Mechanism of stilbene glycosides on apoptosis of SH-SY5Y cells via regulating PI3K/AKT signaling pathway

Objective:To investigate the effects of stilbene glycoside(TSG)on okadaic acid-induced apoptosis in human neuroblastoma cells(SH-SY5Y)via the PI3K/AKT pathway.Methods:The optimal concentration of OA was screened by CCK-8 assay,and SH-SY5Y cells were divided into control group,model group,TSG group,LY294002 group and LY294002+TSG group.The proliferation and apoptosis in each group were detected by CCK-8 and TUNEL assays;Western blotting method and real-time fluorescence quantitative polymerase chain reaction was used to detect the expression of PI3K,P-PI3K(Y607),AKT,P-AKT(Ser473),Bcl-2 and Bax proteins.The relative protein expression was represented by P-PI3K(Y607)/PI3K,P-AKT(Ser473)/AKT and Bcl-2/Bax gray ratio.Results:CCK-8 screened the optimal concentration of OA as 40 nmol/L.Compared with the control group,the model group increased relative cell viability,decreased apoptosis rate,the pathway and apoptotic proteins expression levels of P-PI3K(Y607)/PI3K,P-AKT(Ser473)/AKT and Bcl-2/Bax were decreased,and the mRNA expression levels of PI3K,AKT and Bcl-2 were decreased.Bax mRNA expression level increased(P<0.05);Compared with model group,TSG group increased relative cell viability,decreased apoptosis rate,increased protein expression levels of P-PI3K(Y607)/PI3K,P-AKT(Ser473)/AKT,Bcl-2/Bax,and increased mRNA expression levels of PI3K,AKT,and Bcl-2.Bax mRNA expression decreased(P<0.05),LY294002 group decreased relative cell viability,increased apoptosis rate,P-PI3K(Y607)/PI3K protein expression levels were significantly decreased(P<0.05),P-AKT(Ser473)/AKT and Bcl-2/Bax protein expression levels were significantly decreased,but there was no statistical significance,PI3K,AKT and Bcl-2 mRNA expression levels were decreased,and Bax mRNA expression levels were increased(all P<0.05);Compared with LY294002 group,LY294002+TSG group increased relative cell viability,decreased apoptosis rate,and the protein expression levels of P-PI3K(Y607)/PI3K,P-AKT(Ser473)/AKT and Bcl-2/Bax were increased.The mRNA expression levels of PI3K,AKT,Bcl-2 were increased,Bax was decreased(all P<0.05).Conclusion:Stilbene glycoside may alleviate okadaic acid-induced apoptosis in SH-SY5Y cells by interfering with the PI3K/AKT signaling pathway,which in turn regulates the expression of apoptotic factors such as Bcl-2 and Bax.

电项针对全脑缺血VD模型大鼠PI3K/AKT/GSK-3β信号通路的影响

目的研究电项针对全脑缺血血管性痴呆(vascular dementia,VD)模型大鼠磷脂酰肌醇-3-激酶/丝氨酸-苏氨酸蛋白激酶/糖原合成酶激酶-3β(Phosphatidylinositol-3 kinase/serine-threonine kinase/glycogen synthase kinase-3β,P13K/AKT/GSK-3β)信号通路的影响。方法采用四血管阻断方法制备VD模型大鼠,电项针组取双侧风池穴、供血穴,电针30 min/次,1次/d,治疗14d。采用Y迷宫评价大鼠学习记忆能力;荧光定量PCR(RT-PCR)、Western blot法检测大鼠海马组织中磷酸化蛋白激酶B(phosphorylatedproteinkinaseB,p-AKT)、磷酸化糖原合成酶激酶-3β(Phosphorylated GSK-3β,P-GSK-3β)mRNA和p-AKT、p-GSK-3β蛋白的表达。结果与模型组比较,电项针组可显著提高VD大鼠Y迷宫学习与记忆正确次数(P<0.01)。与模型组比较,电项针组大鼠海马组织中p-AKT、p-GSK-3βmRNA和p-AKT、p-GSK-3β蛋白表达均有不同程度的升高(P<0.01)。结论电项针能够改善VD模型大鼠学习记忆能力,具体机制可能是激活PI3K/AKT/GSK-3β信号通路,发挥抗凋亡作用,起到对缺血海马神经元的保护作用。

Vaspin对HUVECs中TNF-α介导的NF-κB和PI3K/Akt信号通路的影响

目的探讨Vaspin对TNF-α介导的人脐静脉内皮细胞（HUVECs）NF-κB和P13K／Akt信号通路的影响。方法体外分离并培养HUVECs。NF-κB荧光酶报告质粒瞬时转染HUVECs，用不同浓度（0～320wg／L）Vaspin预培养，随后用lO斗∥L的TNF．仪作用于HUVECs，荧光酶报告分析法测定NF-κB的转录活性。Westernblot测定磷酸化的Akt水平。ELISA测定细胞上清液中IL-1及IL-6的浓度。Real．timePCR、Westernblot检测细胞问黏附分子-1（ICAM-1）、血管细胞黏附分子-1（VCAM-1）及单核细胞趋化蛋白-1（MCP-1）的mRNA和蛋白表达水平。结果在HUVECs中，Vaspin抑制TNF．仅介导的NF-κB转录活性（P〈0．05），并且NF-κB下游因子IL-1、IL-6、ICAM-1、VCAM-1和MCP-1的表达降低（P〈0．05）。Vaspin增加了炎性因子TNF-d介导的磷酸化的Akt水平（P〈0．05）。结论Vaspin抑制HUVECs中TNF-Ⅱ介导的NF-κB信号通路，增强TNF-α介导的P13K／Akt信号通路的传导。

基于PI3K/Akt信号通路探讨抗纤抑癌方干预肝癌前病变的作用机制

目的:研究抗纤抑癌方对肝癌前病变大鼠PI3K/Akt信号通路的影响。方法:雄性Wistar大鼠85只,随机分为正常组、模型组、抗纤抑癌低、中、高剂量组及鳖甲软肝组。采用二乙基亚硝胺腹腔注射制备肝癌前病变模型,抗纤抑癌方进行干预,复方鳖甲软肝片作为对照;采用免疫组化法检测GST-Pi的表达,实时荧光定量PCR及Western blot法检测PI3K、Akt mRNA及蛋白的表达。结果:与模型组相比,抗纤抑癌高剂量组GST-Pi阳性表达面积明显减少,染色减轻,MOD值显著降低(P<0.05);与模型组相比,抗纤抑癌低、中、高剂量组及鳖甲软肝组PI3K、Akt mRNA的表达均显著降低(P<0.01),抗纤抑癌低、中、高剂量组PI3K、p-PI3K蛋白表达显著降低(P<0.01或P<0.05),抗纤抑癌中、高剂量组p-Akt蛋白表达显著降低(P<0.01);与鳖甲软肝组相比,抗纤抑癌高剂量组PI3K mRNA及p-PI3K蛋白的表达显著降低(P<0.05)。结论:抗纤抑癌方可通过调控PI3K/Akt信号抑制肝癌前病变。

Novel insights into D-Pinitol based therapies:a link between tau hyperphosphorylation and insulin resistance

Alzheimer’s disease is a neurodegenerative disorder characterized by the amyloid accumulation in the brains of patients with Alzheimer’s disease.The pathogenesis of Alzheimer’s disease is mainly mediated by the phosphorylation and aggregation of tau protein.Among the multiple causes of tau hyperphosphorylation,brain insulin resistance has generated much attention,and inositols as insulin sensitizers,are currently considered candidates for drug development.The present narrative review revises the interactions between these three elements:Alzheimer’s disease-tau-inositols,which can eventually identify targets for new disease modifiers capable of bringing hope to the millions of people affected by this devastating disease.

PI3K-AKT-mTOR signaling in prostate cancer progression and androgen deprivation therapy resistance

Prostate cancer （PCa） is the second most common malignancy among men in the world. Castration-resistant prostate cancer （CRPC） is the lethal form of the disease, which develops upon resistance to first line androgen deprivation therapy （ADT）. Emerging evidence demonstrates a key role for the PI3K-AKT-mTOR signaling axis in the development and maintenance of CRPC. This pathway, which is deregulated in the majority of advanced PCas, serves as a critical nexus for the integration of growth signals with downstream cellular processes such as protein synthesis, proliferation, survival, metabolism and differentiation, thus providing mechanisms for cancer cells to overcome the stress associated with androgen deprivation. Furthermore, preclinical studies have elucidated a direct connection between the PI3K-AKT-mTOR and androgen receptor （AR） signaling axes, revealing a dynamic interplay between these pathways during the development of ADT resistance. Thus, there is a clear rationale for the continued clinical development of a number of novel inhibitors of the PI3K pathway, which offer the potential of blocking CRPC growth and survival. In this review, we will explore the relevance of the PI3K-AKT-mTOR pathway in PCa progression and castration resistance in order to inform the clinical development of specific pathway inhibitors in advanced PCa. In addition, we will highlight current deficiencies in our clinical knowledge, most notably the need for biomarkers that can accurately predict for response to PI3K pathway inhibitors.

Activation of Rac1-PI3K/Akt is required for epidermal growth factorinduced PAK1 activation and cell migration in MDA-MB-231 breast cancer cells

Epidermal growth factor （EGF） may increase cell motility, an event implicated in cancer cell invasion and metastasis. However, the underlying mechanisms for EGF-induced cell motility remain elusive. In this study, we found that EGF treatment could activate Ras-related C3 botulinum toxin substrate 1 （Racl）, PI3K/Akt and p21- actived kinase （PAK1） along with cell migration. Ectopic expression of PAK1 K299R, a dominant negative PAK1 mutant, could largely abolish EGF-induced cell migration. Blocking PI3K/Akt signalling with LY294002 or Akt siRNA remarkably inhibited both EGF-induced PAK1 activation and cell migration. Furthermore, expression of dominant-negative Racl （T17N） could largely block EGF-induced PI3K/Akt-PAK1 activation and cell migration. Interestingly, EGF could induce a significant production of ROS, and N-acetyl-L-cysteine, a scavenger of ROS which abolished the EGF-induced ROS generation, cell migration, as well as activation of PI3K/Akt and PAK, but not Racl. Our study demonstrated that EGF-induced cell migration involves a cascade of signalling events, including activation of Racl, generation of ROS and subsequent activation of PI3K/Akt and PAK1.

Insensitivity of PI3K/Akt/GSK3 signaling in peripheral blood mononuclear cells of age-related macular degeneration patients

Our recent studies with cultured retinal pigment epithelium cells suggested that overexpression of interleukin 17 receptor C（IL-17RC）,a phenomenon observed in peripheral blood and chorioretinal tissues with age-related macular degeneration（AMD）,was associated with altered activation of phosphatidylinositide 3-kinase（PI3K）,Akt,and glycogen synthase kinase 3（GSK3）.We wondered whether or not altered PI3 K,Akt,and GSK3 activities could be detected in peripheral blood mononuclear cells（PBMC） obtained from AMD patients.In the patients＇ PBMC,absent or reduced serine-phosphorylation of GSK3α or GSK3β was observed,which was accompanied with increased phosphorylation of GSK3 substrates（e.g.CCAAT enhancer binding protein a,insulin receptor substrate 1,and TAU）,indicative of enhanced GSK3 activation.In addition,decreased protein mass of PI3K85α and tyrosinephosphorylation of PI3K50α was present in PBMC of the AMD patients,suggesting impaired PI3 K activation.Moreover,abnormally lowered molecular weight forms of Akt and GSK3 were detected in PBMC of the AMD patients.These data demonstrate that despite the presence of high levels of IL-17 RC,Wnt-3a and vascular endothelial growth factor,the PI3K/Akt/GSK3 signaling pathway is insensitive to these stimuli in PBMC of the AMD patients.Thus,measurement of PI3K/Akt/GSK3 expression and activity in PBMC may serve as a surrogate biomarker for AMD.

RegⅣ、EGFR、PI3K蛋白在胃腺癌中的表达及意义

目的：研究再生基因蛋白Ⅳ（ Regenerating gene typeⅣ，RegⅣ）、表皮生长因子受体（ Epider-mal growth factor receptor ，EGFR）及磷脂酰肌醇3-激酶（ Phosphatidylinositol -3-kinase，PI3K）蛋白在胃腺癌中的表达及临床意义。方法应用S-P免疫组化技术检测73例胃腺癌及其相应的癌旁正常组织中RegⅣ、EGFR和PI3K蛋白的表达情况。结果73例胃腺癌组织中RegⅣ、EGFR、PI3K蛋白的阳性表达率分别为50．7％（37/73）、56．2％（41/73）、69．9％（51/73）均高于癌旁正常组织20．5％（15/73）、19．2％（14/73）、21．9％（16/73），差异具有统计学意义（ P＜0．05）；RegⅣ的表达与肿瘤分化程度相关，差异有统计学意义（P＜0．05）；EGFR蛋白的表达与浸润深度、淋巴结转移、临床分期相关，差异有统计学意义（P＜0．05）；PI3K蛋白的表达与肿瘤分化程度、浸润深度、淋巴结转移、临床分期相关有统计学意义（P＜0．05）。 RegⅣ与EGFR、PI3K蛋白在胃腺癌组织中的表达呈正相关（r＝0．343、0．248，P＜0．05），EGFR与PI3K蛋白在胃腺癌组织中的表达呈正相关（ r＝0．384，P＜0．05）。结论 RegⅣ可能通过激活EGFR/PI3K/Akt信号通路在胃腺癌的发生发展中发挥重要作用。

Regulatory Effects of Zuogui Pill on Apoptosis of Follicles in Rats Injured by 60Co-γRays Based on PI3K/Akt/m TOR Signaling Pathway

[Objectives]To explore the protective effects of Zuogui Pill on ^(60)Co-γ-ray-induced premature aging of rats based on phosphatidylinositol 3-kinase/protein kinase B/mammalian target of rapamycin(PI3K/Akt/mTOR)signaling pathway.[Methods]Sixty sexually mature female SD rats were irradiated with ^(60)Co-γ-ray(6.0 Gy,LD 40)for 24 h at one time.These rats were randomly divided into model group,Progynova group[0.18(g·kg)/d],Progynova[0.09(g·kg)/d]+Zuogui Pill high dose[23.625(g·kg)/d)]group,Zuogui Pill high dose[23.625(g·kg)/d)]group,Zuogui Pill medium dose[9.45(g·kg)/d)]group and Zuogui Pill low dose[4.725(g·kg)/d]group.The administration(once a day)lasted 21 d.The rat serum[follicle-stimulating hormone(FSH),luteinizing hormone(LH)and estradiol(E_(2))]were detected by Enzyme-linked immunosorbent assay(ELISA).The morphological changes of ovary were observed by hematoxylin-eosin(HE)staining.The apoptosis rate of granulosa cells was detected by terminal deoxynucleotidyl transferase(TdT)-mediated dUTP nick-end labeling(TUNEL).The protein expression of phosphorylated(p)-PI3K,p-Akt,p-mTOR,B-cell lymphoma-2(Bcl-2),and Bcl-2-associated X protein(Bax)in ovarian tissues were detected by Western blot.[Results]Compared with the normal group,the model group showed significant increase in the serum FSH(P<0.01),significant decrease in serum E_(2)(P<0.05),and decrease in the number of early follicles and luteum in the ovary(P<0.01).Besides,the apoptosis rate of granulosa cells increased significantly(P<0.01);the expression of p-PI3K,p-Akt,p-mTOR and Bcl-2 in ovarian tissue decreased significantly,while the expression of Bax increased significantly(P<0.01).Compared with the model group,the number of early follicles in the ovary increased and the apoptosis rate of granulosa cells decreased after intervention in each administration group.In addition,the protein expressions of p-PI3K,p-Akt,p-mTOR and Bcl-2 increased,while the expression of Bax decreased,especially in Progynova+Zuogui Pill high dose group,the differences were statistically significant(P<0.05,P<0.01).[Conclusions]Zuogui Pill may protect the radiation-injured ovary through activating the expression of PI3K/Akt/mTOR protein in ovarian tissue,increasing the amount of Bcl-2 protein and inhibiting the expression of Bax protein.

Liposomalα-cyperone targeting bone resorption surfaces suppresses osteoclast differentiation and osteoporosis progression via the PI3K/Akt axis

Osteoporosis is a metabolic dysregulation of bone that occurs mainly in postmenopausal women,and the hyperfunction of osteoclasts is the primary contributor to postmenopausal osteoporosis.However,the development of effective therapeutic drugs and precise delivery systems remains a challenge in the field of anti-absorption therapy.Here,we reported theα-cyperone(α-CYP)for anti-osteoporosis and developed a liposome-based nano-drug delivery system ofα-CYP,that specifically targets the bone resorption interface.Firstly,we found that theα-CYP,one of the major sesquiterpenes of Cyperus rotundus L.,attenuated the progression of osteoporosis in ovariectomized(OVX)mice and down-regulated the expression of phosphorylated proteins of phosphoinositide 3-kinase(PI3K)and protein kinase B(Akt),causing down-regulation of osteoclast-related genes/proteins and curbing osteoclast differentiation.Furthermore,α-CYP reversed the activation of osteoclastic differentiation and enhanced osteoporosis-related proteins expression caused by PI3K/Akt agonist(YS-49).More importantly,we adopted the osteoclastic resorption surface targeting peptide Asp8 and constructed the liposome(lipαC@Asp8)to deliverα-CYP to osteoclasts and confirmed its anti-osteoporosis effect and enhanced osteoclast inhibition by blocking PI3K/Akt axis.In conclusion,this study demonstrated thatα-CYP inhibits osteoclast differentiation and osteoporosis development by silencing PI3K/Akt pathway,and the liposome targeting delivery systems loaded withα-CYP might provide a novel and effective strategy to treat osteoporosis.

KRAS对肺癌大鼠PI3K/Akt信号通路与免疫功能的影响

目的分析K-RAS原癌基因(KRAS)对肺癌大鼠PI3K/Akt信号通路与免疫功能的影响。方法选取30只Wistar大鼠,10只作为对照组,另外20只建立肺癌大鼠模型,将建模成功的20只大鼠分为模型组与KRAS沉默组,每组10只,观察各组大鼠一般情况,RT-PCR方法验证转染效率,采集各组大鼠静脉血进行检测免疫功能,HE染色观察各组大鼠肺组织病理学改变,流式细胞仪检测大鼠血清CD3^(+)、CD4^(+)、CD4^(+)/CD8^(+)表达,RT-PCR方法与Western blot方法检测大鼠肺组织中PI3K、AktmRNA与蛋白的表达。结果RT-PCR结果显示,KRAS慢病毒载体转染成功。与模型组相比,抑制KRAS表达可改善肺癌大鼠肺组织病理情况,上调大鼠血清CD3^(+)、CD4^(+)、CD4^(+)/CD8^(+)表达水平,降低肺组织PI3K与AktmRNA与蛋白表达水平。结论沉默KRAS表达,可提高肺癌大鼠免疫功能,其机制可能与抑制PI3K/Akt信号通路有关。

RISK信号通路在β_2-肾上腺素受体激动剂Clenbuterol减轻心肌细胞缺氧/复氧损伤中的作用

目的研究β2-肾上腺素受体激动剂clenbuterol对原代培养的心肌细胞缺氧/复氧损伤的作用及其是否与激活再灌注损伤挽救激酶(reperfusion injury salvage kinase,RISK)信号通路有关。方法将原代培养的新生Wistar大鼠乳鼠心肌细胞分为8组,1正常培养组;2缺氧/复氧(A/R)组;3clenbuterol(1μmol·L-1)+A/R;4ICI118,551(10μmol·L-1)+clenbuterol(1μmol·L-1)+A/R组;5美托洛尔metoprolol(10μmol·L-1)+clenbuterol(1μmol·L-1)+A/R组;6 metoprolol(10μmol·L-1)+A/R组;7 PD98059(20μmol·L-1)+clenbuterol(1μmol·L-1)+A/R组;8LY294002(10μmol·L-1)+clenbuterol(1μmol·L-1)+A/R组。采用MTT法测定各组细胞存活率;比色法检测心肌细胞培养液的乳酸脱氢酶(LDH)含量;Hoechst 33342荧光染色法检测细胞凋亡率;分子探针DCFH-DA检测细胞内活性氧的水平;Western blot检测心肌细胞缺氧/复氧后ERK及p-ERK1/2蛋白的表达水平。结果与A/R组比较,clenbuterol+A/R组明显增高细胞存活率,降低LDH含量,降低细胞凋亡率,ROS产生减少,p-ERK1/2蛋白表达水平增高,而选择性β2受体阻断剂ICI 118,551可取消clenbuterol的上述作用,β1受体阻断剂Metoprolol对clenbuterol的作用无影响,PI3K抑制剂LY294002和ERK1/2抑制剂PD98059可阻断clenbuterol对心肌细胞缺氧/复氧损伤的保护作用。结论clenbuterol能够减轻心肌细胞缺氧/复氧损伤,加入选择性β2受体阻断剂ICI 118,551,PI3K抑制剂LY294002和ERK抑制剂PD98059均使clenbuterol的保护作用取消,表明clenbuterol可通过激动β2肾上腺素受体,激活RISK信号通路发挥抗心肌细胞缺氧/复氧损伤的作用。

LY294002联合姜黄素对人膀胱癌EJ细胞的体外抑制作用

目的研究磷脂酰肌醇-3激酶/蛋白激酶B(PI3K/Akt)特异性抑制剂LY294002与姜黄素联合对体外培养的人膀胱癌EJ细胞的抑制作用。方法用MTT法,检测姜黄素单独或联合PI3K/Akt特异性抑制剂LY294002对人膀胱癌EJ细胞的抑制率;用流式细胞技术及Western blotting法,检测药物单独或联合作用对EJ细胞凋亡的影响。结果联合LY294002能够显著提高姜黄素对EJ细胞的抑制率;联合LY294002能够增加EJ细胞的凋亡水平。结论 LY294002通过促进细胞凋亡有效提高姜黄素对EJ细胞抑制作用的敏感性,通过抑制PI3K/Akt信号转导通路的激活,可明显提高姜黄素对膀胱癌的治疗效果。

参芎化瘀胶囊通过PI3-K/Akt信号通路改善大鼠缺血性脑损伤

目的探讨参芎化瘀胶囊对缺血性脑损伤的治疗作用及其机制。方法 SD大鼠分为假手术组、模型组及参芎化瘀胶囊高、低剂量组。改良的Pulsineli 4血管阻断(4-VO)法制作全脑缺血模型;HE染色观察海马区神经细胞的形态变化;免疫组织化学法检测海马区磷脂酰肌醇3-激酶(PI3-K)、蛋白激酶B(PKB/Akt)的表达;原位缺口末端标记法(TUNEL)检测凋亡细胞;八臂迷宫法测试动物学习记忆功能。结果与假手术组比较,模型组大鼠海马区神经元结构损伤明显,PI3-K、Akt表达增高,凋亡神经细胞数量增加,大鼠的学习记忆功能下降;与模型组比较,参芎化瘀胶囊组海马神经元形态结构损伤减轻,PI3-K、Akt表达进一步增多,凋亡神经细胞数量减少,动物学习记忆功能改善,上述变化在高剂量参芎化瘀胶囊更为明显。结论参芎化瘀胶囊对脑缺血损伤有很好的治疗作用,其机制可能与调控PI3-K/Akt信号途径有关。

低氧诱导因子-1的转录活性调控及其信号传导

低氧诱导因子 1(hypoxia induciblefactor 1,HIF 1)是氧平衡调控相关的转录因子 .依赖HIF 1的基因表达调控系统广泛影响葡萄糖代谢、细胞增殖、凋亡和血管发生 ,与机体低氧适应、胚胎发育、各种缺血性疾病及肿瘤相关 .HIF 1自身活性调节是低氧应答基因表达调控的中心环节 .调控主要发生在源于Ras的两条信号途径 :Ras/Raf/MEK介导的HIF 1反式激活功能调控 ,PI(3)K/Akt依赖的HIF 1alpha蛋白稳定性调控 .这两个信号传导途径分别独立又协调地调控着HIF

PI-3K在大鼠非酒精性脂肪性肝病发病中的作用

目的探讨磷脂酰肌醇-3激酶(PI-3K)在非酒精性脂肪性肝病(NAFLD)发病中的作用。方法30只SD大鼠随机分为3组:正常对照组、非酒精性脂肪肝模型:A组(高脂喂养8周)、B组(高脂喂养16周)。评价各组大鼠肝脂变程度和炎症活动度积分水平;免疫组化观察各组PI-3K(p85α)蛋白表达量;RT-PCR检测PI-3K(p85α)mRNA的表达水平。结果模型组脂肪变性明显,炎症活动度记分均显著高于正常对照组(P<0.01),模型B组炎症活动度记分水平显著高于模型A组(P<0.01)。模型A、B组PI-3K(p85α)蛋白表达量均明显低于正常对照组(P<0.01),且模型B组PI-3K(p85α)蛋白表达量明显低于模型A组(P<0.01);模型A组PI-3K(p85α)mRNA表达水平与正常对照组比较无显著差异(P>0.05);模型B组PI-3K(p85α)mRNA表达水平显著低于模型A组及正常对照组(P<0.01)。结论PI-3K可能是影响NAFLD发病的重要因素。

特异性p^(38)MAPK抑制剂SB203580对乳鼠小脑颗粒神经元的保护作用

目的 研究 p38丝裂原激活蛋白激酶 (MAPK)选择性抑制剂SB2 0 35 80对乳鼠小脑颗粒神经元凋亡的保护作用。方法 SD乳鼠小脑颗粒神经元培养 ,琼脂糖凝胶电泳 ,SAPK/JNK分析试剂盒作激酶分析。结果 PI 3 K的特异性抑制剂LY2 940 0 2诱导小脑颗粒神经元凋亡 ,但SB2 0 35 80通过抑制细胞凋亡而促进小脑颗粒神经元的存活 ,且有浓度依赖性。LY2 940 0 2诱导凋亡的颗粒神经元中c Jun的表达量和磷酸化水平均升高 ,JNK被激活。但是 ,当小脑颗粒神经元生长在含SB2 0 35 80的高钾培养基中 ,c Jun的表达量、磷酸化水平和JNK的活性都明显的降低。结论 SB2 0 35 80通过抑制JNK的活性 ,降低c Jun的表达和磷酸化水平 。

狼疮性肾炎患者外周血单个核细胞PI3-K活性研究

目的 了解磷脂酰肌醇三激酶 (PI3 K)在狼疮性肾炎 (LN)外周血单个核细胞 (PBMC)中的活化情况。方法 2 5例狼疮性肾炎患者被分为活动期 (1 3例 )和非活动期 (1 2例 ) ,并以 1 2例健康人为对照。PBMC提取采用Ficol密度梯度离心法 ,PBMCPI3 K酪氨酸磷酸化产物表达采用免疫沉淀、免疫印迹法。细胞培养上清IgG和抗dsDNA检测采用微量ELISA法。结果 与对照组相比 ,活动性SLE患者PI3 K酪氨酸磷酸化活性增高 (P <0 0 1 ) ,非活动性SLE患者PI3 K酪氨酸磷酸化活性无显著性差异 (P>0 0 5 ) ;相关分析显示 ,PI3 K磷酸化产物表达与狼疮活动指数 (r =0 6 1 7,P <0 0 1 )、血清抗dsFNA(r=0 5 4 4,P <0 0 1 )、免疫球蛋白IgG(r=0 6 91 ,P <0 0 1 )呈明显正相关关系 ,与血清补体C3呈明显负相关关系 (r=- 0 6 1 7,P <0 0 1 )。PI3 K特异抑制剂Wortamannin能抑制CD3mAb诱导的SLEPBMC抗dsDNA和IgG的产生 (P <0 0 1 )。结论 PI3 K过度活化可能参与了SLE自身抗体的产生 ,检测PI3 K活化状况可能有助于临床狼疮活动的判断。

Class I phosphatidylinositol 3-kinase inhibitors for cancer therapy

The phosphatidylinositol 3-kinase(PI3K) pathway is frequently activated in human cancers.Class I PI3 Ks are lipid kinases that phosphorylate phosphatidylinositol 4,5-bisphosphate(PIP2) at the 3-OH of the inositol ring to generate phosphatidylinositol 3,4,5-trisphosphate(PIP3), which in turn activates Akt and the downstream effectors like mammalian target of rapamycin(m TOR) to play key roles in carcinogenesis. Therefore, PI3 K has become an important anticancer drug target, and currently there is very high interest in the pharmaceutical development of PI3 K inhibitors. Idelalisib has been approved in USA and Europe as the first-in-class PI3 K inhibitor for cancer therapy. Dozens of other PI3 K inhibitors including BKM120 and ZSTK474 are being evaluated in clinical trials. Multifaceted studies on these PI3 K inhibitors are being performed, such as single and combinational efficacy, resistance, biomarkers,etc. This review provides an introduction to PI3 K and summarizes key advances in the development of PI3 K inhibitors.