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1.
AIM: To observe the effects of vascular endothelial growth factor (VEGF) on the proliferation and differentiation of neural stem cells (NSCs) of rats in vitro.METHODS: NSCs isolated from the hippocampal gyrus of SD rats were primary cultured and subcultured,and then divided into two groups: (1) the cells in VEGF group were treated with 150 μg/L VEGF in the culture system,and VEGF was removed at the 7 th day;(2) control group (without VEGF treatment).The cellular morphology of two groups was observed by contrast phase microscope.Nestin and NF-200 expressing cells were detected via immunofluorescence method.The percentages of the immunostaining positive cells in each group at the 7 th day and at the 11 th day were determined.RESULTS: At the 7 th day,the percentage of nestin positive cells in VEGF group was 52.19%±7.95%,vs 29.26%±4.12% in control group (P<0.01).The percentage of NF positive cells in VEGF group was 22.33%±4.13%,vs 38.62%±5.31% in control group (P<0.01).At the 3 th day after VEGF was removed,the percentage of NF positive cells in VEGF group was 43.10%±3.70%,vs 30.56%±4.16% in control group (P<0.01).CONCLUSION: VEGF stimulates the proliferation of neural stem cells and inhibits their differentiation.  相似文献   

2.
AIM: To investigate whether Notch1/Hes1 signaling pathway regulates the expression of CCAAT/enhancer binding protein alpha (C/EBPα), thus affecting the proliferation and differentiation of type Ⅱ alveolar epithelial cells (AECⅡ). METHODS: Human AECⅡ were cultured in vitro, and randomly divided into control group, activator group (adding Notch pathway activator Jagged1 protein, 500 μg/L) and inhibitor group (adding Notch inhibitor DAPT, 10 μmol/L). AECⅡ in each group were collected after 24 h of intervention. The expression of Notch1, Hes1 and C/EBPα at mRNA and protein levels was determined by RT-qPCR and Western blot. The cell proliferation ability of the AECⅡ was measured by living cell counting and CCK-8 assay. The cell cycle distribution and differentiation of the AECⅡ were analyzed by flow cytometry. RESULTS: Compared with control group, the mRNA and protein expression levels of Notch1, Hes1 and C/EBPα were significantly increased in activator group (P<0.05), AECⅡ entered G2/M phase from S phase, the proliferation of AECⅡ was increased, and the differentiation of AECⅡ was reduced (P<0.05). The mRNA and protein expression levels of Notch1, Hes1 and C/EBPα were significantly reduced in inhibitor group (P<0.05), the cell cycle of AECⅡ cells was arrested in G0/G1 phase, the proliferation of AECⅡ cells was reduced, and the differentiation of AECⅡ cells was increased (P<0.05). CONCLUSION: Notch1/Hes1 signaling pathway regulates the expression of C/EBPα and affects the proliferation and differentiation of AECⅡ.  相似文献   

3.
AIM:To investigate the effect of Xiaoyaosan (XYS) drug-containing serum on proliferation and differentiation of hippocampus nerve precursor cells (HNPC) under high corticosterone (CORT) concentration condition. METHODS:Serum-free culture in vitro was adopted to culture HNPC. CORT at concentration of 120 μmol/L was used to establish the high CORT concentration condition. XYS is composed of Bupleurum Chinese DC, Chinese angelica root, Paeonia lactiflora Pal1, India bread, Largehead atractylodes rhizome, peppermint herb, fresh ginger and Licorice root. XYS drug-containing serum of different dose were prepared.10% drug-containing serum and 10% serum were adopted. RU38486, an antagonist of CORT, was used as positive control. MTT method was used to detect proliferation rate of HNPC, the methods of immunofluorescence ambi-tagged by BrdU with TUNEL, β-tubulin-Ⅲ and glial fibrillary acidic protein (GFAP) with TUNEL respectively were adopted to detect the proliferation and differentiation of HNPC. RESULTS:CORT at concentration of 120 μmol/L degraded the proliferation rate of HNPC significantly (P<0.01), which was reversed by RU38486 (P<0.05). 10% XYS drug-containing serum of each dose enhanced the proliferation rate (P<0.05 or P<0.01). After 120 μmol/L CORT processed, the staining fluorescence intensity ratio of BrdU/TUNEL of HNPC degraded significantly (P<0.01), while increased after treatment with RU38486 and 10% XYS drug-containing serum of each dose (P<0.01). Under high CORT concentration condition, the apoptotic rates of glial cells and neurons differentiated from HNPC increased significantly (P<0.01). RU38486 and 10% XYS drug-containing serum of each dose inhibited the apoptotic rates of glial cells and neurons (P<0.05 or P<0.01). CONCLUSION:Under high CORT concentration condition, XYS promotes the proliferation of HNPC, and inhibits the apoptotic rates of glial cells and neurons differentiated from HNPC, which act as the effect of anti-damage of CORT to hippocampus in stress state.  相似文献   

4.
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5.
AIM: To investigate the roles of Notch signaling in lipopolysaccharide (LPS)-induced proliferation and secretion of interleukin-6 (IL-6) and chemokine CXCL1 in bone marrow mesenchymal stem cells (BMSCs).METHODS: BMSCs were isolated by whole bone marrow culture. The expression levels of Notch signaling pathway receptors and ligands in the BMSCs treated with LPS were measured by qPCR and Western blot. The proliferation of BMSCs was analyzed by MTT assay and viable cell counting. The secretion levels of IL-6 and CXCL1 induced by LPS were measured by ELISA.RESULTS: Treatment with LPS at 1 mg/L effectively induced the proliferation of BMSCs and the secretion of IL-6. Obvious expression of Notch receptors and ligands in the BMSCs was observed, and LPS had little effect on the mRNA and protein levels of Notch receptors and ligands, but LPS increased the protein levels of Hes1 and Hey1, the target genes of Notch signaling. LPS at 1 mg/L increased the proliferation of BMSCs, whereas DAPT (Notch signal inhibitor) reduced the basal and LPS-induced proliferation of BMSCs (P<0.01). LPS treatment robustly increased the secretion of IL-6 and CXCL1 as assessed by ELISA. However, inhibition of Notch signaling almost completely abolished LPS-induced secretion of IL-6 and CXCL1 (P<0.05).CONCLUSION: Inhibition of Notch signaling reduced not only the proliferation of BMSCs but also IL-6 and CXCL1 secretion induced by LPS.  相似文献   

6.
AIM: To investigate the proliferation and migration of neural stem cells (NSCs) in the subventricular zone (SVZ) on focal cerebral ischemia with curcumin treatment, and to explore the relationship between these effects and Notch signaling. METHODS: The animal model was established by an intraluminal suture method to induce middle cerebral artery occlusion in rats. The rats were randomly divided into sham group, cerebral ischemia/reperfusion (I/R) group, and I/R+curcumin group. The animals were given curcumin for 7 d intraperitoneally. One hour after the model was successfully established, the rats were sacrificed. Immunofluorescence was used to label the NSCs by BrdU and BrdU/DCX, and the migration tendency was observed. The expression of Notch intracellular domain (NICD, the Notch signaling pathway intermediate product) was detected by Western blotting. RESULTS: Compared with I/R group, BrdU-positive and BrdU/DCX double positive cells in I/R+curcumin group were significantly higher than those in I/R group (P<0.05), and more positive cells were on the way of migration to the ischemic lesion zone. The expression level of NICD in I/R+curcumin group was significantly higher than that in I/R group (P<0.05). CONCLUSION: Curcumin promotes NSC proliferation and migration in SVZ after focal cerebral ischemia. The possible mechanism may be that curcumin activates Notch signaling.  相似文献   

7.
AIM: To observe the characterization in neural cells derived from the hippocampus of embryonic rats and to examine the effect of myelin-associated glycoprotein (MAG) on the proliferation, differentiation and neurite growth of neural stem cells (NSCs). METHODS: The hippocampus cells of embryonic rats were isolated and cultured in vitro. The expressions of nestin and doublecortin, the marks of NSCs, were observed by immunocytochemical method. The rate of proliferating cells was examined by BrdU immunocytochemistry. The average neuronal neurite length and the percentage of differentiated neurons were detected by immunocytochemistry staining. RESULTS: The hippocampus cells of 16 days old embryonic rats had the characteristics of NSCs. The percentage of differentiated neurons (β-tubulin Ⅲ-positive cells) was 18.17%±2.79% and the average neuronal neurite length was (136.27±33.66)μm, seven days after the differentiation initiated in vitro in control group. After NSCs were treated with MAG-Fc (200 μg/L), the percentage of differentiated neurons and the average neurite length were decreased, respectively, to 10.05%±3.42% (P<0.01) and (84.87±24.94)μm (P<0.01). The results from proliferation test indicated that the rate of BrdU incorporation did not changed after the treatment of MAG-Fc with different dosages (P>0.05). CONCLUSION: MAG-Fc inhibits the differentiation and neurite growth of the NSCs, but has no effect on the proliferation.  相似文献   

8.
AIM: To observe the changes of Notch1 expression and autophagy in the renal tissues of diabetic mice, and to explore the regulatory effect of Notch1 on tubulointerstitial fibrosis by inhibiting autophagy in diabetic nephro-pathy. METHODS: The mice were randomly divided into normal control group (db/m mice) and diabetes group (db/db mice), with 8 rats in each group. After 12 weeks of feeding, the mice were sacrificed and the corresponding biochemical indexes were measured. The protein expression of Notch1 in the renal tubular epithelial cells was observed by immunohistochemical staining. The protein levels of Notch1, PTEN, p-Akt (Thr308), Akt, p-mTOR (Ser2448), mTOR, LC3, P62, collagen type Ⅰ (Col-Ⅰ) and collagen type Ⅲ (Col-Ⅲ) were determined by Western blot. RESULTS: Compared with the db/m mice, the blood glucose, glycosylated hemoglobin, serum creatinine, triglyceride and total cholesterol were increased in the db/db mice (P<0.01). Renal tubular epithelial cell vacuolar degeneration, renal tubular expansion and interstitial inflammatory cell infiltration in db/db mouse renal tissues with HE staining were observed. The images of Masson staining showed collagenous fiber-like substance deposition in the glomerular capillaries and renal interstitium, and disarrangement of tubular structure in the renal tissues of db/db mice. The protein expression levels of PTEN and LC3-Ⅱ were decreased (P<0.01 or P<0.05), while the protein levels of Notch1, P62, p-mTOR (Ser2448), p-Akt (Thr308), Col-I and Col-III were increased in the db/db mice as compared with the db/m mice (P<0.01). However, no significant change of total mTOR and Akt proteins between the 2 groups was found. CONCLUSION: Notch1 protein expression was increased, PTEN expression was significantly reduced, Akt/mTOR pathway was activated, autophagy was inhibited, and fibrosis was aggravated in the renal tissues of the diabetic mice.  相似文献   

9.
AIM: To investigate the effect of 18 alpha-glycyrrhetinic acid (18α-GA) on delaying the senescent progress and promoting the proliferation in late-passage bone marrow mesenchymal stem cells (BMSCs). METHODS: Late-passage BMSCs were incubated with 2.0 mg /L 18α-GA or the same volume of DMSO for 30 d, and the cells were harvested to determine the proteasome activity. The expression of senescence-related proteins p53, p21 and p16 was detected by senescence-associated β-galactosidase (SA-β-Gal) staining and Western blot. The cell proliferation, the expression level of cell cycle-related proteins and cell cycle distribution of the cells were measured by CCK-8 assay, BrdU incorporation, Western blot and flow cytometry. RESULTS: Compared with DMSO group, the proteasome activity in 18α-GA group increased significantly by about 0.2 times (P<0.01). SA-β-Gal-positive cells in 18α-GA group decreased, and cell staining was lighter. The contents of p53 and p21 in 18α-GA group were decreased (P<0.05). The results of CCK-8 assay showed that the A value in 18α-GA group was 0.3 times higher than that in DMSO group (P<0.01). BrdU incorporation showed the increased proliferation in 18α-GA group compared with DMSO group (P<0.05). The cells in G1 phase in 18α-GA group decreased significantly compared with DMSO group, while the cells in S phase increased significantly (P<0.05). The expression level of cyclin D1 in 18α-GA group was 2.8 times higher than that in DMSO group (P<0.01), and the CDK4 level was 1.4 times higher than that in DMSO group (P<0.05). CONCLUSION: Activation of the proteasome activity by 18α-GA delays the aging process in the BMSCs and promotes the cell proliferation via up-regulation of the cell cycle-related proteins.  相似文献   

10.
AIM:To evaluate whether bone marrow stromal cells (BMSCs) could provide a supportive microenvironment for proliferation and differentiation of neural stem cells (NSCs). METHODS:The proliferation and differentiation of NSCs were compared, in media without growth factors or in BMSCs-conditioned media. RESULTS:The proportion of neurons to total NSCs was significantly increased when NSCs were cultured in BMSCs-conditioned media (41.1%±3.2% vs 23.3%±16.5%, P<0.05), while the proportion of astrocytes was significantly decreased (33.8%±4.9% vs 65.0%±10.4%, P<0.01), as compared with the NSCs cultured in media without growth factors. The proportion of proliferated cells was also significantly increased (74.7%±4.7% vs 51.4%±12.3%, P<0.01). CONCLUSION:These finding indicates that BMSCs support the proliferation and neuronal differentiation of NSCs and suggests that their addition may be useful to improve outcomes of NSCs transplantation.  相似文献   

11.
ZHAO Yan  FU Yu 《园艺学报》2017,33(10):1751-1758
AIM: To investigate the effect of 8-week middle intensity voluntary wheeling exercise on the depressive-like behavior and the circadian rhythmic alterations of plasma hormone and peptide induced by chronic unpredictable mild stress (CUMS) in rats. METHODS: Male Sprague-Dawley rats (n=90) were randomly divided into model group, model+exercise group and control group. Rats in model+exercise group received 8-week voluntary wheel running exercise plus CUMS procedure during the last 3 weeks at the same time. Exploratory locomotor activity was assessed by open field test, the anxiety-like behavior was measured by elevated plus-maze test, and lack of pleasure was detected by sucrose preference test. Blood samples were collected at each of 6 time points (ZT1, 5, 9, 13, 17 and 21 on the 2nd day after behavior testing). Plasma concentrations of corticosterone (CORT), melatonin (MT) and vasoactive intestinal peptide (VIP) were detected by ELISA. The plasma concentration of adrenocorticotropic hormone (ACTH) was measured by radioimmunoassay. The circadian rhythm changes of serum CORT, MT, VIP and ACTH concentrations in each group were compared by cosinor analysis. RESULTS: Compared with control group, locomotor activity, weight gain and sucrose consumption in model group were significantly reduced (P<0.01). The values of the percentage of open-arm time (OT%) and open arm entries (OE%) were obviously lower in model group than those in control group (P<0.01). Eight-week voluntary wheel running exercise may improve the above depression behavior caused by CUMS. The rats in model group showed an obvious disorder in circadian rhythm of plasma ACTH and CORT, including phase advance and decrease in amplitude. There also showed a markedly blunted circadian rhythm and decreased level of plasma MT in model rats compared to control rats. VIP expression was significantly higher than that in control group with 24 h rhythm, but the amplitude was significantly lower than that in control group, peak phase also delayed for 6 h. Eight-week exercise significantly ameliorated the abnormal expression and the disturbance secretion rhythm of ACTH, CORT, MT and VIP in plasma. CONCLUSION: Eight-week voluntary wheeling exercise ameliorates CUMS-induced depressive-like behaviors probably by rescuing the disturbed circadian rhythms and abnormal secretion of these neuroendocrine factors.  相似文献   

12.
AIM: To study the expression of Jagged2/Notch3 signaling molecules in pulmonary vascular wall of pulmonary hypertensive rats induced by monocrotaline. METHODS: SD rats were randomly divided into normal control group (C group,n=15), solvent control group (S group,n=15) and monocrotaline model groups (M group,n=15). The model of pulmonary hypertension was established by a single subcutaneous injection of monocrotaline (50 mg/kg). The rats in S group were given a single subcutaneous injection of the same dose of solvent. After 4 weeks, the pulmonary vascular remodeling was assessed by HE staining, and the mean pulmonary artery pressure (mPAP) and right ventricular systolic pressure (RVSP) were determined by right heart catheterization. The expression of Jagged2/Notch3/Hes5 molecules in the pulmonary vascular wall was detected by immunohistochemical method and real-time PCR. RESULTS: Compared with S group and C group, the percentage of medial wall thickness of smaller arteries in model group increased significantly (P<0.01). The levels of mPAP and RVSP in M group were significantly higher than those in S group and C groups (P<0.01). The results of real-time PCR showed that the expression of Jagged2, Notch3 and Hes5 was significantly increased in M group compared with S group and C group. The data from immunohistochemical detection indicated that Jagged2 mainly expressed in the intima of small lung artery, Notch3 and Hes5 mainly expressed in the medial smooth muscle cells. Compared with S group and C group, the expression of Jagged2 and Notch3 was significantly increased in the lung small arteries of M group. CONCLUSION: The activation of Jagged2/Notch3 signaling pathway might play an important role in the formation of pulmonary hypertension.  相似文献   

13.
AIM：To estimate the neural differentiation efficiency of bone marrow mesenchymal stem cells (MSCs) derived from amyloid precursor protein (APP) transgenic mice and to investigate the correlation with Notch1 signaling and the autophagy activity during the differentiation. METHODS：The MSCs were divided into APP group (MSCs from APP transgenic mice) and WT group (MSCs from wild-type mice). MSCs were treated with β-mercaptoethanol as an inducer for differentiating into neurons. The levels of Aβ40 and Aβ42 were measured using enzyme-linked immunosorbent assay kits. The expression of neural cell-specific markers, neuron-specific enolase (NSE) and microtubule-associated protein 2 (MAP-2), was measured by immunocytochemistry and Western blotting. The expression levels of Notch1, Notch intracellular domain (NICD), Hes5, LC3 and p62 (a selective substrate of autophagy) were also detected by Western blotting. RESULTS：The neural differentiation capacity and the Aβ expression level of the MSCs in APP group were higher than those in WT group, and stronger inhibition of Notch1 signaling pathway in the MSCs from APP group was observed. However, the process of autophagy, which is essential for the survival and function of the neural cells, was impaired in the neural differentiated counterpart of the MSCs in APP group. CONCLUSION:Over-expression of APP might contribute to the high neural differentiation capacity of MSCs by inhibiting Notch1 signaling pathway in vitro. However, autophagy is impaired in the differentiated MSCs from APP transgenic mice.  相似文献   

14.
AIM: To study the effect of Xiaozhong (detumescence)-Zhitong (analgesia) mixture on the function of vascular endothelial cells of rat skin flaps and the expression of VEGF-Dll4/Notch signaling pathway-related proteins. METHODS: Vascular endothelial cells of rat skin flaps were isolated and cultured. The cells were divided into control group, hypoxia group, hypoxia+detumescence analgesia group, hypoxia+detumescence analgesia+axitinib (VEGF receptor inhibitor) group, and hypoxia+detumescence analgesia+MK-0752 (Notch signaling pathway blocker) group. The serum levels of VEGF were measured by ELISA. The number of dead and living cells at 1 d, 2 d and 3 d after hypoxia was determined by cell calcein-AM and PI double staining. The protein expression levels of VEGF-A, Notch and Dll4 in the cells at 24 h and 48 h were detected by Western blot. RESULTS: Compared with control group, the content of VEGF was increased significantly after 24 h and 48 h, and the protein expression of VEGF-A, Notch and Dll4 was increased significantly (P<0.05). Compared with hypoxia group, the content of VEGF was increased significantly after the intervention of Xiaozhong-Zhitong mixture, the death rate was decreased significantly, and the protein expression of VEGF-A, Notch and Dll4 was increased significantly (P<0.05). Compared with Xiaozhong-Zhitong mixture group, the protective effect of Xiaozhong-Zhitong mixture on hypoxia-induced vascular endothelial cell injury was weakened by VEGF receptor inhibitor, the cell mortality was significantly increased, the content of VEGF was decreased, and the protein expression of VEGF-A, Notch and Dll4 was decreased (P<0.05). After intervention with Notch signaling pathway blocker, the cell viability remained unchanged, the expression level of VEGF-A was increased, and the increased Notch and Dll4 protein expression was effectively resisted (P<0.01). CONCLUSION: Xiaozhong-Zhitong mixture improves the function of vascular endothelial cells of rat skin flaps, and its mechanism may be related to the influence of the signal transduction pathway of VEGF-Dll4/Notch.  相似文献   

15.
AIM: To study fibroblast proliferation and collagen synthesis during wound healing in diabetic rats induced by streptozotocin. METHODS: 30 Wistar male rats were randomly divided into control group and model group. 55 mg/kg STZ were given intraperitoneally to model rats. After 3 weeks, a round skin of 2.04 cm2 was excised on all dorsal back of rats. The healing time and healing rate were observed according to re-epithelization. The numbers of fibroblasts and the expression of proliferating cell nuclear antigen (PCNA) were observed by Hematoxylin-Eosin (HE) staining and immuno-histochemistry assay. Collagen Ⅰ and Ⅲ stained by Picric acid-Sirius red were calculated by image analysis. RESULTS: The healing time in model group was (27.13±1.81) days，significantly longer than that in control group ［(15.25±1.67） days， P<0.01］. The healing rates in model group were significantly less than that in control group at day 3, day 7 and day 15 (P<0.01). The amount of fibroblasts and the expression of PCNA in model group were significantly less than those in control group on day 3, day 5, day 7 and day 9, respectively (P<0.05, P<0.01). Even the content of collagen I in the wound of both groups increased with time, the values were much higher than that in model group at different times (P<0.05), respectively. For model group, the ratio of collagen Ⅰ/Ⅲ was less than that in control group 3, 7 and 11 days after wound (P<0.01). CONCLUSION: STZ impaires wound healing in rats, which is possible caused by the disturbance of fibroblast proliferation and collagen synthesis in the wound.  相似文献   

16.
AIM:To study the influence of excitatory amino acids on the proliferation of neural stem cells from the embryonic rat. METHODS:The neural stem cells from subventricular zone (SVZ) of the embryonic SD rat were isolated, cultured and identified in vitro. The effects of two excitatory amino acids, N-methyl-D-aspartate (NMDA) and glutamic acid (Glu), on cell proliferation were detected by MTT colorimetric assay. RESULTS:The cultured cells showed embryo-derived characteristic were neural stem cells. NMDA and Glu significantly decreased the proliferation rate in all these cells. CONCLUSION:The subventriculcr zone cells have the self-renew and multipotent differentiation potential, belong to neural stem cells of central nervous system. Excitatory amino acids, NMDA and Glu, efficiently inhibit the cell proliferation.  相似文献   

17.
AIM: To study the effect of chronic corticosterone (CORT) injection on the depression-like behaviors and the brain glycogen level in mice. METHODS: Male C57BL/6N mice (n=40) were randomly divided into normal control group and model group. The mice in model group were subcutaneously consecutively injected with CORT for 4 weeks. The mouse model of chronic stress depression was constructed. The forced swim test and open field experiment were conducted to prove chronic stress model. The serum level of CORT in the mice was measured by radioimmunoassay. The protein levels of hippocampal synaptophysin (SYP) and brain-derived neurotrophic factor (BDNF) were detected by Western blot. Hippocampus glycogen, glycogen synthase and glycogen phosphorylase were determined by indirect fluorescence measurement. RESULTS: Compared with normal control group, the immobility time of the forced swim test in model group was significantly lengthened (P<0.01), and the ability of spontaneous activity was reduced (P<0.01), indicating that chronic CORT injection induced depression-like behaviors in mice. The CORT level increased significantly (P<0.01) in model group. CORT injection decreased the protein expression of hippocampal SYP and BDNF (P<0.01), reduced hippocampal glycogen level (P<0.05) and glycogen synthase activity (P<0.05), and increased glycogen phosphorylase activity (P<0.05). CONCLUSION: Chronic CORT injection causes hippocampal neuron damage and induces the depression-like behaviors of mice, which may be associated with decreasing hippocampal glycogen level by CORT.  相似文献   

18.
AIM: Retinoblastoma-like protein 2 (Rbl-2) plays an important role in the cell proliferation and DNA methyltransferase (DNMT) may involve in the regulation of differentiation in embryonic stem cells. This study is to investigate the effect of knocking down Rbl-2 by specific siRNA on apoptosis in human cardiac stem cells.METHODS: The siRNA of Rbl-2 (siRbl-2) was transfected into human cardiac stem cells. The mRNA expression of Rbl-2 and DNMT-3B was detected by real-time RT-PCR 48 h after transfection. The DNMT-3B protein expression and the activation of caspase-3 were determined by Western-blotting. The cell apoptosis was analyzed by flow cytometry with annexin V-FITC/PI staining. RESULTS: Knocking down of Rbl-2 gene increased the expression of DNMT-3B in human cardiac stem cells, and induced cell apoptosis. Compared with negative control group, caspase-3 was activated and cleaved caspase-3 was increased in the stem cells transfected with siRbl-2. The cleaved caspase-3 accounted for more proportions of total caspase-3 in transfected cells than that in non-transfected cells (P<0.01). The apoptotic rate was also increased significantly in transfected group (P<0.01).CONCLUSION: Rbl-2 plays an important role in the regulation of survival and apoptosis in human cardiac stem cells. This regulatory mechanism may involve in epigenetic modification, which is mediated by DNMT.  相似文献   

19.
AIM: To explore the effects of metanephric cell microenvironment on inducing embryonic stem cells (ESCs) to differentiate toward renal cells.METHODS: Embryoid bodies (EBs) of D3 mouse embryonic stem cells were prepared by hanging drop culture, and the EBs were co-cultured indirectly with metanephric cells derived from E12.5 d mouse embryo. The EBs cell with spontaneous differentiation was used as the control. The proteins of Pax2 and WT-1 were analyzed by immunofluorescence assay. The mRNA expression of Pax2, WT-1, Lim1, Sall1, Emx2, GDNF, Wnt4, BMP7, Nephl, Nephrin, KSP and CD24 genes was detected by RT- PCR.RESULTS: The genes related to kidney development were expressed in the EBs cells after co-culture on day 3, and the mRNA expression of Pax2, WT-1, Emx2, GDNF, Nephl, Nephrin, KSP and CD24 was stronger than those in control group. Pax2 positive cells were found on day 3 in the co-cultured EBs cells, and the positive cells increased on day 5 and day 7. WT-1 protein positive cells were found in the co-cultured EBs cells on day 5. No Pax2 or WT-1 positive cell was observed in control group.CONCLUSION: Metanephric cell microenvironment promotes ESCs differentiation toward renal cells.  相似文献   

20.
AIM: To investigate the effects of cardiotrophin 1 (CT-1) on differentiation of swine bone marrow mesenchymal stem cells (MSCs) into cardiomyocyte-like cells in vitro.METHODS: MSCs were isolated and proliferated from Tibet miniswine. Adipogenic and osteogenic potentials were identified. MSCs were divided into 4 groups for induction: untreated group, 5-azacytidine (5-Aza) group,CT-1 group and 5-Aza combined with CT-1 group. After induction for 4 weeks, the expression of cardiac cell markers including α-actin and cardiac troponin-T (cTnT) was estimated by immunofluorescence staining. Finally, the rates of red fluorescence positive-staining cells were calculated. RESULTS: The expression of α-actin in the 4 groups by red fluorescence staining was as follows: the differentiation rate of cardiomyocyte-like cells in combination group was 29.90%±4.76%, significantly higher than that in 5-Aza group (17.73%±2.34%, P<0.01), CT-1 group (6.63%±0.55%, P<0.01) and untreated group (1.62%±0.09%, P<0.01). The differentiation rate in 5-Aza group was significantly higher than that in CT-1 group (P<0.01) and untreated group (P<0.05). The differentiation rate in CT-1 group was significantly higher than that in untreated group (P<0.01). The expression of cTnT in the 4 groups was as follows: the differentiation rate of cardiomyocyte-like cells in combination group was 36.50%±4.09%, significantly higher than that in 5-Aza group (14.37%±1.65%, P<0.01), CT-1 group (7.50%±0.61%, P<0.01) and untreated group (1.12%±0.23%, P<0.01). The differentiation rate in 5-Aza group was significantly higher than that in CT-1 group (P<0.01) and untreated group (P<0.01). The differentiation rate in CT-1 group was significantly higher than that in untreated group (P<0.01).CONCLUSION: Appropriate concentrations of 5-Aza (10 μmol/L) and CT-1 (0.1 μg/L) induce swine bone marrow MSCs to differentiate into cardiomyocyte-like cells in vitro. CT-1 combined with 5-Aza significantly increases the differentiation rate.  相似文献   

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