Degeneration and death of dopaminergic neurons in the substantia nigra of the midbrain are the main pathological changes in Parkinson’s disease (PD); however, the mechanism underlying the selective vulnerability of specific neuronal populations in PD remains unclear. Here, we used single-cell RNA sequencing to identify seven cell clusters, including oligodendrocytes, neurons, astrocytes, oligodendrocyte progenitor cells, microglia, synapse-rich cells (SRCs), and endothelial cells, in the substantia nigra of a rotenone-induced rat model of PD based on marker genes and functional definitions. We found that SRCs were a previously unidentified cell subtype, and the tight interactions between SRCs and other cell populations can be improved by icaritin, which is a flavonoid extracted from Epimedium sagittatum Maxim. and exerts anti-neuroinflammatory, antioxidant, and immune-improving effects in PD. We also demonstrated that icaritin bound with transcription factors of SRCs, and icaritin application modulated synaptic characterization of SRCs, neuroinflammation, oxidative stress, and survival of dopaminergic neurons, and improved abnormal energy metabolism, amino acid metabolism, and phospholipase D metabolism of astrocytes in the substantia nigra of rats with PD. Moreover, icaritin supplementation also promotes the recovery of the physiological homeostasis of the other cell clusters to delay the pathogenesis of PD. These data uncovered previously unknown cellular diversity in a rat model of Parkinson’s disease and provide insights into the promising therapeutic potential of icaritin in PD.

Neuropathic pain (NP) is a prevalent clinical problem for which satisfactory treatment options are unavailable. Tetrandrine (TET), a bisbenzylisoquinoline alkaloid extracted from Stephania tetrandra S. Moore, possesses anti-inflammatory and immune-modulatory properties. Chemokine-like factor 1 (CKLF1) is known to play a crucial role in both peripheral and central inflammatory processes. This study aimed to investigate the potential anti-NP effects of TET and the involvement of CKLF1 in the action of TET. A male C57BL/6J mice model of NP caused by spared nerve injury (SNI) was established and mechanical withdrawal thresholds were measured using von Frey filaments. The results showed that TET improved mechanical allodynia in SNI mice and the propofol-induced sleep assay demonstrated that the TET group did not exhibit central inhibition, while the pregabalin (PGB) group showed significant central inhibition. Western blotting and immunofluorescence staining showed that TET significantly inhibited spinal protein expression levels of CKLF1, p-NF-κB/NF-κB, p-IKK/IKK, pro-inflammatory cytokines IL-1β and TNF-α, and increased protein expression levels of the anti-inflammatory cytokine IL-10, while inhibiting the expression levels of microglia and astrocyte markers IBA-1 and GFAP of SNI mice. Moreover, immunofluorescence double-labeling results revealed that CKLF1 was predominantly colocalized with microglia of the spinal cord (SC) in SNI mice. C19 (an antagonism peptide of CKLF1) alleviated SNI-induced mechanical pain hypersensitivity, while C27 (an analog peptide of CKLF1) induced mechanical allodynia in normal mice. TET significantly attenuated mechanical allodynia induced by C27 in mice. TET may effectively alleviate NP by reducing neuroinflammation and decreasing CKLF1.

目的:探究类叶升麻苷(ACT)对1-甲基-4-苯基-1,2,3,6-四氢吡啶(MPTP)诱导的帕金森病(PD)模型小鼠的神经保护作用及其机制.方法:采用腹腔注射MPTP构建PD模型小鼠,实验动物随机分为对照组、模型组和100 mg·kg-1 ACT给药组,通过灌胃(ig)的方式连续给药14 d,给药完成后,行为学评价和生化学检测评价ACT对PD小鼠的神经保护作用.采用双向电泳技术、质谱分析技术和Western blot检测进一步对ACT抗PD机制进行探究.结果:ACT预防性给药后明显缩短MPTP诱导PD小鼠的爬杆时间,提高游泳测试评分,延长悬挂持续时间,并降低后肢张力测试评分和圆柱体实验评分.ACT可以有效改善MPTP诱导PD小鼠黑质纹状体组织中超氧化物歧化酶(SOD)、过氧化氢酶(CAT)和谷胱甘肽过氧化物酶(GSH-Px)的活性,并降低丙二醛(MDA)的含量.双向电泳和Western blot检测发现胆绿素还原酶B(biliverdin reductase B,BLVRB)在MPTP诱导PD小鼠的黑质纹状体中的表达升高,而ACT给药后显著降低此蛋白的表达.结论:ACT对MPTP诱导的PD小鼠具有神经保护作用,其潜在机制可能与ACT降低BLVRB的表达及抗氧化应激相关.

随着益生菌的深入研究及广泛应用,后生元的概念也在被逐渐完善.后生元是对宿主健康有益的无生命微生物和/或其成分,具有抗炎、免疫调节、抗增殖和抗氧化活性等作用.在实际研究与生产过程中,后生元可分为副益生菌与副生元,副益生菌是益生菌的灭活菌体细胞,副生元是益生菌培养的无细胞上清液.近些年来,在关于益生菌与肠道菌群的相关研究中,许多后生元成分已被广泛报道在人类疾病中发挥健康益处,包括在各种代谢性疾病、炎症性疾病、神经退行性疾病、癌症等疾病中.文章结合近年来对后生元的研究成果,综述了副生元与副益生菌各自的主要成分与研究进展,并讨论了副益生菌与副生元俩者的差异.本综述为后生元的标准制定、产品开发提供了一定的参考.

Traditional Chinese medicine (TCM) fundamentally different from Western medicine, has been widely investigated using various approaches and made great contributions to the prevention and treatment of human diseases. In recent years, research about quality control, safety evaluations, and determination of mechanisms of TCMs focused on the single-cell and molecular levels and applied to a new type of analysis method imaging technology. Cellular-or molecular-based imaging can directly get spatial distribution information and relative content of biomolecules on the tissue and mainly contribute to the progress of our understanding of various diseases, diagnosis, and drug delivery, to assess noninvasively disease-specific in cellular and molecular levels of living models in vivo. Will provide new tools to treat patients with TCM, and accelerates new drug discovery from Chinese herbal medicine. Therefore, this article reviews the imaging technology and its application in the research of TCM against cerebrovascular and neurological diseases.

Parkinson's disease (PD) is a progressive neurodegenerative disorder that occurs after Alzheimer's disease. Rotenone is a neurotoxin commonly used in creating PD models. Safflower (Carthamus tinctorius L.) contains some flavonoids that are effective against neurodegenerative diseases, and it has long been used in the treatment of cerebrovascular diseases in China. In this study, we investigated the preventive effect of safflower standardized flavonoid extract (SAFE) on a rotenone-induced PD rat model. The results showed that SAFE (17.5, 35, or 70 mg kg(-1).day(-1)) treatment modified the progressive loss in body weight, alleviated behavioral deficits, and promoted survival, especially in the middle-dose SAFE (35 mg kg(-1).day(-1)) group. SAFE treatment significantly modifies the progressive decrease in the level of DA and its metabolites, DOPAC and HVA, 5-HT and its metabolite 5-HIAA in the St, and levels of TH-positive DA-ergic neurons in the SNpc. SAFE also inhibited the decrease in TH and DA levels and increase in Ach content in the St. SAFE (35 kg(-1).day(-1)) group treatment modifying the rotenone-induced downregulation of JAK2, STAT3, and alpha 7-nAChR, and also modifying the increase in ACh in the hippocampus. SAFE preventive treatment can also partially inhibit changes in the ECS parameters associated with PD. The marker components of SAFE such as Kaempferol 3-O-rutinoside or anhydrosafflor yellow B can bind with TH, JAK2, STAT3, and alpha 7-nAChR based on molecular docking analyses. Current studies have shown that SAFE is a potential candidate for the prevention of PD.

Dl-3-n-butylphthalide (NBP) has been widely used to treat ischemic stroke in China. To investigate the mechanisms underlying NBP activity, we established a permanent middle cerebral artery occlusion (pMCAO) rat model and injected the rats with 4 mg/kg/d NBP for nine days. We then assessed neuroinflammation, neovascularization and nerve regeneration within the brain. Matrix-assisted laser desorption ionization time-of-flight mass spectrometry imaging (MALDI-TOF MSI) was used to determine the phospholipid distribution, while laser ablation-inductively coupled plasma mass spectrometry imaging (LA-ICP MSI) was used to measure Foxp3, Ki-67 and pCREB levels in the brain. Immunohistochemistry was used to investigate the expression of NLR family pyrin domain containing 3 (NLRP3) and its inflammatory products, caspase-1 and interleukin-1β, in brain tissues. NBP attenuated ischemic damage and ameliorated neurological deficits in rats with pMCAO. In the ischemic brain region, NBP reduced phosphatidylethanolamine (18:0), NLRP3, caspase-1 and interleukin-1β levels, but increased levels of Foxp3, Ki-67, pCREB and several phospholipids. In molecular docking analyses, NBP bound to NLRP3, interleukin-1β, caspase-1, Foxp3, and Ki-67. These results demonstrate that NBP reduces neuroinflammation in brain tissues and promotes nerve and blood vessel regeneration, thus protecting neuromorphology and function.

Ela tablets contain an optimized extract of a Chinese herbal prescription.The original prescription has been used to treat erectile dysfunction (ED),while the detailed indications and specific treatment mechanisms of Ela tablets remain unclear.In the present study,we aimed to investigate whether Ela tablets could protect mice from ED caused by repeated restraint stress and to explore the possible mechanisms.Mice were restrained daily in centrifuge tubes with venting holes for 14 consecutive days and administered Ela tablets at 1 h before restraining.On the 14th night,mating experiments were conducted.Thereafter,blood,testicular,and penile samples were collected.Serum testosterone levels were measured using a kit.Testicular tissue morphology was observed by H&E staining.The PDE5 expression in the corpus cavernosum was measured by Western blotting analysis.The results indicated that Ela tablets significantly shortened the sexual arousal time,increased the number of sexual encounters,maintained the normal morphology of testicular tissue,increased the serum testosterone levels,and decreased the PDE5 expression in the corpus cavernosum.Our data showed that Ela tablets elicited protective effects on the sexual ability of mice exposed to repeated restraint stress.Moreover,the underlying mechanism was related to the preservation of testicular tissue morphology,maintenance of testosterone levels,and inhibition of penile PDE5 expression.

目的 本研究以大阪大学和东京大学为例,从改革需求、人才培养规模与特点、未来发展趋势等角度对传统药学教育优势突出的日本国立学校改革进行系统梳理与深入分析,以期为我国高级临床药学人才培养改革提供参考.方法 采用焦点访谈与案例分析法并结合文献调研法.结果 与结论 日本传统药学教育以学术科学为核心,21世纪初开始学习美国药学教育模式,在全国范围内开展以药师职业为导向的6年制药学教育改革,但是公立学校仍然以培养学术型人才为主,与我国药学教育特点具有类似之处.为了顺应市场需求,同时充分发挥传统药学学术人才培养优势,以大阪大学为代表的日本国立学校创新性探索学术型与应用型相结合的复合型药学人才培养模式,其药学教育改革与发展历程对我国药学教育、特别是一流大学临床药学教育发展与改革有重要的参考意义.

Background Notoginseng leaf triterpenes (PNGL) is believed to have neuroprotective effects via the inhibition of inflammatory response and neuronal apoptosis. However, its mechanisms underlying the anti-ischemia/reperfusion (I/R) injury effects on the regulation of small molecule metabolism in rat brain remains unclear. The purpose of this study was thus to explore the mechanisms of PNGL on the regulation of small molecule metabolism in rat brain after I/R injury using matrix-assisted laser desorption ionization-mass spectrometry imaging (MALDI-MSI). Methods As a model of in vivo cerebral I/R injury, male Sprague-Dawley (SD) rats were established with a middle cerebral artery occlusion/reperfusion (MCAO/R) model after PNGL administration with 40 mg·kg-1 through intraperitoneal injection (i.p.) for 7 days. We assessed the neurological behavior, regional cerebral blood flow (r CBF), neuron injury, and spatial distribution of metabolic small molecules. Results Our in vivo results suggested that PNGL increased cerebral blood flow and relieved neurological dysfunction. Furthermore, using MALDI-MSI, we demonstrated that PNGL regulated 16 endogenous small molecules implicated in metabolic networks including tricarboxylic acid (TCA) cycle, adenosine triphosphate (ATP) metabolism, malate-aspartate shuttle, metal ions, and antioxidants underwent noticeable changes after reperfusion for 24 h. Conclusions PNGL is a novel cerebrovascular agent that can improve cerebral blood flow and attenuate adverse neurological disorders. The mechanisms are closely correlated with relative metabolic pathways, which offers insight into exploring new mechanisms in PNGL for the treatment of cerebral I/R injury.

Neuroinflammation, oxidative stress, and mitochondrial dysfunction are all important pathogenic mechanisms underlying motor dysfunction and dopaminergic neuronal damage observed in patients with Parkinson’s disease (PD). However, despite extensive efforts, targeting inflammation and oxidative stress using various approaches has not led to meaningful clinical outcomes, and mitochondrial enhancers have also failed to convincingly achieve disease-modifying effects. We tested our hypothesis that treatment approaches in PD should simultaneously reduce neuroinflammation, oxidative stress, and improve alterations in neuronal energy metabolism using the flavonoid icaritin in the 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) mouse model of PD. Using matrix-assisted laser desorption/ionization–mass spectrometry imaging (MALDI-MSI), coupled with biochemical analyses and behavioral tests, we demonstrate that icaritin improves PD by attenuating the the NOD-like receptor family pyrin domain-containing protein 3 (NLRP3) inflammasome activity and stabilizing mitochondrial function, based on our extensive analyses showing the inhibition of NLRP3 inflammasome, reduction of NLRP3-mediated IL-1β secretion, and improvements in the levels of antioxidant molecules. Our data also indicated that icaritin stabilized the levels of proteins related to mitochondrial function, such as voltage-dependent anion channel (VDAC) and ATP synthase subunit beta (ATP5B), as well as those of molecules related to energy metabolism, such as ATP and ADP, ultimately improving mitochondrial dysfunction. By employing molecular docking, we also discovered that icaritin can interact with NLRP3, VDAC, ATP5B, and several blood–brain barrier (BBB)-related proteins. These data provide insights into the promising therapeutic potential of icaritin in PD.

目的:利用db/db转基因小鼠模型探究二甲双胍联合小檗碱对糖尿病认知功能障碍的改善作用.方法:将db/db小鼠随机分为模型组(M)、二甲双胍给药组(Mt)、小檗碱给药组(Bb)和二甲双胍联合小檗碱给药组(MB),将同窝出生的db/m小鼠设为空白对照组(C).分别灌胃给药二甲双胍(300 mg·kg-1)、小檗碱(135 mg·kg-1)、二甲双胍联合小檗碱(300 mg·kg-1+ 135 mg·kg-1)10周,测定各组db/db小鼠体重、血糖及其相关指标、行为学、氧化应激和炎症因子相关指标;并用苏木素-伊红染色处理脑组织切片,光镜观察病理学改变;利用基质辅助激光解析电离质谱成像(matrix-assisted laser desorption ionization time-of-flight mass spectrometry imaging,MALDI-TOF-MSI)技术观察小鼠脑组织代谢小分子含量的变化.结果:MB组小鼠空腹血糖值明显低于M组,且低于Mt和Bb组小鼠血糖.水迷宫实验中,和M组小鼠比较,MB组小鼠逃避潜伏期和从对侧象限入水找到平台的时间明显变短;90 s内穿台次数和在目标象限中停留时间明显增多.在外周血清ELISA测定中,MB组小鼠对比M组小鼠HbA1c,8-OHdG,PCO,TNF-α,NLRP3和IL-1β水平均明显降低,IL-6水平明显升高.在HE染色实验中,和M组小鼠比较,MB组小鼠脑海马区中神经细胞排列整齐,形态较完整,核染色清晰,细胞间界限清楚.在质谱成像实验中,和M组小鼠比较,MB组小鼠脑天冬氨酸、N-乙酰-天冬氨酸、谷氨酸水平明显上升且高于Mt和Bb组;牛磺酸、葡萄糖水平明显降低且低于Mt和Bb组.结论:本实验证明了二甲双胍联合小檗碱口服给药对糖尿病认知功能障碍有改善作用,且比单独给药二甲双胍或小檗碱效果更明显,其作用机制或与抗炎、抗氧化应激、改善TCA循环以及提高兴奋性氨基酸水平等相关.

Imaging technologies for the analysis of the central nervous system are rapidly developing. Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry imaging, tracer-based magnetic resonance imaging, CLARITY technology and optogenetics can be used to visualize small molecules in brain tissues, the interstitial system of the brain and neuronal circuits in whole-brain samples. These tools serve as powerful technical means to explore the mechanisms underlying disease models and to evaluate the effects of drugs. Here, we review the constituting principles of these imaging techniques and describe their applications in the field of neuroscience.

Safflower (Carthamus tinctorius. L.), a Chinese materia medica, is widely used for the treatment of cardiovascular and cerebrovascular diseases, with flavonoids being the major active components. Multiple flavonoids in safflower bind to Parkinson's disease (PD)-related protein DJ-1. Safflower flavonoid extract (SAFE) improved behavioral indicators in a 6-hydroxydopamine (6-OHDA)-induced rat model of PD; however, the underlying mechanisms remain unclear. We used a 6-OHDA-induced mouse model of PD and a primary neuron-astrocyte coculture system to determine the neuroprotective effects and mechanisms of SAFE. After three weeks of SAFE administration, behavioral indicators of PD mice were improved. SAFE regulated the levels of tyrosine hydroxylase (TH) and dopamine metabolism. It significantly inhibited the activation of astrocytes surrounding the substantia nigra and reduced Iba-1 protein level in the striatum of PD mice. SAFE reduced the plasma content of inflammatory factors and suppressed the activation of nod-like receptor protein 3 (NLRP3) inflammasome. In the coculture system, kaempferol 3-O-rutinoside and anhydrosafflor yellow B significantly improved neuronal survival, suppressed neuronal apoptosis, and reduced IL-1β and IL-10 levels in the medium. Thus, SAFE showed a significant anti-PD effect, which is mainly associated with flavonoid anti-inflammatory activities.

目的:探究紫红獐牙菜(口山)酮提取物对1-甲基-4-苯基-1,2,3,6-四氢吡啶(1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine,MPTP)致帕金森病(Parkinson's disease,PD)小鼠的神经保护作用及其作用机制.方法:小鼠灌胃(ig)给药14d(9.0 mg·kg-1),后5d腹腔注射(ip,30 mg· kg-1)MPTP建立模型,之后进行酪氨酸羟化酶(tyrosine hydroxylase,TH)免疫组织化学染色、非标记定量蛋白质组学法检测和生物信息学分析.结果:紫红獐牙菜(口山)酮提取物可减少MPTP致帕金森小鼠脑黑质部位多巴胺能神经元的丢失;筛选出150个给药组与模型组之间的差异表达蛋白质;其潜在作用信号通路为泛素介导的蛋白水解、核糖体、硫胺素代谢通路,核心差异蛋白有Hspa13,Rplp1,Rplp2和Rps8.结论:紫红獐牙菜(口山)酮提取物对MPTP所致帕金森小鼠具有神经保护作用,其潜在机制可能包括减少α-syn错误折叠,抑制星形胶质细胞过度活化,减少活性氧簇,减轻炎症反应等.

Asthenozoospermia (AS), defined as low-motility spermatozoa in the ejaculate, is a frequent cause of human male infertility. DJ-1 (also known as PARK7), a protein highly associated with male sterility, binds to the mitochondrial complex I subunit to protect mitochondrial function. However, its involvement in spermatogenesis has not been fully elucidated. Previously, the levels of DJ-1 were shown to be significantly decreased in testicular tissues of rats with ornidazole (ORN)-induced AS. Here, we used a rat model to investigate the localization and expression levels of DJ-1 and its interacting NDUFS3 and NDUFA4 mitochondrial complex I subunits, as well as AS-induced metabolic alterations in testicular tissues. ORN significantly reduced the levels of DJ-1 in the nucleus of secondary spermatocytes, while increasing the expression of NDUFS3 in the cytoplasm of primary spermatocytes. Further, NDUFA4 showed higher expression after treatment with ORN. The principal ORN-induced changes in metabolic small molecules related to the accumulation of glucose, glutamine, and N-acetyl aspartate, enhancement of purine pathway, increase of the phosphatidic acid (PA) (18:0/18:1), phosphatidylethanolamine (PE) (16:0/18:1), and PA (18:0/20:4) lipid metabolites, and imbalance in the concentrations of Na+ and K+. However, we did not observe any abnormalities of certain small metabolic molecules and metal ions in semen samples from patients with AS. In conclusion, these results suggest that DJ-1 deficiency in testicular tissues might be closely related to the localization of NDUFS3 and content of NDUFA4, thus causing abnormalities in the mitochondrial energy metabolism and multiple other metabolic pathways.

Thymoquinone is one of the main components present in Nigella sativa seeds and is known to have various biological functions in inflammation, oxidative stress, tumors, aging, and in lowering blood glucose levels. Few studies have focused on its neuroprotective effects and its regulation of small-molecule metabolites during cerebral ischemia reperfusion injury. In this study, transient middle cerebral occlusion (tMCAO) was used to establish the rat model of cerebral ischemia reperfusion injury. We investigated the effects of thymoquinone using matrix-assisted laser desorption ionization mass spectrometry imaging (MALDI-MSI) in a model of ischemia reperfusion injury to explore the changes in small-molecule metabolites in the brain. We found that that thymoquinone significantly improved neurobehavioral scores, reduced the cerebral infarct area, alleviated brain edema, and increased the number of normal neurons following injury. MALDI-MSI revealed that thymoquinone reduced abnormal accumulations of glucose, citric acid, succinate and potassium ions. Thymoquinone also increased the amount of energy-related molecules such as ADP, AMP, GMP, and creatine, antioxidants such as glutathione, ascorbic acid, and taurine, and other metabolism-related molecules such as glutamate, glutamine, aspartate, N-acetyl-L-aspartate, and sodium ions in damaged areas of the brain following cerebral ischemia reperfusion injury. In summary, based on the neuroprotective effect of thymoquinone on cerebral ischemia reperfusion injury, this study revealed the regulation of thymoquinone on energy metabolism and small-molecule substance metabolism.

中枢神经系统疾病极大地影响着患者的生活质量.但是脑病新药的研发进展却陷入困境,在细胞实验中显示出较好活性的药物,在动物实验或临床试验中屡遭失败.近年研究表明,人们对于细胞外间隙的结构及生理条件的认识不充分是其原因之一.有学者提出脑分区稳态假说,认为脑内存在组织液引流的分区屏障,维持着脑内不同分区的稳态,这种假说为神经系统新型给药方式——细胞外间隙给药提供了理论基础.本文拟通过文献检索对脑分区稳态假说以及脑细胞外间隙给药在中枢神经系统新药研发中的应用前景进行综述.

Dear Editor,Methamphetamine（METH） is a highly addictive central nervous system stimulant that has severe physical and psychological side-effects,including loss of appetite,hyperactivity,dysphoria,and depression [1].Due to its illegal production,distribution,sale,and possession it has become a worldwide burden.METH is directly toxic to dopaminergic and serotoninergic neurons,resulting in excitotoxicity,oxidative stress,and other processes [2].Research on biomolecules associated with these processes will be useful for identifying potential markers,exploring the mechanism of METH dependence,and even developing prevention and treatment strategies.