Effects of tricholoma matsutake polysaccharides on 1-methy-4-pehnyl-pyridine ion-induced PC12 cell damage / 解剖学报

Obｊective To investigate the protective mechanism of tricholoma matsutake polysaccharides(TMP) against 1-methy-4-pehnyl-pyridine ion (MPP

Effect of bergapten on glucose and oxygen deprivation induced PC12 cell injury by regulating long non-coding RNA opioid receptor gene expression / 解剖学报

Objective To study whether bergapten (BG) protects PC12 cells from oxygen-glucose deprivation (OGD) induced cell injury by regulating long non-coding RNA (lncRNA) opioid receptor gene (Oprm1) expression. Methods PC12 cells were divided into control (Con) group, OGD group, OGD+ low concentration BG (BG-L) group, OGD+medium concentration BG (BG-M) group, OGD + high concentration BG (BG-H) group, OGD + pcDNA group, OGD+pcDNA-Oprm1 group, OGD+BG+si-NC group, OGD+BG+si-Oprm1 group. The malondialdehyde (MDA) content, superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) activities were measured by the kits. Cell apoptosis rate was analysed by flow cytometry. The expression level of Oprm1 was analysed by Real-time PCR. Results Compared with the Con group, the apoptosis rate and MDA content of PC12 cells in OGD group increased significantly, whereas Oprm1 expression, SOD and GSH-Px activity decreased significantly (P < 0. 05). Compared with the OGD group, the apoptosis rate and MDA content of PC12 cells in the OGD + BG-L group, OGD + BG-M group, OGD + BG-H group were significantly reduced, whereas the Oprm1 expression, SOD and GSH-Px activities increased significantly (P < 0. 05). Compared with the OGD+pcDNA group, the apoptosis rate and MDA content of the PC12 cells in the OGD+pcDNA-Oprm1 group reduced significantly, whereas the SOD and GSH-Px activities increased significantly (P<0. 05). Compared with the OGD+BG+si-NC group, the apoptosis rate and MDA content of PC12 cells in the OGD+BG+si-Oprm1 group increased significantly, whereas the SOD and GSH-Px activities decreased significantly (P < 0. 05). Conclusion Bergapten may alleviate OGD-induced PC12 cell injury, which is correlated to the up-regulation of lncRNA Oprm1 expression.

MicroRNA-486 targeting TRIM10 to inhibit Parkinson’s disease cell model damage / 解剖学报

[Abstract] Objective To study the effect and mechanism of microRNA-486 (miR-486) on 1-methyl-4-phenylpyridine (MPP

Comparison of PC-12 cells’ apoptosis induced by triazine herbicides atrazine, simazine and cyanazine / 西安交通大学学报(医学版)

【Objective】 To compare PC-12 cells’ apoptosis caused by the serotonin herbicides atrazine （ATR）， simazine （SIM） and cyanazine （CYA）. 【Methods】 The rat adrenal medullary pheochromoma PC-12 cell line was selected for routine culture. At the cell logarithmic growth phase， ATR， SIM and CYA were used at a concentration of 200 μmol/L for 24 h， respectively， and the same solvent was added in the control group. The CCK-8 method was used to detect the cell survival rate； the content of reactive oxygen species （ROS） in PC-12 cells was detected； the Real-time PCR and Western blotting methods were used to detect the mRNA and protein expressions of Bax， p53， Bcl-2 and Caspase-3. 【Results】 Compared with that in the control group， the survival rate of the cells in ATR group， SIM group and CYA group was significantly decreased. The intracellular ROS activity of the three groups was significantly increased， and the mRNA and protein expressions of Bax， p53 and Caspase-3 were increased. mRNA and protein expressions of Bcl-2 were significantly reduced （P<0.01）. Compared with that in ATR group， the cell survival rate of SIM group and CYA group was significantly increased， the intracellular ROS activity of the two groups was significantly decreased， the expressions of Bax， p53 and Caspase-3 mRNA and protein were significantly reduced， and the expressions of Bcl-2 mRNA and protein were significantly increased （P<0.01）. Compared with that of SIM group， the cell survival rate of CYA group was significantly increased， while the intracellular ROS activity was significantly decreased； the Bax， p53 and Caspase-3 mRNA and protein expressions were significantly reduced， and Bcl-2 mRNA and protein expressions were significantly increased （P<0.01）. 【Conclusion】 ATR， SIM and CYA can all promote PC-12 cells’ apoptosis； ATR has the strongest effect while CYA has the weakest effect.

Amphetamine causing damage of dopamine cells via inhibiting of protein kinase B / glycogen synthase kinase-3β/ collapsin response mediator protein-2 signal pathway / 解剖学报

Objective To explore the damage mechanism of dopamine cells induced by amphetamine (AMPH). Methods The damage model of dopaminergic cells in mice was established by intraperitoneal injection of AMPH. The mice were randomly grouped into control, saline, amphetamine treatment for 1 day, 7 days, 14 days and 28 days. Each group contained 10 mice. The model of cell injury was established by use of AMPH in PC12 cells. The dopaminergic fibers of corpus striatum and PC12 cells were observed by the immunohistochemistry and immunofluorescence method, and changes of proteins in the protein kinase B (Akt) / glycogen synthase kinase 3β(GSK-3β) / collapsin response mediator protein 2 (CRMP-2) signal pathway were detected by Western blotting. Results AMPH caused the damage of dopaminergic fibers in the mouse corpus striatum and PC12 cells. Meanwhile, AMPH inhibited Akt and GSK-3β phosphorylation levels, and increased phosphorylated CRMP-2 level. Nerve growth factor(NGF), an agonist of Akt, or SB216763, an inhibitor of GSK-3β protected PC12 cells against AMPH-induced toxicity through upregulation of Aat and GSK-3β phosphorylation and downregulated of phosphorylation CRMP-2. Conclusion AMPH causes damage of dopamine cells via inhibition of Akt/ GSK-3β/ CRMP-2 signal pathway.

Protective effect of fluoxetine against hypoxia induced injury on PC12 cells / 中国药理学通报

Aim To study the protective effect of fluoxetine against hypoxia induced injury on PC12 cells. Methods PC12 cells were randomly divided into control group, hypoxia group, and fluoxetine hydrochloride group. The last two groups were put into a hypoxic culture chamber for 18 hours, the cell state was observed under inverted microscope, and cell viability was detected using CCK-8 assay. Reactive oxygen species (ROS) level was evaluated by DCFH-DA. Lactate dehydrogenase (LDH), superoxide dismutase (SOD), catalase (CAT), malondialdehyde (MDA) in cell culture supernatant were evaluated by enzyme labeling method. The expression levels of Bcl-2, Bax and caspase-3 were determined by Western blot. Results Compared with normal group, hypoxia caused obvious damage to PC12 cells. Fluoxetine hydrochloride at 10

Protective effect of α-asarone and β-asarone on Aβ -induced inflammatory response in PC12 cells and its / 浙江大学学报·医学版

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). 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.

Baicalein reduces Aβ25-35-induced PC12 cell damage by inhibiting the JAK2/STAT1 pathway / 药学学报

This study investigated the mechanism by which baicalein protected PC12 cells from Aβ25-35-induced injury. PC12 cells were treated with Aβ25-35 (20 μmol·L-1) and the ability of baicalein to prevent apoptosis was investigated by monitoring changes in cell morphology, Hoechst 33342 staining, and measurement of inflammatory factors. Western blotting was used to detect the expression of the apoptosis-related proteins cysteinyl aspartate specific proteinase-3 (caspase-3), cleaved cysteinyl aspartate specific proteinase-3 (cleaved caspase-3), proteins involved in the Janus kinase 2/signal transducer and activator of transcription 1 (JAK2/STAT1) pathway, and downstream inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2). The results show that baicalein (80 μmol·L-1) can significantly inhibit apoptosis and the release of inflammatory factor IL-8 and TNF-α in Aβ25-35-treated PC12 cells. Western blotting results showed that baicalein can inhibit the phosphorylation of JAK2 and STAT1 and decrease the expression of downstream iNOS and COX-2, thereby inhibiting the JAK2/STAT1 signaling pathway and preventing Aβ25-35-induced PC12 cell damage.

Effects of calycosin on mitochondrial apoptotic pathway in oxygen-glucose deprivation / reoxygenation PC12 cells / 解剖学报

Objective To investigate the effect of calycosin on mitochondrial apoptotic pathway in oxygen-glucose deprivation/ reoxygenation PC12 cells. Methods PC12 cells were randomly divided into four groups: control group, model group, calycosin group and nimodipine group. Except for the control group, the other groups were treated with oxygen and glucose deprivation for 2 hours and compound oxygen and glucose for 24 hours. Calycosin group and nimodipine group were treated with drug-containing medium containing calycosin (0. 07 μmol/ L) and nimodipine (5. 00 μmol/ L) simultaneously with reoxygenation. CCK-8 method was used to detect cell survival rate, flow cytometry was used to detect cell apoptosis rate, immunofluorescence method was used to detect Bax/ Bcl-2 ratio, Western blotting was used to detect the expression of key proteins cytochrome C (Cyt-C), apoptotic protease activating factor-1 (Apaf-1) and Caspase-3 in mitochondrial apoptotic pathway. Results Compared with the control group, the survival rate of cells in model group decreased significantly (P<0. 05), and the apoptotic rate increased significantly (P<0. 05), the ratio of Bax/ Bcl-2 was significantly increased (P<0. 05), and the expression of key proteins Cyt-C, Apaf-1 and Caspase-3 in mitochondrial apoptotic pathway were significantly increased (P<0. 05). Compared with the model group, the cell survival rates of calycosin group and nimodipine group increased significantly (P < 0. 05), apoptotic rate decreased significantly (P<0. 05), the ratio of Bax/ Bcl-2 was significantly decreased (P<0. 05), and the expression of key proteins of mitochondrial apoptotic pathway, Cyt-C, Apaf-1 and Caspase-3 were significantly decreased (P < 0. 05). The difference has statistical significance. Conclusion Calycosin can significantly improve the survival rate of oxygen-glucose deprivation/ reoxygenation PC12 cells and inhibit cell apoptosis. Its mechanism is closely related to the inhibition of the expression of key proteins Cyt-C, Apaf-1 and Caspase-3 in mitochondrial apoptotic pathway by calycosin.

Effect of ligustilide on oxygen and glucose deprivation/reperfusion-induced mitochondria fission in PC12 cells / 中国中药杂志

This study aimed to investigate the effect and mechanism of ligustilide, the main active ingredient in Ligusticum wallichii, on mitochondria fission after PC12 cell injury induced by oxygen and glucose deprivation/reperfusion(OGD/R). In the experiment, an OGD/R model was established in vitro, and PC12 cells were pre-treated with ligustilide for 3 h, and then the cell viability was detected by CCK-8 method. The effect of different concentrations of ligustilide on the morphology of PC12 cells after OGD/R injury was observed under an inverted microscope. Transmission electron microscopy was used to observe the mitochondrial fission of PC12 cells after OGD/R injury. DCFH-DA immunofluorescence staining method was used to detect intracellular reactive oxygen species(ROS) changes. Changes in mitochondria membrane potential(MMP) were detected by flow cytometry. Hochest 33258 was used to observe the apoptosis of PC12 cells. Western blot was used to detect changes in cytochrome C(Cyt C) content in mitochondria and cytoplasm, and mitochondrial fission-related proteins Drp 1 and Fis 1. All results showed that compared with the model group, ligustilide significantly increased the survival rate of PC12 cells and the number of cells. Further experiments showed that ligustilide inhibited the release of ROS and decline of mitochondrial membrane potential in PC12 cells after OGD/R injury. Moreover, ligustilide reduced the release of Cyt C and promoted the expressions of Drp1 and Fis1 in mitochondrial fission proteins. Verification experiments showed that mitochondrial fission inhibitor mdivi-1 decreased cell survival rate and inhibited fission. The results indicated that ligustilide exerted neuro-protective effects by promoting mitochondrial fission and reducing cell damage. It preliminary proves that the mechanism of ligustilide on ischemic brain injury may be related to the promotion of mitochondrial fission and the maintenance of cell homeostasis.

Study on the Protective Effects of Butein on Oxidative Stress Injury of PC 12 Cell and Its Effects on Mitochondrial Function / 中国药房

OBJECTIVE：To study the protective effects of butein on oxidative stress injury of PC12 cell and its effects on mitochondrial function. METHODS：Rats PC12 cells were divided into normal control group，model group，solvent control group（1 ‰ dimethyl sulfoxide），butein high，medium and low concentration groups（2，1，0.5 μmol/L）. The latter 4 groups were given relevant reagent/medicine for intervention；24 h later，other groups were given 100 mU/mL glucose oxidase to induce oxidant stress model except for normal control group. After 4 h culture，cell survival rate，apoptosis rate，the levels or activities of ROS，MDA，SOD，CAT，GSH-Px，ATP，IL-1β and TNF-α as well as the change of MMP were detected. RESULTS：Compared with normal control group，cell survival rate，the levels or activities of SOD，CAT，GSH-Px and ATP were all decreased significantly，and apoptotic rate，the content of ROS，the levels of MDA，IL-1β and TNF-α were all increased significantly（P＜0.05 or P＜0.01），while the MMP was decreased significantly. Compared with model group，above indexes of solvent control group had no significant change （P＞0.05），cell survival rates，the levels or activities of SOD （except for medium and low concentration groups），CAT，GSH-Px（except for medium and low concentration groups），ATP（except for low concentration group）were increased significantly in butein high，medium and low concentration groups，while apoptotic rates，the content of ROS，the levels of MDA，IL-1 β and TNF-α were decreased significantly（P＜0.05 or P＜0.01），while the MMP were increased significantly. CONCLUSIONS：Butein can increase the antioxidant enzyme activity， stabilize mitochondrial function， inhibit oxidative stress and inflammationthus， increase energy generation inhibiting neuronal cell apoptosis ultimately exerting a neuroprotective effect.

Protective Effect and Mechanism of Tetramethylpyrazine Against t-BHP-induced PC12 Cell Injury / 中国实验方剂学杂志

Objective:To study the protective effect of tetramethylpyrazine (TMP) on PC12 cells induced by tert-butyl hydroperoxide (t-BHP) and the regulatory mechanism on signaling pathway of phosphatidylinositol-3-kinases (PI3K)/kinase B (Akt)/mammalian target of rapamycin(mTOR). Method:PC12 cells cultured in vitro were treated with t-BHP (200 μmol·L-1) for 6 h to establish a model of oxidative damage in PC12 cells. The experiment was divided into blank group, model group (200 μmol·L-1t-BHP), TMP group. PC12 cells were pretreated with TMP(25, 50, 100 μmol·L-1) for 12 h, and then treated with t-BHP for 6 h. The cell viability was detected by cell counting kit-8(CCK-8) method, and lactate dehydrogenase (LDH) leakage, malondialdehyde (MDA) content, superoxide dismutase (SOD) activity, reactive oxygen species (ROS) and glutathione peroxidase (GSH-Px) activity were detected by enzyme-linked immunosorbent assay (ELISA). Apoptosis was observed by annexin V-FITC/PI double staining. B-cell lymphoma-2 (Bcl-2), Bcl-2-associated X protein (Bax), total protein kinase B (Akt), and phosphorylated protein kinase B (p-Akt), mTOR and p-mTOR expressions were detected by Western blot. Result:The cell viability of PC12 cells treated with 200 μmol·L-1 t-BHP decreased to about 50%after 6 h. This condition was suitable for the establishment of oxidative damage model. Compared with the model group, TMP (25, 50, 100 μmol·L-1) pretreatment for 12 h significantly increased the survival rate of PC12 cells (PPPPPPP-1) pretreatment group increased significantly (PConclusion:Ligustrazine protects PC12 cell injury induced by t-BHP by activating PI3K/Akt/mTOR signaling pathway.

Effects of puerarin on the expressions of CaM, CaMKⅡ, MECP2, BDNF and Akt in oxygen and glucose deprivation cell models / 西安交通大学学报(医学版)

Objective: To observe the effects of puerarin on the expressions of CaM, CaMKⅡ, BDNF and Akt in vascular dementia cell models induced by oxygen and glucose deprivation (OGD). Methods: The passaged well-differentiated PC12 cells were randomly divided into control group, model group, and low-dose, medium-dose and high-dose intervention groups. Vascular dementia cell model was established by OGD. Suitable OGD time and concentration of puererin were obtained from the cell viability measured by MTT assay. The release of LDH was measured to assess the extent of cell damage and identify cell models. The expressions of CaM, CaMKⅡ, MECP2, BDNF and Akt were detected by Western blot. Results: PC12 cells with OGD prolonged viability decreased in a time-dependent manner, with increased concentrations of puerarin increased in a concentration-dependent manner. Effective intervention of puerarin was 0.1-10 μmol/L and optimal time of OGD was 6 h. Compared with control group, the release of LDH in model group was significantly increased (P0.05). Puerarin could down-regulate the level of CaM protein, increase the expressions of MECP2 and BDNF and the phosphorylation of CaMKⅡ, and also increase the phosphorylation of Akt in addition to the low-dose group (P<0.05). Conclusion: The neuroprotective effect of puerarin may be related to the increase of the autophosphorylation of CaMKⅡ mediated by Ca2+-CaM complex, induce the phosphorylation of MECP2, up-regulate the expression of BDNF and activate the PI3K-Akt pathway to inhibit the expression of apoptotic genes and proteins.

Protective effect of water soluble CoQ10 on rotenone-induced apoptosis in PC12 cells / 西安交通大学学报(医学版)

Objective To investigate the protective effect and the underlying mechanism of water soluble coenzyme Q10 (CoQ10)against rotenone induced injury on PC12 cells model.Methods PC12 cells were cultured with rotenone,water-soluble CoQ1 0 was added to the culture media 3 hours prior to the rotenone incubation.We determined cell viability by CCK8;reactive oxygen species (ROS)was detected by spectrophotometer;and Bcl-2, Bax,active Caspase-3,Caspase-9 and apoptosis-inducing factor (AIF)were measured by Western blotting after 24-hour rotenone incubation.Results After the treatment by rotenone,cell viability decreased significantly (P＜0.01)and ROS level increased (P＜0.01).CoQ10 could improve PC12 cell viability (P＜0.01)and reduce the level of ROS (P＜0.01).Western blotting experiments showed that CoQ10 could reduce rotenone-induced Caspase-9 (P＜0.05),active Caspase-3 (P＜0.05)and Bax (P＜0.01)expressions,increase the expression of Bcl-2 (P＜0.01),and prevent nuclear translocation of AIF (P＜0.05).Conclusion CoQ10 has a protective effect on rotenone-induced apoptosis in PC12 cells,the mechanism of which may be through scavenging ROS in cells;decreasing caspase-9 ,active caspase-3 and Bax expressions;and increasing the expression of Bcl-2 ;and preventing AIF nuclear translocation.

Effect of microenvironment on β-site amyloid precursor protein cleaving enzyme 1 expression in PC12 Cells / 上海交通大学学报（医学版）

Objective • To investigate the effect of microenvironment on the expression of β-site amyloid precursor protein cleaving enzyme 1 (BACE1) in PC12 cells. Methods • The PC12 cells were respectively treated with thapsigargin (Tg), lipopolysaccharide (LPS), H2O2, hypoxia incubator and hypoglycemic medium to produce mild endoplasmic reticulum stress (ERS), inflammatory environment, mild oxidative stress, hypoxia and low glucose conditions. Western blotting and fluorescence quantitative PCR were used to detect the expression of BACE1 protein and BACE1 mRNA. Results • After PC12 cells were treated with Tg (0.13 μmol/L) for 12 h or LPS (0.01 mg/mL) for 36 h, BACE1 protein and mRNA levels were significantly up-regulated (P0.05). Conclusion • Mild ERS and inflammatory condition can up-regulate BACE1 mRNA and protein expression in PC12 cells.

Construction of SERCA Interference Lentivirus Expression Vector and Establishment of Stable PC 12 Cell Line / 中国药学杂志

OBJECTIVE: To construct the SERCA gene interference lentivirus expression vector and establish stable transfected PC 12 cell line. METHODS: The interference sequence targeting at rat SERCA gene was designed and synthesized. pGag/Pol, pRev, and pVSV-G were co-transfected into 293T cells. The lentivirus particles were packaged and generated. The virus titer was detected. PC 12 cells were transfected for establishing the stable cell line; RT-PCR and Western blot were used to detect SERCA gene and protein expression in stable PC 12 cells,and the RESULTS were compared with those in the control group. RESULTS: The lentivirus expression vector targeted at SERCA was successfully constructed and the virus titer was 3×108 U•mL-1. Stable transfected PC 12 cells line was established. The effective interference verification revealed that shSERCA could significantly reduce the mRNA and protein levels of SERCA (P<0.01). CONCLUSION: The shRNA lentiviral expression vector of SERCA gene is successfully constructed and the PC 12 cell line stably interfering with SERCA expresion is established.

Chemical Constituents of Drynaria fortunei and Their Protective Effects on PC12 Cell / 中国药学杂志

OBJECTIVE: To study the chemical constituents from the rhizome of Drynaria fortunei and the protective effects of them on PC12 cells induced by Aβ25-35. METHODS: The compounds were isolated and purified by silica gel, Sephadex LH-20, ODS column chromatography, and their structures were identified on basis of spectroscopic METHODS:, such as MS and NMR. PC12 cells were treated with Aβ25-35 to establish the Alzheimer' s disease models. The compounds of different concentrations were added into culture medium to detect the protection. MTT assay was used to detect cell vitality and to observe the protective effects of compounds on PC12 cells induced by Aβ25-35. RESULTS: Nine compounds were isolated and identified as naringin(1), neoeriocitrin(2), 5,7-dihydroxychromone-7-neohesperidoside(3), (E)-4-O-β-D-glucopyranosyl caffeic acid(4), kaempferol(5), luteolin(6), protocatechoic acid(7), psoralen(8), and β-sitosterol(9). The cell experiments were performed on the compounds 1-8 and the RESULTS: showed they can promote the proliferation of PC12 cells. The cell vitality increase with concentration rising, and the difference is statistically significant (P<0.05). CONCLUSION: Compounds 1-8 play an important role in protecting Aβ25-35-induced injury in PC12 cells and they are the main active components of Drynaria fortunei in the protection of central nervous function.

Protective effects of genistein on Aβ₂₅₋₃₅-induced PC12 cell injury via regulating CaM-CaMKIV signaling pathway / 中国中药杂志

Genistein is a kind of isoflavone compounds， also called phytoestrogens， with clinical effects on cardiovascular disease， cancer and postmenopausal-related gynecological diseases， and also has the potentiality in the prevention and treatment of Alzheimer's disease(AD). In this study， the protective effect of genistein on Aβ₂₅₋₃₅-induced PC12 cell injury and effect on CaM-CaMKIV signaling pathway were observed to investigate its mechanism for AD. PC12 cells were cultured and then the safe concentration of genistein and the modeling concentration and optimal time point of administration of Aβ₂₅₋₃₅ were screened by MTT assay. After being pretreated with different concentrations of genistein(25， 50， 100 μmol·L⁻¹) on PC12 cells， the AD model of PC12 cells was induced by Aβ₂₅₋₃₅. Then the survival rate of cells was detected by MTT assay; morphological change of cells was observed under the inverted microscope， and apoptosis of cells was assessed by AO/EB fluorescence staining; the neuroprotective effects of genistein on AD cell model were observed and the optimal concentration of genistein was determined. Expressions of mRNA and protein levels of CaM， CaMKK， CaMKIV and tau were detected by qRT-PCR and Western blot assay， respectively. The results showed that as compared with the blank group， the cell survival rate was decreased; the cell damage and apoptosis were increased; and the expressions of mRNA and protein levels of CaM， CaMKK， CaMKIV and tau were increased in AD model group. Genistein could significantly improve the cell survival rate， reduce the cell damage and apoptosis of AD cell model， and significantly down-regulate the expressions of mRNA and protein levels of CaM， CaMKK， CaMKIV and tau of AD cell model. These results indicated that genistein has obviously neuroprotective effect on the AD cell model induced by Aβ₂₅₋₃₅， and the mechanism may be related to the down-regulation of CaM-CaMKIV signaling pathway and Tau protein expression.

Preparation of DJ-1 overexpressing cell model by using lentiviral vector / 基础医学与临床

Objective To prepare gene overexpressing cell model of human wild-type DJ-1 and its L166P mutant, and to investigate the role of lentiviral vector in gene overexpressing cell model .Methods Wild type DJ-1 and L166P mutant DJ-1 lentiviral vector plasmids were respectively constructed .After sequencing and comparing cor-rectly, the plasmid was amplified and transfected into HEK 293T cell line.Expression of WT DJ-1 and L166P mu-tant DJ-1 in cell lines was detected by fluorescence and Western blot .After determining the accurate expression of the target protein, a large amount of HEK293T cells was transfected and packaged to produce lentiviral particles. The PC12 cells were infected with the titer of virus supernatant.The fluorescence intensity of GFP and the expres-sion of target protein were observed by fluorescence microscope and Western blot method ,and the infection effi-ciency of the virus was determined .Results Lentiviral vectors carrying wild type DJ-1 and its mutants were suc-cessfully constructed .The virus vector can be transfected into HEK 293T cells and the target protein can be correctly expressed.The viral titers of LV-DJ-1 and LV-DJ-1/L166P were 2×109 TU/mL and 2×108 TU/mL, respectively. Virus supernatant can efficiently infect PC 12 cells, and most cells can express target proteins .The protein expres-sions of exogenous wild-type DJ-1 and L166P mutants were 315% and 285% of endogenous content ,respectively. Conclusions Lentivirus vector can infect cells efficiently , and it is a good way to prepare gene over expressing cell model.A cell model overexpressing DJ-1 or its L166P mutant is successfully prepared .The model can be used for subsequent DJ-1 function research .

Effect of pyrroloquinoline quinone on learning and memory ability of apolexis rats / 中国食品卫生杂志

Objective This study aimed to investigate the role of pyrroloquinoline quinone (PQQ) in repairing oxidative nerve cells,and to study the antioxidant capacity of PQQ on the oxidative damage of rats caused by apolexis,as well as the effects on learning and memory abilities of apolexis rats.Methods Oxidative damage of PC12 was induced by H2O2,and the repairing rate of PQQ on oxidative PC12 cells was tested by methylthiazolyldiphenyl-tetrazolium bromide assay kit.The 18-month-old male SD rats were administered PQQ (0,10,20,40 mg/kg).After 4 weeks,Morris water maze test was used to test the learning and memory ability.After 6 weeks,serum and brain tissue related indicators and antioxidant capacity were recorded.Results The survival rate of PC12 cells increased from 59.1％ to 90.5％ with 200 nmol/L PQQ.Compared with the apolexis model group,the latency of the PQQ group (20,40 mg/kg) was shortened in the Morris water maze experiment,the swimming distance was reduced,pass-through counts were increased,and the first secure platform pass-through was reduced.Meanwhile,the levels of malondialdehyde and lipofuscin in serum and brain tissue of PQQ group decreased,the levels of superoxide dismutase,glutathione peroxidase vitality,antioxidant capacity of PQQ group (20,40 mg/kg) were enhanced.Conclusion PQQ could repair the oxidative damage of nerve cells,and it was confirmed that PQQ could play the same antioxidant effect in body and brain,and increase the learning and memory ability of apolexis rats.