Sodium aescinate induces apoptosis of HeLa cells by inhibiting Akt/ERK signaling pathways and increasing death receptor expression

AIM:To study the effects of sodium aescinate on the apoptosis of cervical cancer HeLa cells and its molecular mechanism. METHODS:MTT assay was used to detect the growth and proliferation of HeLa cells. The morphological alteration was observed under inverted microscope. Annexin V-FITC/PI double staining and DAPI nuclear staining were used to determine the apoptosis of HeLa cells induced by sodium aescinate. The apoptosis-related proteins PARP, cleaved caspase-8 and pro-caspase-3, and the proliferation-associated molecules Akt and ERK, as well as TRAIL receptors DR4 and DR5 were detected by Western blotting. RESULTS:Sodium aescinate inhibited the growth of HeLa cells in a concentration-dependent manner. Treatment with sodium aescinate induced the typical morphology of apoptotic cells and increased the apoptotic rate significantly. The cleaved PARP, cleaved caspase-8 and cleaved caspase-9 protein expression was observed. The expression of DR4 and DR5 was up-regulated. Meanwhile, pro-caspase-3 was decreased, and the levels of p-Akt and p-ERK were down-regulated by sodium aescinate in a dose-dependent manner. CONCLUSION:Sodium aescinate inhibits the proliferation and promotes the apoptosis of HeLa cells by increasing death receptor expression and repressing proliferation-associated signaling pathways.     

Effect of Sonic Hedgehog signaling blockade on growth of hepatocarcinoma cells

AIM: To investigate the effect of Sonic Hedgehog (Shh) signaling blockade on the growth of hematocarcinoma cells and underlying mechanisms. METHODS: The expression of Shh signaling molecules in hematocarcinoma cell lines BEL-7402, Huh7 and HepG2 was detected by RT-PCR. The cell viability was detected by MTT assay. The cell cycle and apoptosis were analyzed by flow cytometry. The expression of apoptosis-related proteins was determined by Western blot. RESULTS: Shh signaling molecules were all expressed in BEL-7402, Huh7 and HepG2 cells. The mRNA expression of Patched (Ptch), Gli1 and Gli2 was down-regulated by anti-Shh antibody. Blockade of Shh signaling pathway inhibited the proliferation of hepatocarcinoma cells with increasing cells in G₀/G₁ phase and induced the apoptosis of hepatocarcinoma cells. Treatment with anti-Shh antibody down-regulated the protein expression of pro-caspase-3, pro-caspase-8 and pro-caspase-9, while up-regulated the protein levels of cleaved caspase-3, cleaved caspase-8 and cleaved caspase-9 in BEL-7402 cells. CONCLUSION: Blockade of Shh signaling pathway inhibits the growth of hepatocarcinoma at different levels by cell cycle arrest and inducing apoptosis of hematocarcinoma cells.     

Pseudolaric acid B induces glioblastoma cell line U87 mitotic arrest and apoptosis

AIM: To investigate the effects of pseudolaric acid B on the growth and apoptosis of glioblastoma cell line U87. METHODS: The cell morphological changes were observed under inverted microscope. The cell viability was evaluated by MTS assay. The cell cycle was analyzed by flow cytometry and Western blot. The cell apoptosis was detected by flow cytometry. The changes of apoptosis-related proteins cleaved PARP, caspase-3, procaspase-9 and caspase-8 were determined by Western blot. RESULTS: Pseudolaric acid B inhibited the viability of U87 cells, arrested U87 cells in mitosis. Apoptosis of U87 cells was induced by pseudolaric acid B. The caspase pathway was activated. CONCLUSION: Pseudolaric acid B induces glioblastoma cell line U87 mitotic arrest and apoptosis.     

Ethyl acetate extract of Pleione bulbocodioides (Franch.) Rolfe induces apoptosis of human leukemia K562 and HL-60 cells through intrinsic mitochondrial apoptosis pathway

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.     

Pim-1 kinase inhibitor SMI-4a inhibits proliferation and induces apoptosis in U937 cells

AIM: To study the growth-inhibiting and proapoptotic effects of Pim-1 kinase inhibitor SMI-4a on human acute myeloid leukemia cell line U937.METHODS: The effect of SMI-4a on U937 cell viability was measured by CCK-8 assay. The apoptotic rate was assessed by flow cytometry with Annexin V-PI staining and by fluorescence microscopy with Hoechst 33342 staining. Methylcellulose was used to assess colony formation ability of the cells. The expression of β-catenin in the cell cytosol and nucleus was detected by Western blot, and the expression of apoptosis-related proteins in the U937 cells was also examined. Intracellular distribution of β-catenin was detected by the method of immunofluorescence.RESULTS: SMI-4a inhibited the viability of U937 cells. Annexin V-PI staining showed that SMI-4a induced apoptosis in dose-and time-dependent manners. Hoechst 33342 staining also verified the apoptosis. SMI-4a significantly inhibited the colony formation capacity of the U937 cells. The results of Western blot demonstrated that SMI-4a upregulated the expression of PARP and Bax, downregulated the expression of Bcl-2 and change the distribution of β-catenin in intracellular compartment. Immunofluorescence observation found that SMI-4a decreased the expression level of β-catenin in the U937 cells.CONCLUSION: SMI-4a induces U937 cell apoptosis through regulating the expression of apoptosis-related genes.     

Down-regulated BARF1 expression induces EBV-positive gastric carcinoma cell apoptosis via activating caspase-dependent mitochondrial pathway

AIM: To investigate the effects of BARF1 down-regulation on EBV-positive gastric carcinoma cell apoptosis, and the molecular mechanisms by BARF1 silencing-mediated apoptosis. METHODS: After NUGC3 and SNU719 cells were transfected with NCsiRNA and siRNA, respectively, the protein levels of BARF1, Bcl-2, Bax, cytochrome C, caspase 3 and capase 9 were detected by Western blot, and the mRNA expression of BARF1, Bcl-2 and Bax was determined by RT-PCR. The cell viability was measured by the method of Trypan blue exclusion and the cell apoptosis was analyzed by flow cytometry analysis with Annexin V-FITC/PI staining. The expression of the apoptosis-related proteins in the cells transfected with siRNA and NCsiRNA was examined by human apoptosis antibody arrays. Mitochondrial membrane potential was determined by flow cytometry. The interaction between Apaf-1 and caspase 9 was confirmed by immunoprecipitation. RESULTS: Compared with untreated and NCsiRNA groups, BARF1 gene silencing significantly inhibited the cell viability, induced apoptosis, and reduced the mitochondrial membrane potential in the NUGC3 and SNU719 cells transfected with siRNA. BARF1 gene silencing up-regulated the expression of pro-apoptotic proteins and down-regulated the expression of anti-apoptotic proteins, and the Bcl-2/Bax ratio was significantly decreased. In BARF1 gene silencing cells, the caspase inhibitor z-VAD-fmk inhibited BARF1 silencing-mediated apoptosis, and significantly increased the levels of cleaved caspase 3 and caspase 9. The concentration of cytochrome C significantly increased as compared with NCsiRNA group, and Apaf-1 interacted with caspase 9 in the cytoplasm. CONCLUSION: BARF1 silencing induces apoptosis via the mitochondrial pathway through regulating the expression of Bcl-2 and Bax proteins in a caspase-dependent manner in the NUGC3 and SNU719 cells.     

Inhibitory effect of poncirin on viability of gastric cancer cells and underlying mechanism

AIM: To explore the effect of poncirin on the growth of AGS gastric cancer cells and the underlying mechanism.METHODS: The effect of poncirin on AGS cell viability was measure by MTT assay. The cell cycle distribution and cell apoptosis were analyzed by flow cytometry. Nuclear staining with DAPI was used to reflect the morphological change of the AGS cells treated with poncirin. The protein levels of extrinsic apoptosis pathway-related proteins such as FasL, caspase-8, caspase-3 and PARP, and mitochondria-mediated intrinsic apoptosis pathway-associated proteins such as Bak, Bcl-xL, Bax and caspase-9 were determined by Western blot.RESULTS: Poncirin inhibited the viability of AGS gastric cancer cells in a time-and concentration-dependent manner (P<0.05). Poncirin induced accumulation of G₁ DNA content and significantly increased total apoptosis in the AGS cells. Nuclear staining showed a dose-dependent increase in the number of apoptotic cells after treated with poncirin.The protein level of FasL was upregulated in a dose-dependent manner by treatment with poncirin. Poncirin significantly activated caspase-8 and caspase-3. Moreover, poncirin significantly induced the cleavage of PARP in a dose-dependent manner (P<0.05). In addition, the protein levels of Bcl-xL, Bax and Bak were unchanged after treated with different doses of poncirin. Furthermore, caspase-9 was not activated by poncirin treatment in the AGS cells.CONCLUSION: Poncirin has the anti-cancer effect via extrinsic apoptosis pathway to inhibit the growth of AGS gastric cancer cells, possibly making it a therapeutic agent for human gastric cancer treatment.     

Effect of ARMCX1 on proliferation and apoptosis of cervical cancer SiHa cells

AIM: To observe the effect of Armadillo repeat-containing X-linked protein 1 (ARMCX1) on the proliferation and apoptosis of human cervical cancer SiHa cells by knock-down of ARMCX1 expression with small interfering RNA. METHODS: After knock-down of ARMCX1 expression, the cell cycle distribution and apoptosis of SiHa cells were detected by flow cytometry. The proliferation of SiHa cells was observed by plate colony formation assay after knock-down of ARMCX1 for 10 d. The protein levels of cell proliferation-and apoptosis-related molecules were determined by Western blot. RESULTS: After knock-down of ARMCX1 expression in the SiHa cells, the cell colony formation ability was significantly inhibited (P < 0.05), the cell cycle was arrested in S phase, and the protein levels of cyclin E and cell division cycle 25A (Cdc25A) in the SiHa cells were decreased. Meanwhile, knock-down of ARMCX1 expression promoted the apoptosis of SiHa cells, significantly reduced the protein expression of Bcl-2, and significantly increased the protein levels of Bax and active caspase-3 (P < 0.05). CONCLUSION: Knock-down of ARMCX1 expression inhibits the proliferation of SiHa cells and induces apoptosis.     

Neuronal apoptosis and the chance of apoptosis-related proteins in ovariectomied rats with the effect of App17-mer peptide

AIM: To observe the expression of apoptosis-related proteins in hippocampal neurons of ovariectomized (OVX) rats and explore the neuroprotective mechanism of the App17-mer peptide. METHODS: Female Wistar rats were randomly divided into three groups. Bilaterally ovariectomized rats with injection of App 17P peptide (3.5 μg in 0.1 mL/per rat, three times a week) formed the experimental group (17P+ OVX group). Anti-AIF, Bcl-2 and Bax antibodies were applied in the immunohistochemistry experiment. TUNEL was employed to detect apoptosis. RESULTS: The number of apoptotic neurons was clearly higher in hippocampal and cortex in OVX group than that in OVX+17P group. Immunohistochemistry demonstrated the increased expression of AIF, Bax in hippocampal neurons of OVX group. OVX group showed a significantly reduced expression of Bcl-2 in hippocampal neurons. Hippocampal tissue from OVX group showed the increased expression of AIF,Bax ,and showed diminished expression of Bcl-2 , treating with App17-mer peptide normalized the expression of these proteins. CONCLUSIONS: The expression of apoptosis-related proteins were abnormal in the OVX rats. App17-mer peptide normalized these changes ; Estrogen deficiency induced neuronal apoptosis. App17-mer peptide diminished apoptosis.     

Effects of baicalin on CA46 cell proliferation inhibition and apoptosis induction

AIM: To investigate the effects of baicalin on proliferation inhibition and apoptosis induction in human Burkitt lymphoma cell line CA46 and to explore its underlying mechanisms. METHODS: CA46 cells were exposed to baicalin at different dosages and its proliferation inhibition was detected by MTT assay. The ability of baicalin to induce CA46 cell apoptosis was examined by Annexin V-FITC/PI double staining analysis, TUNEL labeling method and DNA fragmentation. The mRNA expressions of c-myc and bcl-2 were detected by RT-PCR, and the protein expressions of c-Myc, Bcl-2, caspase-3 precursor (procaspase-3) and poly ADP-ribose polymerase (PARP) were detected by Western blotting. RESULTS: Baicalin remarkably inhibited the CA46 cell proliferation, with an IC50 value of 10 μmol/L. Apoptosis was remarkably induced by baicalin in a dose-dependent manner, and its earlier and later stages were detected by annexin V-FITC/PI double staining analysis, TUNEL labeling method and DNA fragmentation, respectively. Furthermore, RT-PCR showed that the mRNA expressions of c-myc and bcl-2 in treated CA46 cells decreased in a time-dependent manner. Western blotting showed that the protein expressions of c-Myc, Bcl-2, procaspase-3 and PARP (116 kD) in baicalin treated CA46 cells were down-regulated in a time-dependent manner, while the expression of PARP(85 kD) was up-regulated. CONCLUSION: Baicalin efficiently induces proliferation inhibition and apoptosis in CA46 cells, which may be related with the down-regulation of c-Myc and Bcl-2 expressions, as well as the up-regulation of caspase-3 activity.     

Downregulation of DEK induces cell apoptosis via inhibition of NF-κB signaling pathway in gastric carcinoma SGC-7901 cells

AIM: To investigate the effect of DEK downregulation on the apoptosis of gastric carcinoma SGC-7901 cells, and to explore its associations with NF-κB signaling pathway and apoptosis related proteins. METHODS: SGC-7901 cells with different treatments were divided into 3 groups including untreated group, control siRNA group and DEK siRNA group. The expression of DEK at mRNA and protein levels in the SGC-7901 cells was detected by real-time PCR and Western blot. The cell apoptosis was examined by flow cytometry. Furthermore, the activities of caspase-3 and caspase-9 in the SGC-7901 cells were investigated by Caspase-Glo^®-3/9 kit. Finally, the expression of key regulatory protein p65 of NF-κB signaling pathway and apoptosis-related proteins Bcl-2 and Bax in the SGC-7901 cells was investigated by Western blot. RESULTS: Compared with untreated group and control siRNA group, the expression of DEK at mRNA and protein levels was significantly downregulated in DEK siRNA group (P<0.05). In addition, the ratios of early phase apoptosis and total apoptosis in DEK siRNA group were markedly higher than those in untreated group and control siRNA group (P<0.05). Most notably, the decrease in p65 and Bcl-2 proteins, increase in Bax protein and the increases of caspase-3 and caspase-9 activities were observed in DEK siRNA group. CONCLUSION: Downregulation of DEK mediates cell apoptosis of gastric carcinoma may be tightly associated with NF-κB signaling pathway.     

Ginsenoside Rh4 induces apoptosis of human hepatocellular carcinoma HepG2 cells

AIM: To investigate the apoptosis and molecular mechanism of human hepatocellular carcinoma HepG2 cells induced by ginsenoside Rh4. METHODS: Human hepatocellular carcinoma HepG2 cells were treated with ginsenoside Rh4 at doses of 10, 20 and 40 μmol/L, and the inhibitory effect of ginsenoside Rh4 on HepG2 cell viability was measured by MTT assay. The apoptotic rate of HepG2 cells was analyzed by flow cytometry. The morphological changes of the HepG2 cells were observed by Hoechst 33258 and TUNEL staining. The expression of apoptosis-related proteins Bax, Bcl-2, caspase-3 and caspase-9 was determined by Western blot. RESULTS: Ginsenoside Rh4 promoted apoptosis of HepG2 cells in a dose-dependent manner. TUNEL and Hoechst 33258 staining showed that the cells appeared obvious shrinking, swelling and rupture after treated with ginsenoside Rh4 for 24 h. The results of Western blot showed that with the increasing concentrations of ginsenoside Rh4, the expression of pro-apoptotic proteins Bax, cleaved caspase-3 and caspase-9 increased, while anti-apoptotic protein Bcl-2 decreased gradually. CONCLUSION: Ginsenoside Rh4 induces apoptosis of human hepatocellular carcinoma HepG2 cells, and the main mechanism may be related to down-regulation of Bcl-2 and up-regulation of Bax, cleaved caspase-3, and caspase-9.     

Tert-butyl hydroperoxide damages mitochrondria and induces apoptosis in cortical neurons

AIM: To explore the possible mechanism of tert-butyl hydroperoxide (t-BHP)-induced apoptosis in rat cortical neurons. METHODS: Primary cultured rat cortical neurons were performed in vitro and cell viability was measured by MTT assay. DNA fragmentation was used to evaluate cell apoptosis and mitochondrial transmembrane potential (ΔΨm) was determined by flow cytometric assay. Cellular glutathione (GSH) content was measured by spectrophotometer. Bcl-2 and Bax protein, cytosolic cytochrome c, cleaved caspase-3 and poly (ADP-ribose) polymerase (PARP) were detected by Western blotting. RESULTS: After exposure of cortical neurons to tBHP (25-400 μmol/L), the cell viability was reduced. ΔΨm and cellular GSH content were also decreased significantly. The level of Bcl-2 protein was reduced and Bax was elevated. Meanwhile, tBHP exposure resulted in cytochrome c release, caspase-3 and PARP proteolysis, DNA fragmentation and eventually neuron apoptosis. CONCLUSION: Mitochondrial damage may mediate tBHP- induced apoptosis in cortical neurons.     

Mechanism of 13-methyltetradecanoic acid in inducing bladder cancer T24 cell apoptosis in vitro

AIM: To study the effect of 13-methyltetradecanoic acid (13-MTD) on bladder cancer T24 cell apoptosis and the underlying mechanisms. METHODS: T24 cells were treated with 13-MTD at different concentrations. MTT cell proliferation assay was used to observe the inhibitory effect. The cell cycle and cell apoptosis were displayed by FCM. Apoptosis index was investigated by TUNEL assay. Western blotting was used to detect the expression of apoptosis related proteins. RESULTS: p38 and JNK phosphorylation were presented 2 h after stimulated by 13-MTD, but AKT phosphorylation were inhibited. Cytochrome C was released from the intermembrane space of mitochondria 8 h later. No change of FADD and p-FADD was observed. Several downstream caspase substrates (PARP, lamin B, RB) were cleaved in 12 h. CONCLUSION: In the course of T24 cell apoptosis induced by 13-MTD, JNK/SAPK and p38MAPK signaling pathway are activated and the PI3K/AKT signaling pathway is inhibited. The substrates of caspase: PARP, Rb, lamin B are cleavaged by activated caspase enzymes to promote T24 cell apoptosis.     

Expression of miR-625-3p in colorectal carcinoma tissues and cells

AIM: To investigate the expression of microRNA-625-3p (miR-625-3p) in colorectal carcinoma (CRC) and its underlying mechanism. METHODS: Quantitative real-time PCR was employed to detect the levels of miR-625-3p expression in different CRC cell lines, CRC tissues and pair-matched adjacent normal tissues. The relationships between the expression levels of miR-625-3p and the patients' clinicopathological parameters were estimated. The effects of miR-625-3p on the apoptosis and the cell mitotic cycle of CRC cells were analyzed with propidium iodide staining and flow cytometry. The effect of miR-625-3p on the apoptosis-related proteins was analyzed by Western blot. RESULTS: The expression level of miR-625-3p in the CRC tissues was higher than that in the pair-matched adjacent normal tissues (P<0.05). The expression of miR-625-3p in the CRC tumor tissues was significantly correlated with the tumor infiltrative depth, TNM stage and distant metastasis (P<0.05). The expression levels of miR-625-3p in CRC SW620 cells were higher than that in SW480 cells. The CRC cell mitotic cycle was significantly inhibited and cell apoptosis was significantly promoted when the expression of miR-625-3p was inhibited (P<0.05). The expression of Bax protein didn't change and the expression of Bcl-2 protein increased after miR-625-3p mimics were transfected into CRC SW620 cells(P<0.05). CONCLUSION: miR-625-3p may be a promising approach for the treatment of CRC by promoting cell proliferation and inhibiting apoptosis.     

Effects of AZD8055 on autophagy and apoptosis in cholangiocarcinoma cells

AIM: To explore the effects of mammalian target of rapamycin (mTOR) double inhibitor AZD8055 on autophagy and apoptosis of human cholangiocarcinoma cell line HuCCT1. METHODS: The effect of AZD8055 on the viability of HuCCT1 cells was detected by MTT assay. Autophagosome was detected by acridine orange (AO) staining. After treated with AZD8055, the expression levels of apoptosis-related proteins Bcl-2, Bax and cleaved caspase-3 and auto-phagy marker proteins beclin 1, LC3 and p62 were determined by Western blot. Apoptotic rate was analyzed by flow cyto-metry with Annexin V-FITC/PI double staining. RESULTS: AZD8055 significantly inhibited the viability of HuCCT1 cells (P<0.05). AO staining showed that AZD8055 significantly increased orange granules in the cytoplasm. After treated with AZD8055, compared with the control group, the protein level of beclin 1 and the ratio of LC3-Ⅱ/LC3-I were enhanced, while p62 was attenuated (P<0.05). The protein expression level of pro-apoptotic regulator Bax was down-regulated and anti-apoptotic regulator Bcl-2 was increased. The protein level of cleaved caspase-3 was reduced (P<0.05). The results of flow cytometry showed that AZD8055 inhibited cell apoptosis. CONCLUSION: AZD8055 inhibits the viability of cholangiocarcinoma cells, and the mechanism is closely related with autophagy induced by AZD8055.     

Perifosine inhibits cell proliferation and triggers apoptosis in human gastric cancer cell line SGC-7901 cells

AIM: To investigate the effect of perifosine, a novel inhibitor of Akt, on the cell proliferation and apoptosis in human gastric cancer cell line SGC-7901.METHODS: Cell growth inhibition was detected by MTT assay. Cell cycle was analyzed by flow cytometry. Annexin V-FITC apoptosis detection kit was used to determine apoptosis in the cells. Protein expression was examined by Western blotting.RESULTS: Akt phosphorylation was dose-dependently inhibited by perifosine in SGC-7901 cells. The results of MTT and cell cycle analysis indicated that perifosine inhibited the growth of human gastric cancer cells in a dose-dependent manner. Perifosine arrested the cell cycle progression at G₂ phase. Apoptosis induction became more effective with increasing the concentration of perifosine. The caspase cascade and its downstream effector poly(ADP-ribose) polymerase (PARP) were also activated upon perifosine treatment, and the level of Bcl-2 was down-regulated, whereas the protein level of Bax was up-regulated.CONCLUSION: The small molecule Akt inhibitor perifosine shows substantial anti-tumor activity in human gastric cancer cell line SGC-7901. Perifosine induces caspase-dependent apoptosis, and the key regulators include caspase-3, caspase-9 and Bcl-2.     

Resveratrol modulates SDF-1/CXCR4 signaling to inhibit hypoxia-ischemia-induced apoptosis of neurons from the brain of newborn rats

AIM:To investigate the effect of resveratrol (RSV) on apoptosis and stromal cell-derived factor-1 (SDF-1)/CXC chemokine receptor 4 (CXCR4) pathway in hypoxia-ischemia-induced neurons from the brain of newborn rats and its mechanism. METHODS:The cortex neurons from the brain of newborn rats were given oxygen-glucose deprivation (OGD) treatment to mimic neonatal hypoxic-ischemic encephalopathy (HIE). The cortex neurons were randomly divided into 4 groups:control group, HIE model group, HIE+RSV-low (10 μmol/L) group, and HIE+RSV-high (50 μmol/L) group. After OGD treatment for 2 h, the neurons were cultured with indicated dose of RSV for 24 h. The apoptosis was analyzed by flow cytometry. Western blot was used to determine the levels of apoptosis-related proteins, SDF-1 and CXCR4. Real-time PCR was used to detect the mRNA expression of SDF-1 and CXCR4. Additionally, to explore the effects of RSV on cell apoptosis and apoptosis-related proteins after the suppression of SDF-1/CXCR4 signaling, a CXCR4 antagonist AMD3100, and RSV were used to co-treat OGD-injured neurons for 24 h. RESULTS:RSV alleviated OGD-induced neuronal apoptosis, down-regulated cleaved caspase-3 and cytochrome C levels, and up-regulated the ratio of Bcl-2/Bax. Compared with the control group, OGD treatment increased the expression of SDF-1 and CXCR4 (P<0.05). Compared with the HIE model group, RSV further up-regulated the expression of SDF-1 and CXCR4 (P<0.05). AMD3100 reversed the effects of RSV on OGD-induced cell apoptosis. CONCLUSION:RSV suppresses hypoxia-ischemia-induced apoptosis of neurons from the brain of newborn rats via up-regulating SDF-1/CXCR4 signaling pathway.