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Objective To investigate the expression of P2X7 receptor (P2X7R) and the effect of P2X7R agonist 2'-3'-O-(4-benzoyl-benzoyl) ethane adenosine triphosphate three amine salt (BzATP) on cell growth and apoptosis in non-small cell lung cancer A549 cells,and to explore the related mechanism.Methods The expression of P2X7R in A549 cells was detected by immunofluorescence.Cells were treated with different concentrations (150,300,600 μmol/L) of BzATP.Cells untreated with BzATP were used as control group.3-(4,5-dimethyl-2-thiazoly)-2,5-diphenyl-2H-tetrazolium bromide (MTF) assay and Hoest33342 staining were respectively used to detect cell viability and apoptosis.Enzyme-linked immunosorbent assay (ELISA) was uesd to detect the concentration of tumor necrosis factor-α (TNF-α) of cell culture supernatants.The expressions of nuclear factor-κB (NF-κB) p65,inhibitor of α of NF-κB (IκBα) and the phosphorylation of inhibitor of α of NF-κB (phospho-IκBα) were detected by Western blotting.Results P2X7R was expressed on the cell membrane of A549 cells.Survival rate of A549 cell was significantly decreased with the concentrations of BzATP at 300 and 600 μmol/L [(67.87 ± 8.98) ％,(44.73 ± 6.92) ％],compared with the control group (98.60 ± 1.44) ％,the differences were statistically significant (t =4.481,P =0.027;t =3.920,P =0.038).BzATP promoted apoptosis,and increased the concentration of TNF-α of supernatant at 300 and 600 μmol/L [(57.35 ±6.41) pg/ml,(78.63 ± 11.33) pg/ml],compared with the control group (42.56 ±0.37) pg/ml,the differences were statistically significant (t =6.410,P =0.035;t =11.330,P =0.005).In addation,the expressions of NF-κB p65 and IκBα were respectively downregulated and upregulated by BzATP,while the expression of phospho-IκBα was not significantly altered.Conclusion P2X7R is expressed on A549 cell membrane.BzATP can inhibit cell proliferation and induce the apoptosis of A549 cells,and the mechanism of action may be related to promoting the release of TNF-α and inhibition of NF-κB pathway.
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Objective To compare the influence of three kinds of Walnut kernel extracts on learning and memory ability as well as ultrastructural pathology in hippocampus of Alzheimer's disease (AD) rats. Methods AD rat model was established by injection of amyloid-beta protein (Aβ1-40 ) into the nucleus basalis of meynert. The AD rats were randomly divided into Walnut kernel-water group,Walnut kernel-ethanol group,Walnut kernel-acetone group,and the model control group,10 rats in each group. In addition,10 rats of normal control group and 10 rats of sham operation group were selected. The model control group was not treated; the treatment groups were intragastrically given Walnut kernel water extract,ethanol extract,and acetone extract ( the equivalent of pharmacognosy 0. 3 g·mL-1 ),respectively,dose 3 g·kg-1 . The learning and memory ability was studied by Morris water maze,and ultrastructure of neurons was observed under the transmission electron microscopy. Results The time of looking for platform in Walnut kernel-water group,Walnut kernel-ethanol group,and Walnut kernel-acetone group were dropped swiftly at the beginning of 3 days, the third day is (51. 80±4. 37),(61. 20±4. 67),and (59. 63±5. 24),respectively; the model control group is (67. 67±6. 12) s. Compared with the model group,the differences were significance (P<0. 05); However,the acetone extract of Walnut kernel can obviously enhance the learning and memory ability (P<0. 01),and the ultrastructure almost returned to normal. Conclusion The acetone extracts of Walnut kernel have the function of preventing Alzheimer's disease.
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It has been suggested that HCN1 is primarily expressed in hippocampus, however little is known about its effects on spatial learning and memory. In the present study, we investigated the effects of non-specific HCN1 blocker CsCl on spatial learning and memory by using Morris water maze and in situ hybridization in mice. The results showed CsCl 160 mg/kg ip for 4 days, and the mean escape latency was 34 s longer than that of normal control (P＜0.01). In hippocampal tissues, staining for the HCN1 mRNA was stronger in the DG and CA1 region of the hippocampus (P ＜0.05, P＜0.05, when CsCl-administration group was compared with normal group). Our results suggested that CsCl could significantly affect the spatial learning and memory in mice, and HCN channel is involved in the process of learning and memory.
RESUMO
It has been suggested that HCN1 is primarily expressed in hippocampus, however little is known about its effects on spatial learning and memory. In the present study, we investigated the effects of non-specific HCN1 blocker CsCl on spatial learning and memory by using Morris water maze and in situ hybridization in mice. The results showed CsCl 160 mg/kg ip for 4 days, and the mean escape latency was 34 s longer than that of normal control (P<0.01). In hippocampal tissues, staining for the HCN1 mRNA was stronger in the DG and CA1 region of the hippocampus (P <0.05, P<0.05, when CsCl-administration group was compared with normal group). Our results suggested that CsCl could significantly affect the spatial learning and memory in mice, and HCN channel is involved in the process of learning and memory.
