• <tr id="yyy80"></tr>
  • <sup id="yyy80"></sup>
  • <tfoot id="yyy80"><noscript id="yyy80"></noscript></tfoot>
  • 99热精品在线国产_美女午夜性视频免费_国产精品国产高清国产av_av欧美777_自拍偷自拍亚洲精品老妇_亚洲熟女精品中文字幕_www日本黄色视频网_国产精品野战在线观看 ?

    Compound of icariin, astragalus, and puerarin mitigates iron overload in the cerebral cortex of Alzheimer’s disease mice

    2018-05-05 06:47:12YuZhangWeiNaKongXiQingChai

    Yu Zhang, Wei-Na Kong, Xi-Qing Chai, ,

    1 Department of Neurology, the First Hospital of Hebei Medical University, Shijiazhuang, Hebei Province, China

    2 Hebei Chemical and Pharmaceutical College, Shijiazhuang, Hebei Province, China

    Introduction

    Alzheimer’s disease (AD) is an irreversible and progressive neurodegenerative brain disorder, and the most common cause of dementia in the elderly. Its progression is characterized by deterioration of cognitive function and memory. The main histological hallmarks of AD brains are senile plaques,neurofibrillary tangles, and extensive neuronal loss (Katzman and Saitoh, 1991). However, the precise mechanisms underlying AD have not been completely elucidated.

    Previous studies have suggested that iron is involved in progression of AD (El Tannir El Tayara et al., 2006), and that altered iron homeostasis may play an crucial role in disease pathogenesis (Crapper McLachlan et al., 1991). Nevertheless, the mechanism of iron overload in AD brain remains unclear. During the last decade, many studies have shown that metabolic imbalance in iron and the resulting oxidative stress play important roles in disease pathogenesis (Crapper McLachlan et al., 1991). In a state of toxic overload, iron is perceived as an endogenous toxin in traditional Chinese medicine (Lu and Black, 2016). Indeed, abnormally high levels of brain iron constitute an endogenous toxin that is important to AD pathogenesis. Iron accumulation is also thought to be highly correlated with abnormal function of other organs including the kidney, liver, heart, and spleen(Crapper McLachlan et al., 1991; El Tannir El Tayara et al.,2006). Excessive iron levels were previously shown to lead to increased oxidative stress using Fenton chemistry (Rolston et al., 2009). Therefore, therapeutic intervention to decrease iron accumulation may be an effective way of improving AD pathology.

    Figure 1 Chemical structures of icariin,astragaloside IV, and puerarin.

    Iron chelators, such as deferoxamine (DFO), significantly relieve the symptoms of AD patients, and consequently exert neuroprotective effects on the disease (Wan et al.,2006; Hishikawa et al., 2008). However, chemical drugs have drawbacks such as side effects and poor oral availability, and there is a need for research into alternative therapies. In the last decades, many traditional Chinese medicines have been studied, with benefits shown using different AD experimental models and clinical trials (Xian et al., 2012). Accordingly,components of Epimedium, Astragalus, and Puerariae may overcome these disadvantages. Icariin is extracted from Epimedium (Chen et al., 2005), and studies have shown that icariin improves learning and memory abilities in aged rats(Cheng et al., 2007; Hagemeier et al., 2012). Astragalus polysaccharide has antioxidative (Ko et al., 2005), immunoregulatory, and antiviral effects (Huang et al., 2013), antitumor activities (Du et al., 2012), and cardiac protective properties(Dang et al., 2009; Dai et al., 2014). While Puerariae is reported to have anti-inflammatory effects (Lim et al., 2013).Furthermore, these compounds have effects on reducing inflammation, free radical scavenging capacity, altering multiple visceral organ functions (Gao et al., 2013), and reducing brain iron load (Chen and Huang, 2008). Further, previous research in our laboratory suggests that these active components reduce spatial learning and memory impairments in the APP/PS1 mouse model, and inhibit iron overload in the cerebral cortex in a mouse model of AD (Dong et al., 2015),although the mechanism is still not clear.

    Thus, the aim of this study was to investigate the mechanism of the active components extracted from Astragalus,Icariin, and Puerariae on brain iron overload in APP/PS1 transgenic mice.

    Materials and Methods

    Ethics statement and animals

    All animal handling procedures were in accordance with the Guidelines of Animal Experiments from the Committee of Medical Ethics, Ministry of Health of China. Experiments were approved by the Ethics Committee for Animal Experiments of Hebei Medical University of China (approval No.HEBMU-2010-10). Precautions were taken to minimize suffering and the number of animals used in each experiment.Seven male six-month-old C57BL/6J (C57) mice and forty-two male six-month-old APPswe/PS1ΔE9 (APP/PS1)mice (weighing 28.8 ± 5 g) were obtained from Beijing HFK Bioscience Co., Ltd., Beijing, China (SCXK (Jing) 2014-0004). The APP/PS1 transgenic mouse model of AD overexpresses the Swedish (K594M/N595L) mutation of APP, and with presenilin 1 (PS1) deleted in exon 9, on a C57 genetic background. APP/PS1 mice were genotyped by polymerase chain reaction. Mice were housed in specific-pathogen-free conditions on a 12-hour light/dark cycle with free access to water and food.

    Compound preparation

    The compound comprised purified extracts from three different herbs used in Chinese medicine, namely Epimedium, Astragalus, and Puerariae. The individual components extracted from Epimedium, Astragalus, and Puerariae are icariin, astragaloside, and puerarin, respectively (Figure 1). These components were purchased from China Nanjing Zelang Medical Technology Co., Ltd. (Nanjing, China). Drug purity was > 98%. Icariin, astragaloside IV, and puerarin were dissolved in distilled water at a ratio of 3:2:2 (w/w).

    Drug treatment

    APP/PS1 transgenic mice were randomly divided into six groups, with seven mice in each group. Mice in the AD model group were intragastrically administered 1 mL normal saline. Mice in the compound group were intragastrically administered compound containing icariin (120 mg/kg),astragalus (80 mg/kg), and puerarin (80 mg/kg). The DFO group received 30 mg/kg DFO by intraperitoneal injection.The puerarin group received 80 mg/kg puerarin by intragastric administration. The astragalus group received 80 mg/kg astragalus by intragastric administration. The icariin group received 120 mg/kg icariin by intragastric administration.All groups were given treatments once a day for 3 consecutive months. In addition to the six groups, seven male C57 mice were included as a normal control group, and were intragastrically administered 1 mL normal saline.

    Tissue preparation

    After 3 consecutive months, mice were deeply anesthetized with sodium pentobarbital (50 mg/kg) by intraperitoneal injection, and immediately perfused through the heart with 0.9% NaCl. Brains were rapidly removed and divided into separate hemispheres on an ice-cold surface. The cerebral cortex was dissected from one hemisphere and stored at–80°C for enzyme-linked immunosorbent assay (ELISA) and biochemistry. The other cerebral cortex was dried at 110°C overnight and used for detecting iron content by flame atomic absorption spectroscopy.

    Flame atomic absorption spectroscopy

    The dried cerebral cortex was weighed and treated with 1 mL concentrated nitric acid and 0.5 mL perchloric acid, then dissolved and placed in a fume hood at 90°C for 1 hour until the moisture evaporated. Dissolved tissues were diluted with deionized water to 3 mL. Absorbance values were measured at 248.3 nm using the Varian Spectra AA-10 Spectrophotometer(Agilent Technologies, Santa Clara, CA, USA). Standard iron curves were prepared from commercially available standards.

    ELISA

    ELISA was used to determine levels of interleukin-1β (IL-1β)and interleukin-6 (IL-6), and tumor necrosis factor alpha(TNF-α). Tissues were weighed and homogenized in an icecold protease and phosphatase inhibitor cocktail. Homogenates were centrifuged at 13,000 r/min for 20 minutes at 4°C.Supernatants were collected for analysis using mouse IL-1β,IL-6, and TNF-α ELISA kits (Raybio, Norcross, GA, USA).Protein concentration of supernatants was determined by the Lowry method using bovine serum albumin as a standard.IL-1β ELISA assay used an antibody specific for mouse IL-1β, which was coated on 96-well plates. Standards and samples were pipetted into the wells and IL-1β present bound to wellsviaimmobilized antibody. The wells were washed and biotinylated antibody added. After washing away unbound biotinylated antibody, horseradish peroxidase-conjugated streptavidin was added. The wells were washed again, and a tetramethylbenzidine substrate solution added. Color developed in proportion to the amount of bound IL-1β: stop solution changed the color from blue to yellow, and the color intensity was measured at 450 nm. The IL-6 and TNF-α assay procedures were the same as for IL-1β. All assays were performed as outlined by the manufacturer’s protocols.

    Colorimetry of glutathione peroxidase (GSH-Px) and superoxide dismutase (SOD) activities andmalondialdehyde (MDA) content

    Tissues were weighed and 10% (w/v) buffered homogenates prepared. Homogenates were centrifuged at 2500 r/min for 10 minutes at 4°C, and the supernatants used for biochemical analyses. The protein concentration of supernatants was determined by the Lowry method using bovine serum albumin as the standard. SOD activity in homogenates was examined using a commercially available kit (Jian Cheng Biological Engineering Institute, Nanjing, China) based on auto-oxidation of hydroxylamine. The developed blue color of the reaction was measured at 550 nm.

    Activity of GSH-Px was determined by the velocity method using a GSH-Px kit (Jian Cheng Biological Engineering Institute). The reaction was initiated by addition of hydrogen peroxide. A series of enzymatic reactions was activated by GSH-Px in homogenates, which subsequently led to conversion of GSH (reduced glutathione) to GSSG (oxidized glutathione). The change in absorbance during conversion of GSH to GSSG was recorded spectrophotometrically at 412 nm. The procedure was performed according to the manufacturer’s instructions.

    MDA concentration was determined in homogenates using a commercially available kit (Jian Cheng Biological Engineering Institute) based on thiobarbituric acid reactivity.Trichloracetic acid was mixed with homogenate and centrifuged, then the supernatant removed and thiobarbituric acid added. The developed red color of the resulting reaction was measured at 532 nm using a spectrophotometer. The rest of the procedure was performed as outlined by the manufacturer’s instructions.

    Statistical analysis

    Data are expressed as the mean ± SD, and analyzed using repeated measures analysis of variance. For single dependent variables assays, univariate analysis of variance was performed using group as the between-subject factor. Following a significant omnibus analysis of variance, Bonferronipost hoccomparisons were performed. All analyses were performed using the SPSS 17.0 for Windows (SPSS Inc., Chicago, IL, USA). A value ofP< 0.05 was considered statistically significant.

    Results

    Effect of icariin, astragalus, and puerarin on iron levels in the cerebral cortex of APP/PS1 transgenic mice

    To determine whether icariin, astragalus, and puerarin reduce iron levels in APP/PS1 mice, total iron levels in the cortex were detected using a flame atomic absorption spectrometry method. Compared with the normal control group, iron levels were higher in the AD model group (P<0.05). Compared with the AD model group, iron levels were reduced in the compound group and DFO group (P< 0.05).There was no significant difference in iron levels between the compound and DFO groups (P> 0.05). Iron levels in the puerarin, astragalus, and icariin groups were higher compared with the compound group, and lower compared with the AD group (P< 0.05; Figure 2).

    Effect of icariin, astragalus, and puerarin on expression of proinflammatory factors in the cerebral cortex of APP/PS1 transgenic mice

    Figure 2 Total iron levels in the cerebral cortex of APP/PS1 transgenic mice (Flame atomic absorption spectroscopy).

    ELISA was used to investigate proinflammatory factors including IL-1β, IL-6, and TNF-α. Compared with the normal control group, IL-1β, IL-6, and TNF-α expression was higher in the AD model group (P< 0.05). Compared with the AD model group, IL-1β, IL-6, and TNF-α expression was significantly reduced in the compound group (P< 0.05).DFO-treated mice showed a significant reduction compared with AD model mice (P> 0.05). IL-6 expression was higher in the puerarin group than the normal control and lcariin groups. IL-6 expression was increased in the puerarin group compared with the AD model group. There was no difference in IL-1β expression between the puerarin and normal control groups. IL-1β expression was increased in the icariin andPuerarin groups compared with the AD model and DFO groups. Expression of TNF-α was higher in the icariin group than the control group (P< 0.05). TNF-α expression was higher in the astragalus and puerarin groups compared with the AD model group. No significant differences were found in TNF-α expression between the astragalus and puerarin groups and DFO group (Figure 3).

    Effect of icariin, astragalus, and puerarin on MDA content and SOD and GSH-Px activity in the cerebral cortex of APP/PS1 transgenic mice

    Figure 3 Effect of a compound containing icariin, astragalus, and puerarin on IL-6, IL-1β, and TNF-α levels in the cerebral cortex of AD model mice (ELISA assay).

    Figure 4 Effect of a compound containing icariin, astragalus, and puerarin on MDA content and SOD and GSH-Px activity in the cerebral cortex of AD model mice.

    Compared with the normal control group, MDA content was significantly enhanced in the AD model group (P<0.05). While compared with the AD model group, excess iron-mediated enhancement of MDA content was significantly inhibited in the compound group (P< 0.05). Furthermore, SOD and GSH-Px activity was significantly decreased in the AD model group compared with the normal control group (P< 0.05). SOD activity was increased in compoundand DFO-treated mice (P< 0.05). No difference in GSH-Px was determined between effective component-treated and DFO-treated mice (Figure 4). MDA content and SOD activity were increased in the icariin, astragalus, and puerarin groups compared with the compound group. GSH-Px activity was decreased in the icariin and astragalus groups (P< 0.05).

    Discussion

    Abnormal iron accumulation is thought to occur early in the onset of AD, appearing before the formation of neurofibrillary tangles and senile plaques. Brain iron content was previously shown to be associated with degree of dementia,with increasing iron related to more severe disease (Zhu et al., 2009). Excessive brain iron levels may play an important role in progression of AD by initiating a cascade that eventually leads to neuronal death (Zhu et al., 2009). Consequently,iron chelation may be an effective therapeutic intervention for AD (Whitnall and Richardson, 2006). Indeed, the icariin,astragalus, and puerarin compounds may overcome these shortcomings in iron chelation. They scavenge free radicals,decrease inflammation, and alter multiple organ functions,which may ease iron overload on central nervous system function. Here, we have attempted to demonstrate and explain this using contemporary methods and transgenic animal models of AD. In our present study, we first investigated the neuroprotective effect of effective extracts of Epimedium,Astragalus, and Puerariae. We evaluated brain iron levels using flame atomic absorption spectroscopy,and found they were increased in the cerebral cortex of the APP/PS1 mouse model, but could be relieved by compound treatment. Thus,the compound may have a similar mechanism to DFO, and act by chelating and clearing free iron ion to reduce brain deposition (Genlain et al., 2007).

    Oxidative stress induced by iron overload is not only a crucial mechanism in AD, but also in development of neurodegenerative diseases (Qian and Shen, 2001) and cerebral ischemia (Yoo et al., 2016). Oxidative stress produces oxygen free radicals, which promote generation of amyloid-beta(Aβ). Moreover, Aβ promotes generation of oxygen free radicals, which leads to neuronal damage and further positive feedback (Misonou et al., 2000). Iron overload causes an increase in lipid peroxidation (Galleano and Puntarulo, 1997).As a major product of lipid peroxidation (Lee et al., 2011),MDA indirectly reflects the extent of damage (Parks et al.,1994). SOD and GSH are enzymatic antioxidants that scavenge harmful reactive oxygen species, forming the first line of defense against free radicals, which convert toxic superoxide into the less toxic hydrogen peroxide (Geisser, 1997).Our results provide evidence that the icariin, astragalus, and puerarin compounds exert effective protection against lipid peroxidation induced by iron overload. Furthermore, our results show that MDA content is significantly increased in the brain of AD model mice. The response of the cerebral cortex to excess toxic iron was acceleration in lipid peroxidation accompanied by increased MDA content, as well as decreased SOD and GSH activity. Treatment with compounds of purified herbal extracts may increase antioxidant defenses by enhancing GSH and SOD activity, consequently decreasing lipid peroxidation and MDA content. The resulting improvement in antioxidant defenses effectively protects the brain from tissue damage due to free radicals induced by iron overload. This increase in antioxidant activity may be an important mechanism in protective effects of the compound against brain damage.

    During AD, aggregation of Aβ peptide induced by iron overload in the brain usually leads to glial cell activation,which initiates a neuroinflammatory response involving inflammatory cytokines (including IL-6, IL-1β, and TNF-α)(Mucke, 2009; Vetrivel and Thinakaran, 2010; Tang et al.,2013), as well as reactive oxygen intermediates.In vitrostudies have shown that iron deposition on glial cells in the cortex, cerebellum, substantia nigra, and hippocampus is associated with neuroinflammation in AD (Fu et al., 2013).Inflammatory cytokines activate cell apoptosis, stimulate plaque-associated microglia, and increase lipid peroxidation (Griffin, 2006; Hoozemans et al., 2006). Here, we found significantly decreased IL-1β, IL-6, and TNF-α expression in the compound-treated AD group compared with the untreated AD group. In contrast, levels were increased in the AD model group compared with the normal control group.These results suggest that treatment with the effective component of the herbs Astragalus, Puerariae, and Icariin may attenuate damage and slow progression of AD by inhibiting the inflammatory response. It has been shown that increased inflammation contributes to progression of AD (Swardfager et al., 2010). Therefore, it is hypothesized that inflammatory components might be involved in microglial activation. Infl ammation may augment disease progression by facilitating brain iron deposition during progression of AD. Moreover,released inflammatory cytokines can disrupt the blood-brain barrier (Blasko et al., 2001; Qiao et al., 2001; Sastre et al.,2003), increasing vascular permeability, and allowing iron to enter and accumulate in the brain .

    Our results demonstrate that the icariin, astragalus, and puerarin compounds remarkably ameliorate pathological changes in iron overloaded mice. These remarkable protective effects against injury caused by excess iron may be attributed to prevention of iron deposition, inhibition of neuronal apoptosis induced by oxidative stress, and inflammatory factors. Therefore, the compounds are effective in clearing heat, removing toxin, strengthening the brain, and developing intellect. Although there is still a long way to go,icariin, astragalus, and puerarin compounds may have potential benefits for AD treatment.

    Author contributions:YZ provided data and ensured the integrity of the data, analyzed the data and wrote the paper. WNK participated in study concept and design. XQC served as a principle investigator and obtained the funding. All authors approved the final version of the paper.

    Conflicts of interest:None declared.

    Financial support:This study was supported by the National Natural Science Foundation of China, No. 81273983. The conception, design execution, and analysis of experiments, as well as the preparation of and decision to publish this manuscript, were made independent of this funding organization.

    Research ethics:All animal handling procedures were in accordance with the Guidelines of Animal Experiments from the Committee of Medical Ethics, Ministry of Health of China, and experiments were approved by the Ethics Committee for Animal Experiments of Hebei Medical University of China (approval No. HEBMU-2010-10).

    Data sharing statement:Datasets analyzed during the current study are available from the corresponding author on reasonable request.

    Plagiarism check:Checked twice by iThenticate.

    Peer review:Externally peer reviewed.

    Open access statement:This is an open access journal, and articles are distributed under the terms of the Creative Commons Attribution-Non-Commercial-ShareAlike 4.0 License, which allows others to remix, tweak,and build upon the work non-commercially, as long as appropriate credit is given and the new creations are licensed under the identical terms.

    Blasko I, Apochal A, Boeck G, Hartmann T, Grubeck-Loebenstein B,Ransmayr G (2001) Ibuprofen decreases cytokine-induced amyloid beta production in neuronal cells. Neurobiol Dis 8:1094-1101.

    Chen GF, Huang WF (2008) Progress in pharmacological effects of compositions of Astragalus membranaceus. Zhongguo Xinyao Zazhi 17:1482-1485.

    Chen KM, Ge BF, Ma HP, Liu XY, Bai MH, Wang Y (2005) Icariin, a flavonoid from the herb Epimedium enhances the osteogenic differentiation of rat primary bone marrow stromal cells. Pharmazie 60:939-942.

    Cheng S, Qiu F, Wang S, He J (2007) HPLC analysis and pharmacokinetics of icariin in rats. J Sep Sci 30:1307-1312.

    Crapper McLachlan DR, Dalton AJ, Kruck TP, Bell MY, Smith WL, Kalow W, Andrews DF (1991) Intramuscular desferrioxamine in patients with Alzheimer’s disease. Lancet 337:1304-1308.

    Dai H, Jia G, Liu X, Liu Z, Wang H (2014) Astragalus polysaccharide inhibits isoprenaline-induced cardiac hypertrophy via suppressing Ca(2)(+)-mediated calcineurin/NFATc3 and CaMKII signaling cascades.Environ Toxicol Pharmacol 38:263-271.

    Dang SS, Jia XL, Song P, Cheng YA, Zhang X, Sun MZ, Liu EQ (2009)Inhibitory effect of emodin and Astragalus polysaccharide on the replication of HBV. World J Gastroenterol 15:5669-5673.

    Dong XH, Bai JT, Kong WN, He XP, Yan P, Shao TM, Yu WG, Chai XQ,Wu YH, Liu C (2015) Effective components of Chinese herbs reduce central nervous system function decline induced by iron overload.Neural Regen Res 10:778-785.

    Du X, Zhao B, Li J, Cao X, Diao M, Feng H, Chen X, Chen Z, Zeng X(2012) Astragalus polysaccharides enhance immune responses of HBV DNA vaccination via promoting the dendritic cell maturation and suppressing Treg frequency in mice. Int Immunopharmacol 14:463-470.

    El Tannir El Tayara N, Delatour B, Le Cudennec C, Guégan M, Volk A,Dhenain M (2006) Age-related evolution of amyloid burden, iron load,and MR relaxation times in a transgenic mouse model of Alzheimer’s disease. Neurobiol Dis 22:199-208.

    Fu JT, Wang P, Guo C (2013) Disturbance of iron metabolism in brain and Alzheimer’s disease. Jiepou Kexue Jinzhan 19:79-82.

    Galleano M, Puntarulo S (1997) Dietary alpha-tocopherol supplementation on antioxidant defenses after in vivo iron overload in rats. Toxicology 124:73-81.

    Gao J, Inagaki Y, Liu Y (2013) Research progress on flavonoids isolated from traditional Chinese medicine in treatment of Alzheimer’s disease.Intractable Rare Dis Res 2:3-10.

    Geisser P (1997) Iron therapy and oxidative stress. Met Based Drugs 4:137-152.

    Genlain M, Godaux E, Ris L (2007) Involvement of hyperpolarization-activated cation channels in synaptic modulation. Neuroreport 18:1231-1235.

    Griffin WS (2006) Inflammation and neurodegenerative diseases. Am J Clin Nutr 83:470S-474S.

    Hagemeier J, Geurts JJ, Zivadinov R (2012) Brain iron accumulation in aging and neurodegenerative disorders. Expert Rev Neurother 12:1467-1480.

    Hishikawa T, Ono S, Ogawa T, Tokunaga K, Sugiu K, Date I (2008) Effects of deferoxamine-activated hypoxia-inducible factor-1 on the brainstem after subarachnoid hemorrhage in rats. Neurosurgery 62:232-240; discussion 240-241.

    Hoozemans JJ, Veerhuis R, Rozemuller JM, Eikelenboom P (2006) Neuroinflammation and regeneration in the early stages of Alzheimer’s disease pathology. Int J Dev Neurosci 24:157-165.

    Huang WM, Liang YQ, Tang LJ, Ding Y, Wang XH (2013) Antioxidant and anti-inflammatory effects of Astragalus polysaccharide on EA.hy926 cells. Exp Ther Med 6:199-203.

    Katzman R, Saitoh T (1991) Advances in Alzheimer’s disease. FASEB J 5:278-286.

    Ko JK, Lam FY, Cheung AP (2005) Amelioration of experimental colitis by Astragalus membranaceus through anti-oxidation and inhibition of adhesion molecule synthesis. World J Gastroenterol 11:5787-5794.

    Lee CH, Yan B, Yoo KY, Choi JH, Kwon SH, Her S, Sohn Y, Hwang IK,Cho JH, Kim YM, Won MH (2011) Ischemia-induced changes in glucagon-like peptide-1 receptor and neuroprotective effect of its agonist, exendin-4, in experimental transient cerebral ischemia. J Neurosci Res 89:1103-1113.

    Lim DW, Lee C, Kim IH, Kim YT (2013) Anti-inflammatory effects of total isoflavones from Pueraria lobata on cerebral ischemia in rats.Molecules 18:10404-10412.

    Lu Q, Black SM (2016) Iron metabolism, oxidative stress, and neonatal brain injury. Neural Regen Res 11:725-726.

    Misonou H, Morishima-Kawashima M, Ihara Y (2000) Oxidative stress induces intracellular accumulation of amyloid beta-protein (Abeta)in human neuroblastoma cells. Biochemistry 39:6951-6959.

    Mucke L (2009) Neuroscience: Alzheimer’s disease. Nature 461:895-897.

    Parks RR, Huang CC, Haddad J, Jr. (1994) Evidence of oxygen radical injury in experimental otitis media. Laryngoscope 104:1389-1392.

    Qian ZM, Shen X (2001) Brain iron transport and neurodegeneration.Trends Mol Med 7:103-108.

    Qiao X, Cummins DJ, Paul SM (2001) Neuroin flammation-induced acceleration of amyloid deposition in the APPV717F transgenic mouse.Eur J Neurosci 14:474-482.

    Rolston RK, Perry G, Zhu X, Castellani RJ, Dwyer BE, Lee HG, Petersen RB, Smith MA (2009) Iron: A pathological mediator of Alzheimer disease? Agro Food Ind Hi Tech 19:33-36.

    Sastre M, Dewachter I, Landreth GE, Willson TM, Klockgether T, van Leuven F, Heneka MT (2003) Nonsteroidal anti-inflammatory drugs and peroxisome proliferator-activated receptor-gamma agonists modulate immunostimulated processing of amyloid precursor protein through regulation of beta-secretase. J Neurosci 23:9796-9804.

    Swardfager W, Lanct?t K, Rothenburg L, Wong A, Cappell J, Herrmann N (2010) A meta-analysis of cytokines in Alzheimer’s disease. Biol Psychiatry 68:930-941.

    Tang J, Wu L, Huang H, Feng J, Yuan Y, Zhou Y, Huang P, Xu Y, Yu C(2013) Back propagation artificial neural network for community Alzheimer’s disease screening in China. Neural Regen Res 8:270-276.

    Vetrivel KS, Thinakaran G (2010) Membrane rafts in Alzheimer’s disease beta-amyloid production. Biochim Biophys Acta 1801:860-867.

    Wan S, Hua Y, Keep RF, Hoff JT, Xi G (2006) Deferoxamine reduces CSF free iron levels following intracerebral hemorrhage. Acta Neurochir Suppl 96:199-202.

    Whitnall M, Richardson DR (2006) Iron: a new target for pharmacological intervention in neurodegenerative diseases. Semin Pediatr Neurol 13:186-197.

    Xian YF, Lin ZX, Mao QQ, Ip SP, Su ZR, Lai XP (2012) Protective effect of isorhynchophylline against beta-amyloid-induced neurotoxicity in PC12 cells. Cell Mol Neurobiol 32:353-360.

    Yoo DY, Yoo KY, Park JH, Kwon HJ, Jung HY, Kim JW, Choi GM, Moon SM, Kim DW, Yoon YS, Won MH, Hwang IK (2016) Time- and celltype specific changes in iron, ferritin, and transferrin in the gerbil hippocampal CA1 region after transient forebrain ischemia. Neural Regen Res 11:924-930.

    Zhu WZ, Zhong WD, Wang W, Zhan CJ, Wang CY, Qi JP, Wang JZ, Lei T (2009) Quantitative MR phase-corrected imaging to investigate increased brain iron deposition of patients with Alzheimer disease.Radiology 253:497-504.

    岛国在线免费视频观看| 日韩av在线大香蕉| 亚洲精品在线观看二区| 一个人看视频在线观看www免费| 国产精品亚洲av一区麻豆| 老鸭窝网址在线观看| 欧美一区二区国产精品久久精品| 日韩人妻高清精品专区| 男女那种视频在线观看| 成人av在线播放网站| 国产精品精品国产色婷婷| eeuss影院久久| 91久久精品电影网| 国产 一区 欧美 日韩| 欧美乱妇无乱码| 九色成人免费人妻av| 亚洲美女黄片视频| 国产一区二区三区视频了| 国产 一区 欧美 日韩| 观看免费一级毛片| 免费黄网站久久成人精品 | av在线老鸭窝| 国内精品久久久久久久电影| 亚洲最大成人手机在线| 欧美黑人巨大hd| 国产成年人精品一区二区| 亚洲成人免费电影在线观看| 日韩精品中文字幕看吧| 亚洲天堂国产精品一区在线| 日韩欧美精品免费久久 | 51国产日韩欧美| 少妇裸体淫交视频免费看高清| 丰满的人妻完整版| 深夜a级毛片| 日韩欧美精品v在线| 国产极品精品免费视频能看的| 色5月婷婷丁香| 欧美高清性xxxxhd video| 99久久精品国产亚洲精品| 国产中年淑女户外野战色| 午夜老司机福利剧场| 最后的刺客免费高清国语| 日本 欧美在线| 国产精品女同一区二区软件 | 久久亚洲精品不卡| 91av网一区二区| 我的老师免费观看完整版| 99久久精品一区二区三区| 黄色丝袜av网址大全| 首页视频小说图片口味搜索| 国产精品,欧美在线| 人妻制服诱惑在线中文字幕| 激情在线观看视频在线高清| 搞女人的毛片| 看片在线看免费视频| 激情在线观看视频在线高清| 成人特级黄色片久久久久久久| 婷婷亚洲欧美| 欧美绝顶高潮抽搐喷水| 波野结衣二区三区在线| 国产亚洲精品久久久久久毛片| 亚洲最大成人中文| 亚洲电影在线观看av| 国产不卡一卡二| 长腿黑丝高跟| 一区福利在线观看| 九九在线视频观看精品| 日本精品一区二区三区蜜桃| 成人无遮挡网站| 又爽又黄无遮挡网站| av在线老鸭窝| 国产视频内射| 男人的好看免费观看在线视频| 嫩草影院入口| 在现免费观看毛片| 非洲黑人性xxxx精品又粗又长| 特大巨黑吊av在线直播| 中文在线观看免费www的网站| 亚洲三级黄色毛片| 欧美日韩国产亚洲二区| .国产精品久久| 悠悠久久av| 午夜影院日韩av| 波多野结衣高清无吗| 精品久久久久久久久亚洲 | 国产免费一级a男人的天堂| 午夜福利在线在线| 午夜影院日韩av| 久久精品国产清高在天天线| 欧美一区二区国产精品久久精品| 狠狠狠狠99中文字幕| 一本久久中文字幕| xxxwww97欧美| 中文字幕人妻熟人妻熟丝袜美| 中文字幕av在线有码专区| 欧美日韩国产亚洲二区| 欧美日韩黄片免| 日韩精品青青久久久久久| 一级a爱片免费观看的视频| 美女高潮喷水抽搐中文字幕| 精品久久国产蜜桃| 天天躁日日操中文字幕| 国产欧美日韩一区二区精品| 美女高潮的动态| 欧美国产日韩亚洲一区| 窝窝影院91人妻| 成人高潮视频无遮挡免费网站| 久久精品91蜜桃| 97人妻精品一区二区三区麻豆| 中文字幕人成人乱码亚洲影| 欧美精品国产亚洲| 国产精品爽爽va在线观看网站| 99久久无色码亚洲精品果冻| 狂野欧美白嫩少妇大欣赏| 最新在线观看一区二区三区| 久久国产精品人妻蜜桃| 国产免费男女视频| 精品福利观看| 男人舔奶头视频| 高清日韩中文字幕在线| 国产一区二区亚洲精品在线观看| 噜噜噜噜噜久久久久久91| 毛片一级片免费看久久久久 | 日本 av在线| 国产亚洲欧美在线一区二区| 在线观看舔阴道视频| 非洲黑人性xxxx精品又粗又长| 如何舔出高潮| 亚洲美女搞黄在线观看 | 亚洲av一区综合| 午夜精品在线福利| 成人亚洲精品av一区二区| 人妻丰满熟妇av一区二区三区| 天天一区二区日本电影三级| 久久热精品热| 变态另类成人亚洲欧美熟女| 国产爱豆传媒在线观看| 丁香欧美五月| 有码 亚洲区| 成人无遮挡网站| 亚洲人与动物交配视频| 黄色丝袜av网址大全| 91狼人影院| 我的老师免费观看完整版| 久久精品久久久久久噜噜老黄 | 欧美区成人在线视频| 大型黄色视频在线免费观看| 一区福利在线观看| 午夜福利免费观看在线| 18禁裸乳无遮挡免费网站照片| 女人被狂操c到高潮| 99热精品在线国产| 欧美日韩综合久久久久久 | 国产黄片美女视频| 免费看a级黄色片| 久久精品综合一区二区三区| 毛片女人毛片| 99热这里只有是精品在线观看 | 少妇人妻一区二区三区视频| 亚洲av成人av| 日本a在线网址| 1024手机看黄色片| 成人亚洲精品av一区二区| 国产精品野战在线观看| 网址你懂的国产日韩在线| 欧美一区二区精品小视频在线| 午夜影院日韩av| 久久精品综合一区二区三区| 国产亚洲av嫩草精品影院| 蜜桃亚洲精品一区二区三区| 欧美国产日韩亚洲一区| 韩国av一区二区三区四区| 1000部很黄的大片| 91av网一区二区| 国产黄a三级三级三级人| 最后的刺客免费高清国语| 最新在线观看一区二区三区| 精品久久久久久久久久久久久| 免费看光身美女| av在线观看视频网站免费| 哪里可以看免费的av片| 美女cb高潮喷水在线观看| bbb黄色大片| 欧美高清性xxxxhd video| 亚洲无线观看免费| 夜夜看夜夜爽夜夜摸| 国内少妇人妻偷人精品xxx网站| 国产午夜精品久久久久久一区二区三区 | 国产三级在线视频| 色综合亚洲欧美另类图片| 中文字幕熟女人妻在线| 99riav亚洲国产免费| 成熟少妇高潮喷水视频| 18+在线观看网站| 一进一出抽搐gif免费好疼| 欧美日韩乱码在线| 可以在线观看毛片的网站| 搡老岳熟女国产| www.www免费av| 看片在线看免费视频| 欧美bdsm另类| 简卡轻食公司| av天堂中文字幕网| 一个人免费在线观看的高清视频| 亚洲久久久久久中文字幕| 国产免费一级a男人的天堂| 国产精品女同一区二区软件 | 亚洲精品一卡2卡三卡4卡5卡| 国产欧美日韩精品亚洲av| 久久久国产成人精品二区| 中文亚洲av片在线观看爽| 国产一区二区三区在线臀色熟女| 成人鲁丝片一二三区免费| 国产精品亚洲一级av第二区| 色综合站精品国产| 99久久成人亚洲精品观看| 成人国产一区最新在线观看| 亚洲七黄色美女视频| 久久久久久久亚洲中文字幕 | 国产成人啪精品午夜网站| 91久久精品电影网| 久久午夜福利片| 午夜视频国产福利| av欧美777| 99在线视频只有这里精品首页| 国产免费男女视频| 成年女人永久免费观看视频| 精品久久久久久成人av| 自拍偷自拍亚洲精品老妇| 国产精品美女特级片免费视频播放器| 国产伦在线观看视频一区| 国产精品免费一区二区三区在线| 在线免费观看不下载黄p国产 | 热99在线观看视频| 无人区码免费观看不卡| 亚洲国产高清在线一区二区三| 丰满乱子伦码专区| 色视频www国产| 可以在线观看毛片的网站| 一个人免费在线观看电影| 国产69精品久久久久777片| 日本熟妇午夜| 大型黄色视频在线免费观看| 一个人免费在线观看电影| 欧美另类亚洲清纯唯美| 变态另类成人亚洲欧美熟女| 亚洲国产精品999在线| 性色av乱码一区二区三区2| 欧美色欧美亚洲另类二区| 成人一区二区视频在线观看| 99久久九九国产精品国产免费| 老熟妇仑乱视频hdxx| 色综合亚洲欧美另类图片| 一个人观看的视频www高清免费观看| 直男gayav资源| 国产一区二区三区视频了| 天堂√8在线中文| 亚洲五月婷婷丁香| 亚洲不卡免费看| 国产毛片a区久久久久| 村上凉子中文字幕在线| 淫秽高清视频在线观看| 麻豆国产av国片精品| 久久伊人香网站| 成人高潮视频无遮挡免费网站| 国产午夜精品久久久久久一区二区三区 | 人妻夜夜爽99麻豆av| 深夜精品福利| 给我免费播放毛片高清在线观看| 小蜜桃在线观看免费完整版高清| 久久这里只有精品中国| 如何舔出高潮| 日本与韩国留学比较| 久久久色成人| 熟女电影av网| 女人十人毛片免费观看3o分钟| 美女被艹到高潮喷水动态| 国产蜜桃级精品一区二区三区| 99热这里只有是精品在线观看 | 国产中年淑女户外野战色| 国产视频一区二区在线看| 国产黄色小视频在线观看| 欧美潮喷喷水| 一区福利在线观看| 国产精品嫩草影院av在线观看 | 91狼人影院| 久久精品影院6| 欧美3d第一页| 亚洲最大成人中文| 综合色av麻豆| 日韩大尺度精品在线看网址| 亚洲成人免费电影在线观看| 欧美性猛交黑人性爽| 中文在线观看免费www的网站| 深夜a级毛片| 日韩有码中文字幕| 色哟哟哟哟哟哟| 日本 欧美在线| 国产伦在线观看视频一区| 久久精品影院6| 欧美高清性xxxxhd video| 亚洲真实伦在线观看| 99在线视频只有这里精品首页| 一个人看的www免费观看视频| 黄色视频,在线免费观看| 国产色婷婷99| 久久久久久久久久黄片| 欧美国产日韩亚洲一区| 午夜a级毛片| 欧美色欧美亚洲另类二区| 麻豆久久精品国产亚洲av| 午夜福利在线观看吧| 国产探花极品一区二区| 一个人免费在线观看的高清视频| 亚洲在线观看片| 欧美bdsm另类| 久久久久性生活片| 日本免费一区二区三区高清不卡| 午夜免费激情av| 婷婷精品国产亚洲av| 女生性感内裤真人,穿戴方法视频| 一本精品99久久精品77| 美女免费视频网站| 蜜桃亚洲精品一区二区三区| 欧美又色又爽又黄视频| x7x7x7水蜜桃| 国产三级中文精品| 三级毛片av免费| 亚洲精品在线美女| 18禁裸乳无遮挡免费网站照片| 黄色配什么色好看| 欧美成人性av电影在线观看| 在线观看舔阴道视频| 女生性感内裤真人,穿戴方法视频| 久久人妻av系列| 婷婷六月久久综合丁香| 久久精品国产亚洲av天美| 丁香欧美五月| 欧美成人免费av一区二区三区| 午夜免费成人在线视频| 日韩 亚洲 欧美在线| 国产一区二区三区视频了| 成人无遮挡网站| 婷婷亚洲欧美| 亚洲欧美日韩卡通动漫| 天天一区二区日本电影三级| 18禁在线播放成人免费| 欧美日韩国产亚洲二区| 一级毛片久久久久久久久女| 99久久99久久久精品蜜桃| 九色成人免费人妻av| 国产色爽女视频免费观看| 久久99热这里只有精品18| 欧美极品一区二区三区四区| 日本五十路高清| 国产人妻一区二区三区在| 国产一级毛片七仙女欲春2| 成人无遮挡网站| 欧美最新免费一区二区三区 | 成人精品一区二区免费| 欧美性猛交黑人性爽| 99热精品在线国产| 亚洲精品粉嫩美女一区| 国产激情偷乱视频一区二区| 99在线人妻在线中文字幕| 精品一区二区三区av网在线观看| 国产精品免费一区二区三区在线| 尤物成人国产欧美一区二区三区| 国产高清激情床上av| 欧美极品一区二区三区四区| 亚洲精品成人久久久久久| 国产成人欧美在线观看| 免费看a级黄色片| 一级黄片播放器| 91字幕亚洲| 午夜精品久久久久久毛片777| 欧美日韩黄片免| 国产亚洲精品久久久com| 中文亚洲av片在线观看爽| 成人无遮挡网站| 国产精品98久久久久久宅男小说| 欧美黑人巨大hd| h日本视频在线播放| 91久久精品国产一区二区成人| 欧美精品啪啪一区二区三区| 亚洲av熟女| 亚洲最大成人中文| 一个人观看的视频www高清免费观看| 特级一级黄色大片| 亚洲av第一区精品v没综合| 91九色精品人成在线观看| 舔av片在线| 国产精品人妻久久久久久| 露出奶头的视频| 欧美高清成人免费视频www| 我要看日韩黄色一级片| 久久精品国产亚洲av涩爱 | av天堂中文字幕网| 亚洲欧美清纯卡通| 日韩欧美 国产精品| 能在线免费观看的黄片| 人人妻,人人澡人人爽秒播| x7x7x7水蜜桃| 少妇人妻一区二区三区视频| 极品教师在线视频| av国产免费在线观看| 欧美3d第一页| 少妇被粗大猛烈的视频| 直男gayav资源| 精品一区二区免费观看| 国产 一区 欧美 日韩| 在线免费观看的www视频| 男女那种视频在线观看| 99在线人妻在线中文字幕| 国产精品野战在线观看| a级毛片a级免费在线| 久久久久久久久中文| 嫩草影院精品99| 国产伦一二天堂av在线观看| 免费观看人在逋| 免费看光身美女| 欧美zozozo另类| 精品福利观看| 男插女下体视频免费在线播放| 精品国产亚洲在线| 成人av在线播放网站| 国内少妇人妻偷人精品xxx网站| 一进一出抽搐动态| 亚洲乱码一区二区免费版| 国产成人欧美在线观看| 欧美bdsm另类| 色尼玛亚洲综合影院| 国产91精品成人一区二区三区| 国产大屁股一区二区在线视频| 97超视频在线观看视频| 99久久精品一区二区三区| 90打野战视频偷拍视频| 国产午夜精品论理片| 嫩草影院新地址| 老熟妇乱子伦视频在线观看| 在线观看午夜福利视频| 国产免费一级a男人的天堂| 精品人妻熟女av久视频| 日本与韩国留学比较| 国产精品久久久久久久久免 | 免费在线观看成人毛片| 18禁黄网站禁片免费观看直播| 精品日产1卡2卡| 久久精品国产亚洲av香蕉五月| 国产aⅴ精品一区二区三区波| 日本精品一区二区三区蜜桃| 麻豆一二三区av精品| av在线天堂中文字幕| 国产精品久久久久久久电影| a在线观看视频网站| 欧美三级亚洲精品| 一进一出抽搐动态| 97人妻精品一区二区三区麻豆| 蜜桃久久精品国产亚洲av| 高清日韩中文字幕在线| 怎么达到女性高潮| 极品教师在线免费播放| 看十八女毛片水多多多| 毛片女人毛片| 久久欧美精品欧美久久欧美| 如何舔出高潮| 免费电影在线观看免费观看| 日日摸夜夜添夜夜添小说| 亚洲性夜色夜夜综合| 亚洲最大成人中文| 亚洲欧美清纯卡通| 久久精品人妻少妇| 欧美成人免费av一区二区三区| av欧美777| av在线老鸭窝| 一个人免费在线观看的高清视频| 人妻制服诱惑在线中文字幕| 成人无遮挡网站| 每晚都被弄得嗷嗷叫到高潮| 夜夜夜夜夜久久久久| 国产不卡一卡二| 中文字幕人妻熟人妻熟丝袜美| 国产欧美日韩精品亚洲av| 99久久99久久久精品蜜桃| 狠狠狠狠99中文字幕| av福利片在线观看| 麻豆国产av国片精品| av中文乱码字幕在线| 精品久久久久久成人av| 国产高清视频在线观看网站| 色综合婷婷激情| 岛国在线免费视频观看| 精品一区二区三区视频在线观看免费| 可以在线观看的亚洲视频| 免费看a级黄色片| 亚洲久久久久久中文字幕| 精品乱码久久久久久99久播| 在线国产一区二区在线| 两个人视频免费观看高清| 久久香蕉精品热| 欧美色视频一区免费| av中文乱码字幕在线| 成人亚洲精品av一区二区| 日韩欧美三级三区| 国产亚洲精品av在线| 一个人观看的视频www高清免费观看| 亚洲成人免费电影在线观看| 国产中年淑女户外野战色| 色噜噜av男人的天堂激情| 99热这里只有是精品50| 亚洲性夜色夜夜综合| 国产黄片美女视频| 成人性生交大片免费视频hd| 亚洲成人中文字幕在线播放| 午夜激情福利司机影院| 国产主播在线观看一区二区| 99精品在免费线老司机午夜| 亚洲av第一区精品v没综合| 免费av不卡在线播放| 99精品久久久久人妻精品| 亚洲国产精品999在线| 乱码一卡2卡4卡精品| 九九热线精品视视频播放| 亚洲最大成人中文| 久久久色成人| 无遮挡黄片免费观看| 中文在线观看免费www的网站| 久久久久久久午夜电影| 看十八女毛片水多多多| 最近在线观看免费完整版| 亚洲欧美日韩无卡精品| 十八禁国产超污无遮挡网站| 99国产综合亚洲精品| 又黄又爽又免费观看的视频| 亚洲五月婷婷丁香| 国产三级在线视频| 国产 一区 欧美 日韩| 亚洲狠狠婷婷综合久久图片| 黄色日韩在线| 51国产日韩欧美| 亚洲片人在线观看| 国产av麻豆久久久久久久| 亚洲欧美日韩卡通动漫| 成人av一区二区三区在线看| 中出人妻视频一区二区| 亚洲av成人av| 黄色配什么色好看| 亚洲国产色片| 男女之事视频高清在线观看| 天天躁日日操中文字幕| 搡老熟女国产l中国老女人| 99热只有精品国产| 1024手机看黄色片| 国产乱人视频| 欧美色欧美亚洲另类二区| 美女大奶头视频| 国产大屁股一区二区在线视频| 国产不卡一卡二| 亚洲av成人精品一区久久| 色哟哟·www| 国产精品亚洲美女久久久| 日本免费a在线| 久久国产精品影院| 久久久色成人| 亚洲乱码一区二区免费版| 丰满人妻熟妇乱又伦精品不卡| 国产欧美日韩精品亚洲av| 国产午夜精品论理片| 狠狠狠狠99中文字幕| 永久网站在线| 桃色一区二区三区在线观看| 宅男免费午夜| 色5月婷婷丁香| 男插女下体视频免费在线播放| 亚洲专区中文字幕在线| 中文字幕免费在线视频6| 十八禁网站免费在线| 国产午夜精品论理片| 毛片女人毛片| 无遮挡黄片免费观看| 老师上课跳d突然被开到最大视频 久久午夜综合久久蜜桃 | 一本一本综合久久| 国产午夜精品久久久久久一区二区三区 | 在线观看免费视频日本深夜| 非洲黑人性xxxx精品又粗又长| av国产免费在线观看| 啦啦啦观看免费观看视频高清| 欧美性猛交╳xxx乱大交人| 亚洲人成网站在线播放欧美日韩| 可以在线观看的亚洲视频| 男女下面进入的视频免费午夜| 精品国内亚洲2022精品成人| 成人午夜高清在线视频| 99久久久亚洲精品蜜臀av| 亚洲欧美精品综合久久99| 十八禁网站免费在线| 老熟妇乱子伦视频在线观看| 日韩有码中文字幕| 搡女人真爽免费视频火全软件 | 成人av在线播放网站| 一a级毛片在线观看| 国产激情偷乱视频一区二区| 国产高清有码在线观看视频| 两个人的视频大全免费| 成人亚洲精品av一区二区| 久久人人爽人人爽人人片va | 噜噜噜噜噜久久久久久91| 白带黄色成豆腐渣| 一进一出抽搐动态| 99久久精品热视频| 韩国av一区二区三区四区| 久久久久九九精品影院| 亚洲成人精品中文字幕电影| 国产色婷婷99| 欧美在线一区亚洲|