Correction to: Protective Effects of 28-O-Cafeoyl Betulin (B-CA) on the Cerebral Cortex of Ischemic Rats Revealed by a NMR-Based Metabolomics Analysis

Neurochemical Research - In the original version of this article, unfortunately the Fig. 3C and 3F were published with incorrect version. The correct version of the Fig. 3C and 3F...     

Distinct Metabolic Profile of Primary Focal Segmental Glomerulosclerosis Revealed by NMR-Based Metabolomics

Background

Primary focal segmental glomerulosclerosis (FSGS) is pathological entity which is characterized by idiopathic steroid-resistant nephrotic syndrome (SRNS) and progression to end-stage renal disease (ESRD) in the majority of affected individuals. Currently, there is no practical noninvasive technique to predict different pathological types of glomerulopathies. In this study, the role of urinary metabolomics in the diagnosis and pathogenesis of FSGS was investigated.

Methods

NMR-based metabolomics was applied for the urinary metabolic profile in the patients with FSGS (n = 25), membranous nephropathy (MN, n = 24), minimal change disease (MCD, n = 14) and IgA nephropathy (IgAN, n = 26), and healthy controls (CON, n = 35). The acquired data were analyzed using principal component analysis (PCA) followed by orthogonal projections to latent structure discriminant analysis (OPLS-DA). Model validity was verified using permutation tests.

Results

FSGS patients were clearly distinguished from healthy controls and other three types of glomerulopathies with good sensitivity and specificity based on their global urinary metabolic profiles. In FSGS patients, urinary levels of glucose, dimethylamine and trimethylamine increased compared with healthy controls, while pyruvate, valine, hippurate, isoleucine, phenylacetylglycine, citrate, tyrosine, 3-methylhistidine and β-hydroxyisovalerate decreased. Additionally, FSGS patients had lower urine N-methylnicotinamide levels compared with other glomerulopathies.

Conclusions

NMR-based metabonomic approach is amenable for the noninvasive diagnosis and differential diagnosis of FSGS as well as other glomerulopathies, and it could indicate the possible mechanisms of primary FSGS.     

高压氧预处理对大鼠局灶性脑缺血再灌注损伤的保护作用

目的:研究高压氧预处理对大鼠脑缺血再灌注损伤的保护作用。方法:36只SD大鼠随机分为假手术组、模型组及高压氧预处理组,每组12只。高压氧预处理组大鼠在造模前5天给予高压氧预处理。采用线栓法建立大鼠脑缺血再灌注模型,观察高压氧预处理对脑缺血再灌注损伤大鼠神经功能缺损评分、脑梗死面积的影响,检测大鼠缺血脑组织COX-2 mRNA和蛋白的表达以及IL-1β、TNF-α、MDA的含量。结果:高压氧预处理可明显改善脑缺血再灌注大鼠神经功能缺损评分,减少脑梗死面积,降低COX-2m RNA和蛋白表达量,抑制IL-1β、TNF-α的表达,降低MDA水平。结论:高压氧预处理对大鼠脑缺血再灌注损伤具有明显的保护作用,其机制可能与抑制IL-1β、TNF-α、COX-2的表达以及减弱脂质过氧化反应有关。     

白藜芦醇预处理对人鼠脑缺血再灌损伤的保护作用

目的:探讨白藜芦醇预处理对大鼠脑缺血再灌损伤的保护作用及其分子机制.方法:大鼠随机分成假手术组、缺血溶剂组、白藜芦醇预处理组,四动脉阻塞(4-VO)法建立前脑缺血模型,缺血10min/再灌22h,试剂盒检测大鼠海马组织SOD活力及NO、MDA含量变化,RT-PCR法观察GRP78 mRNA的表达.结果:缺血溶剂组海马组织SOD活性明显低于假手术组,NO、MDA含量高于假手术组;缺血前白藜芦醇预处理能显著反转缺血诱导的SOD活力和NO、MDA水平变化,脑缺血能明显上调GRP78 mRNA水平;白藜芦醇预处理能有效抑制缺血诱导的GRP78表达,与缺血组比有显著性差异.结论:白藜芦醇能通过上调SOD活力,减少NO、MDA的生成来抑制缺血后自由基的生成和积累,继而缓解内质网应激、下调GRP78的表达,减轻缺血性脑组织损伤.     

Study of the Cardiotoxicity of Venenum Bufonis in Rats using an 1H NMR-Based Metabolomics Approach

Venenum Bufonis, a well-known traditional Chinese medicine, has been widely used in Asia and has gained popularity in Western countries over the last decade. Venenum Bufonis has obvious side effects that have been observed in clinical settings, but few studies have reported on its cardiotoxicity. In this work, the cardiotoxicity of Venenum Bufonis was investigated using a ¹¹H NMR-based metabolomics approach. The ¹H NMR profiles of the serum, myocardial extracts and liver extracts of specific-pathogen-free rats showed that Venenum Bufonis produced significant metabolic perturbations dose-dependently with a distinct time effect, peaking at 2 hr after dosing and attenuating gradually. Clinical chemistry, electrocardiographic recordings, and histopathological evaluation provided additional evidence of Venenum Bufonis-induced cardiac damage that complemented and supported the metabolomics findings. The combined results demonstrated that oxidative stress, mitochondrial dysfunction, and energy metabolism perturbations were associated with the cardiac damage that results from Venenum Bufonis.     

Protective Effects of Naringenin on Iron-Overload-Induced Cerebral Cortex Neurotoxicity Correlated with Oxidative Stress

Iron is a component of several metalloproteins involved in crucial metabolic processes such as oxygen sensing and transport, energy metabolism, and DNA synthesis. This metal progressively accumulates in the pathogenesis of Alzheimer’s (AD) and Parkinson’s (PD) diseases. Naringenin (NGEN), a natural flavonoid compound, has been reported to possess neuroprotective effect against PD-related pathology, however, the mechanisms underlying its beneficial effects are poorly defined. Thus, the aim of this study is to investigate the potential mechanism involved in the cytoprotection of NGEN against iron-induced neurotoxicity in the cerebral cortex of Wistar rats. Animals that were given repetitive injections of iron dextran for a total of 4 weeks showed a significant increase in lipid and protein markers such as thiobarbituric reactive acid substances, protein carbonyl product content levels, and DNA apoptosis in the cerebral cortex. These changes were accompanied by a decrease of enzymatic antioxidants like superoxide dismutase and catalase and in the levels of nonenzymatic antioxidants like total thiols and ascorbic acid. The activity of glutathione peroxidase remained unchanged in rats. A significant decrease in acetylcholinesterase and Na⁺/K⁺-ATPase activities was also shown, with a substantial rise in the nitric oxide levels. Coadministration of NGEN to iron-treated rats significantly improved antioxidant enzyme activities and attenuated oxidative damages observed in the cerebral cortex. The potential effect of NGEN to prevent iron-induced neurotoxicity was also reflected by the microscopic study, indicative of its neuroprotective effects.     

大鼠肢端缺血预处理对脑缺血后炎症反应的影响

目的通过观察肢端缺血预处理（1imbisehemicpreconditioning,LIP）对大鼠脑缺血性损伤后重要炎症因子表达的影响,探讨LIP诱导的脑缺血耐受与炎症反应之间的关系。方法选取72只SD大鼠,实验组（LIP组）30只、缺血组30只和对照组12只。实验组和缺血组设立5个时间点：6h、12h、24h、48h和72h,每点6只。通过线栓法建立大鼠大脑中动脉阻塞（middlecerebralarteryocclusion,MCAO）的局灶性脑缺血模型及LIP法建立脑缺血耐受模型,采用HE观察每组大鼠的脑组织形态学改变、QRT—PCR和ELISA方法检测脑组织中炎症因子IL-17及IL-6的表达变化。结果实验组脑组织学病理改变明显轻于缺血组。与缺血组相比：实验组的IL-17和IL-6的基因和蛋白表达在整体水平均呈下降趋势;mRNA水平提示实验组在缺血12h、24h和48h后脑组织中IL-17、IL-6的表达量显著减少（P〈0．01）;蛋白水平提示实验组在缺血24h和48h后脑组织中的IL-6以及在缺血12h、24h和48h后脑组织中IL-17的表达量均降低（P〈0．05）。结论LIP诱导脑缺血耐受,可以减轻脑缺血后的炎症反应,对缺血性脑损伤有一定的保护作用。     

Evaluation of Pacific White Shrimp (Litopenaeus vannamei) Health during a Superintensive Aquaculture Growout Using NMR-Based Metabolomics

Success of the shrimp aquaculture industry requires technological advances that increase production and environmental sustainability. Indoor, superintensive, aquaculture systems are being developed that permit year-round production of farmed shrimp at high densities. These systems are intended to overcome problems of disease susceptibility and of water quality issues from waste products, by operating as essentially closed systems that promote beneficial microbial communities (biofloc). The resulting biofloc can assimilate and detoxify wastes, may provide nutrition for the farmed organisms resulting in improved growth, and may aid in reducing disease initiated from external sources. Nuclear magnetic resonance (NMR)-based metabolomic techniques were used to assess shrimp health during a full growout cycle from the nursery phase through harvest in a minimal-exchange, superintensive, biofloc system. Aberrant shrimp metabolomes were detected from a spike in total ammonia nitrogen in the nursery, from a reduced feeding period that was a consequence of surface scum build-up in the raceway, and from the stocking transition from the nursery to the growout raceway. The biochemical changes in the shrimp that were induced by the stressors were essential for survival and included nitrogen detoxification and energy conservation mechanisms. Inosine and trehalose may be general biomarkers of stress in Litopenaeus vannamei. This study demonstrates one aspect of the practicality of using NMR-based metabolomics to enhance the aquaculture industry by providing physiological insight into common environmental stresses that may limit growth or better explain reduced survival and production.     

Functionally Brain Network Connected to the Retrosplenial Cortex of Rats Revealed by 7T fMRI

Functional networks are regarded as important mechanisms for increasing our understanding of brain function in healthy and diseased states, and increased interest has been focused on extending the study of functional networks to animal models because such models provide a functional understanding of disease progression, therapy and repair. In rodents, the retrosplenial cortex (RSC) is an important cortical region because it has a large size and presents transitional patterns of lamination between the neocortex and archicortex. In addition, a number of invasive studies have highlighted the importance of the RSC for many functions. However, the network based on the RSC in rodents remains unclear. Based on the critical importance of the RSC, we defined the bilateral RSCs as two regions of interest and estimated the network based on the RSC. The results showed that the related regions include the parietal association cortex, hippocampus, thalamus nucleus, midbrain structures, and hypothalamic mammillary bodies. Our findings indicate two possible major networks: a sensory-cognitive network that has a hub in the RSCs and processes sensory information, spatial learning, and episodic memory; and a second network that is involved in the regulation of visceral functions and arousal. In addition, functional asymmetry between the bilateral RSCs was observed.     

正常与脑缺血大鼠的脑皮质蛋白质差异分析鉴定

Wistar大鼠随机分为正常组和模型组,采用改进的线栓法制备模型,在规定的时间点快速断头取脑,分离脑皮质组织,提取蛋白质后双向电泳展示,以ImageMaster 2D Elite v301软件对2_DE图谱进行差异表达分析,目标蛋白点用基质辅助激光解析电离质谱测定肽质量指纹图进行鉴定。线粒体应激70蛋白前体、血小板活化因子乙酰基水解酶Ibβ亚单位、ADP核糖基化因子蛋白3、电压依赖性阴离子选择通道蛋白1、泛素C末端水解酶同工酶L1、突触结合蛋白等11个蛋白在模型6h组表达上调,谷胱甘肽S-转移酶omega 1、 谷胱甘肽S-转移酶P、Cu-Zn超氧化物歧化酶、 ATP合酶D链、G蛋白β亚单位1、微管蛋白β链15、苹果酸脱氢酶等15个蛋白在模型6h组表达上调。胆绿素还原酶B、细胞因子A4前体为模型组新出现点,腺苷酸激酶同工酶1在模型组消失,Thiore doxin peroxidase 1在模型组分为2个点。以双向电泳技术得到分辨率较好的电泳图谱,并初步鉴定脑缺血后差异表达蛋白,为深入研究缺血性脑损伤病理机制奠定了基础。     

UPLC-QTOF/MS-Based Metabolomics Reveals the Protective Mechanism of Hydrogen on Mice with Ischemic Stroke

Neurochemical Research - As a reductive gas, hydrogen plays an antioxidant role by selectively scavenging oxygen free radicals. It has been reported that hydrogen has protective effects against...     

Effects of Injury on the Indoleamines in Cerebral Cortex

It was shown previously that focal cortical freezing lesions in rats cause widespread depression of local cerebral glucose utilization (LCGU) in cortical areas of the lesioned hemisphere. This was interpreted as reflecting functional depression. The underlying mechanisms were postulated to involve alterations of biogenic amine systems. Accordingly, levels of serotonin (5-HT), its metabolite 5-hydroxyindoleacetic acid (5-HIAA), and its precursor tryptophan were determined by an HPLC method with electrochemical detection in frontoparietal cortical areas of both hemispheres at 4 h and 1, 3, 6, 8, and 10 days after a unilateral cortical freezing lesion. The 5-HT content was significantly lower than normal in the lesioned hemisphere only at 24 h, whereas the 5-HIAA level peaked at 24 h but was significantly elevated above normal values between 4 h and 6 days after lesioning. No changes were noted in 5-HT and 5-HIAA contents in the hemisphere contralateral to the lesion. These results indicate that cortical 5-HT metabolism is increased throughout the lesioned hemisphere of a focally injured brain. The increase in tryptophan content of the lesioned brain appeared to have a time course more closely related to previously demonstrated changes in cortical LCGU than to the increase in 5-HIAA content.     

Different Expression Patterns of Ngb and EPOR in the Cerebral Cortex and Hippocampus Revealed Distinctive Therapeutic Effects of Intranasal Delivery of Neuro-EPO for Ischemic Insults to the Gerbil Brain

The purpose of this study was to evaluate the neuroprotective effects of intranasally delivered recombinant human neuronal erythropoietin (Neuro-EPO) on brain injury induced by unilateral permanent ischemia in the Mongolian gerbil. Expression of EPO receptor (EPOR) and neuroglobin (Ngb) over 5 weeks after intranasal treatment with Neuro-EPO was determined using immunohistochemistry. Mortality of Neuro-EPO-treated gerbils decreased after surgery, and the sensory and motor function was significantly improved. Histopathological mapping showed that Neuro-EPO significantly reduced delayed neuronal death in the brain. Expression of Ngb was upregulated in the cerebral cortex at most time points (expect for 10 min and 48 hr) and in the hippocampus at 10 min and from 48 hr to 5 weeks, whereas EPOR was almost downregulated or unchanged in the brain (expect for 48 hr). The 10 min and 48 hr seemed to be two time points for the brain to switch the expression of both Ngb and EPOR to early and late recovery phase, respectively. In addition, there were two phases, 10 min to 1 hr and 24 hr to 72 hr, respectively, closing to the “golden hour” of about 60 min and the “silver day” of 1 to 3 days, for the brain to recover from stroke onset with intranasal Neuro-EPO treatment. Therefore, the results suggest that the intranasal administration of Neuro-EPO is effective in the treatment of acute brain ischemia. The different expression patterns of Ngb and EPOR is probably due to ischemic tolerance in the cerebral cortex and ischemic sensitivity in the hippocampus.     

参芎注射液对脑缺血再灌注大鼠的神经元保护作用

目的:观察参芎注射液对大鼠局灶性脑缺血再灌注后神经细胞凋亡及内质网应激相关因子葡萄糖调节蛋白/免疫球蛋白结合蛋白(GRP78/Bip)表达的影响.方法:100只雄性SD大鼠随机分为正常组、假手术组、脑缺血再灌注组、参芎治疗组;后两组根据再灌注时间不同各分为6、12、24、72 h四个亚组;采用大鼠大脑中动脉线栓法制备局灶性脑缺血再灌注模型.TUNEL法观察细胞凋亡情况;免疫组化和RT-PCR法检测各实验组中缺血周围区GRP78/Bip的表达.结果:TUNEL法表明参芎治疗组大鼠大脑神经细胞凋亡程度较缺血再灌注组明显减轻.免疫组化和RT-PCR检测均发现各时间点缺血再灌注组大鼠GRP78/Bip表达高于假手术组及正常组;脑缺血再灌注组及参芎治疗组GRP78/Bip的表达于缺血后12 h最高,72 h恢复至正常水平,且均呈现先升高后降低的趋势;各时间点缺血再灌注组GRP78/Bip表达均高于参芎治疗组.结论:再灌注损伤后12 h内出现GRP78/Bip表达升高.参芎注射液可以下调其表达,从而可能通过减轻内质网应激而减轻缺血再灌注损伤起到神经元保护的作用.     

Short-Term Effects of Thyroid Hormones on Cytoskeletal Proteins Are Mediated by GABAergic Mechanisms in Slices of Cerebral Cortex from Young Rats

Thyroid hormones play important roles in brain function. However, few information is available about the effect of 3,5,3′-triiodo-l-thyronine (T₃) or thyroxine (T₄) on the in vitro phosphorylation of intermediate filament (IF) proteins from cerebral cortex of rats. In this study we investigated the involvement of GABAergic mechanisms mediating the effects of T₃ and T₄ on the in vitro incorporation of ³²P into IF proteins from cerebral cortex of 10-day-old male rats. Tissue slices were incubated with or without T₃, T₄, γ-aminobutiric acid (GABA), kinase inhibitors or specific GABA antagonists and ³²P-orthophosphate for 30 min. The IF-enriched cytoskeletal fraction was extracted in a high salt Triton-containing buffer and the in vitro ³²P incorporation into IF proteins was measured. We first observed that 1 μM T₃ and 0.1 μM T₄ significantly increased the in vitro incorporation of ³²P into the IF proteins studied through the PKA and PKCaMII activities. A similar effect on IF phosphorylation was achieved by incubating cortical slices with GABA. Furthermore, by using specific GABA antagonists, we verified that T₃ induced a stimulatory effect on IF phosphorylation through noncompetitive mechanisms involving GABA_A, beyond GABA_B receptors. In contrast, T₄ effects were mediated mainly by GABA_B mechanisms. In conclusion, our results demonstrate a rapid nongenomic action of T₃ and T₄ on the phosphorylating system associated to the IF proteins in slices of cerebral cortex of 10 day-old male rats and point to GABAergic mechanisms mediating such effects.     

老年和青年大鼠大脑皮质脂肪酸组成及含量的对比分析

大脑功能与脑内脂肪酸成分密切相关,分析研究老年时脑脂肪酸组成及含量有助于阐明不同类型脂肪酸在老年认知能力降低过程中的作用.采用Y型电迷宫测试老年组(22月龄)和青年组(3月龄)大鼠的学习记忆能力,表明老年组大鼠的学习记忆能力显著下降.用气相色谱的方法分析大鼠大脑皮质脂肪酸的组成及含量,显示老年组和青年组大脑皮质均含有16种脂肪酸.以C23:0为内标对各种脂肪酸进行了定量,老年组总脂肪酸含量比青年组降低约15%,各脂肪酸中含量下降的脂肪酸有长链饱和脂肪酸(C14:0、C16:0、C18:0)及多不饱和脂肪酸(C18:3、C20:4、C22:4、C22:6),含量升高的脂肪酸有单不饱和脂肪酸C20:1、C24:1.用峰面积归一法计算了各脂肪酸的相对含量,老年组相对含量下降的脂肪酸有C18:0、C20:4、C22:6,相对含量升高的有极长链饱和脂肪酸(C20:0、C24:0)及单不饱和脂肪酸(C16:1、C18:1、C20:1、C22:1、C24:1).相关性分析显示,大鼠学习能力与脑皮质C22:6、C22:4、C20:4水平呈正相关,与C20:1、C24:1水平呈负相关.上述结果为阐明不同脂肪酸在老年大脑认知功能障碍中的作用提供了实验依据。     

Excitatory Transmitter Amino Acid Release from the Ischemic Rat Cerebral Cortex: Effects of Adenosine Receptor Agonists and Antagonists

The effects of selective adenosine receptor agonists [N6-cyclopentyladenosine (CPA) and N-ethylcarboxamidoadenosine (NECA)] and antagonists [8-cyclopentyl-1,3-dipropylxanthine (DPCPX) and 9-chloro-2-(2-furanyl)-5,6-dihydro-1,2,4-triazolo[1,5-c]quinazoline-5-im ine (CGS-15943A)] on aspartate and glutamate release from the ischemic rat cerebral cortex were studied with the cortical cup technique. Cerebral ischemia (for 20 min) was elicited by four-vessel occlusion. Excitatory amino acid releases were compared from control ischemic rats and drug-treated rats. Basal levels of aspartate and glutamate release were not greatly affected by pretreatment with the adenosine receptor agonists or antagonists. However, CPA (10(-10) M) and NECA (10(-9) M) significantly inhibited the ischemia-evoked release of aspartate and glutamate into cortical superfusates. The ability to block ischemia-evoked release of excitatory amino acids was not evident at higher concentrations of CPA (10(-6) M) or NECA (10(-5) M). The selective A1 receptor antagonist DPCPX also had no effect on release when administered at a low dosage (0.01 mg/kg, i.p.) but blocked the ischemia-evoked release of aspartate and glutamate at a higher dosage (0.1 mg/kg). Evoked release was inhibited by the selective A2 receptor antagonist CGS-15943A (0.1 mg/kg, i.p.). Thus, adenosine and its analogs may suppress ischemia-evoked release of excitatory neurotransmitter amino acids via high-affinity A1 receptors, whereas coactivation of lower-affinity A2 receptors may block (or reverse) the A1-mediated response.     

缺血后处理对大鼠局灶性脑缺血再灌注损伤时TLR4 信号通路表达的影响

目的:观察缺血后处理对大鼠局灶性脑缺血再灌注损伤后TLR4通路表达的影响。方法:成年健康雄性SD大鼠110只,随机分为假手术组(sham组)(n=10)、缺血再灌注组(I/R组)和后处理组(IP组),后两组又依据缺血再灌注6h、12h、24h、48h、72h不同的时间点再分五个亚组。对各组行神经行为学评分,脑组织梗死体积测量,TUNEL技术检测神经细胞凋亡的情况,免疫组织化学技术观察各组大鼠脑组织TLR4、NF-κB和TNF-α蛋白的表达,原位杂交方法检测各组大鼠脑组织TLR4mRNA、NF-κBmRNA的表达。结果:缺血后处理可下调TLR4、NF-κB、TNF-α细胞炎性因子的表达,抑制细胞凋亡、减少脑梗死体积,改善神经行为。结论:后处理可通过抑制TLR4信号通路表达,减少脑梗死体积,改善神经功能。