRE-Luc reporter assay, reserpine treatment repressed the FHRE-Luc reporter activity in a dose dependent manner. Finally, we showed that reserpine treatment induced the phosphorylation of AMPK as well as cell death in MCF-7 cells. These results suggest that AMPK is a potential cellular target of reserpine.
AMP-Activated Protein Kinases*
;
Animals
;
Cell Death
;
Hypertension
;
MCF-7 Cells
;
PC12 Cells
;
Phosphorylation
;
Reserpine*
;
Threonine
8.Establishing the model of kappaB-decoy inhibiting the activity of NF-kappaB in PC12 cells.
Dong-mei WANG ; De-zheng GONG ; Sheng-ming YIN ; De-qin YU ; Hong XU ; Jing XU ; Jie ZHAO ; Yi-ping SUN
Chinese Journal of Applied Physiology 2010;26(2):195-198
OBJECTIVEObserving the time course and establishing the model of kappaB-decoy oligodeoxynucleotides (rcB-decoy) inhibiting the activity of NF-kappaB in the PC12 cells.
METHODSPC12 cells cultivating in the 6 wells plate were divided into 3 groups, experimental group: adding kappaB-decoy complex (6 microg DNA/well), the control group: adding scrambled-decoy complex, the normal group: adding lipid-Lipofectamine 2000, transfer and cultivate 48 h, then lipopolysaccharide (LPS, 200 ng/ml) was added in the cells for 0.5-4 h. The immunocytochemistry and Western blot were used to measure the expression or the activity of NF-kappaB in PC12 cells.
RESULTSIn PC12 cells, compared with normal group, the expression of NF-kappaB enhanced obviously with the time of the stimulation of LPS in scrambled-decoy treated control group (P < 0.01), in 2-4 h the level reached the peak; the expression of NF-kappaB showed the stable level with the time of the stimulation of LPS in kappaB-decoy treated experimental group, compared with the control group, the expression levels were obviously lower than the respective time point of control groups (P < 0.01).
CONCLUSIONkappaB-decoy could reduce the expression of NF-kappaB in the normal PC12 cells and inhibit the activity of NF-cB in the pathologic PC12 cells.
Animals ; Cells, Cultured ; Lipopolysaccharides ; pharmacology ; NF-kappa B ; antagonists & inhibitors ; metabolism ; Oligodeoxyribonucleotides ; pharmacology ; PC12 Cells ; Rats
9.The effects of autophagy on cell survival under different hypoxia.
Yun-Ling HE ; Li-Ying WU ; Xin HUANG ; Tong ZHAO ; Xue-Feng DING ; Kui-Wu WU ; Ming FAN ; Ling-Ling ZHU
Chinese Journal of Applied Physiology 2013;29(3):193-196
OBJECTIVETo investigate the regulation of different hypoxia on cell survival and autophagy.
METHODSPC12 cells were treated with different hypoxia. The cell survival was measured by MTT assay, expressions of LC3 and p62 were marked for autophagy detected by Western Blot, and the level of reactive oxygen species (ROS) was analyzed by flow cytometry.
RESULTSThe cell viability was different under different hypoxia: moderate hypoxia promoted cell viability, and severe hypoxia caused a decrease in cell viability; autophagy marker molecules, p62 and LC3-II expressions were different: moderate hypoxia increased p62 and LC3-II expressions, in contrast, severe hypoxia led to the decrease of p62 and LC3-II expressions; compared to normoxia, moderate hypoxia did not change the levels of ROS, while severe hypoxia increased the levels; 3-MA, the inhibitor of autophagy, elevated the levels of ROS in the three oxygen concentrations, additionally, the increased amplitudes in the moderate and severe hypoxia groups were higher than that in the normoxia group.
CONCLUSIONModerate hypoxia promotes cell survival, severe hypoxia causes the cell death, and the autophagy activity may mediate the effects of different hypoxia.
Animals ; Autophagy ; physiology ; Cell Death ; Cell Hypoxia ; Cell Survival ; PC12 Cells ; Rats ; Reactive Oxygen Species ; metabolism
10.High-throughput microwell plate for cell research.
Gang DU ; Philip Ching Yat WONG ; Qing CHANG ; Zicheng LI
Journal of Southern Medical University 2014;34(1):5-7
We developed a novel simple microwell plate that allows researchers to culture cells on chips easily without the use of tubes and pumps for studies of cell proliferation, apoptosis and pharmacology. Unlike any other previous design, this device can be put into an incubator directly and maintains cells in an environment with stable temperature, humidity, air pressure, oxygen and CO2 concentration. We cultured PC-12 cells in these microwell plates and measured the cell proliferation and apoptosis. The microwell plate is recommended for use in all research including those involving rare samples and expensive reagents.
Animals
;
Apoptosis
;
Cell Culture Techniques
;
instrumentation
;
Cell Proliferation
;
PC12 Cells
;
Protein Array Analysis
;
Rats