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@#Objective To investigate the expression of C-C chemokine ligand 5(CCL5) in head and neck squamous cell carcinoma(HNSCC),and explore the effect of CCL5 on the biological characteristics of laryngeal carcinoma cells.Methods Gene Expression Profiling Interactive Analysis(GEPIA) database was used to investigate the expression of CCL5 in HNSCC.The laryngeal carcinoma cells TU177 were transfected with siRNA(siRNA group),and the control(NC) group was set up.The cell proliferation,migration,cycle and apoptosis of each group were detected by CCK8 assay,cell scratch test and flow cytometry respectively.RT-PCR and Western blot were used to detect the knock-down efficiency of CCL5 and the mRNA transcription and protein expression of multidrug resistance protein 2(MRP2) and bcl-2-associated x protein(Bax).Results The expression of CCL5 in HNSCC was higher than that in normal tissues(P <0.05).Compared with NC group,siRNA showed higher knock-down efficiency(t=12.898 and 22.656 respectively,each P <0.01);siRNA interference with CCL5 inhibited the proliferation and migration of laryngeal carcinoma cells,and promoted the late apoptosis of laryngeal carcinoma cells and the expression of apoptosis protein Bax(t=2.600～11.667,each P <0.05).Conclusion CCL5 was highly expressed in HNSCC,while siRNA interference with CCL5 inhibited the proliferation,migration and promoted apoptosis of laryngeal carcinoma cells TU177 by up-regulating the expression of Bax,which laid a foundation of the possibility of CCL5 as a new target for the treatment of laryngeal carcinoma.
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@#Objective:To explore the mechanism of EYA1 (eyes absent 1) inhibiting the malignant progression of gastric cancer SGC7901 cells through regulating PTEN/PI3K/AKT signaling pathway. Methods: Twenty-nine pairs of gastric cancer tissues and para-cancerous tissues collected at the General Surgery center, Southwest Hospital Affiliated to Military Medical University during June 2016 and June 2018 were used in this study. Wb and RT-PCR assays were used to test the mRNA and protein expressions of EYA1 in gastric cancer tissues and the paired para-cancerous tissues; Transfection with plasmid or siRNAs were used to up-regulate or down-regulate EYA1 or PTEN expression in gastric cancer SGC-7901 cells; MTT, Flow Cytometry, Wound Healing and Transwell assays were carried out to detect cell proliferation, apoptosis, metastasis and invasion abilities, respectively. Results: EYA1 expression was decreased in gastric cancer tissues as compared with the para-cancerous tissues at both mRNA and protein levels (P<0.01); EYA1 over-expression significantly enhanced the proliferation, metastasis and invasion of SGC-7901 cells (all P<0.05), and inhibited cell apoptosis (P<0.05); moreover, its over-expressionsignificantly increased the expression of PTEN, and inhibited the activation of PI3K/AKT pathway (all P< 0.05 or P<0.01). However, the above effects mediated by EYA1 up-regulation were significantly impaired after the knockout of PTEN (all P<0.05 or P<0.01). Conclusion: EYA1 can inhibit the malignant progression of gastric cancer SGC-7901 cells through promoting the expression of PTEN and activating PI3K/AKT pathway.
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Aim: To investigate the effect of salvianolic acid B (Sal B) on liver fibrotic cells in vivo and in vitro from die perspective of apoptosis, as well as the effect on cleaved caspase-9. Methods: Diethylnitrosamine (DEN) was used to induce liver fibrosis in mice for 12 weeks. The padiological changes were detected by HE staining, and the fibrotic lesion area was determined. The cell apoptosis in the fibrotic area was observed by Hoechst 33258 fluorescence staining. The expression of cleaved caspase-9 in fibrotic tissues was detected by Western blot. The apoptotic rate of each group was detected by double standard method AnnexinV-FITC/PI, and the expression of apoptotic protein cleaved caspase-9 in HSC-T6 was detected by Western blot. Results: HE staining suggested that 12 weeks were the period of liver fibrosis in mice. No pseudoplobular structure was formed in group with low and high dose of Sal B, and the degree of fibrosis was lower than that in model group. In the fibrotic lesion area, the fluorescence staining of Hoechst 33258 showed that apoptotic cells significantly increased in group with low Sal B and high dose compared with model group. The results of the AnnexinV-FITC/PI method showed that TGF-β1 inhibited the apoptosis of HSC-T6, and Sal B promoted the apoptosis of HSC-T6 after TGF-β1 intervention and showed a concentration dependence (P <0. 01). Western blot results showed that in fibrotic liver tissues, Sal B increased the expression of cleaved caspase-9 in HSC-T6 cells compared with model group. Compared with TGF-β1 group, Sal B increased cleaved caspase-9 protein expression (P < 0. 01). Conclusions: Sal B can significantly promote apoptosis of liver fibrotic cells in vitro and in vivo, and its pro-apoptosis mechanism may be related to the up-regulation of cleaved caspase-9.
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AIM:To investigate the effect and mechanism of osthole on increasing the cytotoxicity of doxorubi -cin(DOX)to prostate cancer cells.METHODS:MTT assay was performed to evaluate the viability of LNCaP cells trea-ted with osthole and DOX.The protein expression of silent information regulator 1(SIRT1),p53,acetylated p53 and Pu-ma,as well as release of cytochrome C and activation of caspase-9 and caspase-3 in the LNCaP cells treated with osthole and DOX were determined by Western blot.The apoptosis of the LNCaP cells treated with osthole and DOX was analyzed by flow cytometry.RESULTS:Osthole significantly increased the cytotoxicity of DOX against p 53-wildtype prostate cancer cell line LNCaP.Osthole significantly inhibited the expression of SIRT 1 in the LNCaP cells.Transfection with SIRT1 plas-mid decreased the cytotoxicity of osthole and DOX co-treatment against LNCaP cells.Combination with osthole and DOX significantly induced the over-expression and acetylation of p53.Transfection with p53 siRNA significantly decreased the synergistic effect of osthole on cytotoxicity of DOX-treated LNCaP cells.Combination with osthole and DOX significantly in-duced the release of cytochrome C into the cytoplasm from mitochondria,followed by activation of caspase-9 and its down-stream molecule caspase-3,thus leading to cell apoptosis in the LNCaP cells.CONCLUSION:Osthole promotes the p53-dependent apoptosis in DOX-treated prostate cancer LNCaP cells by down-regulating the expression of SIRT1.
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Objective To investigate the effect of different concentrations of vinorelbine on apoptosis ,telomerase ac-tivity and expression of human telomerase-reverse transcriptase gene ( hTERT ) in human epithelial ovarian cancer cells SKOV3.Methods Ovarian cancer cells SKOV 3 were treated with vinorelbine under different concentrations . The cell proliferation was measured by cell counting kit-8 ( CCK-8) assay , and the cell apoptosis was detected by flow cytometry.The telomerase activity of SKOV3 cells was determined by TRAP-PAGE-silver staining;The mRNA expression of hTERT was performed by RT-PCR assay .Results Vinorelbine could significantly inhibited the prolifer-ation of SKOV3 cells,induce cell apoptosis (P<0.01),reduce the telomerase activity and expression of hTERT mRNA (P<0.01), in dependent of a concentration-time manner.Conclusions The detection of telomerase activity and the mRNA expression of hTERT might be vital for predicting the prognosis of patients with epithelial ovarian cancer .
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<p><b>OBJECTIVE</b>To investigate the underlying mechanisms of cyclovirobuxinum D (Cvb-D) on alleviating cardiac hypertrophy in rats.</p><p><b>METHODS</b>Sprague-Dawley rats were randomly divided into 5 groups: control group; levothyroxine-induced cardiac hypertrophy group (model); levothyroxine-induced cardiac hypertrophy + Cvb-D group (Cvb-D); levothyroxine-induced cardiac hypertrophy + captopril group (captopril); levothyroxine-induced cardiac hypertrophy + SB203580 group (SB203580), n=10 for each group. Rats were daily administered the respective drugs continuously for14 days by gastric gavage. A rat model of cardiac hypertrophy was established by intraperitoneal injection of levothyroxine to investigate whether Cvb-D protects against cardiac hypertrophy by inhibiting the p38 mitogen-activated protein kinase (MAPK) signaling pathway and preventing apoptosis of cardiac cells.</p><p><b>RESULTS</b>Treatment with Cvb-D significantly deceased left ventricle hypertrophy, improved the histopathology, hemodynamic conditions, and cardiac function in rats with cardiac hypertrophy. Compared with the normal control group, in rats with cardiac hypertrophy, expression of bax in the heart and phospho-p38 MAPK protein levels were significantly up-regulated (P<0.01 or 0.05), whereas the bcl-2 protein level was down-regulated (P<0.01). In contrast, Cvb-D treatment reversed the changes in bax and phospho-p38 MAPK protein levels but increased the bcl-2 protein level (P<0.01 or 0.05), and these effects were similar to those of captopril and SB203580 (a specific p38MAPK inhibitor) treatment. Furthermore, both Cvb-D, captopril and SB203580 reduced mRNA expression of p38α, p38β, c-fos, and c-jun mRNA, and Cvb-D had a stronger effect (P<0.01).</p><p><b>CONCLUSION</b>These results demonstrate that Cvb-D protects against cardiac hypertrophy, which is possibly mediated by prevention of cardiac cell apoptosis and inhibition of the p38MAPK signaling pathway.</p>
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Objective To investigate the effects of Delta-opioid receptor activator DADLE on the expression of the p38 MAPK and the neuronal apoptosis after global cerebral ischemia-reperfusion and explore the neuroprotective mechanisms of DADLE.Methods Global cerebral ischmemia-reperfusion models in rats were induced by bilateral common carotid artery occlusion combined with hypotension.Rats were randomly divided into sham group (n=10),ischemia-reperfusion group(n=10) and three treatment groups with different doses of DADLE (2 mg group,3 mg group,5 mg group,n=10).TUNEL method and Western blot analysis were used to measure apoptotic neurons and expression levels of p38 MAPK phosphorylation,repectively.Results DADLE treatment significantly reduced neuron apoptosis (P＜0.05).The expression levels of p38 MAPK were increased in ischemia-reperfusion group than in sham group (P＜ 0.05).In DADLE treated groups,the expression levels of p38 MAPK were dose-dependently decreased compared with the ischemia-reperfusion group.Conclusion Delta-opioid receptor activator DADLE can be neuroprotective against global I/R injury.Attenuation of apoptosis and p38 MAPK signal pathway might be involved in the neuroprotective mechanism of DADLE.
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Background The light damage model of retinal pigment epithelium (RPE) cells is a research direction of retinal degeneration diseases,and RPE cell apoptosis induced by light damage and inflammation is an important pathologic basis of light-induced RPE cell damage.However,whether endoplasmic reticulum stress (ERS) paticipates in light-induced RPE cell damage is rarely reported.Objective This study was to explore the effects of ERS on light-induced RPE cell damage.Methods Human RPE cell line (ARPE-19) was cuhured,and light damage models were created by irradiating the cells for 3-,6-,12-and 24-hours with white fluorescent lamp with the intensity of (2 000±500)lx for the selection of optimal irradiating time,and the cells in the normal control group were cultured in the dark environment.The cells were divided into normal control group,light exposure group and 4-phenylb utyric acid (4-PBA) pretreated +light exposure group.The cells from 4-PBA pretreated +light exposure group were cultued firstly with 4-PBA for 30 minutes and followed by light exposure for 12 hours.The apoptisis rate of the cells and intracellular reactive oxygen species (ROS) content were detected by flow cytometry;the concentrations of interleukin 1β (IL-1β) and tumor necrosis factor-α (TNF-α) in the cell supernatant were assyed by ELISA.The relative expressing levels of activating transcription factor 6 (ATF-6),C/enhancer binding protein homologous protein (CHOP) and caspase-12 mRNA and protein in the cells were detected by real-time quantitative PCR and Western blot,respectively.Results The cultured cells showed a long spindle shape,the border was not clear,the cytoplasm was degranulation,and the cell fragments increased.Flow cytometry showed that compared with the normal control group,the ROS content in the cells and the apoptosis rate were evidently increased with the lapse of light exposure time (F=763.00,119.30,both at P＜0.01).ELISA results showed that the concentrations of IL-1β and TNF-α in the cell supernatant were significantly higher in the light exposure 6-hour group than those in the normal control group with the peak value in the light exposure 12-hour group.Compared with the normal control group,the relative expression levels of ATF-6,CHOP and caspase-12 mRNA and protein in the cells were elevated in the light exposure group and peaked in the light exposure 12-hour group.In addition,the relative expression levels of ATF-6 mRNA,CHOP mRNA and caspase-12 mRNA in the cells were significantly reduced in 4-PBA pretreated+light exposure group compared with the light exposure group (F=281.69,473.88,308.45,all at P＜0.01),and their proteins were also significantly reduced (F =47.86,57.93,106.59,all at P ＜ 0.01).The apoptosis rate,concentrations of IL-1β and TNF-α in the cell supernatant were significantly reduced in 4-PBA pretreated+light exposure group compared with the light exposure group (F =88.64,245.47,101.01,all at P＜0.01).Conclusions The light exposure at (2 000 ± 500)lx induces intracellular ROS accumulation and activates the ERS response,which results in apoptosis and inflammatory process of human RPE cells.4-PBA,a inhibitor of ERS,can suppress light-induced ERS response and therefore reduces the apoptosis rate and inhibits inflammatory process.
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Objective To investigate the effects of microRNA (miR)106a on the proliferation, apoptosis, migration and invasion of thyroid cancer cells in vitro.Methods 8505C and CGTH-W3 cell lines were used in the study.Overexpression and inhibition of miR106a were achieved by transfection of lentiviral vectors.The changes of gene expression were detected by quantitative real-time PCR (qRT-PCR) and Western blot analysis.Cell viability and apoptosis were evaluated by MTT assay and flow cytometry analysis, respectively.The caspase-9 activities in parental CGTH-W3 and 8505C cells and transfected sublines were measured.Wound healing and Transwell invasion assays were performed to determine cell migration and invasion.Two-sample t test and one-way analysis of variance were used to analyze the data.Results The level of miR106a in 8505C was up-regulated when compared to that in CGTH-W3 cells (t=10.28, P<0.01).Scrambled control and miR106a(-) were also successfully transfected into cells.Inhibition of miR106a suppressed cell viability, migration and invasion while promoted apoptosis and caspase-9 activity of 8505C cells, with significant differences among 8505C, 8505C-control, 8505C-miR106a(-) cells (F=147.0, 19.2, 100.3, 537.8, 804.3;all P<0.01).Overexpression of miR106a promoted cell viability, migration and invasion while inhibited apoptosis and caspase-9 activity of CGTH-W3 cells, with significant differences among CGTH-W3, CGTH-W3-control, CGTH-W3-miR106a(+) cells(F=9.2, 13.3, 622.8, 12.3, 19.6, all P<0.01).In addition, miR106a may up-regulate the expression of MEKK2 and p-ERK1/2.Conclusion Acting as an onco-miR, miR106a might promote the proliferation, migration and invasion of thyroid cancer cells and inhibit their apoptosis in vitro.
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Objective To investigate the effects of Delta-opioid receptor activator DADLE on the expression of the p38 MAPK and the neuronal apoptosis after global cerebral ischemia-reperfusion and explore the neuroprotective mechanisms of DADLE.Methods Global cerebral ischmemia-reperfusion models in rats were induced by bilateral common carotid artery occlusion combined with hypotension.Rats were randomly divided into sham group (n=10),ischemia-reperfusion group(n=10) and three treatment groups with different doses of DADLE (2 mg group,3 mg group,5 mg group,n=10).TUNEL method and Western blot analysis were used to measure apoptotic neurons and expression levels of p38 MAPK phosphorylation,repectively.Results DADLE treatment significantly reduced neuron apoptosis (P＜0.05).The expression levels of p38 MAPK were increased in ischemia-reperfusion group than in sham group (P＜ 0.05).In DADLE treated groups,the expression levels of p38 MAPK were dose-dependently decreased compared with the ischemia-reperfusion group.Conclusion Delta-opioid receptor activator DADLE can be neuroprotective against global I/R injury.Attenuation of apoptosis and p38 MAPK signal pathway might be involved in the neuroprotective mechanism of DADLE.
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AIM:To investigate the role of microRNA-101-3p (miRNA-101-3p) on the proliferation, apopto-sis and invasion of gastric cancer cells and the possible regulatory mechanisms .METHODS: The expression of miRNA-101-3p in two kinds of gastric cancer cells and a gastric mucosal cell line was detected by real -time PCR.The miRNA-101-3p was overexpressed by Lipofectamine 2000 transfection with miRNA-101-3p mimics.The effects of miRNA-101-3p on cell cycle distribution and apoptosis were analyzed by flow cytometry .The effects of miRNA-101-3p on cell proliferation and migration abilities were detected by CCK-8 assay, trypan blue exclusion test and Transwell assay .The protein expression of enhancer of zeste homolog 2 ( EZH2) was determined by Western blot .RESULTS: The expression of miRNA-101-3p in gastric cancer cells was lower than that in gastric mucosal cells (P<0.05).The gastric cancer cell MGC-803 had the low-est expression level of miRNA-101-3p.The result of flow cytometry showed that the population of S phase was reduced , and the population of G0/G1 phase and the early stage apoptotic rate were increased after the expression of miRNA-101-3p was overexpressed (P<0.05).The results of CCK-8 assay, trypan blue exclusion test and Transwell assay showed that overex-pression of miRNA-101-3p significantly reduced the proliferation and migration abilities of gastric cancer cells (P<0.05). Overexpression of miRNA-101-3p decreased the protein level of EZH2 (P<0.05).CONCLUSION:miRNA-101-3p may suppresses the gastric cancer cell proliferation and migration , and promotes the gastric cancer cell apotosis by down-regula-tion of EZH2.
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Objective To evaluate the role of endoplasmic reticulum stress in ketamine-induced apoptosis in rat neurons.Methods Rat adrenal pheochromocytoma cell line (PC12 cells) was seeded in the culture dishes 100 mm in diameter (10 ml/dish) or in 6-well plates (2 ml/well) at a density of 5 × 105 cells/ml.PC12 cells were divided into 4 groups (n =6 each) using a random number table:control group (group C);ketamine group (group K);endoplasmic reticulum stress inhibitor salubrinal group (group S);ketamine + salubrinal group (group K+S).In group C,the cells were cultured in the plain culture medium.In group K,1.5 mmol/L ketamine was added.In group S,30 mmol/L salubrinal was added.In group K + S,1.5 mmol/L ketamine and 30 mmol/L salubrinal were added.At 24 h of incubation,the cell morphology was observed under light microscope,the expression of Bip and caspase-12 in PC12 cells was detected by Western blot,and the cell apoptosis was measured by flow cytometry.The apoptosis rate was calculated.Results Compared with group C,the expression of Bip and caspase-12 was significantly upregulated,and the apoptosis rate was increased in K and K + S groups (P ＜ 0.05),and no significant change was found in the parameters mentioned above in group S (P＞ 0.05).Compared with group K,the expression of Bip and caspase-12 was significantly down-regulated,and the apoptosis rate was decreased in group K+S (P＜0.05).The degree of damage to PC12 cells was more serious in group K than in group C..The degree of damage to PC12 cells in group K+S was significantly mnilder than that in group K and more serious than that in group C.Conclusion The mechanism by which ketamine induces neuronal apoptosis is related to the enhancement of endoplasmic reticulum stress in rats.
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Objective To investigate the effect of two different odontoblast inducer on the proliferation and apoptosis of rat adiposed-derived stem cells.Methods Adiposed-derived stem cells were collected by enzyme digestion from inguinal fat pads of 4 days post natal mice.Immunocytochemistry was performed to identify the cells.MTT and flow cytometry were tested the prolifera-tion and apotosis of adiposed-derived stem cells by co-cultured with tooth germ cell conditioned medium(TGC-CM)or dentin non-collagenous protein medium (DNCPM).Results Cells displayed a fibroblast-like appearance and positively expressed CD44 and CD105 when cufured to the secend yeneration.After 3 day the cells polarity changed by co-cultured.Count of cells were no obvious change by TGC-CM co-cultured,while that ruduced significantly by DNCPM co-cultured.It confirmed that the proliferation rate of ADSCs in TGC-CM group and control group is higher than DNCPM group(P 0.05).Conclusion TGC-CM may have more advantage as inducer in rat adiposed-derived stem cells differentiate into dentin like cells than DNCPM.
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Objective To further approach the effect of miR-33 to melanoma cells line B16F10 proliferation and apoptosis.Methods Constructing targeted miR-33 over-expression mimics and inhibitor,the same B16F10 cells were divided into five groups,control group,miR-33 mimics group,mimics control group,miR-33 inhibitor group,inhibitor control group,then gene transfer technology was used to transfer corresponding gene into B16F10 cells.The effect of miR-33 on B16F10 cell' s proliferation and apoptosis were analysed.Results The relative miR-33 gene expression of miR-33 mimics group (1773.3±245.83) was higher than that of control group,which had statistical significance (P ＜ 0.01).The gene expression of miR-33 inhibitor group (0.6973±0.1958) was lower than those of control group and inhibitor control group.The cell growth rate of miR-33 mimics group was lower than those of control group and the trend after transfection 48 h (1.1875±0.0502) and 72 h (1.7500±0.0933) was significant (P ＜ 0.05).The cell apoptotic ratio of miR-33 mimics group [(1.8050±0.2050) ％] was higher than that of control group [(1.13000±0.1414) ％] (P ＜ 0.05).Compared with control group the cell proportion ofG1 period in miR-33 mimics group [(62.7000±1.7321) ％]increased,the cell proportion of S period [(23.4000±2.5044) ％] decreased,both of them had statistical significance (both P ＜ 0.05).Conclusion miR-33 over-expression can restrain the proliferation of B16F10 cells line,promote B16F10 cells' apoptosis.
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Objective To investigate the effect of intermedin (IMD) on tubular cells apoptosis induced by renal ischemia/reperfusion (I/R) injury and its associated mechanism.Methods A total of twenty-four male Wistar rats were randomly divided into four groups (control group, I/R group, empty plasmid group and IMD group). One week after the removal of right kidney, ultrasound plasmid was used to transfect empty or IMD plasmid into the left kidney. Renal I/R model was made by clasping the left renal artery for 45 minutes. Tubular cell apoptosis was determined by TUNEL. Expression of Bax, Bcl-2, Fas was detected by semi-quantitative RT-PCR.Activity of caspase-8 and caspase-9 was evaluated with commercially available kits respectively.Protein level of caspase-3 was measured by Western blotting analysis. Results Compared with control group, apoptosis of tubular epithelial cells, expression of Bax and Fas, activities of caspase-8 and caspase-9, as well as protein level of caspase-3 were all significantly increased in I/R group (all P＜0.05). IMD pre-treatment significantly inhibited all these effects (all P＜0.05). There were no differences of above parameters between empty plasmid group and I/R group. Conclusion IMD pre-treatment protects against renal I/R injury by inhibion of tubular epithelial cell apoptosis.
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Objective To investigate the influence of iodine excess on expression of TRAIl/TRAIL-sR1 in NOD and Balb/c mice and to study the effect of TRAIl/TRAIL-sR1 on the pathogenesis of experimental autoimmune thyroiditis(EAT). Methods Both Balb/c and NOD mice were divided randomly into control and iodine excess group by feeding with water containing no NaI or 0.05% Nal. The mice were sacrificed after 8 weeks. TRAIL and TRAIL-sR1 mRNA levels were detected by RT-PCR. The function, morphology and apoptosis of thyroids were also observed by ELISA and Tunnel stain. Results Treated by HI, enlarged follicles and flattened epithelium by accumulation of colloid were found in thyroids of both NOD and Balb/c mice. But significant lymphoid cell infiltration and local fibrosis were only found in thyroids of NOD HI group. The relative weight of thyroids of NOD mice in HI group[(104.8±14.5)mg/kg]was heavier than that of control group [(71.8±20.4)mg/kg]. The level of TT4 declined in HI group[(30.77±3.59)mmol/L]compared with control group[(36.43±2.66)mmol/L], meanwhile, the level of TSH was higher in HI group[(6.98±0.66)μg/L]than that in control group [(5.55±0.56)μg/L]. The difference being statistically significant(t=7.773,-9.526,-4.458, all P < 0.05). The relative weight of thyroids of Balb/c mice of HI group[(155.8±20.8)mg/kg]also heavier than that of control group [(105.1±22.0) mg/kg]. The level of TT4 droped in HI group [(19.75±3.32) mmoL/L]was higher than that in control group[(23.46±6.21)mmoL/L], the level of TSH in HI group[(4.14±1.71)μg/L]was higher than that in control group[(3.55±1.41)μg/L], the difference being statistically significant(t=7.554,-7.239,3.140, all P< 0.05). A great deal of apoptotie ceils observed in NOD (3.97±0.91) and Balb/c mice (1.05±0.45) by Tunnel stain were greater than control groups (0.21±0.15, 0.10±0.03), the difference being statistically significant in beth of the two species(t=-7.167,-17.772, both P < 0.05). The apoptosis index of thyroid follicular epithelium in NOD was obviously higher than Balb/c(t=-7.625, P<0.05). The level of TRAIL mRNA did not remarkably change in Balb/c between control group(0.000 59±0.000 39) and HI group(0.001 24±0.000 46, t=-1.940, P>0.05), but it increased apparently in NOD mice HI group(0.018 88±0.005 77) than that of control group(0.009 61± 0.00591, t=-2.71, P<0.05). The level of the expression of TRAIL-sR1 mRNA increased in HI groups of NOD (0.000 53±0.000 15) and Balb/c mice(0.000 42±0.000 09) than that in control groups of NOD(0.000 28± 0.000 05) and Balb/c mice (0.000 17±0.000 06) and the differences were statistically significant between the two species(t=3.050,3.990, all P<0.05). The differences of the expression of TRAIL and TRAIL-sR1 mRNA between the two species were significant(t=-3.37,-4.76, all P<0.05). Conclusions Iodine excess induces colloid goiter in beth species of mice and thyroiditis in NOD mice. The increase of TRAIL and TRAIL-sR1 influenced by iodine excess is one of the molecular bases of follicular epithelium apoptosis and inflammation in thyroids. Genetic factor is a key factor in the pathogenesis of thyroiditis.
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Objective To investigate the role of serum tissue polypeptide-specific antigen (TPS)detection in the evaluation of hepatocyte apoptosis in patients with alcoholic hepatitis and to further evaluate its application in the differential diagnosis of alcoholic hepatitis and fatty liver. Methods The automatic biochemistry analyzer was employed to detect common markers for liver injury (AST,ALT, GGT and bilirubin) ; electrochemiluminescence assay was performed to measure serum TPS level; immunohistochemistry staining was done to evaluate M30 immunoreactivity in hepatic tissue biopsy specimens and the apoptotic score was acquired. Then the correlations were analyzed between various indices and the apoptotic scores. Results The serum TPS level was positively correlative to the hepatocyte apoptotic score in alcoholic hepatitis group; and their correlation coefficient was the largest among various liver functional indices and the apoptotic scores. The correlation coefficient between TPS level and apoptotic score was significantly higher in alcoholic hepatitis group than that in fatty liver group (P＜0.05). Conclusion Serum TPS may be used as a good hepatocyte apoptosis marker for alcoholic hepatitis, which contributes to the differential diagnosis of alcoholic hepatitis and fatty liver.
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Objective To investigate the effects of exogenous IGF- Ⅰon apoptosis, bax,bcl-2 and caspase-3 gene mRNA transcription in intestinal mucosal epithelial cell of SAP rats. Method Seventy-two male Wistar rats were randomly divided into sham operation group (SO),SAP group(SAP) and IGF-Ⅰ treatment (SAP + IGF-Ⅰ) group.Every group was randomly divided into 3 time units (6,12,24 h),8 rats as each time unit. SAP was induced in the rats by injecting adversely 5.0 % sodium taurocholate into biliary-pancreafic duct. The SO rats were infused with NS by the same way. The rats in IGF-Ⅰgroup were injected with IGF-Ⅰ by subcutano at half an hour before operation and three hours after operation,respectively. Animals in each group were killed separately at 6,12 and 24 hours after operation.The apoptosis in mucesal cells of small intestine was detected by TUNEL, and histo pathological changes of the small intestine was observed. The expressions of bax and bcl-2 and caspase-3 mR-NA gene in small intestine were measured by reverse transcription polymerase chain reaction(RT-PCR). Results Compared with the SAP group,the serum amylase were lower in IGF-Ⅰ group,and there existed significant at 12 h and24 h (P < 0.05).The pathological score of small intestinal was significantly reduced in IGF-Ⅰ group com-pared with SAP group,and there were statistical differences at 12 h and 24 h.ln IGFo-Ⅰ group,the apoptosis index of intestinal epithelial decreased significantly compared with SAP group[6 h: (13.88±1.73) vs. (19.00±2.78) ;12h:(10.13±1.55) vs. (17.63±.60);24 h:(9.50±1.07) vs. (17.25±2.76)] (P <0.05); the histopathdogical changes were more improved compared wit SAP group under the electronic microscope; the expres-sion of bax mRNA [6 h:(1.35±0.18) vs.(0.85±0.12);12 h:(1.21±0.21) vs. (0.86±0.24);24 h:(1.14±0.24) vs. (0.95±0.22)] and caspese-3 mRNA[6 h:(0.78±0.01) vs. (0.55±0.04);12 h:(0.79±0.04) vs. (0.57±0.05) ;24 h: (0.81±0.06) vs. (0.55±0.01) (P < 0.01)] were higher in three time units in SAP group than those in SO group (P < 0.01) ,and in IGF-1 group it was weakened significantly compared with the SAP group at each time point (P <0.05). bcl-2 mRNA expression was weak and have no difference between the SO group and SAP group (P > 0.05), but increased signifycantly in the IGF-± group at each time point [6 h:(0.65±0.07) vs. (0.54±0.04) vs. (0.57±0.06);12 h:(0.69±0.04) vs. (0.56±0.05) vs. (0.53±0.05);24h:(0.72±0.05) vs. (0.54±0.07) vs. (0.58±0.08)] (P <0.05). Conclusions Exogenous IGF-Ⅰ could rivalry SAP induced apoptosis to mucosal cells of small intestine , then could alleviate SAP induced injury to intestinal mucosal, It may be associated with the mechanisms that IGF-Ⅰ could improve the expression of bcl-2 mRNA and inhibit the expression of bax,caspase-3 mRNA.
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Objective To investigate the effects of propofol on neuronal apoptosis in anterior horn of spinal cord in rabbits with spinal cord ischemia-reperfusion (IR) injury. Methods Sixty New Zealand white rabbits aged 4-6 months weighing 2.0-2.5 kg were randomized to receive normal saline (group C), 10% intralipid (group F) and propofol 30 mg/kg (group P1 ), 40 mg/kg (group P2), 50 mg/kg (group P3) and60 mg/kg (group P4 ). 10% intralipid was added to propofol solution to make the fluid infused equal in volume between the 6 groups ( n = 10 each). Spinal cord ischemia was induced by occlusion of abdominal aorta distal to the left renal arteries combined with simultaneous occlusion of bilateral common iliac arteries for 30 min. A catheter was inserted into abdominal aorta close to the site of occlusion via left femoral artery. Normal saline, 10% intralipid or different doses of propofol was infused through the catheter as soon as aorta was clamped at the rate of 12 ml·kg-1·h-1 for 30 min. The aorta and bilateral iliac arteries were then declamped. The L4-6 of spinal cord was removed at 48 h of reperfusion for microscopic examination and the total number of normal motor neurons in the anterior horn of spinal cord was counted. The total number of neurons and apoptosis neurons in the anterior horn of spinal cord was counted by TUNEL and the apoptosis index of neurons was calculated. The expression of caspase-3 in the anterior horn of spinal cord was determined by immunohistochemical technique. Results The number of normal motor neurons was significantly higher, and the apoptosis index and expression of caspase-3 were significantly lower in group P1-4 than in group C and F ( P < 0.05). Compared with group P1, the number of normal motor neurons was significantly increased and the apoptosis index was significantly decreased in group P2-4 and the expression of caspase-3 was down-regulated in group P3 and P4 ( P < 0.05). Compared with group P2, the number of normal motor neurons was significantly increased in group P3 while decreased in group P4, and the apoptosis index was significantly decreased and the expression of caspase-3 was down-regulated in group P3 and P4 ( P < 0.05). Compared with group P3, the number of normal motor neurons was significantly decreased and the apoptosis index was significantly increased and the expression of easpnse-3 was up-regulated in group P4 ( P < 0.05) . Conclusion Propofol 30-60 mg/kg infused through aorta during occlusion can inhibit the neuronal apoptosis and attenuate IR injury to spinal cord dose-dependently in rabbits. The underlying mechanism may be related to the down-regulation of caspase-3 expression.
RESUMEN
Objective To detect the expression of Livin in gastric cancer tissues and cells, and investigate its relation to tumorigenesis and progression of gastric cancer cells. Methods (1) The expression of Livin in gastric cancer tissues was located by immunocytochemisty. (2) Livin mRNA and protein expression in adjacent tissues of carcinoma, gastric ulcers and gastric cancer tissues were detected by RT-PCR and Western blot. Results The expression of Livin in the nucleus and cytoplasm of gastric cancer cell line SGC-7901 was observed. The positive rate of Livin expression in the gastric cancer tissues was 67% (20/30). There were no expression of Livin-α and Livin-β in the adjacent tissues of carcinoma and the gastric ulcers. The positive rate of Livin-α expression in the gastric cancer tissues was positively correlated with TNM stage of gastric cancer (x2=11.719, P<0.05). The expression rate of Livin in the differentiation and location of gastric cancer, age and gender of patients had no statistical difference (x2=1.607, 0.000, 0.156, 0.419, P>0.05). Conclusion Correlation of high Livin-α expression and pathologic staging of gastric cancer indicates that Livin-α may play an important role in the carcinogenesis and progression of gastric cancer.