Suppressing high mobility group box-1 release alleviates morphine tolerance via the adenosine5'-monophosphate-activated protein kinase/heme oxygenase-1 pathway

Opioids,such as morphine,are the most potent drugs used to treat pain.Long-term use results in high tolerance to morphine.High mobility group box-1(HMGB1) has been shown to participate in neuropathic or inflammatory pain,but its role in morphine tolerance is unclear.In this study,we established rat and mouse models of morphine tolerance by intrathecal injection of morphine for 7 consecutive days.We found that morphine induced rat spinal cord neurons to release a large amount of HMGB1.HMGB1 regulated nuclear factor κB p65 phosphorylation and interleukin-1β production by increasing Toll-like receptor 4receptor expression in microglia,thereby inducing morphine tolerance.Glycyrrhizin,an HMGB1 inhibito r,markedly attenuated chronic morphine tole rance in the mouse model.Finally,compound C(adenosine 5’-monophosphate-activated protein kinase inhibitor) and zinc protoporphyrin(heme oxygenase-1 inhibitor)alleviated the morphine-induced release of HMGB1 and reduced nuclear factor κB p65 phosphorylation and interleukin-1β production in a mouse model of morphine tolerance and an SH-SY5Y cell model of morphine tole rance,and alleviated morphine tolerance in the mouse model.These findings suggest that morphine induces HMGB1 release via the adenosine 5’-monophosphate-activated protein kinase/heme oxygenase-1 signaling pathway,and that inhibiting this signaling pathway can effectively reduce morphine tole rance.

Electroacupuncture improves neuropathic pain Adenosine, adenosine 5'-triphosphate disodium and their receptors perhaps change simultaneously

Applying a stimulating current to acupoints through acupuncture needles–known as electroacupuncture–has the potential to produce analgesic effects in human subjects and experimental animals. When acupuncture was applied in a rat model, adenosine 5-triphosphate disodium in the extracellular space was broken down into adenosine, which in turn inhibited pain transmission by means of an adenosine A1 receptor-dependent process. Direct injection of an adenosine A1 receptor agonist enhanced the analgesic effect of acupuncture. The analgesic effect of acupuncture appears to be mediated by activation of A1 receptors located on ascending nerves. In neuropathic pain, there is upregulation of P2X purinoceptor 3 （P2X3） receptor expression in dorsal root ganglion neurons. Conversely, the onset of mechanical hyperalgesia was diminished and established hyperalgesia was significantly reversed when P2X3 receptor expression was downregulated. The pathways upon which electroacupuncture appear to act are interwoven with pain pathways, and electroacupuncture stimuli converge with impulses originating from painful areas. Electroacupuncture may act via purinergic A1 and P2X3 receptors simultaneously to induce an analgesic effect on neuropathic pain.

白花蛇舌草提取物通过AMPK/ATG5信号通路对急性胰腺炎大鼠肺损伤的保护作用机制研究

目的:通过5’-单磷酸腺苷活化蛋白激酶(AMPK)/自噬相关蛋白5(ATG5)信号通路,探讨白花蛇舌草提取物减轻急性胰腺炎(AP)大鼠肺损伤的机制。方法:建立AP肺损伤大鼠模型,随机分为模型组、提取物(白花蛇舌草提取物)组、3-MA(自噬抑制剂3-甲基腺嘌呤)组、AICAR(AMPK激活剂)组、提取物+AICAR组,每组15只,另取15只大鼠作为假手术组;ELISA检测血清淀粉酶(AMY)、IL-1β、IL-6、IL-18水平;检测大鼠腹水量及肺湿/干重比(W/D);HE观察胰腺及肺组织病理损伤;Western blot检测溶酶体相关膜蛋白2(LAMP2)、自噬底物p62、IL-1β前体蛋白(pro-IL-1β)、胰蛋白酶原活化肽(TAP)、AMPK、p-AMPK、ATG5、自噬标志物LC3-Ⅱ/Ⅰ、泛素特异性蛋白酶10(UPS10)表达。结果:与假手术组相比,模型组大鼠腹水量、肺W/D、胰腺和肺组织病理损伤评分、血清AMY、IL-1β、IL-6、IL-18水平、胰腺组织p62、TAP、pro-IL-1β表达、肺组织pAMPK/AMPK、ATG5、LC3-Ⅱ/Ⅰ、UPS10、pro-IL-1β表达均升高(P<0.05),胰腺和肺组织LAMP2蛋白表达降低(P<0.05);模型大鼠经白花蛇舌草提取物或自噬抑制剂3-MA干预后,上述指标均得到显著改善(P<0.05),且白花蛇舌草提取物的改善效果优于3-MA(P<0.05);而AMPK激活剂AICAR可削弱白花蛇舌草提取物对AP大鼠肺损伤的改善作用(P<0.05)。结论:白花蛇舌草提取物可通过抑制AMPK/ATG5信号通路减轻炎症反应、降低自噬水平改善AP大鼠肺损伤。

Diabetes and inflammatory diseases:An overview from the perspective of Ca^(2+)/3'-5'-cyclic adenosine monophosphate signaling

A large amount of evidence has supported a clinical link between diabetes and inflammatory diseases,e.g.,cancer,dementia,and hypertension.In addition,it is also suggested that dysregulations related to Ca^(2+)signaling could link these diseases,in addition to 3'-5'-cyclic adenosine monophosphate(cAMP)signaling pathways.Thus,revealing this interplay between diabetes and inflammatory diseases may provide novel insights into the pathogenesis of these diseases.Publications involving signaling pathways related to Ca^(2+)and cAMP,inflammation,diabetes,dementia,cancer,and hypertension(alone or combined)were collected by searching PubMed and EMBASE.Both signaling pathways,Ca^(2+)and cAMP signaling,control the release of neurotransmitters and hormones,in addition to neurodegeneration,and tumor growth.Furthermore,there is a clear relationship between Ca^(2+)signaling,e.g.,increased Ca^(2+)signals,and inflammatory responses.cAMP also regulates pro-and anti-inflammatory responses.Due to the experience of our group in this field,this article discusses the role of Ca^(2+)and cAMP signaling in the correlation between diabetes and inflammatory diseases,including its pharmacological implications.As a novelty,this article also includes:(1)A timeline of the major events in Ca^(2+)/cAMP signaling;and(2)As coronavirus disease 2019(COVID-19)is an emerging and rapidly evolving situation,this article also discusses recent reports on the role of Ca^(2+)channel blockers for preventing Ca^(2+)signaling disruption due to COVID-19,including the correlation between COVID-19 and diabetes.

In Vitro Functional Study of Rice Adenosine 5'-Phosphosulfate Kinase

Sulfate can be activated by ATP sulfurylase and adenosine 5'-phosphosulfate kinase(APSK) in vivo. Recent studies suggested that APSK in Arabidopsis thaliana regulated the partition between APS reduction and phosphorylation and its activity can be modulated by cellular redox status. In order to study regulation of APSK in rice(Os APSK), Os APSK1 gene was cloned and its activity was analyzed. Os APSK1 C36 and C69 were found to be the conserved counterparts of C86 and C119, which involved in disulfide formation in At APSK. C36A/C69 A Os APSK1 double mutation was made by site directed mutagenesis. Os APSK1 and its mutant were prokaryotically over-expressed and purified, and then assayed for APS phosphorylation activity. Os APSK1 activity was depressed by oxidized glutathione, while the activity of its mutant was not. Further studies in the case that oxidative stress will fluctuate in vivo 3'-phosphoadenosine-5'-phosphosulfate content, and all APSK isoenzymes have similar regulation patterns are necessary to be performed.

Electrochemical Studies of Effect of Eu^(3+) on Adenosine-5′-Diphosphate at Mercury Electrode

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The Adenosine Receptor Agonist 5’-N-Ethylcarboxamide-Adenosine Increases Mouse Serum Total Homocysteine Levels, Which Is a Risk Factor for Cardiovascular Diseases

An increase in total homocysteine (Hcy) levels (protein-bound and free Hcy in the serum) has been identified as a risk factor for vascular diseases. Hcy is a product of the methionine cycle and is a precursor of glutathione in the transsulfuration pathway. The methionine cycle mainly occurs in the liver, with Hcy being exported out of the liver and subsequently bound to serum proteins. When the non-specific adenosine receptor agonist 5’-N-ethylcarboxamide-adenosine (NECA;0.1 or 0.3 mg/kg body weight) was intraperitoneally administered to mice that had been fasted for 16 h, total Hcy levels in the serum significantly increased 1 h after its administration. The NECA treatment may have inhibited transsulfuration because glutathione levels were significantly decreased in the liver. After the intraperitoneal administration of a high dose of NECA (0.3 mg/kg body weight), elevations in total Hcy levels in the serum continued for up to 10 h. The mRNA expression of methionine metabolic enzymes in the liver was significantly reduced 6 h after the administration of NECA. NECA-induced elevations in total serum Hcy levels may be maintained in the long term through the attenuated expression of methionine metabolic enzymes.

Changes in Adenosine Metabolism in Asthma. A Study on Adenosine, 5&#39-NT, Adenosine Deaminase and Its Isoenzyme Levels in Serum, Lymphocytes and Erythrocytes

Background: Adenosine deaminase (ADA) and 5'-nucleotidase (5'-NT) play a crucial role in adenosine metabolism in healthy individuals. Adenosine is an inflammatory mediator of asthma. Changes in adenosine metabolism and role of ADA and 5'-NT in regulating adenosine level in asthmatics and correlation of these changes with severity of asthma are not clearly understood. Methods: In this study, we screened 5217 patients, of which 2416 were diagnosed with asthma. Further, of 2416 asthmatics, only 45 patients who strictly fulfilled the selection criteria were enrolled in the study. The patients were classified into mild, moderate and severe persistent groups;each group consisted of fifteen patients. Fifteen healthy subjects served as controls. Adenosine levels and activities of 5'-NT, total ADA, ADA1 and ADA2 in serum, lymphocytes and erythrocytes were determined. The data were analysed statistically and p vice versa.

SYNTHESIS OF ADENOSINE DERIVATIVES WITH CARBOXYALKYL SIDE CHAIN AT 2',3'OR 5'POSITION

Some nucleoside carhoxylic acid derivatives,such as(±)ethyl ester of griseolic acid (4)and 9-(2'-deoxy-2'-(benzyloxycarbonyl)-methylene-β-D-rihohept-2'-enofuranosyluranate)adenine (11),were synthesized.The formation of lactone of 5'-deoxy-adenosineacetic acid(AAA,3)was investigated by using different reagents for lactonization from AAA,but all of the efforts failed, and sone unexpected compounds were obtained.

The Adenosine Receptor Agonist 5’-<i>N</i>-Ethylcarboxamide-Adenosine Increases Glucose 6-Phosphatase Expression and Gluconeogenesis

Intraperitoneal administration of the non-selective adenosine receptor agonist 5’-N-ethylcarboxamide-adenosine (NECA) (0.1 or 0.3 mg/kg) increased fasting serum glucose levels in mice. To clarify the mechanism responsible for this, the expression of liver glucose 6-phosphatase (G6Pase: a gluconeogenic enzyme) was analyzed, and it was found that G6Pase mRNA was increased by NECA treatment. Administration of 0.3 mg/kg NECA resulted in elevated serum glucose levels at 1 h and were further elevated at 6 h. Administration of 0.1 mg/kg NECA increased serum glucose levels at 1 h and had returned to control levels by 6 h. The increase in fasting serum glucose levels induced by NECA are thought to be caused, in part, by elevated G6Pase expression.

腺苷A2a受体/Krüppel样因子5对缺血再灌注损伤大鼠心肌的影响

目的:探讨腺苷A2a受体/Krüppel样因子5(KLF5)对缺血再灌注损伤大鼠心肌的影响。方法:42只250~300 g雄性SD大鼠随机分为4组:假手术组(Sham组,n=6)、缺血再灌注组(I/R组,n=12)、缺血再灌注+腺苷A2a受体特异性激动剂CGS21680组(I/R+CGS组,n=12)、缺血再灌注+腺苷A2a受体拮抗剂ZM241385组(I/R+ZM组,n=12)。采用结扎左冠状动脉前降支再灌注的方法制备大鼠心肌缺血再灌注损伤模型。I/R+CGS组于再灌注前5 min静脉注射CGS21680后持续泵注60 min,CGS+ZM组于再灌注前5 min静脉注射ZM241385。于造模前,缺血5 min,再灌注10 min、45 min及120 min时分别记录各组大鼠的心率(HR)、平均动脉压(MAP)以及心率与收缩压乘积(RPP)。再灌注结束后取血液,酶联免疫吸附测定法检测血清心肌钙蛋白I(c TnI)和成纤维细胞生长因子21(FGF21)水平。再灌注结束后再次结扎左冠状动脉前降支,采用TTC染色法确定心肌梗死面积。处死大鼠,采用Western blot法检测缺血区心肌组织中的肿瘤坏死因子-α(TNF-α)、白细胞介素(IL)-1β和KLF5的蛋白表达水平。结果:各组造模前、Sham组各时间点HR、MAP及RPP差异无统计学意义。与Sham组比较,I/R组缺血5 min时HR、MAP及RPP降低;与I/R组比较,I/R+CGS组再灌注10 min和45 min时HR、MAP及RPP降低,I/R+ZM组再灌注10 min和45 min时HR、MAP及RPP升高(P均<0.05)。与Sham组比较,I/R组心肌梗死面积增大,血清cTnI水平升高,FGF21水平降低;与I/R组比较,I/R+CGS组心肌梗死面积减小,血清cTnI水平降低,FGF21水平升高,I/R+ZM组心肌梗死面积增大,血清c TnI水平升高,FGF21水平降低(P均<0.05)。与Sham组比较,I/R组心肌组织中TNF-α、IL-1β和KLF5的蛋白表达水平均明显升高;与I/R组相比,I/R+CGS组心肌组织中TNF-α、IL-1β和KLF5的蛋白表达水平均明显降低,I/R+ZM组心肌组织中TNF-α、IL-1β和KLF5的蛋白表达水平均明显升高(P均<0.05)。结论:激活腺苷A2a受体可抑制缺血心肌细胞中KLF5表达,降低心肌梗死再灌注损伤程度,缓解心肌细胞缺氧状态,促进心肌细胞损伤后修复,对缺血再灌注损伤大鼠心肌起保护作用。

Autophagy occurs within an hour of adenosine triphosphate treatment after nerve cell damage:the neuroprotective effects of adenosine triphosphate against apoptosis

After hypoxia, ischemia, or inflammatory injuries to the central nervous system, the damaged cells release a large amount of adenosine triphosphate, which may cause secondary neuronal death. Autophagy is a form of cell death that also has neuroprotective effects. Cell Counting Kit assay, monodansylcadaverine staining, flow cytometry, western blotting, and real-time PCR were used to determine the effects of exogenous adenosine triphosphate treatment at different concentrations （2, 4, 6, 8, 10 mmol/L） over time （1, 2, 3, and 6 hours） on the apoptosis and autophagy of SH-SY5Y cells. High concentrations of extracellular adenosine triphosphate induced autophagy and apoptosis of SH-SYSY cells. The enhanced autophagy first appeared, and peaked at 1 hour after treatment with adenosine triphosphate. Cell apoptosis peaked at 3 hours, and persisted through 6 hours. With prolonged exposure to the adenosine triphosphate treatment, the fraction of apoptotic cells increased. These data suggest that the SH-SY5Y neural cells initiated autophagy against apoptosis within an hour of adenosine triphosphate treatment to protect themselves against injury.

直肠癌组织中ADAM12、A2aR表达水平与神经脉管侵犯及5-FU化疗敏感性的关系

目的探究直肠癌组织中ADAM12、A2aR表达水平与神经脉管侵犯及5-FU化疗敏感性的关系。方法选取62例新型辅助放化疗治疗前活检石蜡标本,采用免疫组化法检测直肠癌组织中ADAM12、A2aR表达水平,采用直肠癌消退分级标准对直肠癌组织化疗敏感性进行评价。结果A2aR蛋白的表达与肿瘤大小、患者性别、年龄大小、神经脉管侵犯程度无关(P>0.05),与TNM分期、浸润程度有关(P<0.05)。ADAM12蛋白的表达与肿瘤大小、患者性别、年龄大小、神经脉管侵犯程度无关(P>0.05),与浸润程度、TNM分期有关(P<0.05)。直肠癌化疗敏感性与浸润程度、TNM分期、A2aR和ADAM12表达强度有关(P<0.05)。直肠癌组织中A2aR表达与5-FU化疗敏感性呈正相关(γ=0.352,P<0.001)。直肠癌组织中ADAM12表达与5-FU化疗敏感性呈正相关(γ=0.276,P<0.030)。结论直肠癌组织中ADAM12、A2aR阳性表达率较高,对神经脉管侵犯度不高,与5-FU化疗敏感性呈现正相关。

基于“肾脑相济”理论探讨艾灸对阿尔茨海默病大鼠海马AMPK/mTOR信号通路的影响

目的观察艾灸对阿尔茨海默病(Alzheimer’s disease,AD)大鼠腺苷酸活化蛋白激酶(adenosine 5-monophosphate activated protein kinase,AMPK)/雷帕霉素靶蛋白(mammalian target of rapamycin,mTOR)信号通路相关递质表达的影响,探讨艾灸治疗AD的作用机制。方法将SD大鼠按照随机数字表法分为正常组8只、模型组32只,采取侧脑室注射β淀粉样蛋白(amyloidβ-protein,Aβ)_(25-35)的方法建立大鼠AD模型。将模型复制成功的大鼠随机分为模型组、药物组、艾灸组,每组8只。艾灸组大鼠用艾条灸“百会”“肾俞”“三阴交”,每次15 min,同时按3 mg/kg灌胃蒸馏水;药物组大鼠按3 mg/kg灌胃盐酸多奈哌齐;对照组及模型组大鼠按3 mg/kg灌胃蒸馏水。采用Morris水迷宫法检测大鼠行为学表现,苏木精—伊红染色法观察大鼠海马病理组织改变,Western blot法检测大鼠海马磷酸化雷帕霉素靶蛋白(phosphorylated mammalian target of rapamycin,p-mTOR)、核糖体蛋白S6激酶(ribosomal protein S6 kinase p70,P70S6K)、自噬相关基因5(autophagy-related gene 5,ATG5)、磷酸化腺苷酸活化蛋白激酶(phosphorylated adenosine 5-monophosphate activated protein kinase,p-AMPK)、微管相关蛋白1轻链3B(microtubule associated protein light chain 3B,LC3B)-Ⅱ/LC3B-Ⅰ的表达水平。结果苏木精—伊红染色结果表明,模型组海马神经元萎缩明显,与模型组比较,药物组和艾灸组海马神经元形态及分化程度均有明显改善。与正常组比较,模型组大鼠的逃避潜伏期显著延长(P<0.05),p-mTOR及P70S6K表达水平均显著升高(P<0.05),ATG5、LC3B-Ⅱ/LC3B-Ⅰ、p-AMPK表达水平均显著降低(P<0.05)。与模型组比较,药物组和艾灸组大鼠的逃避潜伏期均显著缩短(P<0.05),p-mTOR及P70S6K表达水平均显著下降(P<0.05),ATG5、LC3B-Ⅱ/LC3B-Ⅰ、p-AMPK表达水平均显著上升(P<0.05)。与药物组比较,艾灸组大鼠逃避潜伏期显著缩短(P<0.05);p-mTOR及P70S6K表达水平显著降低(P<0.05),ATG5、LC3B-Ⅱ/LC3B-Ⅰ、p-AMPK表达水平均显著上升(P<0.05)。结论艾灸能够调控AMPK/mTOR信号通路,诱导细胞自噬,阻断脑内Aβ表达,从而改善认知功能。

氧化石墨烯-DNA纳米探针用于三磷酸腺苷的检测与药物递送

癌细胞内三磷酸腺苷(ATP)浓度异常与肿瘤发生发展过程密切相关,因此,快速、准确地检测细胞内外ATP水平具有重要意义。盐酸阿霉素(DOX)是一种广泛使用的抗癌药物,能嵌入DNA碱基对,并通过抑制DNA复制和转录诱导细胞凋亡。氧化石墨烯(Graphene oxide, GO)由于具有毒性低、比表面积大和易功能化,可以有效、稳定地负载DNA纳米探针进入细胞等优点而被广泛应用。然而,复杂环境中的生物分子容易通过物理吸附竞争性结合到GO表面,导致假阳性信号。基于此,提出了一种新型的GO-DNA纳米探针,并将其应用于ATP的检测与抗癌药物DOX靶向递送。以ATP的核酸适配体与其互补链杂交形成双链DNA(dsDNA),并通过G-C碱基对负载DOX,利用互补链延伸的poly A序列吸附到GO表面构建了GO-dsDNA-DOX纳米探针,能极大程度地降低复杂环境中物理吸附引起的干扰,减少假阳性信号产生。ATP与适配体特异性结合会导致DOX释放,根据其荧光“off-on”实现ATP的定量分析,DOX荧光强度与ATP含量在0.08~8.0 mmol/L范围内呈现良好的线性关系,线性方程为IF=3.0897c+129.08,检测限(3σ/k)为0.059 mmol/L,且方法具有良好的选择性和抗干扰能力。细胞毒性及荧光成像结果表明,负载DOX的纳米探针在人乳腺癌细胞(MCF-7)内有明显的药物释放,显著诱导细胞凋亡。该研究建立了一种免修饰、简单和快速的ATP含量检测分析方法,并利用癌细胞内高浓度ATP实现靶向药物递送,降低了对正常细胞的毒副作用,为癌症的治疗提供了新思路。

硫酸铈对5′-腺嘌呤及5′-鸟嘌呤核苷酸的水解断裂的催化作用

本文用核磁共振 (NMR)波谱和化学定磷法研究了 Ce(SO4) 2 对 5′-腺嘌呤核苷酸 (5′- AMP)及5′-鸟嘌呤核苷酸 (5′- GMP)的水解断裂作用。结果表明 :Ce(SO4) 2 在 37℃ ,酸性条件下使 5′- AMP断裂为腺嘌呤核苷 (A)及无机磷 ,使 5′- GMP断裂为鸟嘌呤核苷 (G)及无机磷 ,SO2 -4 浓度及酸强度对 5′- AMP及 5′- GMP的水解断裂程度有很大影响 ,并对其水解断裂机制进行了研究。

肝脏疾病诊断中腺苷脱氨酶、前清蛋白、α-L-岩藻糖苷酶、胆碱脂酶和5′-NT价值的临床评价

目的了解腺苷脱氨酶(ADA)、α-L-岩藻糖苷酶(AFU)、胆碱脂酶、前清蛋白和5′-核苷酸酶(5′-NT)在肝脏疾病诊断中的意义。方法选择332例肝脏疾病患者,其中,急性肝炎150例,重型肝炎25例,肝硬化40例,酒精性肝炎35例,慢性肝炎82例;选择同期90例健康体检者作为对照。采用Bayer ADVIA 2400全自动生化分析仪检测其血清ADA、AFU、前清蛋白、胆碱脂酶和5′-NT,绘制受试者工作特征曲线(ROC)并计算曲线下面积(AUC),评价5项指标在不同肝脏疾病中的诊断意义。结果重型肝炎、肝硬化患者血清ADA、AFU、5′-NT显著增高,前清蛋白、胆碱脂酶显著降低;ADA、AFU、5′-NT的异常值比例分别超过80%、65%、90%。急性肝炎、酒精性肝炎患者血清ADA、AFU、5′-NT轻度升高,前清蛋白、胆碱脂酶轻度下降;急性肝炎患者血清胆碱脂酶、5′-NT的异常值比例分别为80.0%、54.67%。血清前清蛋白、胆碱脂酶的AUC在重型肝炎患者中分别为0.022、0.000,在肝硬化组患者中分别为0.019、0.000;上述两组患者血清ADA、5′-NT的AUC均大于0.950,AFU的AU均大于0.800。在急性肝炎和酒精性肝炎组中,患者血清ADA、5′-NT的AUC均大于0.800,AFU的AUC分别为0.814、0.637。332例患者血清前清蛋白、胆碱脂酶的AUC分别为0.175、0.181,血清ADA、AFU、5′-NT的AUC分别为0.850、0.743、0.714。结论血清ADA、AFU、前清蛋白、胆碱脂酶和5′-NT检测对肝脏疾病的诊断具有重要价值。

蛋白质ADP核糖基化对秦川牛肉品质的影响

为研究蛋白质ADP核糖基化对宰后成熟初期秦川牛肉线粒体功能及肉品质的影响,以20µmol/L Rucaparib(ADP核糖聚合酶1(poly(ADP-ribose)polymerase 1,PARP1)抑制剂)处理的秦川牛背最长肌为研究对象,测定贮藏0 h、6 h、12 h、2 d、4 d、8 d对照组和抑制剂处理组样品的线粒体相关指标及肌原纤维小片化指数(myofibril fragmentation index,MFI)、剪切力、pH值等品质指标,并采用免疫印迹(Western Blot)法检测PARP1、肌间线蛋白表达水平。结果表明:0 h~8 d(12 h除外)抑制剂组活性氧含量显著低于对照组(P<0.05),宰后0~12 h抑制剂组Caspase-3活性、MFI显著低于对照组(P<0.05);在2~4 d抑制剂组线粒体膜电位较对照组高,4~8 d抑制剂处理组琥珀酸脱氢酶(succinate dehydrogenase,SDH)活性显著高于对照组(P<0.05)。说明抑制表征ADP核糖基化反应进行的PARP1后,线粒体膜电位下降变缓,SDH活性升高,这一定程度上维持了线粒体功能,使MFI下降、肌间线蛋白降解变缓,从而延缓肉品嫩化。

ADA、5’-NT、TRF、TBA、LAP和PA在肝病诊断中的临床评价

目的探讨腺苷脱氨酶(ADA)、5’-核苷酸酶(5’-NT)、转铁蛋白(TRF)、总胆汁酸(TBA)、亮氨酸氨基肽酶(LAP)、前白蛋白(PA)6项指标在肝病诊断和治疗中的价值。方法使用日立7600全自动生化分析仪测定41例急性肝炎、78例慢性肝炎、62例肝硬化、50例肝癌患者和53例健康对照者血清的上述6项指标,并对检测结果进行统计学分析。结果与正常对照组相比,急性肝炎组与肝癌组ADA、5’-NT、LAP显著升高(P<0.01),TRF和PA显著降低(P<0.01),TBA升高(P<0.05);慢性肝炎组中ADA、5’-NT、TBA、LAP与正常组相比有显著升高(P<0.01),而TRF、PA显著降低(P<0.01);肝硬化组中ADA、TBA与正常组相比有显著升高(P<0.01),而TRF、PA显著降低(P<0.01),5’-NT升高(P<0.05),但是肝硬化组的LAP与正常组无差异(P>0.05)。急性肝炎组受试者工作特性曲线(ROC)下面积5’-NT最大,慢性肝炎组曲线下面积ADA和PA较大,肝硬化组ADA、TBA和PA曲线下面积较大,肝癌组ADA、5’-NT和PA曲线下面积较大,LAP在慢性肝炎、肝硬化和肝癌组的曲线下面积较小。结论上述六项指标中,ADA和PA对肝病的诊断价值较高,可作为各实验室的常规项目开展,而除LAP外的其他5项指标对肝硬化均有较高临床应用价值。此外,上述6项指标与常规指标联合应用,可对临床医师在诊断、治疗、评估肝功能状况时起到重要的参考作用。

快眼动睡眠剥夺对抑郁模型大鼠下丘脑5-HT和腺苷的影响

目的了解快速眼动睡眠剥夺改善抑郁情绪的可能机制。方法将大鼠分为正常对照组(A组)、抑郁模型组(B组)、抑郁模型+72 h睡眠剥夺组(C组),在建立慢性轻度不可预见性应激的抑郁模型后,采用小平台水环境法对大鼠进行72 h快眼动睡眠剥夺,以强迫游泳实验检测大鼠的不动时间,下丘脑5-HT、腺苷浓度分别用高效液相色谱-荧光检测法和高效液相色谱-紫外检测法进行测定。结果 21 d慢性轻度不可预见性应激后,B组和C组大鼠的强迫游泳不动时间显著延长(P<0.05),快眼动睡眠剥夺后C组大鼠强迫游泳不动时间显著缩短(P<0.01);B组大鼠下丘脑5-HT浓度显著低于A组(P<0.01),C组大鼠下丘脑5-HT浓度显著高于B组(P<0.01);B组大鼠下丘脑腺苷浓度显著低于A组(P<0.05),C组大鼠下丘脑腺苷浓度显著低于B组(P<0.01)。结论快眼动睡眠剥夺可以逆转大鼠的抑郁样行为,并增加大鼠下丘脑5-HT浓度,其可能在睡眠剥夺抗抑郁过程中发挥了重要作用,腺苷可能参与了睡眠剥夺的抗抑郁过程。