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1.
AIM: To study the effects of metoprolol (Meto) on the apoptosis of neonatal rat cardiomyocytes and the phosphorylation of connexin 43 (Cx43) induced by norepinephrine (NE). METHODS: Neonatal SD rat cardiomyocytes were divided into the following five groups (n=6 in each group): (1) control (Con) group: no treatment; (2) NE group: treatment with NE at 0.1 μmol/L for 24 h; (3) NE+Meto group: simultaneous treatment with NE and Meto both at 01 μmol/L for 24 h; (4) NE+Meto+PD98059 group: pretreatment with extracellular signal-regulated kinase (ERK) phosphorylation inhibitor PD98059 at 10 μmol/L for 30 min and then treatment with NE and Meto both at 01 μmol/L for 24 h; (5) NE+PD98059 group: pretreatment with PD98059 at 10 μmol/L for 30 min and then treatment with NE at 01 μmol/L for 24 h. The beating rates of cardiomyocytes in various groups were calculated, and the viability of cardiomyocytes was assayed by MTT method. The Cx43 mRNA expression was detected by RT-PCR, and the protein expression of phosphorylated Cx43 (p-Cx43), phosphorylated ERK1/2 (p-ERK1/2) and cleaved caspase-3 was detected by Western blotting. RESULTS: (1) Separate NE treatment could significantly increased the beating rate of cardiomyocytes and reduced cell viability, while Meto showed the opposite effects. PD98059 treatment had no significant effect on cardiomyocyte beating rate, but suppressed Meto to improve cell viability to some extent. (2) Compared with Con group, separate NE treatment significantly increased the Cx43 mRNA expression (P<001). Compared with NE group, Meto or PD98059 intervention could significantly inhibited Cx43 mRNA expression (both P<001), and simultaneous treatment with Meto and PD98059 could further suppress Cx43 mRNA expression up-regulated by NE (P<001). (3) Compared with NE group, Meto significantly inhibited the increased p-Cx43, p-ERK1/2 and cleaved caspase-3 expression induced by NE (P<001), and simultaneous treatment with Meto and PD98059 could further enhance the inhibition of p-Cx43, p-ERK1/2 and cleaved caspase-3 expression by Meto (P<001). PD98059 treatment had no significant effect on the increased p-Cx43 and cleaved caspase-3 expression induced by NE (P>005). CONCLUSION: The inhibitory effect of Meto on NE-induced cardiomyocyte apoptosis is related to the inhibition of Cx43 phosphorylation, which may be partly mediated via ERK1/2 pathway.  相似文献   

2.
AIM: To investigate whether gap junction participates in transforming growth factor β1(TGF-β1)-induced proliferation of spontaneous hypertensive rat (SHR) vascular smooth muscle cells (VSMCs). METHODS: The thoracic aorta of the rats were sampled. The primary SHR VSMCs were isolated and cultured in vitro. The cells were divided into 4 groups: control group, TGF-β1 group,18α-glycyrrhetinic acid(18α-GA) group and TGF-β1+18α-GA group. The proliferation of SHR VSMCs was observed by the methods of MTT and flow cytometry. The protein expression and co-localization of connexin(Cx)43 and Cx40 in SHR VSMCs were detected by immunofluorescence staining. The protein levels of Cx43 and Cx40 in the cells were also measured by Western blotting. The method of molecular dye transfer (scrape dye transfer method) was applied to detect the function of gap junction in SHR VSMCs. RESULTS: The protein expression of Cx43 and Cx40 in SHR VSMCs was positive and co-localized in the cytoplasm. Compared with control group, the percentage of S-phase detected by cell cycle and A value detected by MTT in TGF-β1 group were obviously increased (P<0.05), indicating that the proliferation of the cells was enhanced. However, the proliferation of the cells decreased in 18α-GA group (P<0.05). Compared with TGF-β1 group, the percentage of S-phase and A value in TGF-β1+18α-GA group were both significantly decreased (P<0.05), indicating that the proliferation of the cells decreased. Compared with control group, the protein expression of Cx43 in TGF-β1 group was increased (P<0.05), whereas the protein expression of Cx40 was not changed (P>0.05), and the protein expression of Cx43 and Cx40 in 18α-GA group were decreased (P<0.05). Compared with TGF-β1 group, the expression of Cx43 in TGF-β1+18α-GA group was significantly decreased (P<0.05),but no difference of the Cx40 protein levels between the two groups was observed. Compared with control group, the function of gap junction detected by scrape dye transfer method in TGF-β1 group was enhanced (P<0.05), and weakened in 18α-GA group (P<0.05). Compared with the TGF-β1 group, the function of gap junction in TGF-β1+18α-GA group was significantly attenuated (P<0.05). CONCLUSION: TGF-β1 enhances the function of gap junction to stimulate the proliferation of SHR VSMCs through the expression of Cx43 protein. The expression of Cx40 protein may not play a major role in this process.  相似文献   

3.
AIM: To study the effect of meglumine cyclic adenylate (MCA) on the differentiation of bone marrow mesenchymal stem cells (BMSCs) into cardiomyocytes in vitro. METHODS: The whole bone marrow adherent culture method was used to isolate, culture and amplify the BMSCs. The surface markers of BMSCs were determined by flow cytometry analysis. MCA at concentrations of 10-2 mol/L, 10-3 mol/L, 10-4 mol/L, 10-5 mol/L, 10-6 mol/L and 10-7 mol/L was added to the culture medium containing the second generation of BMSCs.5-Azacytidine(5-Aza) was used as a positive control. The cell viability was measured by MTT method.The cAMP content in BMSCs was detected by ELISA. The mRNA expression of GATA-4, Cx43 and β-MHC in MCA group and MCA+H89 (a PKA inhibitor) group was measured by SYBR-RT-PCR. The differentiation effects of MCA and 5-Aza were compared by flow cytometry. RESULTS: Most of the BMSCs expressed CD44 and CD71, and did not express CD45. MCA inhibited the viability of BMSCs in a time-and dose-dependent manner, and MCA atthe concentration of 10-2 mol/L showed particularly remarkable effect. MCA significantly increased intracellular cAMP level in BMSCs in a concentration-dependent manner. The mRNA expression of GATA-4, β-MHC and Cx43 in MCA group were significantly higher than that in blank group (P<0.05), and the highest effect was under the condition of MCA induction at the concentration of 10-3 mol/L for 3 days. The mRNA expression of GATA-4, β-MHC and Cx43 in MCA group was higher than that in 5-Aza group and H89+MCA group (both P<0.05). Differentiation rate in MCA group was slightly higher than that in 5-Aza group (20.24%±1.02% vs 18.39%±0.58%, P<0.05). CONCLUSION: MCA stimulates BMSCs to increase intracellular cAMP production and inhibits the viability of BMSCs, thus promoting the mRNA expression of GATA-4, β-MHC and Cx43 through the cAMP/PKA signaling pathway.  相似文献   

4.
AIM: To investigate the effects of crude extracts of Cordyceps gunnii (CGE), Lepista lentinus (LLE), Cordyceps sinensis (CSE) and Lentinus striguellus (LSE) on the proliferation of high glucose-treated human umbilical vein endothelial cells (HUVECs). METHODS: The cultured HUVECs were divided into normal control group (treated with M199 culture medium alone), high glucose group (treated with M199 culture medium containing 33 mmol/L glucose) and 4 crude extracts of edible-medicinal fungi (CGE, LLE, CSE and LSE) intervention groups (treated with the crude extract of edible-medicinal fungus at concentrations of 12.5, 25, 50, 100 mg/L in high glucose M199 culture medium). The cell proliferation was evaluated by MTT assay. The cell cycle and ROS level were measured by flow cytometry. RESULTS: Compared with normal control group, the MTT absorbance value and the percentage of G0/G1 stage of the HUVECs in high glucose group were significantly decreased (P<0.05), while the percentage of S+G2/M and ROS level were significantly increased (P<0.05). Compared with high glucose group, treatment with the crude extracts of Lepista lentinus and Lentinus striguellus decreased the cell absorbance value (P<0.05), and the inhibitory effect was enhanced in a dose-dependent manner. However, Cordyceps gunnii had no effect (P>0.05). The crude extracts of Cordyceps sinensis (12.5~50 mg/L) significantly enhanced the proliferative activity of HUVECs, decreased the percentage of G0/G1 stage of HUVECs, increased the percentage of S+G2/M of HUVECs, and reduced the intercellular ROS level (P<0.05). CONCLUSION: Only Cordyceps sinensis crude extract effectively protects the HUVECs in high glucose-induced injury, which might be due to promoting more cells to enter to the cell cycle and down-regulating oxidative stress.  相似文献   

5.
AIM: To investigate the effect of decorin (DCN) on the proliferation and apoptosis of human pterygium fibroblasts (HPF) in vitro , and to compare the effect of mitomycin C (MMC) in order to search for a new method to prevent the recurrence after pterygium surgery. METHODS: Human pterygium fiborblasts were isolated from the caudomedial part of pterygium tissues in pterygium patients and then cultured in vitro using tissue inoculation method. The cells were treated with DCN and MMC at concentrations of 0.01, 0.1, 1, 5 and 10 mg/L. The morphological alterations of HPF were observed after 24 h, 48 h or 72 h of treatment. MTT method was used to assay the effects of the 2 drugs at different doses after 12 h, 24 h and 48 h on the proliferation of the cells. The expression of proliferating cell nuclear antigen (PCNA) in each group treated with different doses of DCN was detected by the method of immunohistochemistry after 48 h. The cell cycle distribution was determined by flow cytometry analysis. RESULTS: After administration of 10 mg/L DCN or 1 mg/L MMC for 12 h, the proliferation of HPF was significantly inhibited by both drugs in a dose- and time-dependent manner (P<0.05). After treated with 1~10 mg/L DCN for 48 h, the percentage of HPF in G0/G1 phase was increased, while the percentage of HPF in G2/M phase and S phase (G2/M%+S%) was decreased after treated with 5~10mg/L DCN for 48 h (P<0.05). The late-apoptotic cells were not found in DCN group and MMC group. DCN dose-dependently inhibited the expression of PCNA in HPF (P<0.05). CONCLUSION: Decorin significantly inhibits the proliferation of HPF, and blocks the cells in G1 phase.  相似文献   

6.
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7.
AIM: To study the role of gap junction(GJ) in the pathogenesis of epilepsy. METHODS: The expression of connexin(Cx) 32 and Cx43 was evaluated in penlylenetetrazol(PTZ)-induced epilepsy rats at different time points with immunohistochemistry and real-time RT-PCR. The uncoupler of gap junction,carbenoxolone (CBX), and anti-epilepsy drug,carbamazepine (CBZ), were introduced to investigate the role of gap junction in epilepsy. RESULTS: The expression of Cx32 in the cortex and hippocampus of rats increased 2 h after PTZ administration, and was more obvious 8 h later, so was the expression of Cx43. CBX not only notably inhibited the expression of Cx32 and Cx43, but also suppressed the seizures. CBZ had no obvious effect on Cx32/43 expression. Real-time RT-PCR examination showed that the level of Cx32 mRNA went up rapidly 2 h after seizures, and began to decrease 10 h later. The level of Cx43 mRNA in PTZ group was higher than that in control group from 2 h to 5 h. CBX inhibited both the seizures and the increase in the mRNA expression of Cx32/43, while CBZ only suppressed the seizures and did not affect the expression of Cx32/43. CONCLUSION: GJ between neurons is reinforced after epileptic activities and takes part in the pathophysiological mechanism of synchronization and epilepsy. CBZ has no effect on the expression of Cx32 and Cx43, indicating that the anti-epileptic mechanism of CBZ is independent of GJ.  相似文献   

8.
AIM: To study the reverse effects of saikoside (SS) on the multidrug resistance (MDR) of human leukemic cell line K562/ADM and to investigate the related mechanism. METHODS: K562 cells and K562/ADM cells in the culture were treated with SS at the concentrations of 1~100 mg/L. The inhibitory rate of the cell proliferation was measured by MTT assay. Non-cytotoxic dose of SS was determined. K562/ADM cells were treated with SS at non-cytotoxic doses of 1.25, 2.5 and 5.0 mg/L with different concentrations of adriamycin (ADM,0.05~100 mg/L). The 50% inhibitory concentration (IC50) and the reversal index in all groups were determined. The cell morphology was observed after treated with SS+ADM. The effects of SS on ADM accumulation in K562/ADM cells, the cell cycle profile and apoptosis were examined by flow cytometry. RESULTS: The inhibitory rates were significantly increased in a dose-dependent manner when the cells were treated with different doses of SS (1~100 mg/L). The available reversal concentration of SS was 5.0 mg/L and the reversal index was 21.5 folds for K562/ADM cells. After treated with SS+ADM, the number of tumor cells was decreased and apoptotic cells were increased in a dose-response relationship. ADM accumulation in K562/ADM cells treated with SS was significantly higher than that in control cells (P<0.05). SS may significantly enhanced the apoptosis of K562/ADM cells treated with ADM (P<0.05). K562/ADM cells treated with SS were blocked in the stage of G0/G1. CONCLUSION: SS has effect on proliferation inhibition and MDR reversal in K562/ADM cell line. The reversal mechanisms of SS may be due to increasing the accumulation of chemo therapeutics in the cell, inducing the cell apoptosis and arresting the cells in G0/G1 phase.  相似文献   

9.
AIM: To investigate the effect of propofol on the viability, invasion ability and apoptosis of colorectal cancer cells.METHODS: Propofol at 10, 25, 50 and 100 μmol/L was used to treat LoVo cells for 72 h, and propofol at 100 μmol/L was used to treat the LoVo cells for 12, 24, 48 and 72 h. The cell viability was measured by CCK-8 assay. The invasion ability of the LoVo cells treated with propofol at 100 μmol/L for 72 h was detected by Transwell assay. The cell cycle distribution and cell apoptotic rate were analyzed by flow cytometry. The protein levels of matrix metalloproteinase (MMP)-2, MMP-9, cleaved caspase-3, Notch1 and hairy and enhancer of split 1 (Hes1) were determined by Western blot.RESULTS: Propofol inhibited LoVo cell viability. The cell invasion ability, S stage cells, and the protein levels of MMP-2, MMP-9, Notch1 and Hes1 in propofol group were significantly lower than those in control group, and the apoptotic rate, G0/G1 cells and the protein level of cleaved caspase-3 were significantly higher than those in control group (P<0.01).CONCLUSION: Propofol inhibits the viability and invasion ability of colorectal cancer LoVo cells, blocks cell cycle and induces apoptosis. The mechanism is related to down-regulation of Notch1 signaling pathway.  相似文献   

10.
AIM: To investigate the differentiation of neural stem cells (NSCs) after transplanted into vitreous and the effects on the regeneration of retina ganglion cells (RGCs) after optic nerve microcrushed.METHODS: After optic nerve microcrushed in adult rat,2×104/2 μL NSCs or 2 μL 0.1 mol/L PBS was injected into vitreous.Animals were divided into control group (MC group,MC+PBS group) and experiment group (MC+NSCs).Animals in each group were allowed to survive for 3,4,5 weeks,respectively.The regenerating RGCs were labeled retrogradely with granular blue,and the numbers of regenerating RGCs in each retina were observed under fluorescent microscope.In addition after 5 animals in MC+NSCs group survived for 4 weeks,rat eyeballs were removed and prepared as freezing microtome sections for observing the migration of NSCs and NF,GFAP,CNP immumodetection.RESULTS: Compared the mean numbers of regenerating RGCs between experiment group and control group at 3,4,5 weeks,the difference was significant (P<0.01).NSCs expressed NF,GFAP and CNP at 4 weeks and were not found to incorporate into retina.CONCLUSION: It suggests that NSCs enhance the RGCs regeneration after ON microcrushed and differentiate into neurons,astrocytes and oligodendrocytes.  相似文献   

11.
AIM: To observe the effect of platelet-activating factor (PAF) on cultured neuronal viability and glial fibrillary acidic protein (GFAP) expression in cultured astrocytes. METHODS: Neurons and astrocytes obtained from the brain cortex of the embryo and newborn mice respectively were cultured and purified, and they were divided into the control and experimental groups. PAF was added into the experimental groups at concentrations of 4, 8 and 16 μmol/L. Each group was cultured for 4 h, 24 h and 72 h, respectively. MTT method and immunohistochemistry were used to observe the neuronal viability and GFAP expression in astrocytes, respectively. RESULTS: During different time after adding PAF at different concentrations into cultured neurons and astrocytes, respectively, neuronal viability declined, and the number of astrocytes decreased, but GFAP expression in survival astrocytes increased. The effects were shown to be in a concentration-dependent manner. CONCLUSION: PAF decreases the neuronal viability directly and influences the neuronal survival indirectly by astrocytes.  相似文献   

12.
AIM: To investigate the effect of all-trans retinoic acid (ATRA) on the viability of gastric cancer cell SGC-7901 and the sensitivity to radiotherapy. METHODS: MTT assay was used to examine the cell viability. Radio-sensitivity and cell cycle were determined by colony formation assay and flow cytometry, respectively. The mRNA levels of Bax, Bcl-2, survivin and NF-κB in the cells were measured by RT-qPCR. RESULTS: ATRA inhibited the viability of SGC-7901 cells in a concentration-dependent manner. The maximal inhibition was at concentration of 8 μmol/L. Colony formation assay revealed that the combination of ATRA with X-ray treatment significantly reduced the values of D0 and Dq, and shifted down the fitting survival curve, as compared with radiotherapy alone. Moreover, ATAR markedly decreased the percentage of G2/M phase in the SGC-7901 cells (P<0.05). In addition, following ATRA treatment, the mRNA levels of Bcl-2 and survivin were decreased (P<0.05), whereas the mRNA levels of Bax and NF-κB were increased (P<0.05). CONCLUSION: ATRA enhances the sensitivity of SGC-7901 cells to radiotherapy, inhibits G2/M arrest and regulates the mRNA expression of Bax, Bcl-2, survivin and NF-κB.  相似文献   

13.
AIM:To investigate the role of NF-κB in pentetrazole-induced repeated seizure in developing rats with the inhibitor of NF-κB pyrrolidine dithiocarbamate (PDTC). METHODS:10-day-old Wistar rats (n=72) were prepared for epilepsy model and divided into three groups at random: the PTZ group, the PDTC+PTZ group and the control group. The behavioral changes, the cells morphology and neurons counts in hippocampus, the expression of NF-κB, BrdU (5-bromo, 2-deoxyuridine) immunoreactive cells in hippocampus and the mossy fiber sprouting were observed.RESULTS:(1) The NF-κB expressed in PTZ group was significantly higher than that in PDTC+PTZ group and control group (P<0.01). (2) The dentate gyrus granule cell count in PTZ group was significantly higher than that in control group (P<0.05). In PDTC+PTZ group cell counts in CA1, CA3 and hilar region were significantly lower than those in PTZ group (P<0.05). (3) The BrdU-immunoreactive cells counts in dentate gyrus in PTZ group and PDTC+PTZ group were significantly higher than those in control group (P<0.01), but in PDTC+PTZ group BrdU-immunoreactive cell count was significantly lower than that in PTZ group (P<0.01). Correlate analyzes between NF-κB expression and BrdU-immunoreactive cell counts/granule cell counts showed positive correlation (P<0.01). (4) The mossy fiber sprouting in both PTZ and PDTC+PTZ group was observed. However, the degrees of sprouting showed no significant differences between two groups. CONCLUSION:NF-κB plays a crucial role in epilepsy of developing rats. It encourages neurogenesis and protects neurons in hippocampus, but has no significant effect on mossy fiber sprouting.  相似文献   

14.
AIM:To investigate the effects of ethyl acetate (EtOAc) extract of Pleione bulbocodioides (Franch.) Rolfe on proliferation and apoptosis of human leukemia K562 and HL-60 cells and the possible apoptosis pathway. METHODS:Human leukemia cell lines were treated with EtOAc extract of Pleione bulbocodioides at different concentrations. XTT method was used to evaluate the viability of K562 cells and HL-60 cells. The cell growth inhibition was calculated by Trypan blue exclusion test. The percentage of apoptotic cells was determined by flow cytometry, and 4',6-diamidino-2-phenylindole (DAPI) was used to observe morphological changes of the cells. The cell cycle was observed by propidium iodide (PI) staining. The protein expression of Bcl-2, Bax, cleaved poly(ADP-ribose) polymerase (PARP), cleaved caspase-3, cytochrome C and apoptosis-inducing factor (AIF) wase determined by Western blot. RESULTS:The cell viability and proliferation were inhibited by EtOAc extract of Pleione bulbocodioides with IC50 of (42.14±2.54) mg/L for HL-60 cells and (51.28±3.12) mg/L for K562 cells at 24 h. The results of Annexin V-FITC/PI and DAPI staining showed that EtOAc extract of Pleione bulbocodioides induced cell apoptosis in a dose-dependent manner. The apoptotic rate was increased compared with control group (P<0.05). The G2 phase increased with typical cell apoptosis-induced morphological changes. The levels of pro-apoptotic proteins Bax, cleaved PARP and cleaved caspase-3 were increased, while Bcl-2 was down-regulated (P<0.05). Cytochrome C and AIF in cytosol, characteristic proteins of intrinsic mitochondrial apoptosis pathway, also increased with the concentration of EtOAc extract of Pleione bulbocodioides increasing (P<0.05). CONCLUSION:EtOAc extract of Pleione bulbocodioides significantly inhibits cell proliferation and induces cell apoptosis in human leukemia cell lines HL-60 and K562 through intrinsic mitochondrial apoptosis pathway.  相似文献   

15.
AIM: To explore the effects of ephrin-A3 expression on the proliferation of astrocytes induced by lipopolysaccharide (LPS).METHODS: Astrocytes from cerebral cortex of newborn rats were cultured and purified.The cells were exposed to LPS at the concentration of 10 mg/L and divided into 6 groups at random: control group (without LPS) and the groups of exposure to LPS for 30 min, 6 h, 24 h, 48 h and 72 h.The morphological changes of the astrocytes were observed under inverted phase-contrast microscope.The survival rate and apoptotic rate of astrocytes were measured by MTT method and AO/EB staining,respectively.The expression of ephrin-A3 and glial fibrillary acidic protein (GFAP) was measured by fluorescein staining with CY3.The changes of the inflammatory factors TNF-α and IL-6 in the culture supernatant were detected by ELISA.RESULTS: Compared with control group, the survival rates were notably increased, the apoptotic rates were notably decreased, the expression of ephrin-A3 and GFAP was significantly up-regulated and the inflammatory cytokines were significantly increased in the cells exposed to LPS at the concentration of 10 mg/L for 24 h, 48 h and 72 h.CONCLUSION: LPS promotes the proliferation of astrocytes.The expression of ephrin-A3 increases during the proliferation of astrocytes induced by LPS in rats.The mechanism may be related to inflammatory stimulation by cytokines.  相似文献   

16.
AIM: To detect the expression of WNT5B in normal breast epithelial cells and different breast cancer cell lines, and to investigate the effects of WNT5B over-expression on the viability and apoptosis of human breast cell line MCF-7.METHODS: The mRNA expression of WNT5B was detected by RT-PCR in different breast cancer cells. MCF-7 cells were transfected with plasmid pcDNA3.1/WNT5B or pcDNA3.1, and the expression of WNT5B at mRNA and protein levels was examined in the 2 groups by real-time PCR and Western blotting, respectively. Subsequently, the changes of cell viability and cell apoptosis were analyzed by CCK-8 assay and flow cytometry, respectively. RESULTS: The expression of WNT5B in the breast cancer cell lines was lower than that in MCF10A cells. The WNT5B expression in the MCF-7 cells in experimental group was significantly higher than that in vector group (P<0.05). However, the cell viability in experimental group decreased significantly as compared with vector group (P<0.05). The number of the cells in S-phase obviously increased, while the percentage of the cells in G1-phase and G2/M-phase decreased compared with vector group. The number of apoptotic cells in WNT5B group was significantly higher than that in vector group.CONCLUSION: The expression of WNT5B is decreased in breast cancer cells. WNT5B over-expression significantly inhibits the cell growth and promotes the cell apoptosis in breast cancer MCF7 cells.  相似文献   

17.
AIM: To investigate the effect of vorinostat(suberoylanilide hydroxamic acid, SAHA), a histone deacetylase inhibitor, on seizure-induced brain damage in developing rats and its mechanism. METHODS: Male Sprague-Dawley rats(n=32) were randomly divided into control group, pentylenetetrazole(PTZ) group, PTZ+10 mg/kg SAHA group and PTZ+50 mg/kg SAHA group. Intraperitoneal injection of PTZ was used to induce rat seizure. SAHA was injected intraperitoneally 2 h before PTZ injection. The rats in different seizure stages were counted and mean seizure score was analyzed at 30~60 min after PTZ injection. Hippocampal tissues were sampled at 24 h after seizures. The expression of TLR4, MYD88, NF-κB P65 and IL-1β at mRNA and protein levels was detected by RT-qPCR and Western blot, respectively. The pathological changes of the brain tissues were observed by HE staining. The apoptotic neurons were observed by TUNEL staining. RESULTS: The mRNA and protein levels of TLR4, MYD88, NF-κB P65 and IL-1β, the apoptosis of neurons, the inflammation reaction and mean seizure score significantly increased after PTZ treatment(P<0.05), and these effects were attenuated by treatment with SAHA. Compared with PTZ+10 mg/kg SAHA group, PTZ+50 mg/kg SAHA group showed more significant protective effect against seizure-induced brain damage. CONCLUSION: Histone deacetylase inhibitor SAHA suppresses seizure-induced TLR4/MYD88 signaling and reduces apoptosis of neurons, suggesting a protective effect against brain damage associated with seizure in developing rats.  相似文献   

18.
AIM: To investigate the synergistic effect of decitabine (DCA) and valproic acid (VPA) on apoptosis and cell cycle arrest at G0/G1 phase in gastric cancer MGC-803 cells. METHODS: Gastric cancer MGC-803 cells were used in the study and divided into the following groups according to the treatment with different drugs for 72 h: DCA 1.5 μmol/L,DCA 3.0 μmol/L, VPA 1.5 mmol/L, DCA 1.5 μmol/L+VPA 1.5 mmol/L and DCA 3.0 μmol/L+VPA 1.5 mmol/L. The early and late apoptotic rates were detected by annexin V and PI staining. The cell cycle was also determined by flow cytometry. The relative nm23-H1 mRNA expression level was measured by real-time quantitative PCR. RESULTS: The apoptotic rates in VPA 1.5 mmol/L+DCA 1.5 μmol/L group (early: 33.58%±3.88%; late: 31.52%±4.20%) and VPA 1.5 mmol/L+DCA 3.0 μmol/L group (early: 42.61%±4.23%; late: 38.01%±3.86%), the percentages of the cells in G0/G1 phase in VPA 1.5 mmol/L+DCA 1.5 μmol/L group (61.55%±2.38%) and VPA 1.5 mmol/L+DCA 3.0 μmol/L group (66.75%±2.48%), and the relative nm23-H1 mRNA expression levels in VPA 1.5 mmol/L +DCA 1.5 μmol/L group (1.84±0.46) and VPA 1.5 mmol/L+DCA 3.0 μmol/L group (3.02±0.36) were all significantly higher than those in the corresponding concentrations of single drug treatment groups (P<0.01). CONCLUSION: Synergistic effect of VPA and DCA on apoptosis and cell cycle arrest in gastric cancer MGC-803 cells is possibly via inactivation of nm23-H1 gene expression.  相似文献   

19.
AIM: To explore the effects of decorin on procollagen type I (PcI), mRNA expression,collagen type I synthesis and proliferation of synovial type B cells of stiff knee joint synovial membrane. METHODS: Type B cells of synovial membrane were isolated from the stiff knee joint synovial membrane and cultured in vitro. The cells were treated with decorin at concentrations of 0.1 mg/L, 5 mg/L and 10 mg/L. After cultured for 24 h, 48 h and 72 h, the cell proli-feration rates were measured by MTT colorimetric determination. Cell cycle distribution and apoptosis were analyzed by flow cytometry. The mRNA level of Pc I was detected by RT-PCR, while collagen type I was measured by Western blot. RESULTS: The proliferation of synovial type B cells was significantly inhibited, the percentage of synovial type B cells at G1 phase was significantly increased by 5 mg/L and 10 mg/L decorin (P<0.05), and PcⅠmRNA expression and collagen type I synthesis were significantly decreased. The cells with late apoptosis were not found in control group and experimental groups. CONCLUSION: Recombinant human decorin inhibits synovial type B cell proliferation and decreases PcⅠmRNA expression and collagen type I synthesis in synovial type B cells of stiff knee joint synovial membrane in vitro, suggesting that decorin potentially contributes to the therapy of human knee stiffness.  相似文献   

20.
AIM: To explore the mechanism of microRNA-301a-3p (miR-301a-3p) regulating connexin 43 (Cx43) expression in the rat astrocytes. METHODS: miR-301a-3p agomir, miR-301a-3p antagomir and microRNA negative control (miR-NC) were synthesized and transfected into the astrocytes. The protein expression of Cx43 was determined by Western blot. The recombinant vectors wt-pEZX-MT05-Cx43 and mut-pEZX-MT05-Cx43 were constructed. The target gene of miR-301a-3p was validated by dual-luciferase reporter assay. The expressional vector pcDNA3.1-Cx43 was constructed and the biological activity regulation of miR-301a-3p on astrocytes was analyzed by restore experiment. RESULTS: The results of Western blot showed that the Cx43 expression was inhibited by miR-301a-3p agomir transfection (P<0.05). The results of dual-luciferase reporter assay showed that miR-301a-3p bound to 3′-UTR of Cx43, thus regulating expression of Cx43 negatively. Transfection with recombinant vector pcDNA3.1-Cx43 without 3′-UTR of Cx43 into astrocytes resulted in the restoration of the negative function of miR-301a-3p on Cx43 expression, also induced the apoptosis of astrocytes. CONCLUSION: miR-301a-3p binds to 3′-UTR of Cx43 mRNA in the rat astrocytes, thus regulating expression of Cx43 negatively.  相似文献   

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