肝细胞缺氧预处理细胞保护作用中热休克蛋白70表达的调控机制

目的　研究热休克蛋白 70 (HSP70 )表达与肝细胞缺氧预处理的关系及其调控机制。方法　建立肝细胞缺氧预处理模型 ,应用蛋白激酶C(PKC)抑制剂、激动剂和丝裂原激活的蛋白激酶 (MEK)的抑制剂 ,通过检测PKC磷酸化水平、p44 /4 2丝裂原激活蛋白激酶 (p44 /4 2MAPKs)、HSP70表达量、细胞存活率 ,同时在透射电镜下观察肝细胞超微结构改变 ,研究缺氧预处理与HSP70表达的关系及HSP70表达的影响因素。对相关数据进行统计学处理。结果　和缺氧复氧组 (HR)比较 ,预处理组 (HP)和PKC激动剂组的HSP70和 p44 /4 2MAPKs的表达量显著增加 ,同时PKC磷酸化水平显著增高 ,三者分别为每分钟 (4 2 .63± 4.73 )、(10 9.42± 16.0 9)、(15 2 .47± 19.5 9)pmol/g(P均 <0 .0 1) ,肝细胞结构改变较小 ;和缺氧预处理组相比 ,PKC抑制剂组相应的观察指标呈相反的变化 ,PKC磷酸化活性显著降低 ,为每分钟 (65 .2 8± 5 .3 6) pmol/g(P <0 .0 1) ;MEK抑制剂组HSP70表达量和磷酸化激活的p44 /4 2MAPKs表达量降低 ,肝组织细胞结构出现较明显的改变。结论　效应保护蛋白HSP70在缺氧预处理中参与对肝细胞的保护 ,HSP70受到PKC介导p44 /4 2MAPKs信号通路的调控。PKC处在p44 /4 2MAPKs上游 ,PKC对 p44 /4 2MAPKs起正性调控作用 ;p44 /4 2MAP     

热休克蛋白70对肝硬化大鼠肝脏缺血再灌注损伤保护作用的研究

目的 探讨热休克蛋白70（HSP70）对肝硬化大鼠肝脏缺血再灌注损伤的保护作用。方法Wistar雄性大鼠用四氯化碳皮下注射制成肝硬化模型。分两组:IP组（缺血预处理组）,用 Pringle’s法阻断肝门15min后,恢复血供,关腹;C组（对照组）,只予以开、关腹。48h后,再次阻断肝门30min,恢复血供。用Western blotting法检测IP后6、24、48h肝组织中HSP70的表达水平;测再灌注1h血清生化酶（ALT、AST、LDH）水平;计算术后一周生存率,并行肝脏病理组织学检查。结果 IP组在缺血预处理后24-48hHSP70表达显著增强,呈高水平;而C组中在各时点HSP70均无表达增强。再灌注1h,IP组的ALT、AST、LDH水平显著低于C组（P<0.01或P<0.05,n=7）。术后一周生存率IP组（93.10％,n=29）明显高于C组（73.33％,n=30）（P<0.05）。缺血再灌注后1h,IP组的肝细胞损伤明显轻于C组。结论 HSP70能够减轻肝硬化大鼠肝脏缺血再灌注损伤,提高术后生存率。     

肝缺血预处理细胞保护效应中P44/42MAPKs信号通路的作用

缺血预处理肝脏保护作用机制的研究报道较少，P44／42丝裂原蛋白活化激酶(mitogen activated protein kinases，MAPKs)信号通路是否参与缺血预处理肝脏保护作用尚未定论，为获得可为临床肝脏外科借鉴的资料，本实验拟建立肝细胞体内和体外的缺血(氧)预处理模型，对P44／42 MAPKs的作用进行初步的探讨。     

肝脏及肢体缺血预处理抗大鼠肝缺血再灌注损伤的实验研究

目的研究肝脏缺血预处理(经典缺血预处理IPC)的第一保护窗(FW)与肢体缺血预处理(远端缺血预处理RPC)的第二保护窗(SW)及两者联合应用对大鼠肝脏缺血再灌注(I/R)损伤的保护作用及可能机制。方法大鼠随机分成5组:I/R组不行预处理;IPC组以肝缺血5 min行预处理;RPC组以双后肢缺血5 min,反复3次行预处理;RPC+IPC组先行RPC,24 h后行IPC作预处理;S组仅行开腹,不行其他处理。3个预处理组及I/R组均行肝缺血1 h再灌注3 h。取血用于血清谷丙转氨酶(ALT)与血清谷草转氨酶(AST)检测。切取肝组织用于测定肿瘤坏死因子α(TNF-α)和热休克蛋白70(HSP70)的表达、湿干比(W/D)及观察显微、超微结构的变化。结果与I/R组比较,IPC组,RPC组及RPC+IPC组ALT,AST,W/D及TNF-α阳性表达均明显降低(P0.01),HSP70表达量明显增加(P0.01),肝脏的显微及超微结构损伤减轻;IPC,RPC,RPC+IPC组3组间各项指标差异无统计学意义(P0.05)。结论IPC的FW,RPC的SW及两者联合应用对大鼠肝脏I/R损伤均有明显的保护作用,三者在保护强度上无明显差异。其机制可能与抑制TNF-α的产生、诱导保护性蛋白HSP70的表达、减轻肝脏水肿、改善肝组织微循环有关。     

肝缺血预处理对肝再灌注期间热休克蛋白70的表达及肝脏病理改变的影响

评价缺血预处理对大鼠肝缺血再灌注后肝细胞热休克蛋白70(HSP70)的表达和肝组织形态学的影响。方法:大鼠78只分为3组:缺血预处理组、肝血流持续阻断组及假处理组。用免疫金银法动态观察再灌注24h内HSP70蛋白的表达,同时观察肝脏的病理改变。结果:肝缺血预处理能较早诱导HSP70的表达,肝细胞损伤也相应较轻。结论:肝缺血预处理可增加组织对缺血再灌注损伤的耐受性,该保护作用可能与早期诱导HSP70蛋白的产生有关。     

缺血预处理对大鼠肝脏缺血再灌注损伤的保护机制

目的验证缺血预处理(IPC)对大鼠肝脏缺血再灌注损伤(I/R)的保护作用,探讨一氧化氮(NO)与蛋白激酶C(PKC)在IPC过程中的作用.方法在原位灌流的大鼠肝脏缺血再灌注模型上,观察IPC的保护作用.同时经肠系膜上静脉注射NO前体L-精氨酸和蛋白激酶C特异性激动剂1,2-二辛酸甘油(DOG)以及两者的特异性阻滞剂N-硝基-L-精氨酸甲酯(NAME)和多粘菌素B,来检测NO和PKC在IPC中的关系.结果预处理可阻止血清谷丙转氨酶(ALT)[(200.86±40.30)U/Lvs.(257.65±20.18)U/L],谷草转氨酶(AST)[(211.06±13.59)U/Lvs.(309.17±24.79)U/L],乳酸脱氢酶(LDH)[(824.73±127.11)U/Lvs.(1118.60±82.21)U/L]及脂质过氧化物(LPO)[(0.414±0.069)mmol/mgvs.(0.531±0.054)mmol/mg]水平升高(P＜0.05),而使组织超氧化歧化酶(SOD)保持在较高水平[(10.33±0.88)U/mgvs.(6.01±0.91)U/mg],(P＜0.05).NO与PKC均可模拟预处理的保护效应.结论缺血预处理对大鼠肝脏I/R有明确的保护作用,NO与PKC发挥IPC保护作用的途径不同.     

缺血预处理和Caspase抑制剂对缺血再灌注损伤保护作用的比较

目的　比较缺血预处理和Caspase抑制剂治疗对大鼠肝缺血再灌注的保护作用及其机制。方法　SD大鼠随机分为 6组 :( 1)缺血再灌注1 (IR1 )组 ;( 2 )IR2 组 ;( 3 )缺血预处理1 (IP1 )组 ;( 4 )IP2 组 ;( 5 )Caspase抑制剂治疗1 (T1 )组 ;( 6)T2 组。比较各组大鼠 70 %肝脏 60min或 12 0min缺血 ,在再灌注 3h时的肝组织Caspase 3活性 ,肝细胞凋亡率和血清AST和ALT水平 ,及实验动物 7d存活率。结果　在 60min缺血及 3h再灌注时间 ,IP1 组和T1 组的保护作用相同 ,在 12 0min缺血及 3h再灌注时 ,T2 组对Caspase活性和肝细胞凋亡的抑制优于IP2 组 (P <0 .0 1) ,但两者的AST和ALT水平及动物 7d存活率均无显著性差异。结论　缺血预处理和Caspase抑制剂治疗对鼠肝缺血再灌注损伤都有保护作用 ,两者的保护效果无显著性差异。缺血预处理对缺血再灌注损伤的保护更简便、经济、安全 ,临床应用前景十分广阔。     

缺血预处理对大鼠肝脏缺血再灌注损伤的延迟性保护作用及其机制的初步探讨

目的探索减轻肝脏缺血再灌注损伤的有效措施及其机制.方法健康Wistar大鼠36只随机分为两组A组为缺血预处理组;B组为对照组.A组行缺血预处理,B组仅行开关腹.48 h后,再次开腹,先检测肝组织HSP70的表达,接着阻断肝左、中叶入肝血流45 min,再灌注90 min后检测肝组织TNF-α表达,分别于缺血前、缺血再灌注后检测血清ALT及镜下肝组织改变.结果 A组ALT水平明显低于B组.肝组织病变较B组轻.A组肝细胞HSP70表达明显,而B组HSP70表达不明显,A组肝脏TNF-α表达明显弱于B组.结论缺血预处理能减轻肝脏缺血再灌注损伤,这种保护作用是通过诱导肝细胞HSP70产生所致,HSP70能抑制肝组织TNF-α的产生.     

乙醇预处理与HSP70的诱导及对肝脏缺血再灌流损伤的影响

目的探讨乙醇预处理对肝脏缺血再灌流损伤的影响以及与热休克蛋白70诱导的关系。方法雄性成年Wistar大鼠232只,胃饲乙醇浓度40%,剂量为5 g/kg体重,随机分为5组:正常对照组(N组)、胃饲乙醇组(E组)、缺血组(IR组)、生理盐水预处理组(NPC组)、胃饲乙醇预处理组(EPC组)。动物手术采用门静脉转流下的肝脏缺血模式,肝门阻断时限为90 min,于再灌流0、13、、6、12、24、72 h活杀留取血液及肝脏标本。结果EPC组3、6 h血清ALT(1 230.88±132.50、888.88±126.67)IU/L、AST(1 866.38±61.77)(、1 433.88±42.74)IU/L均明显低于IR组及NPC组,肝脏病理改变较轻,而肝组织HSP70含量高于后者。结论乙醇预处理可以减轻大鼠肝脏90 min的缺血再灌流损伤,其肝脏保护作用与肝组织HSP70升高相一致,HSP70可能是其发挥肝脏保护的物质基础之一。     

药物和缺血预处理对肝硬化肝脏缺血再灌注损伤的协同保护作用

目的探讨阿霉素和缺血联合预处理对肝硬化肝脏缺血再灌注损伤的保护作用及其可能机制方法(1)诱导大鼠肝硬化模型；(2)缺血再灌注损伤前3组肝硬化大鼠分别行阿霉素预处理、缺血预处理、阿霉素 缺血联合预处理，比较3组和对照组AST、ALT、LDH和盯、TNF-α、NO、热休克蛋白70(HSP0)差异有显著性。结果阿霉素预处理、缺血预处理、阿霉素 缺血联合预处理能明显抑制AST、ALT、LDH水平升高，其中以阿霉素 缺血联合预处理作用最显著；缺血预处理能显著降低ET、TNF-α水平；阿霉素预处理和缺血预处理使N0显著升高；阿霉素预处理能使肝细胞HSP70显著增加。结论阿霉素和缺血预处理都能减轻肝硬化肝脏缺血再灌注损伤程度；阿霉素 缺血联合预处理对月十硬化肝脏缺血再灌注损伤有协同保护作用。     

The inhibitory effects of sevoflurane on angiotensin II- induced, p44/42 mitogen-activated protein kinase-mediated contraction of rat aortic smooth muscle

Sevoflurane dilates blood vessels and reduces arterial blood pressure in a dose-dependent manner. Angiotensin II (Ang II) is one of the primary regulators of vascular tension and arterial blood pressure, and the p44/42 mitogen-activated protein kinases (p44/42 MAPK) are involved in Ang II-mediated vascular smooth muscle contraction. We designed this study to examine the effects of sevoflurane on Ang II-induced, p44/42 MAPK-mediated contraction of rat aortic smooth muscle. The effects of the p44/42 MAPK kinase (MEK1/2) inhibitor, PD 098059 (10(-5) molar [M], 5 x 10(-5) M and 10(-4) M), and sevoflurane (1.7%, 3.4%, and 5.1%) on Ang II-induced contraction and p44/42 MAPK phosphorylation were tested in rat aortic smooth muscle, using isometric force measurement and Western blot analysis, respectively. Ang II induced both a transient contractile response and phosphorylation of p44/42 MAPK, which were significantly attenuated by PD 098059 (P < 0.05-0.01). Sevoflurane inhibited Ang II-induced contractile response in a dose-dependent manner (P < 0.05 and 0.01 in response to 3.4% and 5.1% sevoflurane, respectively). Sevoflurane also dose-dependently depressed Ang II-elicited p44/42 MAPK phosphorylation (P < 0.01 in response to 3.4% and 5.1% sevoflurane). These results suggest that the inhibitory effect of sevoflurane on Ang II-induced vasoconstriction is, at least in part, caused by the inhibition of the p44/42 MAPK-mediated signaling pathway. IMPLICATIONS: The present study demonstrates that sevoflurane can dose-dependently inhibit both angiotensin II (Ang II)-induced contraction and p44/42 MAPK phosphorylation of rat aortic smooth muscle. These data suggest that sevoflurane-produced inhibition of Ang II-induced vasoconstriction is, at least in part, caused by depression of the p44/42 MAPK-mediated signaling pathway.     

PKCδ在GH3细胞ghrelin/pERK1/2信号通路中的作用及其机制

目的 观察在垂体腺瘤GH3细胞ghrelin/pERK1/2信号通路中发挥关键作用的蛋白激酶C(PKC)亚型,探讨该途径的详细信号转导机制.方法 4种不同PKC亚型抑制剂分别作用于GH3细胞,Western blot检测pERK1/2表达,初步判定在上述信号通路中发挥重要作用的PKC亚型;然后设计、合成、转染针对初步筛选出的PKC亚型的特异性siRNA,检测pERK1/2表达,从而明确介导该信号途径的关键PKC亚型.免疫共沉淀研究与该亚型可能相互作用的蛋白激酶.结果 综合分析4种抑制剂对pERK1/2表达的影响,发现nPKC在该信号通路中起关键作用.PKCδ沉默后,ghrelin诱导的pERK1/2高表达明显降低(P＜0.05),而PKCε、PKCθ沉默后对ghrelin诱导的pERK1/2高表达则无明显影响(P＞0.05).ghrelin作用后,与PKCδ结合的raf-1蛋白明显增加.结论 PKCδ为调控GH3细胞ghrelin/pERK1/2信号通路的关键信号分子,并且可能在raf-1水平与受体酪氨酸激酶/ERK1/2信号通路发生"交叉通讯".     

Activation pattern of mitogen-activated protein kinases in early phase of different size liver isografts in rats.

Mitogen-activated protein kinases (MAPK) play a pivotal role in ischemia reperfusion injuries of heart and liver, but the activation pattern of MAPKs in the early phase of different size liver isografts remains unclear. The experiment is designed to investigate the activation pattern and role of MAPKs in isografts of the rat with different size liver transplantation. The animal models of different size graft liver transplantation (whole graft, 50% size, or 30% size, respectively) were established and the sham operation group served as a control. The recipients were sacrificed at 0.5-, 2-, 6-, and 24-hour time points after transplantation to harvest the graft specimens and blood samples. The serum aspartate amino transferase (AST), alanine amino transferase (ALT) and tumor necrosis factor-alpha (TNF-alpha) levels, and histological findings were evaluated. The expressions of the total and phosphorylated p46/p54 JNKs, p38 MAPK, and p42/p44 ERKs were detected by Western blot. The serum ALT and AST levels increased significantly at the 0.5-hour time point and maintained high with the peak levels at the 6-hour time point after liver transplantation. The different sizes of liver isografts did not change the expressions of total p46/p54JNKs, p38MAPK, and p42/p44 ERKs. While the expressions of phosphorylated p46/p54JNKs, p38 MAPK, and p42/p44 ERKs were either negative or mildly up-regulated in the sham operation group, they were significantly activated in the transplanted liver at the 0.5-hour time point, especially in the 30% size liver transplantation group. In conclusion, the activation of three MAPKs in liver isografts correlates with graft size and the JNK and p38 MAPK are responsible for the graft injury while the ERK signal pathway maybe participate in the regulation of cell growth and differentiation after small-for-size liver transplantation.     

Involvement of Ca2+ sensitization in ropivacaine-induced contraction of rat aortic smooth muscle

BACKGROUND: The mechanisms of amino-amide local anesthetic agent-induced vasoconstriction remain unclear. The current study was designed to examine the roles of the protein kinase C (PKC), Rho kinase, and p44/42 mitogen-activated protein kinase (p44/42 MAPK) signaling pathways in calcium (Ca2+)-sensitization mechanisms in ropivacaine-induced vascular contraction. METHODS: Endothelium-denuded rat aortic rings, segments, and strips were prepared. The cumulative dose-response relations of contraction and intracellular Ca2+ concentration to ropivacaine were tested, using isometric force transducers and a fluorometer, respectively. The dose-dependent ropivacaine-induced phosphorylation of PKC and p44/42 MAPK and the membrane translocation of Rho kinase were also detected using Western blotting. RESULTS: Ropivacaine induced a dose-dependent biphasic contractile response and an increase in intracellular Ca2+ concentration of rat aortic rings, increasing at concentrations of 3 x 10 m to 3 x 10 m and decreasing from 10 m to 3 x 10 m, with a greater tension/intracellular Ca2+ concentration ratio than that induced with potassium chloride. The contraction was attenuated in a dose-dependent manner, by the PKC inhibitors bisindolylmaleimide I and calphostin C, the Rho-kinase inhibitor Y 27632, and the p44/42 MAPK inhibitor PD 098059. Ropivacaine also induced an increase in phosphorylation of PKC and p44/42 MAPK, and membrane translocation of Rho kinase in accordance with the contractile responses, which were also significantly inhibited by bisindolylmaleimide I and calphostin C, Y 27632, and PD 098059, correspondingly. CONCLUSION: These findings demonstrated that PKC-, Rho kinase-, and p44/42 MAPK-mediated Ca2+-sensitization mechanisms are involved in the ropivacaine-induced biphasic contraction of rat aortic smooth muscle.     

Basic fibroblast growth factor induces expression of VEGF receptor KDR through a protein kinase C and p44/p42 mitogen-activated protein kinase-dependent pathway.

Vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF) are angiogenic molecules whose combined mitogenic activity is potently synergistic. However, the molecular mechanism underlying this synergy is incompletely understood. We examined whether VEGF and bFGF affect expression of each other or alter expression of the VEGF receptor KDR in retinal capillary endothelial cells. In addition, we investigated the intracellular signaling mechanisms involved in this response. VEGF-induced [3H]thymidine uptake was tightly correlated with KDR mRNA and protein concentrations, suggesting that increased KDR expression might account for VEGF's synergistic activity in the presence of bFGF. bFGF (10 ng/ml) induced KDR mRNA expression within 4 h and attained a 4.0-fold increase after 24 h. KDR protein expression was increased 7.5-fold after 48 h. VEGF (= 50 ng/ml) did not alter bFGF, VEGF, or KDR mRNA expression under serum-deprived conditions. In contrast, VEGF increased KDR mRNA expression 87% under growth conditions and 2.9-fold under serum-deprived conditions in the presence of bFGF. The protein kinase C (PKC) agonist phorbol myristate acetate (PMA) induced KDR mRNA expression 5.1-fold at 100 nmol/l. bFGF increased p44/p42 mitogen-activated protein kinase (MAPK) phosphorylation within 5 min, reaching a maximum within 15 min and remaining significantly elevated for >6 h. bFGF-induced MAPK phosphorylation and KDR mRNA expression were almost completely inhibited by 5 micromol/l GFX, a non-isoform-selective PKC inhibitor. MAPK inhibitor PD98059 reduced KDR mRNA expression 72% at concentrations that inhibited bFGF-induced MAPK phosphorylation 100%, suggesting that pathways in addition to MAPK might also be involved. Inhibitors of the beta isoform of PKC (LY333531), protein kinase A (PKA) (H89), and phosphotidylinositol (PI) 3 kinase (wortmannin) had no significant effect. These data suggest that bFGF stimulates KDR expression through a PKC and p44/p42 MAPK-dependent pathway not primarily involving the beta isoform of PKC, PKA, or PI-3 kinase. Since bFGF induces VEGF expression and since increased KDR expression potentiates VEGF action, resulting in additional KDR expression and marked mitogenic activity, these data provide a novel mechanistic explanation for the angiogenic synergy between VEGF and bFGF.     

Phosphatidylinositol 3-kinase-dependent activation of akt, an essential signal for hyperthermia-induced heat-shock protein 72, is attenuated in streptozotocin-induced diabetic heart

We tested the hypothesis that phosphatidylinositol 3-kinase (PI 3-kinase)-dependent activation of Akt is essential for the expression of cardiac heat-shock protein 72 (HSP72) and that this pathway is impaired in the streptozotocin (STZ)-induced diabetic heart. STZ-induced male diabetic rats were treated with insulin (STZ-insulin group, n = 26) or vehicle (STZ-vehicle group, n = 61) for 3 weeks. Whole-body hyperthermia (43 degrees C for 20 min) was applied, and the heart was isolated 24 h later. Compared with control heart, hyperthermia-induced HSP72 expression and phosphorylation of Akt were attenuated in the STZ-vehicle heart. Pretreatment with wortmannin attenuated hyperthermia-induced HSP72 expression and phosphorylation of Akt. In isolated perfused heart experiments, the hyperthermia-treated STZ-vehicle heart showed poor left ventricular functional recovery during reperfusion after no-flow global ischemia compared with hyperthermia-treated control heart. Insulin treatment restored HSP72 expression and reperfusion-induced functional recovery. In cultured neonatal rat cardiomyocytes, hyperthermia-induced HSP72 expression was enhanced by insulin, together with tolerance against hypoxia-reoxygenation injury. Wortmannin and LY294002 inhibited hyperthermia-induced HSP72 expression and phosphorylation of Akt. Our results indicate that activation of Akt, in a PI 3-kinase-dependent manner, is essential for hyperthermia-induced HSP72 expression in association with cardioprotection, suggesting impairment of this signaling pathway in the STZ-induced diabetic heart, probably due to insulin deficiency.     

Sphingosine 1-phosphate induces heat shock protein 27 via p38 mitogen-activated protein kinase activation in osteoblasts.

We previously showed that sphingosine 1-phosphate acts as a second messenger for tumor necrosis factor alpha-induced interleukin-6 synthesis in osteoblast-like MC3T3-E1 cells and that the synthesis by sphingosine 1-phosphate is dependent on p42/p44 mitogen-activated protein (MAP) kinase activation. In the present study, we investigated the effect of sphingosine 1-phosphate on the induction of heat shock protein 27 (HSP27) in MC3T3-E1 cells. Not C2-ceramide, but sphingosine and sphingosine 1-phosphate significantly induced HSP27 accumulation dose dependently in the range between 1microM and 30 microM. DL-threo-dihydrosphingosine, an inhibitor of sphingosine kinase, markedly inhibited the sphingosine-induced HSP27 accumulation. Sphingosine 1-phosphate induced increase in the levels of the mRNA for HSP27. Sphingosine 1-phosphate stimulated the phosphorylation of p38 MAP kinase. The sphingosine 1-phosphate-induced HSP27 accumulation was dose dependently suppressed by SB203580, an inhibitor of p38 MAP kinase, but not PD98059, an inhibitor of the upstream kinase that activates p42/p44 MAP kinase. SB203580 reduced the sphingosine 1-phosphate-induced increase of mRNA for HSP27. These results strongly suggest that sphingosine 1-phosphate-stimulated HSP27 induction is mediated via p38 MAP kinase activation in osteoblasts.     

HCV NS3对人肝细胞磷酸化酪氨酸相关蛋白表达的影响

目的:研究HCV NS3对人肝细胞磷酸化酪氨酸相关蛋白表达的影响。方法:通过脂质体稳定转染分别建立表达HCV NS3蛋白的pRcHCNS3/QSG细胞，空白质粒转染细胞pRcCMV/QSG。用Western Blot和免疫细胞化学分别检测磷酸化p44/42，磷酸化p38，磷酸化SAPK/JNK在pRcHCNS3/QSG细胞和pRcCMV/QSG细胞及未转染的QSG7701细胞的表达差异及细胞内的定位;免疫共沉淀实验检测HCV NS3对磷酸化酪氨酸蛋白表达水平的影响。结果:pRcHCNS3/QSG细胞，pRcCMV/QSG细胞，未转染的QSG7701细胞抽提的总蛋白中均可检测出磷酸化p44/42，磷酸化p38，磷酸化SAPK/JNK的表达，阳性信号定位于细胞核。上述3种蛋白在pRcHCNS3/QSG细胞的表达较pRcCMV/QSG细胞和未转染的QSG7701细胞表达增强，其磷酸化酪氨酸蛋白的表达也较其他两对照组增强。结论:HCV NS3可能通过影响酪氨酸蛋白激酶的表达及活性，使一系列受酪氨酸蛋白激酶调节的蛋白质如MAPK分子异常磷酸化而活化，激活下游信号转导通路，导致肝细胞无限增殖和肿瘤形成。