The activation of the microglia plays an important role in the neuroinflammation induced by different stimulations associated with Alzheimer’s disease (AD). Different stimulations, such as pathogen-associated molecular patterns (PAMPs), damage-associated molecular patterns (DAMPs) and cytokines, trigger a consequence of activation in the microglia with diverse changes of the microglial cell type response in AD. The activation of the microglia is often accompanied by metabolic changes in response to PAMPs, DAMPs and cytokines in AD. Actually, we do not know the distinct differences on the energetic metabolism of microglia when subject to these stimuli. This research assessed the changes of the cell type response and energetic metabolism in mouse-derived immortalized cells (BV-2 cells) induced by a PAMP (LPS), DAMPs (Aβ and ATP) and a cytokine (IL-4) in mouse-derived immortalized cells (BV-2 cells) and whether the microglial cell type response was improved by targeting the metabolism. We uncovered that LPS, a proinflammatory stimulation of PAMPs, modified the morphology from irregular to fusiform, with stronger cell viability, fusion rates and phagocytosis in the microglia accompanied by a metabolic shift to the promotion of glycolysis and the inhibition of oxidative phosphorylation (OXPHOS). Aβ and ATP, which are two known kinds of DAMPs that trigger microglial sterile activation, induced the morphology from irregular to amoebic, and significantly decreased others in the microglia, accompanied by boosting or reducing both glycolysis and OXPHOS. Monotonous pathological changes and energetic metabolism of microglia were observed under IL-4 exposure. Further, the inhibition of glycolysis transformed the LPS-induced proinflammatory morphology and decreased the enhancement of LPS-induced cell viability, the fusion rate and phagocytosis. However, the promotion of glycolysis exerted a minimal effect on the changes of morphology, the fusion rate, cell viability and phagocytosis induced by ATP. Our study reveals that microglia induced diverse pathological changes accompanied by various changes in the energetic metabolism in response to PAMPs, DAMPs and cytokines, and it may be a potential application of targeting the cellular metabolism to interfere with the microglia-mediated pathological changes in AD.

Adult neurogenesis plays a crucial role in cognitive function and mood regulation, while aberrant adult neurogenesis contributes to various neurological and psychiatric diseases. With a better understanding of the significance of adult neurogenesis, the demand for improving adult neurogenesis is increasing. More and more research has shown that traditional Chinese medicine (TCM), including TCM prescriptions (TCMPs), Chinese herbal medicine, and bioactive components, has unique advantages in treating neurological and psychiatric diseases by regulating adult neurogenesis at various stages, including proliferation, differentiation, and maturation. In this review, we summarize the progress of TCM in improving adult neurogenesis and the key possible mechanisms by which TCM may benefit it. Finally, we suggest the possible strategies of TCM to improve adult neurogenesis in the treatment of neuropsychiatric disorders.

IntroductionIsorhynchophylline is one of the main active ingredients from Uncaria rhynchophylla, the effects and mechanisms of isorhynchophylline on stress-induced emotional disorders and cognitive impairment remain unclear.MethodsLong-term potentiation (LTP) in vivo was used for synaptic plasticity evaluation; chronic unpredictable mild stress (CUMS) model was used to evaluate the effect of isorhynchophylline on stress induced emotional disorders and cognitive impairment; sucrose preference test (SPT), open field test (OFT), and elevated plus maze (EPM) were used to evaluate emotional disorders; morris water maze (MWM) test was used to evaluate cognitive impairment; Western blotting (WB) was used to the expression of proteins; high performance liquid chromatography (HPLC) was used to quantify neurotransmitters; Nissl staining was used to identify pathological changes induced by stress.ResultsIn this study, we found that isorhynchophylline improved corticosterone-induced in vivo LTP impairment significantly, indicating positive effects on stress. Therefore, 28-day CUMS model was adopted to evaluate the anti-stress effects of isorhynchophylline. The results showed that isorhynchophylline improved CUMS-induced weight loss, anxiety- and depression-like behaviors, and spatial memory impairment. Isorhynchophylline reduced CUMS-induced corticosterone elevation. N-methyl-D-aspartic acid (NMDA) receptors play an important role in the process of emotion and memory. Glutamate and the expression of GluN2B increased in the CUMS mice, while D-serine and the expression of serine racemase (SR) decreased significantly, and isorhynchophylline restored these changes to normal level.ConclusionThese results indicated that isorhynchophylline ameliorated stress-induced emotional disorders and cognitive impairment, modulating NMDA receptors might be one of the underlying mechanisms.

Sulfur mustard (SM), an extremely reactive alkylating toxicant, which poses a continuing threat to both military and civilian populations. SM targets three major organs including skin, eyes and lungs. In recent years, more and more clinical findings have shown that cognitive and emotional disorders in veterans intoxicated with SM, such as anxiety, depression, apathy, cognitive decline and so on, which indicated the long time toxic effects on mental and neurological health of SM. The experimental studies in animal and cell models have also found neurotoxicity which are similar to clinical results. However, these neuropsychological problems are not studied well in victims of SM and the mental and neurological complications are often not subjected to treatment or undertreated. Until now, the exact mechanism of the action of SM toxicity has not been elucidated and no specific therapy for its poisoning exists. Therefore, the studies on neurotoxicity of SM should be strengthened. This review summarizes the main progress of clinical and experimental researches on neurotoxicity of SM for the past few years.

Here, we described the generation of human induced pluripotent stem cells (iPSC) from peripheral blood mononuclear cells (PBMCs) of a 87-year-old female patient with sporadic Alzheimer's disease (sAD) having APOE3 (ε3/ε3) genotype. iPSC line were generated from PBMCs with four factors of OCT4, SOX2, c-MYC and KLF4 using episomal system. The pluripotency of the iPSC line was assessed by embryoid body (EB) formation. Flow cytometry analyses revealed >97% cells positive for the pluripotency markers NANOG, OCT4 and SSEA4. Furthermore, the iPSC line displayed a normal karyotype (46, XX). The iPSC line may provide valuable tools for the study of sAD pathogenesis.

The cell-type-specific response of neural cells to oxidative stress, a crucial mechanism for accelerating aging and cognitive dysfunction in Alzheimer's disease (AD), is still far from understood. Here, we employed human-induced pluripotent stem cells (hiPSCs)-derived neural stem cells (hiPSC-NSCs), neurons (hiPSC-Neurons), and microglia-like cells (hiPSC-MGLs) from sporadic AD (sAD) patients, age-matched cognitive normal controls (CNCs), and young subjects to observe human neural cell-type response to H2O2 stimulation. Without H2O2 exposure, reactive oxygen species (ROS) cannot be detected in hiPSC-NSCs from all three groups, but the viability of hiPSC-NSCs from AD patients was significantly lower than those of CNCs and young subjects. There were no significant differences in ROS, viabilities, neurite length, and neurite branch points in hiPSC-Neurons among three groups. No significant differences in viabilities, phagocytosis, and secretion of cytokines were observed in hiPSC-MGLs among three groups, but higher ROS levels in sAD hiPSC-MGLs. Under H2O2 exposure, the viability, neurite length, and neurite branch points of hiPSC-Neurons from AD patients reduced more significantly accompanied by more ROS release. H2O2 exposure caused hiPSC-MGLs from AD patients to release more ROS, cytokines, and stronger phagocytosis. Nevertheless, H2O2 exposure had no effect on viability of hiPSC-NSCs. Our results showed hiPSC-Neurons and hiPSC-MGLs were more sensitive to H2O2 than hiPSC-NSCs, which indicated the different response styles of hiPSC-NSCs, hiPSC-Neurons, and hiPSC-MGLs to oxidative stress. HiPSC-derived neural cells from AD patients suffered more severe injury from H2O2 than those of CNCs and young subjects, indicating that the vulnerability to oxidative stress of AD patients can be recapitulated in hiPSCs.

目的 研究β淀粉样蛋白1-42寡聚体(oAβ)对认知正常老年人(CNC)及晚期散发性阿尔茨海默病(AD)患者诱导性多能干细胞(hiPSC)源性的小胶质细胞(hiMGL)炎症及氧化应激反应的影响.方法 按照本实验室前期建立的hiPSC向hiMGL诱导分化的方法,首先将CNC和AD患者来源的hiPSC诱导分化为造血祖细胞(hiHPC),用流式细胞术检测其表达hiHPC标志物CD34和CD43的阳性率;进而将hiHPC诱导分化为小胶质细胞(简称CNC hiMGL和AD hiMGL),用免疫荧光法检测其表达离子钙接头蛋白分子1(IBA1)和跨膜蛋白119(TMEM119)的阳性率.用oAβ1μmol·L-1孵育hiMGL 24 h后,用CCK-8法检测hiMGL细胞存活,中性红实验检测细胞吞噬功能,荧光探针法检测hiMGL细胞内活性氧(ROS)水平,Luminex技术检测hiMGL培养上清细胞因子〔白细胞介素(IL)-10,IL-1β,IL-6,肿瘤坏死因子α,CC趋化因子配体7和CXC趋化因子配体10〕水平.以oAβ1μmol·L-1孵育hiMGL 1.5 h,分别于终止孵育后0,3,6和9 h将细胞裂解,ELISA检测细胞内oAβ剩余含量.结果 CNC hiHPC和AD hiHPC表达CD34和CD43,且阳性率均>90％;CNC hiMGL和AD hiMGL表达IBA1和TMEM119,且阳性率均>90％.oAβ1μmol·L-1孵育24 h后,oAβ组CNC hiMGL和AD hiMGL组细胞存活、吞噬能力、上述细胞因子分泌水平和ROS水平均显著高于各自对照组(P<0.01),且oAβ组AD hiMGL上述指标均显著高于同组CNC hiMGL(P<0.01).hiMGL终止孵育oAβ后0 h,AD hiMGL内oAβ摄入含量高于CNC hiMGL(P<0.01);终止孵育后0～9 h,oAβ含量在CNC hiMGL内下降较快,9 h时AD hiMGL内oAβ剩余含量显著高于CNC hiMGL(P<0.01).结论 本研究诱导分化的CNC hiMGL和AD hiMGL 90％以上表达小胶质细胞特异性标志蛋白IBA1和TMEM119,且二者无明显差异.oAβ刺激后,AD hiMGL和CNC hiMGL均出现了炎症反应和氧化应激反应,前者反应更强,而降解oAβ的能力较弱.

Microglia constitute the majority of innate immune cells in the brain, and their dysfunction is associated with various central nervous system diseases. Human microglia are extremely difficult to obtain experimentally, thereby limiting studies on their role in complex diseases. Microglia derived fr om human stem cells provide new tools to assess the pathogenesis of complex diseases and to develop effective treatment methods. This study aimed to develop a reliable method to derive human microglial-like cells (iMGLs) from induced pluripotent stem cells (iPSCs) expressing microglia-specific markers IBA1 and TMEM119 and respond to lipopolysaccharide (LPS) stimulation. Thereafter, we compared iMGL functions from Alzheimer's disease (AD) patients and cognitive normal controls (CNCs). AD-iMGLs displayed stronger phagocytic ability with or without stimulation. High LPS concentrations (>2μg/ml) caused death in CNC-iMGLs, while AD-iMGLs did not display significant cell death. Cytokine analysis revealed that TNF-α, IL-6, and IL-10 secreted by AD-iMGLs were significantly increased upon LPS stimulation compared to those in CNC-iMGLs. The present results indicate that AD-iMGLs exhibit significant inflammatory characteristics and can reflect some pathological changes in microglia in AD, thereby providing new valuable tools to screen candidate drugs for AD and to elucidate the mechanisms underlying AD pathogenesis.

Early, reliable and non‐invasive diagnosis of Alzheimer disease (AD) is a great challenge, which makes the prognosis and therapeutic interventions quite difficult. We aimed to investigate the expressions of let‐7g, miR‐197, miR‐126 and miR‐29a which were screened from 877 microRNAs in 60 serum samples with a microarray platform in prior study in serum of cognitively normal controls (CNC), mild cognitive impairment (MCI), a potential preliminary stage of AD, and AD and identify microRNA panel for predicting and diagnosing AD. Quantitative reverse‐transcriptase polymerase chain reaction assay was applied to evaluate the expressions of let‐7g, miR‐197, miR‐126 and miR‐29a with two independent cohorts including 202 participants CNC, MCI and AD. Logistic regression model based on microRNA panel was constructed using a training cohort (n=150) and then validated using an independent cohort (n=52). Area under the receiver operating characteristic curve (AUC) was used to evaluate diagnostic accuracy. The expressions of the four microRNAs were significantly decreased in MCI and AD versus CNC, and positively correlated with mini mental state examination (MMSE) score. Then, we identified a microRNA panel with the four microRNAs that demonstrated good diagnostic performance for MCI (AUC=0.788 and 0.815 for training and validation data set, respectively) and AD (AUC=0.769 and 0.823 for training and validation data set, respectively). When combined with MMSE score, the diagnostic performance of the microRNA panel was further improved. These results suggested that the serum microRNA panel we found has considerable clinical value in diagnosing AD, and the four microRNAs are potential circulating biomarkers.Support or Funding InformationThe project supported by the National Science and Technology Ministry (2011ZX09102‐101‐07, 2012ZX09301003‐002‐001, 2012BAI29B07)

Objective: Alzheimer's disease (AD) causes progressive hippocampus dysfunctions leading to the impairment of learning and memory ability and a reduced uptake rate of glucose in the hippocampus. LW-AFC is a new formula derived from the classical traditional Chinese medicinal prescription Liuwei Dihuang decoction. In this study, we investigated the beneficial and protective effects of LW-AFC in learning-memory ability and brain glucose metabolism in a widely used AD model, the senescence-accelerated prone mouse (SAMP8). Methods: SAMP8 mice were administrated LW-AFC (1.6 g/kg) for 2 months. The Morris water maze (MWM) test, shuttle box test, and novel object recognition test, were all used to evaluate cognition abilities. Fluorodeoxyglucose positron emission tomography (FDG-PET) was employed to detect the level of glucose uptake in the mouse brain. The concentrations of ATP, ADP and AMP were determined by HPLC. Results: The results from the MWM, shuttle box, and novel object recognition tests showed impairments in object recognition memory and spatial learning and memory, and active avoidance in SAMP8 mice. Administration of LW-AFC ameliorated the impairments in learning and memory abilities. The FDG-PET results showed a decrease in the uptake of glucose in the brains of SAMP8 mice with increasing age. LW-AFC enhanced glucose uptake in the brain of SAMP8 mice. Decreases in the concentrations of ATP, ADP and AMP were also found in the brains of SAMP8 mice. Administration of LW-AFC increased the concentration of all energy substances. Conclusion: LW-AFC could ameliorate deterioration of cognition in SAMP8 mice by improving glucose metabolism.

A series of poly(ADP-ribose)polymerase (PARP)-1 inhibitors containing a novel scaffold, the 1H-thieno[3,4-d]imidazole-4-carboxamide moiety, was designed and synthesized. These efforts provided some compounds with relatively good PARP-1 inhibitory activity, and among them, 16l was the most potent one. Cellular evaluations indicated that the anti-proliferative activities of 16g, 16i, 16j and 16l against BRCA-deficient cell lines were similar to that of olaparib, while the cytotoxicities of 16j and 16l toward human normal cells were lower. In addition, ADMET prediction results indicated that these compounds might possess more favorable toxicity and pharmacokinetic properties. This study provides a basis for our further investigation.

Early studies with first-generation poly (ADP-ribose) polymerase (PARP) inhibitors have already indicated some therapeutic potential for sulfur mustard (SM) injuries. The available novel and more potential PARP inhibitors, which are undergoing clinical trials as drugs for cancer treatment, bring it back to the centre of interest. However, the role of PARP-1 in SM-induced injury is not fully understood. In this study, we selected a high potent specific PARP inhibitor ABT-888 as an example to investigate the effect of PARP inhibitor in SM injury. The results showed that in both the mouse ear vesicant model (MEVM) and HaCaT cell model, PARP inhibitor ABT-888 can reduce cell damage induced by severe SM injury. ABT-888 significantly reduced SM induced edema and epidermal necrosis in MEVM. In the HaCaT cell model, ABT-888 can reduce SM-induced NAD + /ATP depletion and apoptosis/necrosis. Then, we studied the mechanism of PARP-1 in SM injury by knockdown of PARP-1 in HaCaT cells. Knockdown of PARP-1 protected cell viability and downregulated the apoptosis checkpoints, including p-JNK, p-p53, Caspase 9, Caspase 8, c-PARP and Caspase 3 following SM-induced injury. Furthermore, the activation of AKT can inhibit autophagy via the regulation of mTOR. Our results showed that SM exposure could significantly inhibit the activation of Akt/mTOR pathway. Knockdown of PARP-1 reversed the SM-induced suppression of the Akt/mTOR pathway. In summary, the results of our study indicated that the protective effects of downregulation of PARP-1 in SM injury may be due to the regulation of apoptosis, necrosis, energy crisis and autophagy. However, it should be noticed that PARP inhibitor ABT-888 further enhanced the phosphorylation of H2AX (S139) after SM exposure, which indicated that we should be very careful in the application of PARP inhibitors in SM injury treatment because of the enhancement of DNA damage.

We have developed a series of substituted 4-(thiophen-2-ylmethyl)-2H-phthalazin-1-ones as potent PARP-1 inhibitors. Preliminary biological evaluation indicated that most compounds possessed inhibitory potencies comparable to, or higher than AZD-2281. Among these compounds, 18q appeared to be the most notable one, which displayed an 8-fold improvement in enzymatic activity compared to AZD-2281. These efforts lay the foundation for our further investigation.

Alzheimer's disease(AD),the most common form of dementia among the elderly,is a chronic,progressive and degenerative disorder of the brain with a loss of memory and cognition.Because the pathogenesis of AD is complicated and relates to the abnormality and dysfunction of multisystem,there is still no ideal drug for preventing and curing AD till now.Thus the discovery and selection of drug targets is very important for drug research and exploitation.Recently,further studies of pathogenesis of AD provide many new targets for the screen and research of new drugs that may prevent or cure AD.In this review it will be discussed briefly.

目的:研究快速老化模型小鼠(senescence accelerated mice,sAM)的快速老化亚系SAM-prone/8(SAMP8)及同龄抗快速老化亚系SAM-resistance/1(SAMRl)代谢产物的差别,并观察六味(Liuwei Dihuang decoction,LW)及八味地黄汤(Bawei Di-huang decoction,BW)对SAMP8代谢产物的影响,比较二方剂的作用机理.方法:以12月龄雌性和雄性SAMR1及SAMP8为动物模型,灌胃给予LW和BW1个月后,采集血清,应用代谢组学的主要研究技术-核磁共振方法测定核磁共振谱,检测血清中的代谢产物,主成分分析法处理代谢组学数据.结果:SAMR1和SAMP8的血清代谢产物谱能够相互区分,其中SAMP8血清中的乳酸、极低密度脂蛋白、饱和脂肪酸和甘油三脂含量明显高于SAMR1,葡萄糖、磷脂酰胆碱和胆碱、不饱和脂肪酸、低密度脂蛋白和高密度脂蛋白的含量明显低于SAMR1.给予LW和BW后均可影响SAMP8的血清代谢产物谱,SAMP8的代谢状态得到了很大改善.给予LW后,SAMP8血清代谢物中葡萄糖的含量有了明显的增加,胆碱和低密度脂蛋白的含量有所升高;极低密度脂蛋白的含量则有所减少.给予BW后,SAMP8血清代谢物中葡萄糖和磷脂酰胆碱的含量明显升高,而胆碱含量则较低.结论:SAMR1和SAMP8的血清代谢产物谱存在明显差异,主要在于乳酸、葡萄糖、磷酸胆碱和胆碱及脂类含量的差别.LW和BW均能够对SAMP8血清代谢物产生影响,二方剂可能主要通过对葡萄糖、胆碱和脂类的含量的调节而改善了SAMP8能量代谢障碍和学习记忆能力下降.上述结果提示,SAMP8可能存在糖酵解代谢障碍,葡萄糖代谢下降而引起的能量代谢障碍,胆碱能代谢低下及脂类代谢紊乱.给予LW和BW均能够对SAMP8的血清代谢产物产生影响,提示LW和BW能够在代谢网络的多个节点上对SAMP8的代谢紊乱起到调节和改善作用.这些LW和BW的反应代谢产物为深入研究LW和BW的作用机制及肾阴虚、肾阳虚的现代生物学基础提供了新的线索和依据.

Objective To investigate the effect of Huanglian Jiedutang (HLJDT) on hippocampal protein expressions in senescence accelerated mouse-prone/8 (SAMP8). Method The 12-month-old senescence accelerated mice (SAM) were divided into three groups: SAM-resistance/1 (SAMR1), SAM-prone/8 (SAMP8) and SAMP8 treated with HLJDT. The effect of HLJDT on expressions of hippocampal proteins was analyzed by two dimensional electrophoresis (2DE) and matrix-assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF-MS). Result Compared with same age SAMR1, there were 29 differential expressed hippocampal proteins in SAMP8. After treated with HLJDT, the expressions of 38 hippocampal proteins of SAMP8 were changed significantly. 12 reactive proteins of HLJDT were chosen to be identified by MALDI-TOF-MS and the results were searched in MASCOT database. Among 12 reactive proteins, the expressions of 4 hippocampal proteins which expressed differentially between SAMR1 and SAMP8 could be improved by HLJDT. Conclusion HLJDT may improve the aging of SAMP8 by regulating the expressions of proteins related with energy metabolism, signal transduction, cytoskeletal, amino acid metabolism and so on.

To investigate mechanisms of the deficits of learning and memory related with aging, the differentially expressed hippocampal proteins from 6- and 12-month-old SAM-prone/8 (SAMP8) and age-matched SAM-resistance/1 (SAMR1) were analyzed and compared. In comparison with the same age SAMR1, 15 proteins expressions in hippocampus of 6-month-old SAMP8 increased, 5 proteins expressions decreased significantly; 12 proteins expressions in hippocampus of 12-month-old SAMP8 increased, 2 proteins expressions decreased significantly and 2 proteins only expressed in SAMP8 hippocampus; and 22 proteins with significant changes were identified by MALDI-TOF-MS and the results were searched in MASCOT database. These identified proteins could be devided into four categories according to their functions: (1) energy metabolism; (2) mitochondrion function (3) signal transduction; (4) other proteins. The results show that there were significant differences in hippocampus protein expressions between SAMP8 and SAMR1, and some differentially expressed proteins were correlated with the deficits of learning and memory in SAMP8 with aging, and these proteins could provide clues for the study and discovery of new protein targets for improving intelligence drugs.