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1.
AIM:To express recombinant hCD154-GST fusion protein, to prepare anti-hCD154 monoclonal antibody, and to investigate the effect of anti-hCD154 monoclonal antibody on graft rejection. METHODS AND RESULTS: Total RNA was prepared from human peripheral blood mononuclear cell (PBMC) activated with 10ng/mL PMA and 1 μg/mL PHA for 8h, the total RNA was reversetranscribed to cDNA. The entire coding region and a part of the 3'non-coding regions were amplified by PCR using a pair of primers designed and synthesized according to the sequence of human CD154 gene from gene bank. The amplified product, a 820bp DNA fragment was cloned into pGEX-4T-1 plasmid expressing glutathione S-transferase(GST). The cloned insert was identified by double digestion of the cloned pGEX-4T-1 plasmid with retriction enzymes BamHⅠand EcoRⅠ.The fusion protein expression plasmid of PGEX-4T-1/hCD154 was constructed, then transformed to E coli BL21. The human CD154-GST fusion protein expression was induced by IPTG in BL21. The expression of recombinant 26kD GST and 55kD human CD154-GST fusion protein were confirmed by SDS-PAGE. CONCLUSION: We have express the recombinant human CD154-GST fusion protein. The expressed hCD154-GST fusion protein will be used to prepare anti-hCD154 monoclonal antibody, to investigate the role of anti-CD154 monoclonal antibody on graft rejection. 相似文献
2.
AIM: To construct the eukaryotic expression vector CD80-IgG by fusing the cDNA encoding extracellular portion of murine CD80 to the 5'-terminus of cDNA encoding Fc fragment of murine immunoglobulin G1 and to express the fusion protein in Chinese hamster ovary (CHO) cells. METHODS: The two cDNAs was amplified by PCR respectively from plasmid pcDNA/B7 containing the full-length cDNA of murine CD80 from murine spleen cells, and cloned to the eukaryotic expression vector pcDNA3.0 by directional cloning. The resultant recombinant plasmid pcDNA/CD80-IgG was transfected into CHO cells with liposome transfection reagent. The stably expressing cells were obtained by G418 screening. Western blot, Dot ELISA, and flow cytometry were used to detect the expression of the fusion protein and its immunological activity. RESULTS: DNA sequencing verified the correction of the construction of recombinant plasmid pcDNA/CD80-IgG. The expressed fusion protein was detected in the supernatant of transfected CHO cells and the molecular weight of the protein was similar to what we expected. Its immunological activity was also established. CONCLUSION: The recombinant plasmid pcDNA/CD80-IgG was successfully constructed and it expressed the fusion protein CD80-IgG. 相似文献
3.
AIM: To study the expression of hepatitis B virus core gene in Pichia pastoris and to obtain high-level expressed recombinant HBcAg with good immunoreactivity and high specificity. METHODS: HBV core gene was amplified by PCR from plasmid pHBV1 which contained HBV whole DNA sequence. The PCR product was cloned into pGEM-T vector by TA cloning strategy. After confirmed by DNA sequence analysis, the gene of interest was inserted into the yeast expression vector pPIC9. The recombinant plasmid pPIC9-cAg was constructed and transformed into GS115 by electroporation. The recombinant yeast GS115 was induced by 0.5% methanol. The expressed product was analysed by SDS-PAGE,Western blot and ELISA. RESULTS: The restriction analysis and DNA sequence analysis proved that HBV core gene had already been cloned to yeast expression plasmid pPIC9. The expressed HBcAg existed in SDS-PAGE. Good immunoreactivity and high specificity of the recombinant HBcAg have been proved by ELISA and Western blot. The titre of the recombinant HBcAg in the cell lysate was 1∶12 800. CONCLUSION: The recombinant plasmid pPIC9-cAg was successfully constructed. The recombinant HBcAg with good immunoreactivity and high specificity was successfully expressed in Pichia pastoris expression system and can be applied to further developing HBcAb immunoassay. 相似文献
4.
Construction of an immunoconjugate targeting tissue factor and its effect on colorectal cancer cells
XU Xi RAO Ben-qiang LIANG Qi-wen LIU Dan TAN Huo WANG Jian-ping HUANG Zhen-qian 《园艺学报》2012,28(3):420-426
AIM: To construct a recombinant eukaryotic expression vector pcDNA3.1(+)-hFVII-LC+hIgG1-Fc, and to produce and purify the immunoconjugate hFVII-LC+hIgG1-Fc protein. METHODS: The target sequences were amplified by RT-PCR from hepatic tissue and lymphocyte RNA, and cloned into eukaryotic expression vector pcDNA3.1(+). After confirmed by restriction endonuclease digestion and DNA sequencing, the recombinant plasmid was transfected into CHO-K1 cells by lipofectamine 2000. The transfectant clones were selected by G418 screening. The positive monoclonals were grown in CHO-K1 serum-free medium Excel 301 and the culture medium was collected. The hFVII-LC+hIgG1-Fc protein was purified by affinity Ni-NTA resin. The immunoconjugate was identified by ELISA with tissue factor (TF) affinity and specificity. Induction of NK cell-mediated antibody-dependent cell cytotoxicity(ADCC) was examined in HT-29 colorectal cancer cell line. RESULTS: Human liver tissue and lymphocytes from Han population were used as template for amplification of hFVII-LC and hIgG1-Fc DNA fragments, which were confirmed by sequencing and were exactly the same as those GenBank reported. The eukaryotic expression vector pcDNA3.1(+)-hFVII-LC+hIgG1-Fc was successfully constructed, and 1.3 mg of hFVII-LC+hIgG1-Fc protein could be prepared from 1 liter of Excel 301 serum-free culture medium through Ni-NTA affinity chromatography. The immunoconjugate was specially bound to TF and induced a significant ADCC response in HT-29 cells. CONCLUSION: The human hFVII-LC+hIgG1-Fc recombinant plasmid and the hFVII-LC+hIgG1-Fc immunoconjugate are obtained, which provide the basis for further study of cancer-targeted therapy. 相似文献
5.
AIM: To clone and express the metalloproteinase domain of human von Willebrand factor-cleaving protease (vWF-cp). METHODS: The metalloproteinase domain of human vWF-cp, amplified from the plasmid containing the vWF-cp cDNA gene by using polymerase chain reaction, was cloned into pUC18, and its accuracy was verified by sequencing. Then the domain was inserted into the multiclone site of pET28a(+) and included a 6×His Tag at its amino terminal. After induced by IPTG, the recombinant protein was purified by using a Ni-NTA column and confirmed by Western blot. RESULTS: Comparison of the nucleotide sequence of our cloned domain with the GenBank sequence revealed no difference. High-level expression of the recombinant protein was yielded after 5-hour induction, which amounted to 28% of total bacteria protein in inclusion body. Western blot demonstrated that it possessed high specificity. CONCLUSION: The metalloproteinase domain of vWF-cp was high efficiently expressed in Escherichia coli. This might contribute to the further study of the relationship between its structure and function. 相似文献
6.
AIM: To study the function of activin receptor-like kinase Ⅰ (ALK1) gene in vascular endothelium. METHODS: The human umbilical vein endothelial cells (HUVECs) were cultured, and the change of expression of ALK1, ALK5 in activation of HUVECs was analyzed. The full-leng coding sequence of ALK1 was cloned into pcDNA3.1+using standard protocols. The constructed pcDNA3.1+ALK1 plasmid were transfected into HUVECs. The proliferation and migration of HUVECs were detected by boyden champer and flow cytometry. RESULTS: The expression of ALK1 was up-regulated in resolution. ALK1 promoted the proliferation and migration of HUVECs. CONCLUSION: ALK1 has an important function in remodeling by promoting the proliferation and migration of endothelial cells. 相似文献
7.
AIM:The current study was designed to construct eukaryotic expression vector containing NK4 gene and transfect it into human pancreatic cancer cell lines.METHODS:The recombinant of pcDNA3/hNK4 was digested by restriction enzyme,the NK4 gene was cloned into a high effective eukaryotic expressing plasmid which contains CMV2 immediate early gene promoter and then transiently introduced into the pancreatic cancer cell line SW1990 by lipo fectamine and clonal cel lines that secrete high levels of NK4 protein were isolated.The expression of NK4 was observed by RT-PCR and Western blot,in vitro the vascular endothelial cell proliferation inhibiting activity of NK4 was examined by 3-[4,5-dimethylthiazolzyl]-2,5-diphenyl tetrazolium bromide(MTT)method.RESULTS:A specific expression of NK4 gene mRNA by lipofectamine-mediated transfer exhibited only in SW1990/NK4 cells,Western blot analysis demonstrated that there was positive expression of NK4 protein(50 kD).The NK4 inhibited proliferation of the vascular endothelial cellsin vitro.CONCLUSION:The recombinant of pRC/CMV2-hNK4 is a high effective expressing eukaryotic vector.The bio-engineering product of the NK4 is an angiogenesis inhibitor and may play an important role in the gene therapy for tumor. 相似文献
8.
AIM: To isolate a gene encoding mouse ING4, construct pcDNA3.0-ING4 recombinant eukaryotic expression plasmid and investigate its effects on HeLa cells in vitro. METHODS: The mouse ING4cDNA was amplified by RT-PCR from mouse liver. The eukaryotic expression vector pcDNA3.0-ING4 was constructed by DNA recombination technique. The recombinant plasmid pcDNA3.0-ING4 was identified by PCR, restriction enzyme digestion and DNA sequence analysis, then was transfected into HeLa cells by lipofectamine. The expression was determined by RT-PCR. Apoptosis was detected by fluorescence microscope with Hoechst33258 staining and laser scanning confocal microscope. Cell cycle distribution was measured with flow cytometry. RESULTS: RT-PCR product was about 750 bp specific fragment. Analysis by restricting enzyme digestion and PCR of pcDNA3.0-ING4 recombiant plasmid showed that results were about 750 bp, DNA sequencing revealed that ING4 cloning were successful. With Hoechst fluorescence staining, we found that the percentage of apoptotic rate in HeLa cells transfected with pcDNA3.0- ING4 (21.25%) was higher than that in HeLa cells transfected with pcDNA3.0 (8.91%,P<0.01). Apoptosis was also detected by laser scanning confocal microscope. Cell cycle analysis reavealed the cell number in S phase of HeLa cells transfected with pcDNA3.0- ING4 increased. CONCLUSION: The gene encoding mouse ING4 and construction of pcDNA3.0- ING4 eukaryotic expression vector were successfully obtained, ING4 could enhance apoptosis in HeLa cells. 相似文献
9.
AIM:To construct a eukaryotic expression vector expressing outer membrane lipoprotein LipL41 of Leptospira lai and express it in mammalian cell. METHODS:LipL41 gene was amplified by PCR from genome of Leptospira lai 017 strain, and was subcloned into vector pGEX-4T-1. After sequencing, LipL41 gene digested by restriction endonuclease and cloned into vector pcDNA3. After confirming the correctness of the eukaryotic recombinant vector by restrication enzyme digestion, it was transfected into COS7 cells by liposome. Its expression was analyzed by RT-PCR. RESULTS:A fragment of 1 011 bp was amplified, and sequence analysis showed it had a 98% homology with Leptospira kirschneri. The analysis of restriction enzyme indicated that the eukaryotic recombinant vector was correctly constructed. A specific amplified fragment was showed in the cells transfected with recombinant plasmid by RT-PCR, but the cell transfected with blank plasmid did not show this band. CONCLUSIONS:The LipL41 gene of Leptospira lai was successfully inserted into eukaryotic expression plasmid and the recombinant plasmid expressed the LipL41 mRNA. 相似文献
10.
AIM: To construct the recombination plasmid pcDNA3.1-hERβ with the human estrogen receptor 2 (ESR2) full length cDNA and transfect it into hormone-independent prostate cancer PC-3M cell line, and to study the effects of ESR2 on proliferation in transfected cells. METHODS: The complete cDNA of ESR2 was obtained from human ovary tissue by RT-PCR technique and cloned into eukaryotic expression vector pcDNA3.1 by using gene recombination technique to construct the pcDNA3.1-hERβ recombination plasmid. The plasmid was detected by endonuclease digestion and DNA sequencing and was transfected into PC-3M cells. MTT and FCAS assay were used to test the effects of ESR2 on the ability of proliferation in PC-3M cells. RT-PCR and Western blotting were used to detect the expressions of cyclinD1 and P21Cip1. RESULTS: The results of sequencing and endonuclease digestion demonstrated that the construction of pcDNA3.1-hERβ recombination plasmid was successful. The sequence analysis suggested that the ESR2 sequence detected by PCR was identical to that published in GenBank, and the product of endonuclease was as long as the complete human ESR2 gene. 48 h after transfected the pcDNA3.1-hERβ into PC-3M cells, the expression of ESR2 mRNA and protein levels increased significantly detected by RT-PCR and Western blotting. Compared to the cells transfected with vector as control, the PC-3M cells transfected with pcDNA3.1-hERβ showed that cell population decreased and proliferation activity degraded. FCAS showed that the cells in G0/G1 stage increased and in S stage or G2/M stage decreased. RT-PCR and Western blotting showed that the expression of cyclinD1 gene reduced and expression of P21Cip1 increased. CONCLUSION: The recombination of plasmid pcDNA3.1-hERβ is constructed and transfected into the PC-3M cells successfully. The activity of cell proliferation is inhibited after pcDNA3 transfection.1-hERβ. It is possible that ESR2 inhibits cell proliferation by the expression of proliferation related genes cyclinD1 and P21Cip1. 相似文献
11.
AIM: To construct a prokaryotic expression plasmid containing Epstein-Barr viral (EBV) capsid antigen BFRF3 gene and to observe the application of recombinant BFRF3 protein in the serological diagnosis of nasopharyngeal carcinoma (NPC).METHODS: DNA extracted from the B95-8 cells was used as the templates. Polymerase chain reaction (PCR) was used to generate a DNA fragment of BFRF3 gene, and a 531-bp DNA fragment was inserted into a PGEX-5X-1 vector. The recombinant plasmid was transformed into E.coli BL21 (DE3). The expression of GST/BFRF3 fusion protein was induced by IPTG, identified by both SDS-PAGE and Western blotting, and then purified by glutathione-sepharose beads. The purified recombinant protein was coated to microplate for ELISA detection of EBV-IgA antibody in NPC patients.RESULTS: The GST/BFRF3 fusion protein was successfully expressed in E. coli. The molecular weight of the product was approximately 44 kD. The recombinant fusion protein GST/BFRF3 showed good immunoreactivity. A novel ELISA was established using GST/BFRF3 protein. Serum samples collected from the NPC patients and healthy controls were tested by this ELISA. The sensitivity and specificity of GST/BFRF3 tests for NPC patients were 65％ and 87％, respectively.CONCLUSION: The recombinant protein GST/BFRF3 is expressed in E.coli, and it has diagnostic value for screening of NPC patients. 相似文献
12.
AIM：To investigate the role of epidermal growth factor-like domain 7 (EGFL7) in the pathogenesis and progress of laryngeal carcinoma via constructing a lentiviral expression vector for RNA interference (RNAi) of human EGFL7 gene and assessing the gene-silencing effect of the vector in human laryngeal epidermoid carcinoma (HEp-2) cells. METHODS：Specific RNAi target sequences were designed focused on human EGFL7 gene sequence. The double-stranded oligonucleotides were cloned into the pcDNA6.2-GW/EmGFP-miR plasmid after synthesis and annealing. A positive clone was subcloned into the pLenti6.3-MCS/V5-DEST vector after sequence analysis. The recombinant lentivirus was harvested from 293T cells co-transfected with the positive recombinant plasmid and lentiviral packing materials. HEp-2 cells were infected with the recombinant lentivirus and the cells with stable EGFL7 knockdown were screened by blasticidin selection. EGFL7 mRNA expression in the cells was determined by real-time fluorescence quantitative PCR. RESULTS：A recombinant lentiviral vector expressing short hairpin RNAs (shRNAs) against EGFL7 gene was obtained and confirmed by DNA sequencing. The virus titer was 5×1011 TU/L, and the silencing efficiency was 97%. CONCLUSION：A lentiviral vector targeting human EGFL7 gene, capable of stable EGFL7 gene knockdown in HEp-2 cells, has been successfully constructed, which provides a basis for further study of the relationship between human laryngeal carcinoma and EGFL7 protein. 相似文献
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14.
MaWRKY1是从香蕉果实中克隆出的一个转录因子基因。为进一步研究该基因的功能,制备了MaWRKY1多克隆抗体。选取MaWRKY1基因全长中N端第168 ~ 400个氨基酸之间的包括两个WRKYGQK保守域的cDNA序列,构建了原核表达载体pET-MaWRKY1,并转化到大肠杆菌BL21中诱导表达菌体蛋白。SDS-PAGE电泳检测结果表明,His-MaWRKY1融合蛋白成功获得了高效表达,分子量在26 kD左右。His-MaWRKY1融合蛋白经过Ni-NTA琼脂糖凝胶树脂纯化,SDS-PAGE制备胶割胶富集,电洗脱法纯化后得到的纯化蛋白浓度达到0.5 mg • mL-1。经对新西兰兔进行5次免疫,获得了多克隆抗血清,采用免疫吸附方法对抗血清进行了纯化。将纯化后的抗体通过间接酶联免疫（ELISA）和蛋白质印迹（Western blot）分析,表明所制备的抗体具有很好的效价,效价比为1︰160 000,同时具有良好的灵敏度和特异性。进一步提取香蕉不同组织总蛋白,Western blot检测显示,在分子量26 kD左右处出现特异的蛋白质条带,证明所制备的抗体可以与香蕉WRKY蛋白特异性结合,并且低温可以诱导香蕉果实中MaWRKY1蛋白表达,暗示MaWRKY1蛋白表达可能与果实耐冷性有一定的关系。 相似文献
15.
AIM: To construct pcDNA3.1+/MAGE-3 DNA vaccine and investigate the antigen-specific antitumor immune responses induced by pcDNA3.1+/MAGE-3 DNA vaccine in vivo. METHODS: C57BL/6 mice challenged with B16/MAGE-3 cells were immunized by intramuscular injection of pcDNA3.1+/MAGE-3 DNA vaccine every 10 days. pcDNA3.1+ plasmid and PBS were used as controls. After three cycles of immunization, murine splenic lymphocytes, serum, and tumor were obtained for cytotoxity assay, detections of cytokines (IL-2 and IFN-γ), measurement of MAGE-3 antibody, and tumor inhibitory rates, respectively. RESULTS: The pcDNA3.1+/MAGE-3 DNA vaccine immunized murine lymphocytes induced specific cytotoxicity against B16/MAGE-3 cells. Significantly increased secretions of IL-2 and IFN-γ were detected. The titres of antibody against MAGE-3 were 1∶1 and 1∶20, while controls were negative. The tumor inhibitory rate in pcDNA3.1+/MAGE-3 group was significantly different from that in controls. CONCLUSION: The pcDNA3.1+/MAGE-3 DNA vaccine was constructed successfully. pcDNA3.1+/MAGE-3 DNA vaccine activates both cellular and humoral immune responses, and induces antigen-specific antitumor immune responses in vivo. 相似文献
16.
AIM:To further study the anti-tumor effect of angiostatin, an anti-human angiostatin monoclonal antibody was prepared and identified.METHODS:The hybrodoma techniques were used. The BALB/C mice were immunized with angiostatin. The supernatant of cell culture were collected and screened by ELISA and double immunodiffusion.RESULTS:There cell lines which steadily secreted the anti-angiostatin monoclonal antibody were identified by ELISA and double immunodiffusion. The antibody was IgG1 and specifically recognized angiostatin without crossing reactions to rhIL-2, rhTNF-α, rhIFN-α and serum proteins.CONCLUSION:The antibodies secreted by three hybridoma cell lines identified by several methods were specific antibodies of angiostatin. 相似文献
17.
XIONG Mao-lin SONG Chang LUO Rong-cheng LUO Chao-quan LI Min-you LI Xiu-ying ZHU Zhen-yu 《园艺学报》2005,32(1):154-158
AIM: To construct human GPI-B7-1 fusion protein and investigate the therapeutic potentials in the treatment of tumors. METHODS: The chimeric GPI anchored-B7-1 gene was obtained by overlap PCR and inserted into expressing vector pcDNA3.1, named pc3.1/GPI-B7-1. pc3.1/GPI-B7-1was transfected into CHO cells by lipofectamine 2 000 reagent. The CHO cells, expressing GPI-B7-1 on membranes, were obtained after selecting by G418. That was confirmed by flow cytometry, SDS-PAGE and Western blot. RESULTS: Recombinant vector pc3.1/GPI-B7-1 was successfully constructed and sequence result indicated that it was identical with reference sequence. The protein on transfected CHO cell membrane selected by G418 was confirmed to be GPI-anchored protein by flow cytometry, and GPI-B7-1 approximately 60 kD was conformed by SDS-PAGE and Western blot. CONCLUSION: A large amount of GPI-B7-1 fusion protein was obtained and will be further studied in the treatment of tumors. 相似文献
18.
柑橘溃疡病致病基因PthA核定位信号肽抗血清制备及其抑菌的研究 总被引：1，自引：0，他引：1
用PCR法扩增位于柑橘溃疡病菌致病基因pthA C-末端的3个核定位信号序列,并将其克隆到原核表达载体PET32a(+)上,经双酶切及核酸序列测定重组质粒(PthA-NLS),其序列与GenBank中pthA的相关序列有99.9%的同一性。重组质粒转化大肠杆菌BL21(DE3)后诱导了重组多肽的表达,并用Ni2+-NTA纯化柱得到了48kD的纯化重组多肽。把重组多肽注入免疫Balb/c小白鼠,制备了相应的抗血清,Western Blotting和ELISA分析结果表明,抗血清可特异地结合重组多肽,亦可识别溃疡病菌PthA天然蛋白,获得的抗血清可以用于柑橘溃疡病的检测。利用抗血清与溃疡病菌混合接种离体冰糖橙叶片,发现抗血清能推迟溃疡病菌的致病过程,且病斑比对照小,但未能达到抗病的程度。pthA基因末端核定位信号序列的克隆、原核表达及抗血清的制备为进一步研究pthA的致病机理和研发溃疡病快速分子检测技术奠定了基础。 相似文献
19.
ZHANG Wen-feng ZHANG Ju-feng SHAO Hong-wei WANG Jun-wei MOU Yan-fang HUANG Shu-lin 《园艺学报》2008,24(3):527-531
AIM: To construct the recombinant dicistronic eukaryotic expression vector pDC315-TCA-12-2-TCB-7.1, which containing T cell antigen receptor (TCR) genes TCA-12-2 and TCB-7.1, and to transfer this recombinant vector into 293 cells to investigate the expression of TCA-12-2 and TCB-7.1. METHODS: The TCA-12-2 was obtained by RT-PCR from the T cells and the TCB-7.1 was amplified by PCR from plamid pcDNA3.1-TCB-7.1 that we constructed before. TCA-12-2 and TCB-7.1 was cloned into vector pIRES2-AcGFP1 firstly, then subcloned into vector pDC315. The recombinant plasmid pDC315-TCA-12-2-TCB-7.1 was verified by restriction enzyme digestion and sequencing, the positive recombinant plasmid was transferred into 293 cells using Lipofectamine 2000. The expressions of gene TCA-12-2 and TCB-7.1 were identified by RT-PCR and flow cytometry. RESULTS: Both TCA-12-2 and TCB-7.1 genes were constructed into eukaryotic expression vector pDC315 and the expressions of genes in 293 cells were detected successfully with RT-PCR and flow cytometry. CONCLUSION: The dicistronic expression vector pDC315-TCA-12-2-TCB-7.1 is successfully constructed and expressed. 相似文献
20.
AIM: To investigate the effects of silent information regulator 1 (SIRT1) over-expression on the apoptosis and the level of reactive oxygen species (ROS) in high glucose-induced H9c2 cardiomyocytes. METHODS: H9c2 cardiomyocytes were transfected with empty plasmid (pcDNA3.1-NC) and SIRT1 over-expression plasmid (pcDNA3.1-SIRT1), and then stimulated by high glucose. The H9c2 cells were divided into control group, high glucose group, high glucose + pcDNA3.1-NC group and high glucose + pcDNA3.1-SIRT1 group. The expression of SIRT1 at mRNA and protein levels in each group was determined by qPCR and Western blot. The viability of the cells was measured by MTT assay. The apoptotic rate was analyzed by flow cytometry. The protein levels of phosphatidylinositol 3-kinase (PI3K), phosphorylated PI3K, AKT and phosphorylated AKT were examined by Western blot. RESULTS: SIRT1 was significantly decreased in high glucose-induced H9c2 cardiomyocytes, the cell viability was significantly decreased compared with control group, while the ROS levels and apoptotic rate were increased, and the phosphorylated PI3K and AKT protein levels were down-regulated (P<0.05). Over-expression of SIRT1 significantly promoted the viability of H9c2 cardiomyocytes induced by high glucose, decreased the ROS levels and apoptotic rate, and up-regulated phosphorylated PI3K and AKT protein levels (P<0.05). CONCLUSION: SIRT1 over-expression reverses the decrease in the viability of high glucose-stimulated H9c2 cardiomyocytes, and the increases in apoptotic rate and oxidative stress by regulating PI3K/AKT signaling pathway. 相似文献