RG50864对PDGF诱导的肺动脉平滑肌细胞酪氨酸磷酸化蛋白表达的影响

目的 :研究酪氨酸蛋白激酶抑制剂 RG5 0 86 4对 PDGF诱导的肺动脉平滑肌细胞 (PASMC)酪氨酸磷酸化蛋白表达的影响。方法 :用原代培养的小牛肺动脉平滑肌细胞 ,采用 Western Blot方法分析 RG5 0 86 4对 PDGF诱导的PASMC酪氨酸磷酸化蛋白的定性表达。结果 :RG5 0 86 4 (0 .1 μmol/ L)能显著抑制 PDGF诱导的 PASMC酪氨酸磷酸化蛋白的表达 ,尤以 85～ 1 80 KDa明显 ,当 RG5 0 86 4浓度达 1 0 0 μmol/ L 时 ,则能完全抑制磷酸化蛋白的产生 ,其作用呈明显的剂量效应关系。结论 :RG5 0 86 4能显著地抑制 PDGF诱导的 PASMC酪氨酸磷酸化蛋白的表达     

EFFECT OF PHORBOL ESTER ON cAMP-DEPENDENT PROTEIN KINASE ACTIVITY IN CARDIOMYOCYTES

Cardiomyocytes isolated from neonatal rats were treated with phorbol-12-myristate-13-acetate(PMA) ranging from 10^-11 to 10^-7mol/L for 20 min,causing cytosol protein kinase A (PKA) activity to decrease while particulate PKA activity increase in a concentration-dependent manner.The change of PKA activity induced by PMA was abolished completely by pretreatment of polymyxin B or depletion of protein kinase C (PKC).Type Ⅱ PKA activity in particulate fraction was enhanced remarkably,while that of type I PKA was not altered when the cells were treated with 100 nmol/L PMA.The results suggested that subcellular distribution and activity of PKA in cardiomyocytes may be regulated by PKC.     

Paeoniflorin Promotes Angiogenesis in A Vascular Insufficiency Model of Zebrafish in vivo and in Human Umbilical Vein Endothelial Cells in vitro

Objective: To investigate the pro-angiogenic effects of paeoniflorin (PF) in a vascular insufficiency model of zebrafish and in human umbilical vein endothelial cells (HUVECs). Methods: In vivo, the pro-angiogenic effects of PF were tested in a vascular insufficiency model in the Tg(fli-1:EGFP)y1 transgenic zebrafish. The 24 h post fertilization (hpf) embryos were pretreated with vascular endothelial growth factor (VEGF) receptor tyrosine kinase inhibitor Ⅱ (VRI) for 3 h to establish the vascular insufficiency model and then post-treated with PF for 24 h. The formation of intersegmental vessels (ISVs) was observed with a fluorescence microscope. The mRNA expression of fms-like tyrosine kinase-1 (flt-1), kinase insert domain receptor (kdr), kinase insert domain receptor like (kdrl) and von Willebrand factor (vWF) were analyzed by real-time polymerase chain reaction (PCR). In vitro, the pro-angiogenic effects of PF were observed in HUVECs in which cell proliferation, migration and tube formation were assessed. Results: PF (6.25–100 μmol/L) could rescue VRI-induced blood vessel loss in zebrafish and PF (25–100 μmol/L) , thereby restoring the mRNA expressions of flt-1, kdr, kdrl and vWF, which were down-regulated by VRI treatment. In addition, PF (0.001–0.03 μmol/L) could promote the proliferation of HUVECs while PF stimulated HUVECs migration at 1.0–10 μmol/L and tube formation at 0.3 μmol/L. Conclusion: PF could promote angiogenesis in a vascular insufficiency model of zebrafish in vivo and in HUVECs in vitro.     

Experimental study on anti-neoplastic activity of epigallocatechin-3-gallate to digestive tract carcinomas

Background Epigallocatechin-3-gallate (EGCG) has been demonstrated to have anti-neoplastic activity, but the effective concentration of EGCG and its possible mechanisms are uncertain. The study on the killing effects of EGCG on different digestive tract cancer cell lines can find target sites of its anti-neoplastic effect and provide a theoretical basis for its clinical application in the treatment of cancers.Methods Methyl thiazolyl tetrazolium (MTT) analysis was made to detect the differential sensitivities of eight digestive tract cancer cell lines to EGCG. The effect of EGCG on cell cycle distribution of sensitive cancer cell line was measured by flow cytometry. By polymerase chain reaction (PCR)-enzyme linked immunosorbent assay (ELISA) protocol, the influence of EGCG on telomerase activity of sensitive cancer cell line was also investigated. RT-PCR method was employed to detect the influence of EGCG on the expressions of hTERT, c-myc, p53 and mad1 genes in sensitive cancer cell line. Results EGCG exhibited dose-dependent killing effects on all eight disgestive tract cancer cell lines. The 50% inhibitory concentration (IC50) of SW1116, MKN45, BGC823, SGC7901, AGS, MKN28, HGC27 and LoVo cells were 51.7 μmol/L, 55.9 μmol/L, 68.5 μmol/L, 79.1 μmol/L, 83.8 μmol/L, 119.8 μmol/L, 183.2 μmol/L and 194.6 μmol/L, respectively. There were no apparent changes in cell cycle distribution of sensitive cancer cell line MKN45 48 hours after incubating with three different concentrations of EGCG compared with the controls. It was found that EGCG could suppress the telomerase activity of MKN45 cells, and the effects were dose- and time-dependent. After EGCG administration, the expression of hTERT and c-myc genes in MKN45 cells was decreased, that of the mad1 gene increased, and that of the p53 gene unchanged. Conclusions EGCG has dose-dependent killing effects on different digestive tract cancer cell lines. Administration of EGCG has no obvious effect on cell cycle distribution of sensitive cancer cell line MKN45. The anti-neoplastic activity of EGCG might be due to the inhibition of telomerase activity by means of its influence on hTERT and the up-stream regulation genes.     

Role of protein kinase C in advanced glycation end products-induced epithelial-mesenchymal transition in renal proximal tubular epithelial cells

The role of protein kinase C （PKC） activation in advanced glycation end products （AGEs）-induced epithelial-mesenchymal transition in renal proximal tubular epithelial cells was investigated. HKC cells were divided into three groups： normal group, AGE-BSA group （100 mg/L AGE-BSA） and AGE-BSA＋PKC inhibitor （10 μmol/L chelerythrine chloride） group. PKC activity was measured by PKC assay kit. The expression of Vimentin, and phosphorylated β-catenin was detected by using Western blotting, and the content of TGF-β1 was examined by ELISA method. The intracellular disposition of Vimentin was observed by fluorescence microscopy. As compared with normal group, PKC activity was increased significantly in AGE-BSA group. The expression of Vimentin, phosphorylated β-catenin, and TGF-β1 was enhanced significantly in AGE-BSA group. The expression of Vimentin, phosphorylated β-catenin, and TGF-β1 was significantly blocked by chelerythrine chloride. High expression of Vimentin, phosphorylated β-catenin, and TGF-β1 induced by AGE-BSA may be mediated via the activation of PKC signal transduction pathway.     

雷公藤甲素对大鼠体内环磷酰胺药代动力学的影响：对细胞色素P3A4可能的抑制作用

Objective:To evaluate the effect of a 10-day course of triptolide(TP) on rat cytochrome(CY)P3A4 activity,and on the pharmacokinetics of cyclophosphamide(CPA).Methods:In the pharmacokinetics experiment,rats were orally given 0.9%NaCI solution(n=5) and TP[1.2(mg/kg·d)]for 10 days and a single dose of CPA was administered intravenously(100 mg/kg) to rats on day 11.Blood samples were collected up to 4 h at predetermined time intervals,the plasma concentration of CPA was determined by high performance liquid chromatography(HPLC) and pharmacokinetic parameters were determined.In the in vitro CYP3A4 activity inhibition research,rat blank liver microsomes were divided into 3 groups:a control group,a TS(5 μ L,200 μmol/L) with TP(5 μL,12.5 μmol/L) group,a TS with ketoconazole(5 μL,1 μmol/L) group.Concentration of 6 β-hydroxylated testosterone(6 β-OHT) in liver microsomes was measured by HPLC and the activity of CYP 3A4 was calculated through the following formula:E_(inhibitor)/E_(control)×100%=C_(inhibitor)/C_(control)× 100%.Results:Compared with the control group,the area under the plasma concentration-time curve(AUC_(0-∞)) of CPA was significantly increased by 229.05%pretreated with TP(P0.01).Peak plasma concentrations(C_(max))of CPA was significantly increased and plasma half-life was correspondingly extended.The CYP3A4 activity was significantly inhibited by ketoconazole 93.5%±0.2%and TP 84.6%±0.3%compared with the control group(P0.01 and P0.05,respectively).Conclusion:Our results strongly suggested that long-term oral intake of TP can distinctly inhibit the CYP3A4 activity and this inhibition evidently decrease the formation of toxic metabolites of CPA.     

Paeoniflorin Promotes Angiogenesis in A Vascular Insufficiency Model of Zebrafish in vivo and in Human Umbilical Vein Endothelial Cells in vitro

Objective: To investigate the pro-angiogenic effects of paeoniflorin(PF) in a vascular insufficiency model of zebrafish and in human umbilical vein endothelial cells(HUVECs). Methods: In vivo, the pro-angiogenic effects of PF were tested in a vascular insufficiency model in the Tg(fli-1:EGFP)y1 transgenic zebrafish. The 24 h post fertilization(hpf) embryos were pretreated with vascular endothelial growth factor(VEGF) receptor tyrosine kinase inhibitor Ⅱ(VRI) for 3 h to establish the vascular insufficiency model and then post-treated with PF for 24 h. The formation of intersegmental vessels(ISVs) was observed with a fluorescence microscope. The m RNA expression of fms-like tyrosine kinase-1(flt-1), kinase insert domain receptor(kdr), kinase insert domain receptor like(kdrl) and von Willebrand factor(v WF) were analyzed by real-time polymerase chain reaction(PCR). In vitro, the pro-angiogenic effects of PF were observed in HUVECs in which cell proliferation, migration and tube formation were assessed. Results: PF(6.25–100 μmol/L) could rescue VRI-induced blood vessel loss in zebrafish and PF(25–100 μmol/L), thereby restoring the m RNA expressions of flt-1, kdr, kdrl and v WF, which were down-regulated by VRI treatment. In addition, PF(0.001–0.03 μmol/L) could promote the proliferation of HUVECs while PF stimulated HUVECs migration at 1.0–10 μmol/L and tube formation at 0.3 μmol/L. Conclusion: PF could promote angiogenesis in a vascular insufficiency model of zebrafish in vivo and in HUVECs in vitro.     

Ginsenoside Rb1 Ameliorates Autophagy of Hypoxia Cardiomyocytes from Neonatal Rats via AMP-Activated Protein Kinase Pathway

Objective: To investigate whether ginsenoside-Rb1(Gs-Rb1) improves the CoCl2-induced autophagy of cardiomyocytes via upregulation of adenosine 5'-monophosphate-activated protein kinase(AMPK) pathway. Methods: Ventricles from 1-to 3-day-old Wistar rats were sequentially digested, separated and incubated in Dulbecco's modified Eagle's medium supplemented with 10% fetal bovine serum for 3 days followed by synchronization. Neonatal rat cardiomyocytes were randomly divided into 7 groups: control group(normal level oxygen), hypoxia group(500 μmol/L CoCl_2), Gs-Rb1 group(200 μmol/L Gs-Rb1 + 500 μmol/L CoCl_2), Ara A group(500 μmol/L Ara A + 500 μmol/L CoCl_2), Ara A+ Gs-Rb1 group(500 μmol/L Ara A + 200 μmol/L Gs-Rb1 + 500 μmol/L CoCl_2), AICAR group [1 mmol/L 5-aminoimidazole-4-carboxamide ribonucleotide(AICAR) + 500 μmol/L CoCl_2], and AICAR+Gs-Rb1 group(1 mmol/L AICAR + 200 μmol/L Gs-Rb1 + 500 μmol/L CoCl_2). Cel s were treated for 12 h and cell viability was determined by methylthiazolyldiphenyl-tetrazolium bromide(MTT) assay and cardiac troponin I(cTnI) levels were detected by enzyme-linked immunosorbent assay(ELISA). AMPK activity was assessed by 2',7'-dichlorofluorescein diacetate(DCFH-DA) ELISA assay. The protein expressions of Atg4 B, Atg5, Atg6, Atg7, microtubule-associated protein 1 A/1 B-light chain 3(LC3), P62, and active-cathepsin B were measured by Western blot. Results: Gs-Rb1 significantly improved the cell viability of hypoxia cardiomyocytes(P0.01). However, the viability of hypoxia-treated cardiomyocytes was significantly inhibited by Ara A(P0.01). Gs-Rb1 increased the AMPK activity of hypoxia-treated cardiomyocytes. The AMPK activity of hypoxia-treated cadiomyocytes was inhibited by Ara A(P0.01) and was not affected by AICAR(P=0.983). Gs-Rb1 up-regulated Atg4B, Atg5, Beclin-1, Atg7, LC3B Ⅱ, the LC3BⅡ/Ⅰ ratio and cathepsin B activity of hypoxia cardiomyocytes(P0.05), each of these protein levels was significantly enhanced by Ara A(all P0.01), but was not affected by AICAR(all P0.05). Gs-Rb1 significantly down-regulated P62 levels of hypoxic cardiomyocytes(P0.05). The P62 levels of hypoxic cardiomyocytes were inhibited by Ara A(P0.05) and were not affected by AICAR(P=0.871). Conclusion: Gs-Rb1 may improve the viability of hypoxia cardiomyocytes by ameliorating cell autophagy via the upregulation of AMPK pathway.     

Antioxidant and α-glucosidase inhibitor activities of natural compounds isolated from Quercus gilva Blume leaves

Objective: To isolate and investigate antioxidant and α-glucosidase inhibitor compounds in the leaves of Quercus gilva Blume(Q. gilva).Methods: Dry leaves of Q. gilva were extracted with methanol and the methanolic extract was further separated by silica gel column chromatography using several solvents with increasing polarity. The antioxidant activities of the isolated compounds were evaluated using various in vitro assays: 1,1-diphenyl-2-picrylhydrazyl radical scavenging activity, hydrogen peroxide radical scavenging activity, β-carotene bleaching assay, and reducing power assay. The α-glucosidase inhibitory assay was conducted against α-glucosidase from Saccharomyces cerevisiae.Results: Three compounds were isolated and their structures were identii ed as catechin(1), epicatechin(2), and tiliroside(3) using an instrumental analysis. Compound 2 had higher antioxidant activity with inhibitory concentrations(IC50) of(22.55 ± 2.23) μmol/L than that of quercetin, which was used as the standard, with an IC50 of(28.08 ± 2.39) μmol/L, followed by compound 1 with IC50 of(40.86 ± 3.45) μmol/L. On the other hand, compound 3 had the lowest antioxidant activity with an IC50 of(160.24 ± 8.15) μmol/L. However, compound 3 had the highest α-glucosidase inhibitory activity with an IC50 of(28.36 ± 0.11) μmol/L, followed by compounds 1 and 2 with(168.60 ± 5.15) and(920.60 ± 10.10) μmol/L, respectively.Conclusions: The results obtained for the antioxidant activities and α-glucosidase inhibitory activities in a methanolic extract from the leaves of Q. gilva coni rmed the potential of this plant as a source of natural antioxidants and antidiabetic medicine.     

Abl-PTK酪氨酸激酶区基因的克隆与表达

目的 克隆、表达出PTK重组蛋白，以鉴定酪氨酸激酶（PTK）的活性，筛选酪氨酸激酶的抑制剂，方法 从PTK重组克隆载体上切下目的基因片段Abl-PTK使其连接到表达载体pGEX4T-2上，转化大肠杆菌DH5a，筛选鉴定出正确的转化子。转化菌株经IPTG诱导后进行表达并进行SDS－PAGE分析。结果 经酶切和诱导表达鉴定，重组质粒pGEX4T-2-PTK构建成功，并高效表达了58KD的GST－PTK融合蛋白。结论 PTK基因被成功地重组至融合蛋白表达载体中，并在大肠杆菌中获得高效表达。该研究为进一步纯化、鉴定PTK的活性，筛选PTK的抑制剂奠定了基础。     

准分子激光治疗性角膜切削术治疗角膜浅层病变的临床研究

目的：探讨准分子激光治疗性角膜切削术（phototherapeutic keratectomy,PTK)的最佳适应证,治疗参数及评价其有效性。方法：对角膜浅层病变30眼,包括角膜营养不良,12眼,感染后角膜瘢痕7眼,钱币状角膜炎6眼,外伤性角膜瘢痕3眼和角膜带状变性2眼行PTK治疗,于术后1周,1,3,6周分别检查裸眼视力（UCVA）,最佳矫正视力（BCVA）和屈光状态等并回顾分析其疗效。结果：术后6个月BCVA与术前比较提高21眼,不变4眼,下降5眼,术后6个月平均远视移动＋1．52D,Haze 0.5级5眼,1级2眼和0级23眼,原发病无1例复发,结论：PTK治疗角膜浅层病变,简单,安全,有效,视不同疾病选择去除和非去除上皮PTK或PTK＋PRK。适应证是角膜病变不超过前1／3角膜深度,以角膜营养不良和带状变性的疗效为佳。     

PTK7在食管鳞状细胞癌中的表达及临床意义

目的研究酪氨酸蛋白激酶-7(PTK7)在食管鳞状细胞癌、癌旁组织中的表达及其与食管鳞状细胞癌的发展、侵袭、淋巴结转移的关系,分析其在食管鳞癌中的诊断价值及预后变化。方法收集食管鳞状细胞癌标本80例及癌旁组织63例,采用免疫组织化学链霉菌抗生物素蛋白-过氧化物酶连结(SP)法检测PTK7蛋白的表达,结合临床相关因素进行χ2检验及Kaplan-meier等统计学分析。结果 80例患者食管鳞状细胞癌组织中有45例PTK7蛋白表达阳性(χ2=50.17,P<0.01),食管癌旁非典型增生及正常鳞状上皮中PTK7蛋白表达均阴性。PTK7表达与食管鳞癌的年龄、性别、分化程度、浸润深度、淋巴结转移、浸润长度、临床分期等比较差异无统计学意义(P>0.05);与5年生存率比较差异亦无统计学意义(χ2=0.2,P>0.1);当肿瘤组织PTK7蛋白表达综合免疫组化评分≥5分时,5年生存率有下降趋势(χ2=3.35,P=0.06)。结论 PTK7表达与食管鳞癌有关,当PTK7综合免疫评分≥5分时,5年生存率有下降趋势利用检测的表达有助于综合判断食管鳞癌的相关性和临床预后     

蛋白激酶C和蛋白酪氨酸激酶活性对缺血再灌注心肌细胞凋亡影响的实验研究

为探讨缺血预适应（Ｉｓｃｈｅｍｉｃ Ｐｒｅｃｏｎｄｉｔｉｏｎｉｎｇ,ＩＰ）早期以及蛋白激酶Ｃ（Ｐｒｏｔｅｉｎ Ｋｉｎａｓｅ Ｃ,ＰＫＣ）及蛋白酷氨酸激酶（Ｐｅｏｔｅｉｎ Ｔｙｒｏｓｉｎｅ Ｋｉｎａｓｅ,ＰＴＫ）激动剂和抑制剂对缺血再灌注（Ｉ／Ｒ）心肌细胞凋亡的影响以及ＩＰ效能发挥通路中央ＰＫＣ和ＰＴＫ的关系。用ＴＵＮＥＬ法检测Ｉ／Ｒ心肌细胞凋亡。结果显示：１、ＩＰ早期能显著降低Ｉ／Ｒ心肌细胞凋亡（     

蛋白酪氨酸激酶活性变化在大鼠胃粘膜损伤修复中的作用及机理

目的 探讨大鼠胃粘膜损伤后蛋白酪氨酸激酶（ＰＴＫ）活性的变化及通过影响细胞核蛋白质磷酸化和转录因子激活蛋白（Ａｃｔｉｖａｔｉｎｇ ｐｒｏｔｅｉｎ １，ＡＰ－１）的活性对胃粘膜修复的作用及意义。方法 以２０ｍｍｏｌ／Ｌ去氧胆酸钠（ＤＯＣ）给ＳＤ大鼠灌胃，损伤胃粘膜，损伤后３小时取胃，抽提分离细胞膜和细胞核组分，以膜结合法检测细胞膜ＰＴＫ活性和细胞核蛋白质的磷酸化，以凝胶电泳漂移实验检测ＡＰ－１活性，     

二甲双胍对慢性暴露于高糖及高游离脂肪酸的HIT-T15细胞胰岛素受体酪氨酸蛋白激酶活性的影响

目的 观察二甲双胍(MF)对慢性高糖和高脂处理后的HIT-T15细胞(β细胞系)胰岛素受体(IRc)酪氨酸蛋白激酶(TPK)活性的影响,探讨MF对β细胞糖脂毒性,即β细胞胰岛素抵抗的改善作用机制.方法 实验分为对照组、对照 MF组、高糖组、高糖 MF组、高脂组、高脂 MF组.将HIT-T15细胞分别接种于含有5.5 mmol/L、16.7 mmol/L葡萄糖(G)及0.5 mmol/L软脂酸的培养液中,培养48 h后,加入2.5 μg/mL MF干预24 h.用放射性酶分析法测定β细胞IRc TPK活性.结果 高糖[(52.5±18.6) pmol/(min·μg), P＜0.01],且与对照组相比差异无统计学意义.MF对对照组HIT-T15细胞IRc TPK活性无明显影响.结论 高糖和高脂可抑制β细胞IRc TPK活性.MF能明显改善受高糖及高脂所抑制的HIT-T15细胞IRc TPK活性,且恢复到接近正常水平.提示MF对糖脂毒性所致β细胞胰岛素抵抗的改善作用可能与增加IRc TPK活性有关.     

姜黄素衍生物对K562细胞酪氨酸激酶活性的影响

目的研究姜黄素（Cur）衍生物对K562细胞酪氨酸酶激活性的影响,从中筛选酪氨酸激酶（PTKs）抑制剂。方法以PGT[聚谷氨酸钠∶酪氨酸（4∶1）]为激酶反应底物包被96孔酶标板,以磷酸化酪氨酸特异性单克隆抗体为第一抗体,应用酶联免疫吸附测定法（ELISA）检测反应体系中加入的K562细胞裂解液PTKs对反应底物PGT的酪氨酸残基磷酸化的程度,以此测定K562细胞裂解液中的PTKs活性。在此反应体系中加入不同浓度的Cur衍生物,观察对PTKs活性的影响。结果对Cur衍生物进行筛选,得到4种PTKs抑制剂,分别为Cur衍生物FM0815、FM0817、FM0822及FM0824。4个Cur衍生物对PTKs活性的抑制均明显强于Cur,且呈量效、时效关系。结论 Cur衍生物FM0815、FM0817、FM0822及FM0824具有显著的PTKs活性抑制作用。     

芳酰基类蛋白酪氨酸激酶抑制剂的合成及其活性研究

目的：以Ex-Rad为先导化合物,设计并合成具有蛋白酪氨酸激酶（PTK）抑制活性的芳酰基类化合物。方法分别以1-[（4-氟苯基）氨基甲酰基]环丙烷羧酸、1-苯基咪唑烷-2-酮为原料合成中间体3a~3d,将中间体与羧酸通过酰氯法合成目标化合物T1~T7。用酶联免疫吸附法（ELISA）测定PTK抑制活性,计算抑制率,筛选出具有抑制PTK活性的化合物。结果合成芳酰基类新化合物7个,结构经1H NMR确证。活性初筛发现化合物T2、T6的抑制活性强于先导化合物。结论合成方法简单,原料价廉易得。ELISA法测定结果表明T2、T6的PTK的抑制活性较强。     

新型酪氨酸激酶抑制剂的合成及其活性评价

目的 以L029为先导化合物,设计合成具有蛋白酪氨酸激酶(PTK)抑制活性的化合物.方法 以L029为原料,通过还原和(或)在其活泼H等位点接入侧链3-二甲基氨基-1-丙烷、乙酸甲酯、丙酸甲酯等基团,设计合成了一系列L029衍生物,并用酶联免疫吸附法(ELISA)测定其PTK活性,计算抑制率.结果 成功合成5个目标化合物,结构经1H NMR和MS确证.其中3个化合物T2、T3、T5具有较强的PTK抑制活性.结论 本文研究的目标化合物合成路线方法简单,反应条件温和,3个目标化合物具有较强PTK抑制活性,为进一步开展此类分子的设计合成提供了参考.     

抗ABL胞内抗体对K562细胞酪氨酸激酶活性的影响

目的:观察人慢性粒细胞白血病(CML)细胞转导抗ABL胞内抗体基因后,对其酪氨酸激酶活性的影响。方法:构建逆转录病毒载体MSCV-ib-IRES-eGFP,转导K562细胞,分选eGFP 的细胞,用RT-PCR检测胞内抗体mRNA的表达,观察细胞内BCR/ABL、c-ABL酪氨酸激酶活性及细胞总蛋白酪氨酸激酶活性的变化。结果:获得表达胞内抗体的K562细胞:K562-ib-eGFP。RT-PCR证实胞内抗体mRNA在K562-ib-eGFP细胞内表达。与对照组相比,K562-ib-eGFP细胞内BCR/ABL和c-ABL蛋白酪氨酸激酶活性及总蛋白酪氨酸激酶活性明显受抑制。结论:转导抗ABL胞内抗体能有效降低CML细胞内ABL酪氨酸激酶活性,为进一步研究打下基础。