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1.
Journal of Experimental Hematology ; (6): 1050-1055, 2021.
Article in Chinese | WPRIM | ID: wpr-888517

ABSTRACT

OBJECTIVE@#To investigate the effect of sulforaphane (SFN) on G@*METHODS@#KG1a and KG1cells were treated by different concentrations of SFN for 48 h. Flow cytometry (FCM) was used to analyze the phase distribution of cell cycle. High-throughput sequencing was used to detect the effect of SFN on the expression of cell cycle related genes in KG1a cells. The mRNA expression of P53, P21, CDC2 and CyclinB1 were detected by qPCR. The protein expression of P53, CDC2, P-CDC2 and CyclinB1 were detected by Western blot.@*RESULTS@#Cells in the G@*CONCLUSION@#SFN induces leukemia cells to block in G


Subject(s)
Humans , Cell Cycle , Isothiocyanates/pharmacology , Leukemia, Myeloid, Acute , Mitosis , Sulfoxides
2.
Chinese Journal of Natural Medicines (English Ed.) ; (6): 732-740, 2021.
Article in English | WPRIM | ID: wpr-922756

ABSTRACT

Physalin B (PB), one of the major active steroidal constituents of Solanaceae Physalis plants, has a wide variety of biological activities. We found that PB significantly down-regulated β-amyloid (Aβ) secretion in N2a/APPsw cells. However, the underlying mechanisms are not well understood. In the current study, we investigated the changes in key enzymes involved in β-amyloid precursor protein (APP) metabolism and other APP metabolites by treating N2a/APPsw cells with PB at different concentrations. The results indicated that PB reduced Aβ secretion, which was caused by down-regulation of β-secretase (BACE1) expression, as indicated at both the protein and mRNA levels. Further research revealed that PB regulated BACE1 expression by inducing the activation of forkhead box O1 (FoxO1) and inhibiting the phosphorylation of signal transducer and activator of transcription 3 (STAT3). In addition, the effect of PB on BACE1 expression and Aβ secretion was reversed by treatment with FoxO1 siRNA and STAT3 antagonist S3I-201. In conclusion, these data demonstrated that PB can effectively down-regulate the expression of BACE1 to reduce Aβsecretion by activating the expression of FoxO1 and inhibiting the phosphorylation of STAT3.


Subject(s)
Humans , Alzheimer Disease , Amyloid Precursor Protein Secretases/metabolism , Amyloid beta-Peptides/metabolism , Aspartic Acid Endopeptidases/metabolism , Down-Regulation , Forkhead Box Protein O1/genetics , Phosphorylation , STAT3 Transcription Factor/metabolism , Secosteroids
3.
Chinese Journal of Zoonoses ; (12): 793-799, 2017.
Article in Chinese | WPRIM | ID: wpr-659159

ABSTRACT

To compare the discriminatory ability of multilocus sequence typing and serotype of animal-derived Salmonella and find its distribution in Shandong Province,78 chicken-origin,56 duck-origin and 20 swine-origin Salmonella were separated from some regions of Shandong Province.Seven conserve sequences of Salmonella were PCR-amplified for MLST and slide agglutination test for serotyping.Results showed that by serotyping,6 serotypes were identified from chicken-origin Salmonella,including 88.5%0 S.enteritidis,5.1% S.indiana,2.6% S.thompson,1.3% S.typhimurium,1.3% S.senftenberg,1.3% S.agama.Two serotypes were identified from duck-origin Salmonella,including 67.9% S.enteritidis,32.1% S.ty phimurium.Three serotypes were identified from swine-origin Salmonella,including 65% S.typhimurium,20% S.derby,and 15% S.enteritidis.By MLST typing,seven ST types were identified from chicken-origin Salmonella:ST11,ST19,ST26,ST128,ST14,ST17 and Newl.Three ST types were identified form duck-origin Salmonella:ST11,ST19 and New2.Three ST types were identified from swine-origin Salmonella:ST34,ST40 and ST3007.Overall,the types identified with two methods were closed,so MLST and serotype have similar discriminatory ability.

4.
Chinese Journal of Zoonoses ; (12): 793-799, 2017.
Article in Chinese | WPRIM | ID: wpr-657274

ABSTRACT

To compare the discriminatory ability of multilocus sequence typing and serotype of animal-derived Salmonella and find its distribution in Shandong Province,78 chicken-origin,56 duck-origin and 20 swine-origin Salmonella were separated from some regions of Shandong Province.Seven conserve sequences of Salmonella were PCR-amplified for MLST and slide agglutination test for serotyping.Results showed that by serotyping,6 serotypes were identified from chicken-origin Salmonella,including 88.5%0 S.enteritidis,5.1% S.indiana,2.6% S.thompson,1.3% S.typhimurium,1.3% S.senftenberg,1.3% S.agama.Two serotypes were identified from duck-origin Salmonella,including 67.9% S.enteritidis,32.1% S.ty phimurium.Three serotypes were identified from swine-origin Salmonella,including 65% S.typhimurium,20% S.derby,and 15% S.enteritidis.By MLST typing,seven ST types were identified from chicken-origin Salmonella:ST11,ST19,ST26,ST128,ST14,ST17 and Newl.Three ST types were identified form duck-origin Salmonella:ST11,ST19 and New2.Three ST types were identified from swine-origin Salmonella:ST34,ST40 and ST3007.Overall,the types identified with two methods were closed,so MLST and serotype have similar discriminatory ability.

5.
Journal of Experimental Hematology ; (6): 1725-1729, 2016.
Article in Chinese | WPRIM | ID: wpr-332621

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the effects of arsenic trioxide (AsO) on K562 cell proliferation by regulating cell cycle protein D1 and cyclin-dependent kinase inhibitor p27kip1.</p><p><b>METHODS</b>MTT was used to detect the effect of AsOon K562 cell proliferation, so as to screen out the appropriate drug concentration. Furthermore, the K562 cell apoptosis was observed by microscopy. The expression of CyclinD1 and p27kip1 in K562 cells treated with AsOwas analyzed by reverse transcription-polymerase chain reaction(RT-PCR), immunohistochemistry and Western blot.</p><p><b>RESULTS</b>AsOcould inhibit the proliferation of K562 cells in a dose- and time- dependent manner (r= 0.967). And the apoptosis cell number in AsOgroup was significantly higher than that in the control group(P<0.05). AsOcould markedly inhibit the expression of CyclinD1 in K562 cells(P<0.05), but the expression of P27kip1 was not significantly changed after AsOtreatment.</p><p><b>CONCLUSIONS</b>AsOcan induce K562 cell apoptosis and inhibit K562 cell proliferation by regulating the expression of CyclinD1.</p>

6.
Journal of Experimental Hematology ; (6): 517-521, 2015.
Article in Chinese | WPRIM | ID: wpr-357324

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the effects of mannan-binding lectin (MBL) on the maturation and cytokine secretion of human dendritic cells (DC) induced by Candida albicans (C. albicans).</p><p><b>METHODS</b>The plastic-adherent mononuclear cells were prepared from the blood of healthy adult volunteers. The human peripheral blood mononuclear cells-derived dendritic cells (MNC-DC) were induced by 5-day-culture in medium supplemented with rhGM-CSF and rhIL-4, and then cultured for 2 days in presence or absence of C. albicans at varying concentration of human MBL ranging from 1 to 20 mg/L. DC's shape and characters were observed under inverted microscopy, the expression of CD83 and CD86 on DC was analyzed by FACS. The levels of TNF-α and IL-6 were detected by ELISA. FACS also was used to investigate the interaction of MBL with immature DC(imDC) and C. albicans. Western blot was used to detect C. albicans-induced IκBα phosphorylation and p65/NF-κB translocation in DC.</p><p><b>RESULTS</b>MBL at higher concentrations (10-20 mg/L) down-regulated the expression of CD83 and CD86 on the monocyte-derived dentritic cells(MoDC) induced by C. albicans, and inhibited the production of TNF-α and IL-6 induced by C. albicans. FACS showed that MBL could not only bind to C. albicans but also bind to imDCs in a Ca2+-dependent manner. Western blot showed that MBL could decrease the phosphorylation of IκBα and the nuclear translocation of p65/ NF-κB.</p><p><b>CONCLUSION</b>MBL may inhibit TNF-α and IL-6 production induced by C. albicans in DC through NF-κB signaling pathways, suggesting that MBL can play some roles in the regulation of C. albicans-induced immune response.</p>


Subject(s)
Humans , Candida albicans , Cell Differentiation , Cytokines , Dendritic Cells , Mannose-Binding Lectin , NF-kappa B , Protein Transport
7.
Journal of Experimental Hematology ; (6): 770-774, 2013.
Article in Chinese | WPRIM | ID: wpr-332696

ABSTRACT

The study was aimed to investigate the mechanism of mannan-binding lectin (MBL) on bacterial lipopolysaccharide (LPS)-induced human peripheral blood monocyte-derived dendritic cell (DC) maturation. The monocytes were prepared from the peripheral blood of healthy adult volunteers. The immature dendritic cells (imDC) were induced by 5-day-culture in medium supplemented with rhGM-CSF and rhIL-4. FACS was used to investigate the interaction of MBL with imDC and the impact of MBL on LPS binding to imDC. ELISA and Western blot was used to analyze the interaction of MBL with soluble TLR4 ectodomain protein (sTLR4); Western blot was used to detect LPS-induced NF-κB translocation in imDC. The results showed that MBL could directly bind to imDC in the presence of calcium. sTLR4 protein or LPS could competitively inhibit the binding of MBL to imDC. ELISA and Western blot showed that MBL could evidently bind to sTLR4 protein in a concentration-dependent manner. FACS showed that MBL could competitively inhibit the binding of LPS to imDC by binding to imDC directly. Western blot showed that MBL decreased LPS-induced NF-κB translocation in imDC. It is concluded that MBL may competitively inhibit the binding of LPS to imDC by binding to TLR4 expressed on imDC, resulted in inhibition of LPS-induced DC maturation, suggesting that MBL can regulate DC maturation through ligand-binding. This study provides the good foundation to clarify the mechanism of MBL inhibiting the LPS-induced DC maturation.


Subject(s)
Humans , Cell Differentiation , Cells, Cultured , Dendritic Cells , Cell Biology , Metabolism , Ligands , Lipopolysaccharides , Mannose-Binding Lectin , Pharmacology , Monocytes , Cell Biology , Metabolism , Toll-Like Receptor 4 , Metabolism
8.
Chinese Journal of Integrated Traditional and Western Medicine ; (12): 398-402, 2010.
Article in Chinese | WPRIM | ID: wpr-231513

ABSTRACT

<p><b>OBJECTIVE</b>To investigate, from cytoprotein and molecular levels, the action mechanism of astragalus injection (ASI) on the signal conduction of human renal tubular cells (HK-2) injury induced by neonatal postasphyxial-serum (NPS), whether it is through activating the nuclear factor kappaB (NF-kappaB) signaling pathway.</p><p><b>METHODS</b>Taking HK-2 as the target cell and the 20% NPS as the attacking factor, the experiment was conducted by dividing the target cells into two groups before attacking, the blank control group and the ASI pretreated group. The nuclear translocation of NF-kappaB was detected by confocal microscopy with indirect immunofluorescence stain, and the amount of NF-kappaB inhibitor subunit (I-kappaBalpha) was detected by Western blot before attacking. The detections were repeated at various time points in the experiment, i.e., 15 min, 1 h and 2 h after attacking, respectively.</p><p><b>RESULTS</b>Before attacking, the nuclear translocation of NF-kappaB and the amount of I-kappaBalpha were not different in the two groups. But the former increased and the latter decreased significantly in the ASI group at all the time points after attacking with the topmost changes presented at 1 h after attacking, and significantly different to those in the control group at corresponding time</p><p><b>CONCLUSION</b>ASI pretreatment could inhibit the activation of NF-kappaB induced by postasphyxial-serum.</p>


Subject(s)
Humans , Infant, Newborn , Apoptosis , Asphyxia Neonatorum , Blood , Astragalus Plant , Cell Line , I-kappa B Proteins , Metabolism , Kidney Tubules , Cell Biology , Metabolism , NF-KappaB Inhibitor alpha , Signal Transduction
9.
Chinese Journal of Contemporary Pediatrics ; (12): 633-636, 2008.
Article in Chinese | WPRIM | ID: wpr-317373

ABSTRACT

<p><b>OBJECTIVE</b>To study the role of serum from asphyxiated neonates in the inducement of human renal proximal tubular epithelial cells (HK-2) adhesion to neutrophils and possible mechanisms.</p><p><b>METHODS</b>HK-2 cells were cultured randomly with 20% serum from neonates (1, 3, and 7 days after asphyxia), pyrrolidine dithiocarbamate (PDTC) or placebo. The activity of myeloperoxidase (MPO), an indicator of adhesion ability of HK-2 cells to neutrophils in suspensions, was detected by the biochemistry assay. Intercellular adhesion molecule-1 (ICAM-1) and nuclear factor-kappaB (NF-kappaB) of HK-2 cells were examined with the immunohistochemical staining.</p><p><b>RESULTS</b>The expression of MPO in the post-asphyxial 1-day serum treatment group were significantly higher than that in the PDTC treatment and the control groups as well as the post-asphyxial 3 and 7-day serum treatment groups (P<0.01). The expression of ICAM-1 and NF-kappaB in the post-asphyxial 1-day serum treatment group was also significantly higher than that in the other groups (P<0.01).</p><p><b>CONCLUSIONS</b>Serum from asphyxiated neonates can induce HK-2 cell adhesion to neutrophils, possibly through activating NF-kappaB and increasing the synthesis and expression of ICAM-1 on the surface of renal tubular epithelial cells.</p>


Subject(s)
Humans , Infant, Newborn , Asphyxia Neonatorum , Blood , Cell Adhesion , Cells, Cultured , Intercellular Adhesion Molecule-1 , Kidney Tubules , Pathology , NF-kappa B , Metabolism , Neutrophils , Physiology
10.
Journal of Southern Medical University ; (12): 731-735, 2008.
Article in Chinese | WPRIM | ID: wpr-280108

ABSTRACT

<p><b>OBJECTIVE</b>To obtain highly purified tetanus toxin fragment C (TTC) with good immunogenicity.</p><p><b>METHODS</b>The gene fragment encoding TTC was amplified from Clostridium tetani plasmid DNA by PCR, inserted into the vector pET43.1a (+) and expressed in E. coli BL21(DE3)plysS. After purification using Ni2+-chelate affinity chromatography, the expressed fusion protein was digested by thrombin and the resultant TTC protein was purified with Ni2+-chelate affinity chromatography followed by identification with SDS-PAGE and Western blotting. The purifed TTC protein was then used to immunize mice to test its immunogenecity.</p><p><b>RESULTS</b>The 1373-bp gene fragment encoding TTC was obtained, and the constructed recombinant expression vector pET43.1a (+)-TTC was successfully expressed in E. coli BL21(DE3)plysS. SDS-PAGE identified a recombinant fusion protein with relative molecular mass (Mr) of 117 000, which accounted for 22% of the total bacterial protein. The TTC protein with Mr of 50 000 was obtained after purification of the thrombin digestion products of the fusion protein, with a purity reaching 95.5%. Both the fusion protein and TTC protein could be recognized by anti-tetanus toxin antibody as shown by Western blotting. The titer of the anti-serum from mice immunized with the TTC protein was 1:25 600, and the anti-serum could specifically bind to tetanus toxin.</p><p><b>CONCLUSION</b>Highly purified and immunogenetic TTC protein has been successfully obtained, which provides a good model antigen for studying antigen presentation and immune responses in vivo.</p>


Subject(s)
Cloning, Molecular , Electrophoresis, Polyacrylamide Gel , Escherichia coli , Genetics , Metabolism , Genetic Vectors , Peptide Fragments , Genetics , Allergy and Immunology , Recombinant Fusion Proteins , Genetics , Allergy and Immunology , Tetanus Toxin , Genetics , Allergy and Immunology , Tetanus Toxoid , Allergy and Immunology
11.
Chinese Journal of Contemporary Pediatrics ; (12): 559-562, 2007.
Article in Chinese | WPRIM | ID: wpr-325671

ABSTRACT

<p><b>OBJECTIVE</b>Salvia miltorrhiza Bunge (SMB) is a traditional Chinese herb, which is considered to promote blood flow and remove blood stasis. This study examined whether SMB can alleviate injury induced by hypoxia/reoxygenation (H/R) in human kidney proximal tubular cells-2 (HK-2 cells).</p><p><b>METHODS</b>There were 3 experimental groups: control, H/R injury and SMB-treated H/R injury. H/R injury of HK-2 cells was induced by first covering the cells with and then removing liquid paraffin wax. Different concentrations of compound SMB solution (0.05%, 0.10%, 0.15% or 0.20%) were administered to the SMB-treated H/R injury group before the hypoxic injury. After 4, 12 and 24 hrs of hypoxia and 4, 12, 24 and 48 hrs of reoxygenation, morphologic changes of HK-2 cells were observed under an inverted microscope. Cell viability was measured by the MTT method. Lactate dehydrogenase (LDH) activity in the culture supernatants was assayed using biochemical methods; TNF-alpha levels were determined by radioimmunoassay (RIA).</p><p><b>RESULTS</b>The number of HK-2 cells was significantly reduced in the H/R injury group after hypoxia, and reached a nadir 24 hrs after hypoxia treatment. Various concentrations of SMB-treated groups showed significantly greater number of HK-2 cells than the H/R injury group. SMB solution (0.10%) produced the best effect. The levels of LDH and TNF-alpha in the H/R injury group were significantly increased, and reached a peak between 24 hrs of hypoxia and 4 hrs of reoxygenation when compared to the control group. Pre-treating with 0.10% SMB resulted in significantly lower levels of LDH and TNF-alpha than in the untreated H/R injury group at various time points of H/R.</p><p><b>CONCLUSIONS</b>SMB has protective effects against H/R injury of HK-2 cells, possibly through inhibition of inflammatory cytokines.</p>


Subject(s)
Humans , Calcium , Metabolism , Cell Hypoxia , Cells, Cultured , Cytoprotection , Drugs, Chinese Herbal , Pharmacology , Kidney Tubules, Proximal , Pathology , L-Lactate Dehydrogenase , Metabolism , Plant Extracts , Pharmacology , Salvia miltiorrhiza , Tumor Necrosis Factor-alpha
12.
Journal of Applied Clinical Pediatrics ; (24)2006.
Article in Chinese | WPRIM | ID: wpr-639769

ABSTRACT

Objective To explore the protective effects of astragalus on injury of renal tubular cells(HK-2) induced by postasphyxial-serum from neonate.Methods Human renal proximal tubular cell line HK-2 cells were taken as subjects.The experiment was designed as:control group,model group,astragalus pretreatment group.The astragalus pretreatment group had 5 subgroups,which were pretreated for 24 hours.The serum of neonates who suffured asphyxia for 1 day,which concentration were 200 mL/L(volume fraction),were applied as attacking factor.The injury of morphology were observed under inverted microscope.The cell viability was measured by methyl thiazolyl tetrazolium methods.The leakage rate of lactate dehydrogenase(LDH) was determined by biochemical methods.Results Compared with control group,cell morphous changed,leakage rate of LDH elvated and cell survival rate decreased in model group,the variances were obviously significant(Pa

13.
Journal of Applied Clinical Pediatrics ; (24)2006.
Article in Chinese | WPRIM | ID: wpr-638889

ABSTRACT

Objective To investigate the role of postasphyxial-serum of neonate in inducing injury of human renal proximal tubular cells(HK-2 cells).Methods HK-2 cells were used as target cell.The neonatal different concentration postasphyxial-serum of 1,3,7 days after asphyxia were used as attacking means.The experimental groups were divided into 15 groups:the 2.5%,5.0%,10.0%,(20.0%) attacking concertration groups of 1,3,7 day after asphyxia and control group of each concertration.The culture medium and concertration of the control group and the experimental groups were the same.The changes of morphology were observed under inverted microscope,the cell viability was measured by 3-(4,5-dimethyl-2-thiazoly1)-2,5-diphenyl-2H-tetrazolium bromide(MTT) method and the leakage rate of lactate dehydrogenase(LDH) was determined by biochemical methods.Results Compared with control group,the changes in morphology of HK-2 were most serious and obvious,the cell viability were obviously decreased(all P

14.
Journal of Applied Clinical Pediatrics ; (24)2004.
Article in Chinese | WPRIM | ID: wpr-638630

ABSTRACT

Objective To investigate the role and intracellular signal transduction mechanism in the injury of renal tubular cells induced by postasphyxial-serum in neonate.Methods Human renal proximal tubular cell(HK-2 cell) was used as target cell. The experiment was designed as:control group, asphyxia group ,and pyrrolidine dithiocarbamate (PDTC)blocking group. The attacking concentration of serum was 20%, and the apoptosis rate of HK-2 cells was detected by flow cytometer.Results Compared with controls[(13.3?1.70)%],after being stimulated with postasphyxial-serum, the apoptosis rate of HK-2 cells of asphyxia group [(46.73?3.68)%] and PDTC blocking group [(31.19?2.79)%]were significantly increased(P

15.
Journal of Applied Clinical Pediatrics ; (24)1992.
Article in Chinese | WPRIM | ID: wpr-639903

ABSTRACT

Objective To explore the influence of erythropoietin(EPO)on signal conduction mechanism of injury of renal tubular cells(HK-2) induced by postasphyxial-serum of neonates.Methods Heavy term asphyxia neonate were selected which amounted to 50 cases(28 male and 22 female)who had no obvious infectious diseases and immune suppression preparation,and their family consented to drawing their venous blood 5 mL after asphyxia 1 day,and put on centrifuge for 10 minutes with 2 500 r/min,then in aqueous bath for 30 minutes to inactivate complement at 56 ℃,and undertook filtration sterilization by micropore filter.The attacking concentration of serum was 200 mL/L which was prepared by adopting Dulbecco′s Modified Eagle Medium/Nutrient Mixture F12 culture fluid.HK-2 was used as the target cel1.Before attack,the experiment was designed as:blank control group,group of pretreatment with EPO(5?104 IU/L EPO medium preconditioned for 24 hours).After attack,at the 15 minutes,1 hour,2 hours,experiment was designed as:attacking control group and attacking group preconditioned with EPO at each time point.The nuclear translocation of nuclear factor ?B(NF-?B) was detected using indirect immunofluorescence.The amount of I-?B? change by NF-?B restraining was detected by Western blot,the results were scanned into computer,and image analysis software of Image-Pro Plus was used to analyze the strap.The strap integral optical density value represented the quantity of protein.Results Before attack,compared with bland control group,the nuclear translocation of NF-?B increased and amount of I-?B? decreased in group of pretreatment with EPO(Pa

16.
Journal of Applied Clinical Pediatrics ; (24)1992.
Article in Chinese | WPRIM | ID: wpr-639565

ABSTRACT

Objective To explore the effect of postasphyxial-serum in neonate on expression of serine protease Omi/HtrA2 in renal tubular cells(HK-2).Methods Human renal proximal tubular cell line HK-2 cell was used as target cell.The cultural cells in orifice were divided into control group and asphyxia-serum attacking group.Blood was cowected from asphyxia newborns by means of femoral venous puncture,then the serum was garthered,anticoagulated by liquemie,3 000 r/min centrifuged 20 min,abstracted serum,thermostatic waterbathed the serum at 56 ℃,so that to inactivate addiment,filtered germ by micropore filte,the attacking concentrtion of serum was 200 mL/L,the cells of the asphyxia-serum attacking group were attacked by asphyxia-serum,and the cells of control group were cultivated with normal nutritive medium when the cells was needed.After 24 hours,the cells were tixed,then the expression of Omi/HtrA2 in cytoplast was detected by the use of immunohistochemical method.Results Omi/HtrA2 was inaurate or yellow brown and localized to the cytoplast.The rate of the cell expressed Omi/HtrA2 was(9.0?2.5)% in control group,after stimulated with postasphyxial-serum,in asphyxia group the rate of the cell expressed Omi/HtrA2 was(25.15?3.5)%,there was significant difference between 2 groups(t=-15.322 P

17.
Journal of Applied Clinical Pediatrics ; (24)1992.
Article in Chinese | WPRIM | ID: wpr-639124

ABSTRACT

Objective To investigate the role of erythropoietin(EPO)in relieving the injury of human renal tubular cells (HK-2) induced by postasphyxial-serum of neonates.Methods Human renal proximal tubular cell(HK-2) was used as the target cell.The experiment was designed as control group, asphyxia group,and group of pretreatment with EPO. The attacking concentration of serum was 200 mL/L,then the changes of morphology were observed under inverted microscope,and the cell viability was measured by 3-(4,5-dimethy lthiazcl-2-yl)-2,5-diphenyl-tetazolium bromide(MTT) methods,and the leakage rate of lactate dehydrogenase(LDH) was determined by biochemical methods.Results Compared with control group,the change in morphology of HK-2 was most serious and obvious,and the leakage rate of LDH increased significantly,and the cell viability decreased obviously in asphyxia group.But compared with asphyxia group,the change in morphology of HK-2 was obviously improved,and the leakage rate of LDH decreased and the cell viability increased in group of pretreatment with EPO in a dose-dependent manner except the group of 1 IU/mL.Conclusion EPO can play the role in relieving the injury of renal tubular cells induced by postasphyxial-serum in neonates.

18.
Journal of Applied Clinical Pediatrics ; (24)1986.
Article in Chinese | WPRIM | ID: wpr-639498

ABSTRACT

Objective To explore expression of hypoxia inducible factor-1?(HIF-1?)in kidney cells during hypoxia/reoxygenation injury,and to study the effect of Danshen on prevention of the hypoxia/reoxygenation injury to cells.Methods Human renal proximal tubular cells(HK-2)were used as the target cell.There were 3 groups:control group,model group,Danshen group.hypoxia/reoxygenation models after neonatal asphyxia were established with liquid paraffin covering method.Expression of HIF-1? were detected with strcp avidin biotin complex(SABC)immunohistochemistry at following different time points:hypoxia 1,4,8,12,24 h,which means 1,4,8,12,24 h respectively after hypoxia;and hypoxia/reoxygenation 1,4,8,12,24 h,which means 1,4,8,12,24 h respectively after hypoxia/reoxygenation.Results HIF-1? was expressed mainly in HK-2's nucleus.There had low expression of HIF-1? in HK-2 cells under the normal culture,and its expression level kept rising quickly during hypoxic/reoxygenatic culture,until 4 h after the beginning of reoxygenation.Compared with the study group,the expression level of HIF-1? in HK-2 cells of the Danshen group were significantly lower at different time points(Pa

19.
Journal of Applied Clinical Pediatrics ; (24)1986.
Article in Chinese | WPRIM | ID: wpr-639497

ABSTRACT

Objective To explore the effect of postasphyxial-serum in neonate on the expressions of Bcl-2-antagonist of cell death(BAD)and Bcl-2-associated X protein(BAX)in renal tubular cells(HK-2).Methods HK-2 cells were used as target cells.The experiment were divided into control group,asphyxia group and pyrrolodine dithiocarbamate(PDTC)blocking group.Control group:DMEM culture fluid was not contained asphyxia blood serum in every group;asphxia group:DMEM culture fluid contained 20 mL/L asphyxia blood serum in every group;PDTC blocking group:DMEM culture fluid contained 20 mL/L asphyxia blood serum and 40 ?mol/L PDTC in every group.The expressions of both BAD and BAX on cytoplast were detected by immunohistochemical method.Results Calculated Points according to HSCORE,compared with controls group[(1.97?0.26)and(1.77?0.11)],after stimulated with postasphyxial-serum,the expressions of both BAD and BAX of HK-2 cells of asphyxia group[(2.73?0.20)and(2.44?0.13)] and PDTC blocking group[(2.38?0.13)and(2.17?0.08)] significantly increased[F(BAD)=28.61,F(BAX)=15.51 Pa

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