MicroRNA-10 negatively regulates inflammation in diabetic kidney via targeting activation of the NLRP3 inflammasome

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JAK2/STAT3信号通路在应激性溃疡大鼠胃黏膜炎症反应中的作用研究

目的：探讨Janus激酶2（JAK2）/信号转导与转录激活子3（STAT3）信号通路在应激性溃疡（SU）的胃黏膜炎症反应过程中的作用。方法：选用雄性清洁级SD大鼠96只,随机分为4组,分别为：正常对照组（NC组）、应激性溃疡组（SU组）、抑制剂组（AG490组）、二甲基亚砜组（DMSO组）。建立水浸束缚SU大鼠模型。4组大鼠分别于实验开始后2h、4h、8h、16h各取6只,测定胃黏膜血流量（GMBF）,计算溃疡指数（UI）。观察胃黏膜组织学变化,测定TNF-α、IL-1β、IL-6以及JAK2、p-JAK2、STAT3、p-STAT3的表达量。结果：SU组GMBF明显低于NC组,而UI明显高于NC组（P〈0.01）;AG490组GMBF明显高于SU组,而UI明显低于SU组（P〈0.01）。通过造模前给予抑制剂AG490,胃黏膜病理学变化明显改善。SU组TNF-a、IL-1β、IL-6以及p-JAK2、p-STAT 3表达量明显高于NC组（P〈0.01）,而AG490组较SU组显著降低（P〈0.01）。结论：JAK2/STAT 3信号通路参与了SU大鼠胃黏膜炎症反应过程,使用JAK2特异性抑制剂AG490可以减轻SU大鼠胃黏膜炎症反应。     

On the role of plasminogen activator inhibitor-1 in adipose tissue development and insulin resistance in mice

OBJECTIVES: To investigate the role of plasminogen activator inhibitor-1 (PAI-1) in adipose tissue development and insulin metabolism. METHODS: Aged male wild-type (WT) or transgenic mice with adipose tissue overexpression of PAI-1 (45-55 weeks) in 50% C57Bl/6: 50% Friend Virus B-strain (FVB) genetic background, kept on normal chow, were used without or with administration of a synthetic low molecular weight PAI-1 inhibitor (PAI-039) to the food (1 mg g(-1)) for 4 weeks. RESULTS: The PAI-1 transgenic mice showed somewhat lower body weight and adipose tissue mass than WT mice, whereas fasting insulin levels were higher. Glucose and insulin tolerance tests did not reveal significant differences between both genotypes. Addition of PAI-039 to the food did not significantly affect total body fat, weight of the isolated s.c. and gonadal fat territories or their adipocyte size and blood vessel composition in either genotype. Fasting glucose levels and glucose tolerance tests were, for both genotypes, comparable with those without inhibitor treatment. Insulin levels and insulin tolerance tests in WT, but not in PAI-1 transgenic mice, suggested a higher insulin sensitivity after inhibitor treatment (insulin level 30 min after glucose injection of 2.0 +/- 0.17 ng mL(-1) vs. 3.2 +/- 0.48 ng mL(-1) without inhibitor treatment; P = 0.028). CONCLUSIONS: In this model, overexpression of PAI-1 moderately impaired adipose tissue formation without affecting glucose or insulin tolerance. Administration of a synthetic PAI-1 inhibitor for 4 weeks did not affect adipose tissue development in WT or PAI-1 transgenic mice, but induced a higher insulin sensitivity in WT mice.     

SIRT1/FOXO1信号通路对小鼠脑缺血再灌注损伤的调控作用

目的 探讨沉默信号调节器1(SIRT1)/叉头转录因子(FOXO1)信号通路对小鼠脑缺血再灌注损伤的调控作用。方法 24只清洁级C57雄性小鼠随机分为假手术组、缺血/再灌注组(I/R组)、SIRT1FOXO1信号通路抑制组(实验组),每组8只小鼠。I/R组与实验组采用夹闭、开放双侧颈总动脉法建立脑缺血再灌注损伤小鼠模型,同时实验组给予EX527(SIRT1抑制剂)腹腔注射,I/R组和假手术组腹腔注射等剂量的生理盐水。比较三组的神经功能缺损评分、脑组织含水量、感觉功能和行走协调能力评分,血浆中白介素-6(IL-6)、白细胞介素1(IL-1β)、细胞间黏附分子-1(ICAM-1)和肿瘤坏死因子-α(TNF-α)水平,SIRT1、FOXO1、NF-E2相关因子2(Nrf2)和血红素加氧酶1(HO-1)蛋白表达量,B淋巴细胞瘤-2(Bcl-2)、半胱氨酸蛋白酶-3(Caspase-3)、BCL-2关联蛋白X(Bax)和半胱氨酸蛋白酶-9(Caspase-9) mRNA表达,超氧化物歧化酶(SOD)、一氧化氮(NO)、丙二醛(MDA)和谷胱甘肽(GSH)含量。结果与假手术组相比,I/R组经功能缺损评分、行走协调能力评分和感觉功能、脑组织含水量、炎症因子水平、FOXO1蛋白表达、BAX、Caspase-3和Caspase-9 mRNA表达以及氧化应激指标NO和MDA含量均显著增加(P <0. 05),Nrf2、SIRT1和HO-1蛋白表达、Bcl-2 mRNA表达以及SOD和GSH含量均显著降低(P <0. 05);与I/R组相比,实验组所有指标变化更显著。结论 抑制SIRT1/FOXO1信号通路后,脑缺血/灌注小鼠神经功能降低,脑损伤程度均加重,由此推测SIRT1/FOXO1信号通路的激活与抑制脑缺血/灌注后的炎症反应、减轻氧化应激作用以及减缓细胞凋亡进程相关。     

Anti-obesity effects of traditional and standardized meju in high-fat diet-induced obese C57BL/6J mice

The aim of the study was to evaluate the anti-obesity effects of two types of meju in diet induced obese C57BL/6J mice. Animals were randomly divided into 4 dietary group (n = 10); normal diet, high fat diet with 30% soybean, high fat diet with 30% traditional meju, high fat diet with 30% standardized meju. After 16 weeks, after animals were sacrificed. It was observed that the high fat diet with 30% traditional meju and high fat diet with 30% standardized meju significantly reduced body weight gain, epididymal fat weight, serum triglyceride along with serum insulin and leptin levels compared to the high fat diet with 30% soybean. And also, the expression levels of hepatic lipid anabolic genes were significantly decreased in the high fat diet with 30% traditional meju and high fat diet with 30% standardized meju compared to the high fat diet with 30% soybean. In conclusion, the assessment of all the obesity markers strongly advocate the anti-obesity effect of traditional as well as standardized meju in diet induce obesity conditions.     

肠三叶因子介导PI3K/Akt信号通路保护胃黏膜上皮细胞紧密连接

目的：探讨肠三叶因子对胃黏膜上皮细胞紧密连接的保护作用,并研究PI3K/Akt信号通路在其中的作用机制。方法：体外培养GES-1细胞,分别设正常对照组,LPS组,ITF组,LPS＋ITF组,LPS＋ITF＋LY294002组,ITF＋LY294002组。正常对照组：正常培养;LPS组：加入浓度为10mg/L的LPS;ITF组：加入浓度为100μg/L的ITF;LPS＋ITF组：加入浓度为10mg/L的LPS,同时加入100μg/L的ITF;LPS＋ITF＋LY294002组：加入浓度为10mg/L的LPS、100μg/L的ITF,同时加入PI3K/Akt信号通路的抑制剂LY294002（15μM）;ITF＋LY294002组：加入100μg/L的ITF,同时加入15μM的LY294002。培养48h,采用Western blot检测ITF对PI3K/Akt信号通路的作用,采用免疫荧光和Western blot检测细胞紧密连接蛋白Occludin和ZO-1的变化情况。结果：Western blot检测结果说明,与对照组相比,ITF提高了pAkt蛋白的表达水平,而LY294002抑制了ITF激活的pAkt蛋白的表达,说明ITF可以通过激活PI3K/Akt信号通路来调控GES-1细胞的生理活动。免疫荧光和Western blot结果显示LPS导致GES-1细胞的紧密连接遭到破坏,降低紧密连接蛋白Occludin和ZO-1的表达水平,而ITF可以通过激活PI3K/Akt信号通路来保护GES-1细胞的紧密连接的完整性。结论：ITF保护胃黏膜上皮细胞紧密连接的完整性,提高紧密连接蛋白的表达水平,其发挥作用的主要分子机制是通过激活PI3K/Akt信号通路来实现的。     

11β-Hydroxysteroid dehydrogenase type 1-driven cortisone reactivation regulates plasminogen activator inhibitor type 1 in adipose tissue of obese women

BACKGROUND: Plasminogen activator inhibitor type 1 (PAI-1) is the main inhibitor of the fibrinolytic system and contributes to an increased risk of atherothrombosis in insulin-resistant obese patients. In adipose tissue, we have shown that PAI-1 is synthesized mainly in the visceral stromal compartment and is positively regulated by glucocorticoids. We have demonstrated that adipose tissue expression of 11beta-hydroxysteroid dehydrogenase type 1 (11beta-HSD-1), an enzyme that catalyzes the conversion of inactive cortisone to active cortisol, is exaggerated in obese patients. OBJECTIVES: We hypothesized that increased action of 11beta-HSD-1 in adipose tissue of obese subjects may contribute to PAI-1 overproduction. PATIENTS AND METHODS: Using in situ hybridization, we studied the expression of the mRNAs coding for PAI-1 and 11beta-HSD-1 in the stromal compartment of visceral adipose tissue obtained from obese women. The regulation of PAI-1 secretion from in vitro incubated tissue explants was also investigated. RESULTS: Regression analysis showed a significant positive linear relationship between PAI-1 and 11beta-HSD-1 mRNAs expression. In vitro incubation of adipose tissue explants demonstrated that cortisone stimulated PAI-1 gene expression and secretion, and that these effects were inhibited by co-incubation with the 11beta-HSD inhibitor, glycyrrhetinic acid. CONCLUSIONS: Our data demonstrate that 11beta-HSD-1-driven cortisone reactivation regulates adipose PAI-1 synthesis and secretion. They suggest that the increased PAI-1 synthesis and secretion observed in obese patients can be also related, at least in part, to an increased local conversion of cortisone to cortisol. Therefore, local cortisol metabolism in adipose tissue may be involved in increasing the risk of cardiovascular disease in obese subjects.     

LINC00941 promoted in vitro progression and glycolysis of laryngocarcinoma by upregulating PKM via activating the PI3K/AKT/mTOR signaling pathway

BackgroundLINC00941 has been proved to be related to various tumors, but its relationship with laryngocarcinoma remains vague.MethodsLINC00941 expression in laryngocarcinoma tumor and laryngocarcinoma cells was determined by real time‐quantitative polymerase chain reaction (RT‐qPCR). Besides, the five‐year survival of laryngocarcinoma patients with different LINC00941 expression was analyzed with Kaplan–Meier survival analysis, and the clinical characteristics of laryngocarcinoma patients were also recorded. After transfection, cell viability, cell proliferation, apoptosis, cell cycle, migration, and invasion were detected by cell counting kit‐8 (CCK‐8), colony formation, flow cytometry, cell scratch, and Transwell assays, respectively. Glycolysis was assessed by the colorimetric method. Expressions of proliferation‐associated proteins, migration‐associated proteins, glycolysis‐associated proteins, and phosphatidylinositol 3‐kinase (PI3K)/AKT/mammalian target of rapamycin (mTOR) signal pathway‐associated proteins were detected by Western blot.ResultsIn laryngocarcinoma tumor tissues and cells, LINC00941 was highly expressed. High expression of LINC00941 decreased the 5‐year survival of laryngocarcinoma patients, and it was positively related to lymph node metastasis and clinical stages. LINC00941 overexpression decreased apoptosis but promoted cell viability, proliferation, cell‐cycle progression, migration, and invasion, and glucose consumption and lactate production in laryngocarcinoma cells. Moreover, LINC00941 overexpression elevated expressions of Ki‐67, PCNA, MMP2, N‐Cadherin, HK2, PFKFB4, and PKM, activated the PI3K/AKT/mTOR signal pathway but reduced E‐Cadherin expression, while LINC00941 silencing had the opposite effects. PKM overexpression reversed the effects of LINC00941 silencing on cellular and glycolytic phenotypes.ConclusionLINC00941 promoted in vitro progression and glycolysis of laryngocarcinoma cells by upregulating PKM via activating the PI3K/AKT/mTOR signaling pathway.     

辛伐他汀对人急性单核细胞白血病SHI-1细胞PI3K/AKT通路的影响

本研究探讨羟甲基戊二酸单酰辅酶A还原酶抑制剂辛伐他汀对人急性单核细胞白血病株(SHI-1)细胞增殖和凋亡的影响及PI3K/AKT通路变化。取对数生长期细胞,实验分为阴性对照组和辛伐他汀处理组(终浓度分别为5、10、15μmol/L),培养24、48、72 h。采用MTT法观察SHI-1细胞增殖能力;流式细胞术测定SHI-1细胞凋亡指标变化;PCR芯片研究SHI-1细胞PI3K/AKT通路84个特异性基因mRNA的差异表达。结果表明,辛伐他汀对SHI-1细胞有明显抑制增殖和促凋亡作用,呈时间与剂量依赖性。15μmol/L辛伐他汀处理SHI-1细胞24、48、72h,细胞增殖抑制率分别为26.82%、47.09%、63.92%,细胞早期凋亡率分别为5.73%、13.25%、15.59%。与对照组相比,15μmol/L辛伐他汀处理SHI-1细胞48 h组中有39个基因表达发生改变,其中26个基因表达下调、13个基因表达上调。结论:辛伐他汀能抑制SHI-1细胞增殖并诱导其凋亡,其诱导凋亡机制可能与辛伐他汀调节PI3K/AKT通路相关基因的表达有关。     

电针对APP/PS1小鼠皮质区磷脂酰肌醇-3激酶/糖原合成酶激酶-3α通路相关蛋白表达和老年斑沉积的影响

目的 观察电针对APP/PS1双转基因小鼠皮质区磷脂酰肌醇-3激酶/糖原合成酶激酶-3α (PI3K/GSK3α)信号通路上相关蛋白的分布表达,以及老年斑沉积的影响。 方法 基因鉴定后,3月龄雄性转基因小鼠随机分为模型组和电针组,野生型C57小鼠为对照组,每组6只。电针组电针百会(GV20)和双侧肾俞(BL23)14 d。治疗后,Western blotting和免疫组化法检测各组小鼠皮质区老年斑数量,PI3K亚单位P85α和P110α,以及GSK3α和pS^21-GSK3α等蛋白的分布和表达。 结果 模型组皮质区老年斑数较对照组显著升高(P < 0.001);电针组老年斑数较模型组显著降低( P < 0.001)。各相关蛋白主要表达于皮质神经元胞质。与对照组相比,模型组pS ^21-GSK3α、P110α和P85α蛋白表达明显降低(P < 0.01),GSK3α蛋白表达升高( P < 0.05);与模型组相比,电针组pS ^21-GSK3α、P110α和P85α的蛋白表达明显升高(P < 0.01),GSK3α蛋白表达降低( P < 0.05)。 结论 电针能调节APP/PS1双转基因小鼠皮质区PI3K/GSK3α通路相关蛋白表达,减少老年斑沉积。     

Elabela通过α7nAChR/JAK2/STAT3信号通路改善心肌缺血/再灌注损伤的机制研究

目的 探讨Elabela在心肌缺血/再灌注损伤中的作用机制。方法 检测急性ST段抬高型心肌梗死（STEMI）患者体内Elabela的浓度, 通过获取临床血样和体外建立氧化应激模型来模拟缺血再灌注损伤, 评估Elabela在大鼠心肌细胞（H9C2）中的表达和作用。结果 STEMI患者血浆中Elabela的浓度和经过氧化氢（H₂O₂）处理的H9C2细胞中Elabela表达均升高（P均< 0.001）。Elabela可降低H9C2细胞凋亡相关蛋白Bax水平、心肌细胞凋亡率、活性氧阳性细胞数、乳酸脱氢酶水平和丙二醛水平;增加经H₂O₂处理的H9C2细胞的超氧化物歧化酶活性和Bcl-2表达（P均< 0.05）。特异性抑制剂α-银环蛇毒素对α7nAChR/JAK2/STAT3信号通路的抑制抵消了部分Elablela的保护作用。结论 Elabela通过激活α7nAChR/JAK2/STAT3信号通路改善H₂O₂诱导的H9C2细胞损伤。     

长链非编码RNA-PVT1介导JAK/STAT3信号通路对肺纤维化的研究

目的 探讨长链非编码RNA-浆细胞瘤变异易位基因1/蛋白质酪氨酸激酶/信号转导和转录激活因子3（lnc-PVT1/JAK/STAT3）信号通路参与肺纤维化的作用机制。方法 采用不同浓度脂多糖（LPS）作用于人正常肺上皮细胞BEAS-2B,用CCK-8法检测细胞活性、蛋白免疫印迹法检测α-SMA的蛋白相对表达量,对LPS...     

基于Notch1信号通路探讨杭白菊总黄酮治疗心肌肥厚小鼠的机制研究

目的探讨杭白菊总黄酮对异丙肾上腺素（ISO）诱导的心肌肥厚小鼠的保护作用及其机制。 方法将40只雄性小鼠分为对照组、总黄酮组、模型组和干预组,每组各10只。总黄酮组和干预组给予总黄酮（100 mg/kg）灌胃处理14 d,对照组和模型组给予相同体积的等渗NaCl溶液灌胃处理。模型组和干预组于第8天皮下多点注射ISO（5 mg/kg,0.5 mL）连续7 d以建立小鼠心肌肥厚模型,对照组和总黄酮组给予皮下相同体积等渗NaCl溶液多点注射。计算所有小鼠的左室射血分数（LVEF）、左室短轴缩短率（LVFS）、左心室质量指数（LVMI）、全心质量指数（HMI）、心脏质量/胫骨长度（HW/TL）比值。检测心肌组织中超氧化物歧化酶（SOD）、丙二醛及活性氧的含量;并且采用实时定量逆转录聚合酶链反应（qRT-PCR）法检测Collagen-1、Notch1、Hes1、心肌肌球蛋白重链β（β-MHC）、心房利钠肽（ANP）、Nox4及Nrf2的mRNA表达水平。 结果4组小鼠间LVEF、LVFS、LVMI、HMI、HW/TL、SOD、丙二醛及活性氧含量的比较,差异均有统计学意义（F = 62.114、18.814、59.824、66.375、48.362、59.677、46.195、48.257,P均< 0.001）。且与对照组相比,模型组小鼠的LVEF、LVFS及SOD水平均明显降低,LVMI、HMI、HW/TL、丙二醛及活性氧含量均显著升高（P均< 0.05）,而干预组小鼠的LVEF[（56 ± 6）% vs.（38 ± 5）%]、LVFS [（32 ± 5）% vs.（22 ± 4）%]、SOD水平均显著高于模型组,LVMI [（4.1 ± 0.4）mg/g vs.（5.8 ± 0.5）mg/g]、HMI [（5.3 ± 1.1）mg/g vs.（6.5 ± 0.6）mg/g]、HW/TL [（9.8 ± 1.2）mg/mm vs.（12.4 ± 1.3）mg/mm]、丙二醛及活性氧含量较模型组均显著降低（P均< 0.05）。4组小鼠间Collagen-1、Notch1、Hes1、β-MHC、ANP、Nox4及Nrf2的mRNA表达水平的比较,差异均有统计学意义（F = 85.372、43.865、69.841、36.358、23.964、48.354、57.781,P均< 0.001）。且与对照组相比,模型组小鼠Collagen-1、Notch1、Hes1、β-MHC、ANP、Nox4的mRNA表达水平均明显上调,Nrf2的mRNA表达水平明显下调（P均< 0.05）,而干预组小鼠Collagen-1、Notch1、Hes1、β-MHC、ANP及Nox4的mRNA表达水平较模型组均显著下调,Nrf2的mRNA表达水平较模型组明显上调（P均< 0.05）。 结论杭白菊总黄酮可以明显减轻ISO诱导产生的心肌肥厚,其作用与阻断Notch1信号通路,从而减轻心肌氧化应激损伤及抑制心肌组织纤维化的进程有关。     

Dihydromyricetin enhances the osteogenic differentiation of human bone marrow mesenchymal stem cells in vitro partially via the activation of Wnt/β‐catenin signaling pathway

Substantial evidence has demonstrated that the decreased osteogenic differentiation of bone mesenchymal stem cells (BMSCs) is closely related to bone metabolic diseases. Thus, it is very important to develop several potentially useful therapeutic agents to enhance BMSC osteogenesis. Flavonoids show promise in enhancing bone mass. Dihydromyricetin (DMY), a type of flavonoid, has not yet been investigated regarding its effects on BMSC osteogenesis. To investigate the effects of DMY on osteogenesis, human BMSCs were induced with or without DMY. We found that DMY (0.1–50 μm ) exhibited no cytotoxic effect on proliferation, but increased alkaline phosphatase activity, osteoblast‐specific gene expression, and mineral deposition. It also enhanced active β‐catenin expression and reduced dickkopf‐1(DKK1) and sclerostin expression. The Wnt/β‐catenin signaling pathway inhibitor (DKK1 and β‐catenin‐specific siRNA) decreased the enhanced bone mineral formation caused by DMY. Taken together, these findings reveal that DMY enhances osteogenic differentiation of human BMSCs partly through Wnt/β‐catenin in vitro.     

Downregulation of MMP1 functions in preventing perineural invasion of pancreatic cancer through blocking the NT‐3/TrkC signaling pathway

BackgroundPancreatic cancer (PC) is a fatal malignancy that frequently involves perineural invasion (PNI). This study aims to investigate the function and underlying mechanisms of matrix metalloproteinase‐1 (MMP1) in PNI of PC.MethodsHuman pancreatic cancer PANC‐1 cells were co‐cultured with dorsal root ganglion in vitro. The expression of MMP1, epithelial–mesenchymal transition (EMT) markers, Schwann cell markers, neurotrophic factors, NT‐3, and TrkC was measured by qRT‐PCR or Western blot. Transwell assay was performed to evaluate cell migration and invasion. In vivo model of PNI was established via inoculating PANC‐1 cells into mice. PANC‐1 cells and mice were also treated with LM22B‐10 (an activator of TrkC) to confirm the mechanisms involving NT‐3/TrkC in PNI of PC both in vivo and in vitro.ResultsThe expression of MMP1 was significantly higher in PDAC tissues than non‐cancerous tissues, which was positively associated with PNI. MMP1 knockdown repressed the migration and invasion of PANC‐1 cells. Except for E‐cadherin, the expression of EMT markers, Schwann cell markers, neurotrophic factors, NT‐3, and TrkC was inhibited by MMP1 silencing. The same effects of MMP1 knockdown on the above factors were also observed in the PNI model. Moreover, MMP1 knockdown elevated the sciatic nerve function and reduced PNI in the model mice. LM22B‐10 partially abolished the effects of MMP1 knockdown both in vivo and in vitro.ConclusionsSilencing of MMP1 prevents PC cells from EMT and Schwann‐like cell differentiation via inhibiting the activation of the NT‐3/TrkC signaling pathway, thus alleviating the PNI of PC.     

巨刺法对脑缺血再灌注损伤大鼠运动功能、脑影像结构及PI3K/AKT信号通路的影响

目的:观察巨刺法对脑缺血再灌注损伤模型大鼠运动功能及影像学结构的影响,并探讨其可能的作用机制。方法:将30例健康雄性SD大鼠随机分为假手术组、模型组及巨刺组,各10只。模型组及巨刺组大鼠接受改良局灶性脑缺血再灌注大脑中动脉栓塞(MCAO)模型制备,假手术组仅接受切开皮肤+分离血管+皮肤缝合处理。巨刺组进行巨刺曲池穴及足三里穴7d,模型组及假手术组仅接受模拟捉拿及相应穴位点触。各组采用神经行为学评分评估大鼠运动功能,H反射评估大鼠肌张力状况,TTC染色及MRI观察脑梗死体积,Western Blot检测PI3K、AKT、pAKT水平,免疫组化检测Caspase-3的表达,Tunel检测神经元细胞调亡情况。结果:巨刺法可明显降低MCAO大鼠的神经行为学评分、H反射的潜伏期、脑梗死体积、神经元凋亡率以及Caspase-3表达,增加H反射的波幅、PI3K、p-AKT表达水平。结论:巨刺法可明显改善脑缺血再灌注损伤大鼠的运动功能,其作用机制可能与介导PI3K/AKT信号通路有关。     

高强度间歇运动训练上调APP/PS1转基因阿尔兹海默病小鼠海马线粒体自噬的研究

目的:观察高强度间歇运动训练干预是否能改善APP/PS1转基因AD小鼠病理表现,并探讨线粒体自噬在其间的生物学效应。方法:8月龄雄性APP/PS1转基因小鼠分为转基因模型安静组(SED-Tg)和转基因模型+高强度间歇运动训练组(HIIT-Tg),C57BL/6野生型小鼠纳入野生型安静组(SED-Wt)。HIIT-Tg组动物进行12周高强度间歇运动训练。避暗被动回避实验检测学习记忆能力,JC-1荧光探针检测海马线粒体膜电位,二氯荧光素探针检测海马线粒体ROS,Western Blot法检测脑海马Aβ-42、BDNF、AMPK、PINK1、Parkin、Bnip3蛋白表达。结果:(1)SED-Tg组与SED-Wt组比较,逃避潜伏期、线粒体膜电位、BDNF、AMPK、PINK1、Parkin和Bnip3蛋白表达显著降低(P<0.05—0.01),线粒体ROS产生速率和Aβ-42蛋白表达显著升高(P<0.01)。(2)HIIT-Tg组与SED-Tg组比较,逃避潜伏期、线粒体膜电位、BDNF、AMPK、PINK1和Parkin蛋白表达显著升高(P<0.01),线粒体ROS产生速...     

CIB1 deficiency results in impaired thrombosis: the potential role of CIB1 in outside-in signaling through integrin αIIbβ3

Summary.  Background: Agonist-induced inside-out signaling activates platelet integrin α_IIbβ₃, rendering it to bind plasma fibrinogen (Fg). Fg binding induces outside-in signaling that culminates in platelet aggregation, leading to physiological hemostasis and pathological thrombosis. How outside-in signaling through α_IIbβ₃ regulates hemostasis and thrombosis is not well understood. We have previously shown that CIB1 is involved in regulating α_IIbβ₃ function. Objective: To determine the in vivo role of CIB1 in the process of hemostasis and thrombosis. Methods and Results: Genetic ablation of Cib1 significantly increased mouse tail bleeding time. Greater than 50% of the Cib1 null mice showed a rebleeding phenotype. Time taken for complete occlusion of carotid artery upon 10% FeCl₃-induced injury was significantly delayed in the absence of Cib1. This was also associated with unstable thrombus formation. The inside-out signaling appears normal as ADP-, collagen- and PAR4 peptide-induced aggregation and fibrinogen binding was unaffected. The absence of Cib1 also affected the ability of platelets to spread on immobilized Fg, but not filopodia formation. Spreading could be restored in Cib1 null platelets by the addition of exogenous ADP. Outside-in signaling-dependent tyrosine phosphorylation of the integrin β₃ subunit was significantly reduced in the absence of Cib1 as determined by Western blot analysis. Conclusion: Using gene knockout mice, we show for the first time that lack of Cib1 results in impaired thrombosis. CIB1 regulates these processes by affecting platelet spreading, but not platelet filopodia formation. These in vivo and in vitro results clearly show that CIB1 is a key regulator of thrombosis.     

CKS1B promotes cell proliferation and invasion by activating STAT3/PD‐L1 and phosphorylation of Akt signaling in papillary thyroid carcinoma

ObjectiveTo investigate role of GKS1B and its relationship between STAT3/PD‐L1 and p‐Akt in papillary thyroid carcinoma (PTC).MethodsExpression of GKS1B and PD‐L1 was determined in PTC cell lines. GKS1B was overexpressed or knocked down by transfection with overexpression plasmids or si‐CKS1B. STAT3 inhibitor WP1066 was used to suppress STAT3, and PD‐L1 inhibitor Pembrolizumab was used to block PD‐L1. Cell viability and invasion were evaluated by MTT and transwell assay, respectively. The expression of STAT3, p‐STAT3, Akt, and p‐Akt was measured using Western blotting.ResultsBoth protein levels and mRNA levels of CKS1B and PD‐L1 were remarkably up‐regulated in PTC cell lines. Knockdown of CKS1B significantly inhibited cell viability and invasion of PTC cells and suppressed STAT3/PD‐L1 signaling and Akt phosphorylation, while overexpression of CKS1B led to opposite results. Inhibition of STAT3 or PD‐L1 reversed the effects of overexpressed CKS1B on PTC cells.ConclusionThe overexpression of CSK1B could promote cell viability and invasion of PTC cells through activation of STAT3/PD‐L1 signaling and Akt phosphorylation.