观察匙羹藤对胰岛素抵抗(IR)KKAy小鼠脂肪组织中蛋白激酶B表达及磷酸化的影响以及调节机制。方法:将18只胰岛素抵抗KKAy小鼠按体质量随机分为模型组(DM)和匙羹藤水提物组(GS),并选取9只C57BL/6J小鼠为正常对照组(NC),连续灌胃给药8周。实验结束后取血测定空腹血糖(FPG),空腹血清胰岛素(Fins),并计算胰岛素敏感指数(ISI),测定附睾脂肪组织中磷酸激酶依赖的激酶1(PDK1)、蛋白激酶B(PKB)、P-PKB(Ser473),P-PKB(Thr308)蛋白相对含量,及磷酸酶和张力蛋白同源物基因(PTEN)mRNA相对含量。结果:与DM组相比,GS组小鼠FPG、Fins降低,ISI升高,P-PKB(Ser473)蛋白相对含量及其磷酸化水平升高,P-PKB(Thr308)蛋白相对含量升高,PDK1蛋白相对含量降低,PTEN mRNA表达量降低。结论:匙羹藤可通过增加脂肪组织中P-PKB(Ser473)蛋白相对含量,增强Ser473位点磷酸化水平,以及增加P-PKB(Thr308)蛋白相对含量激活PKB,进而改善IR。

This article was aimed to study effects and mechanisms of Gymnema sylvestre on protein kinase B (PKB) and its phosphorylation in adipose tissues of KKAy mice which were mainly characterized by insulin resistance (IR). A total of 18 KKAy mice were randomly divided into the diabetes model (DM) group and Gymnema sylvestre (GS) group according to body weight levels. And 9 normal C57BL/6J mice were used as the normal control (NC) group. Intragastric administration of medication was given to mice for 8 weeks. At the end of the experiment, all animals were tested for fasting plasma glucose (FPG) and fasting insulin level (Fins) for evaluation of insulin sensitivity index (ISI). Expressions of phosphoinositide-dependent kinase-1 (PDK1), PKB, P-PKB (Ser473), P-PKB (Thr 308) in adi-pose tissues of epididymis were determined. The expression of phosphatase and tensin homolog (PTEN) mRNA was also determined. The results showed that compared with the DM group, the GS group showed lower FPG and Fins,

目的 通过观察COPD大鼠中磷脂酰肌醇-3激酶(phosphoinositide-3 kinases,PI3K)、蛋白激酶B(protein kinase B,PKB)与γ-谷氨酰半胱氨酸合成酶(gamma glutamylcysteine synthetase,γ-GCS)的表达,研究PI3K、PKB通路和γ-GCS的关系及其在COPD中可能的参与机制.方法 30只健康雄性Wistar大鼠随机分COPD组和对照组.采用每日熏香烟和两次气管内注入脂多糖法制作COPD大鼠模型.检测大鼠肺组织中γ-GCS活性,原位杂交检测肺组织中γ-GCS mRNA的表达,免疫组织化学分析肺组织中PI3K、PKB与γ-GCS蛋白水平.结果 COPD组大鼠肺组织中γ-GCS差异(P＜0.01).PI3K、PKB与γ-GCS免疫组织化学在COPD组可见肺泡、支气管活性明显高于对照组,组间差异有统计学意义(P＜0.01).原位杂交示COPD组大鼠支气管,肺泡上皮细胞与小动脉平滑肌细胞γ-GCSmRNA广泛表达,与对照组有显著性壁细胞及小血管平滑肌细胞胞浆中皆有蛋白阳性信号表达;图象定量分析示大鼠COPD组PI3K、PKB与γ-GCS蛋白表达显著高于对照组(P＜0.01).直线相关分析得出PI3K、p-PKB蛋白表达与γ-GCS活性、mRNA及蛋白表达呈正相关.结论 COPD大鼠肺组织γ-GCS蛋白和mRNA表达增高,PI3K、p-PKB蛋白也有相应高表达,提示PI3K、PKB和γ-GCS可能在C

Objective To investigate the expression and relationship levels of PI3K,p-PKB and γ-glutamylcysteine synthetase (γ-GCS) in lung of rats with chronic obstructive pulmonary disease.Methods Thirty male Wistar rats were randomly divided into COPD model and control groups.The rat COPD model was established by intratracheal instillation of lipopolysaccharide (LPS 200μg/200μl) twice and exposed to cigarette smoke daily.The level of the activity of γ-GCS in rat lung tissue was measured,the levels of γ-GCS mRNA expression in rat lung tissue was measured by in site hybridization (ISH).The protein expression of PI3K,p-PKB and γ-GCS were observed by immunohistochemistry.Results The levels of the activity of γ-GCS were higher in COPD group (21.608±3.112) U than that in control group [(13.256±2.351) U,P ＜0.01].The ISH showed that the levels of γ-GCS mRNA expression in COPD group (0.369 ± 0.035) were significantly higher than that in the control group (0.231 ± 0.026,P ＜ 0.01)

目的：探讨磷脂酰肌醇3-激酶/蛋白激酶B( PI3K/PKB)信号激活对妊娠滋养细胞疾病( GTD)进展及相关干细胞转录因子FoxD3、Nanog、Stat3、Oct4和Sox2表达的影响。方法：选取20例正常胎盘、38例良性转归葡萄胎( HM-regressed)和13例恶性进展葡萄胎( HM-progressed)、6例绒癌的石蜡包埋组织，采用免疫组化检测p-PKBSer473表达。实时荧光定量PCR( RT-PCR)和Western blot法检测绒癌细胞株JEG3和JAR中FoxD3、Nanog、Stat3、Oct4和Sox2 mRNA和蛋白表达。 CCK8和Transwell小室检测细胞增殖、迁移/侵袭能力。结果：p-PKBSer473表达的免疫组化评分由高到低依次为绒癌、HM-pro-gressed、HM-regressed和正常胎盘，两两比较差异均有统计学意义( P<0.05)。 PI3 K抑制剂LY294002阻遏了PKB的活化，显著减弱了绒癌细胞的增殖、迁移和侵袭能力，降低了FoxD3、Nanog、Stat3的mRNA和蛋白表达，差异均有统计学意义( P<0.05)。结论：PI3 K/PKB信号通道激活与GTD的进展和侵袭表型有关，可通过增强调控滋养细胞的干细胞特性而发挥作用。

Objective:To explore whether the activation of phosphoinositide-3-kinase protein kinase B (PI3K/PKB) signaling affects the progress of gestational trophoblastic disease (GTD) and the expressions of stem cell related transcriptional factors FoxD3,Nanog,Stat3, Oct4 and Sox2. Methods:The phosphorylated active form of PKB(p-PKBSer473) was explored by immunohistochemistry in paraffin block tissues including 20 cases of normal first trimester pla-centas,38 cases of HM regressed,13 cases of HM progressed,and 6 cases of choriocarcinomas. The expressions of mRNAs and proteins in choriocarcinoma cell lines JEG3 and JAR were de-tected by quantitative RT-PCR and Western blot. The capacities of proliferation and motility choriocarcinoma cells were evaluated by CCK8 and transwell assays. Results:The expressions immunoscore of p-PKBSer473 in these tissues were in such order:choriocarcinoma>HM-progressed>HM-regressed>normal placenta,and the differences between the groups were all significant

目的研究异氟醚如何影响乙酰胆碱诱导的细胞内信号传递,探讨挥发性麻醉药影响认知功能的分子机理。方法培养的神经元样PC12细胞随机分成对照组和异氟醚组。对照组正常培养,异氟醚组用1.2%异氟醚处理2 h,分别于处理后0 min、1 h、3 h加入碘化乙酰胆碱(ACh)刺激2 min后收集细胞,采用Westem blot技术检测基础水平和乙酰胆碱诱导下细胞外信号调节激酶1/2(ERK1/2)的磷酸化水平和蛋白激酶B(PKB)的活性。结果单纯ACh刺激可导致ERK1/2磷酸化水平显著增加(P0.01),异氟醚处理后0 min和1 h乙酰胆碱诱导的ERK1/2磷酸化水平明显降低,3 h时恢复到正常水平。与对ERK的影响效应不同,单独异氟醚处理可迅速增加基础状态下PKB的磷酸化水平(P0.05),但1 h后即恢复正常水平。结论异氟醚可以长时程减弱乙酰胆碱诱导的ERK活化,从而干扰胞内信号传递,这可能与术后认知功能障碍发生密切相关。

Objective To investigate the effect of isoflurane on acetylcholine-induced intracellular signaling, and to explore the molecular mechanism underlying cognitive deficits caused by volatile anesthetics exposure. Methods PC12 cells were exposed to 1. 2% isoflurane for 2 h. subsequently,acetylcholine iodide was applied to the cells for 2 min at 0 min,1 h and 3 h separately. Western blot was used to assay basal phosphorylation and acetylcholine-induced phosphorylation of extracellular signal-regulated kinase 1/2 ( ERK) 1/2 and protein kinase B ( PKB) . Results ACh stimulation alone led to significant increase of ERK1/2 phosphorylation levels. After isoflurane exposure,acetylcholine-induced ERK1/2 phosphorylation was persistently reduced at 0 min and 1 h,until 3 h after exposure,acetylcholine-in-duced ERK1/2 phosphorylation had returned to control levels. In contrast,immediately after isoflurane exposure,basal PKB phosphorylation significantly augmented but had returned to normal levels at 1

目的探讨热量限制(caloric restriction,CR)对C57小鼠学习、记忆能力及胰岛素信号通路相关蛋白的影响。方法雄性C57小鼠30只,采用数字表法随机分为3组:正常对照组(n=10)、高能量组(n=10)、低能量组(n=10)。6个月后通过Morris水迷宫检测各组小鼠的学习、记忆能力,每组各抽取5只应用免疫组织化学染色的方法,测定小鼠脑内胰岛素信号转导通路相关蛋白:依赖于烟酰胺腺嘌呤二核苷酸的组蛋白脱乙酰酶(silent mating type information regulation 2 homolog-1,SIRT1)、胰岛素样生长因子-1(insulin-like growth factor 1,IGF-1)、胰岛素受体(insulin receptor,IR)、胰岛素受体底物-1(insulin receptor substrate-1,IRS-1)、磷脂肌醇3激酶(phosphatidylinositol 3-kinase,PI3K)、蛋白激酶B(protein kinase B,Akt/PKB)、磷酸化应答原件结合蛋白(c AMP-response element binding protein,p-CREB)的表达。结果低能组小鼠的逃避潜伏期缩短,表明学习和记忆能力较前增高,SIRT1、IGF-1、IR、IRS-1、PI3K、Akt/PKB、p-CREB表达均明显降低(P0.01)。结论热量饮食提高C57小鼠的学习记忆能力,可能与降低小鼠脑内胰岛素信号通路相关蛋白的表达有关。

Objective To investigate the effect of caloric restrictionon(CR) on learning and memory function and insulin signaling related protein in C57 mice. Methods Thirty male C57 mice were randomly divided into 3 groups: normal control group, high-energy group and low energy group, each group consisted of 10 mice, learning and memory function were measured by Morris test at the sixth month. Five mouse of each group were selected to measurethe insulin signaling pathway related protein by immunochemistry. Results The escape latency of low-energy group was shorter than normal control group, which indicates the learning and memory ability of low-energy C57 mice improved. The expression of Silent mating type information regulation 2 homolog- 1(SIRT1), insulin-like growth factor 1 (IGF-1), insulin receptor(IR), insulin receptor substrate-1 ( IRS-1), phosphatidylinositol 3-kinase( PI3K), protein kinase B( Akt/PKB), cAMP-response element binding protein(p-CREB)in brain decreased significant