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Upregulation of CDGSH iron sulfur domain 2 attenuates cerebral ischemia/reperfusion injury

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Neural Regeneration Research 2023年第7期18卷 1512-1520页

作者： Miao Hu Jie Huang Lei Chen Xiao-Rong Sun Zi-Meng Yao Xu-Hui Tong Wen-Jing Jin Yu-Xin Zhang Shu-Ying Dong department of Pharmacology School of PharmacyBengbu Medical CollegeBengbuAnhui ProvinceChina Key Laboratory of Cardiovascular and Cerebrovascular Diseases Bengbu Medical CollegeBengbuAnhui ProvinceChina Anhui Engineering technology Research Center of biochemical pharmaceutical BengbuAnhui ProvinceChina

CDGSH iron sulfur domain 2 can inhibit ferroptosis,which has been associated with cerebral ischemia/reperfusion,in individuals with head and neck cancer.Therefore,CDGSH iron sulfur domain 2 may be implicated in cerebral ischemia/reperfusion injury.To validate this hypothesis in the present study,we established mouse models of occlusion of the middle cerebral artery and HT22 cell models of oxygen-glucose deprivation and reoxygenation to mimic cerebral ischemia/reperfusion injury in vivo and in vitro,respectively.We found remarkably decreased CDGSH iron sulfur domain 2 expression in the mouse brain tissue and HT22 cells.When we used adeno-associated virus and plasmid to up-regulate CDGSH iron sulfur domain 2 expression in the brain tissue and HT22 cell models separately,mouse neurological dysfunction was greatly improved;the cerebral infarct volume was reduced;the survival rate of HT22 cells was increased;HT22 cell injury was alleviated;the expression of ferroptosis-related glutathione peroxidase 4,cystine-glutamate antiporter,and glutathione was increased;the levels of malondialdehyde,iron ions,and the expression of transferrin receptor 1 were decreased;and the expression of nuclear-factor E2-related factor 2/heme oxygenase 1 was increased.Inhibition of CDGSH iron sulfur domain 2 upregulation via the nuclear-factor E2-related factor 2 inhibitor ML385 in oxygen-glucose deprived and reoxygenated HT22 cells blocked the neuroprotective effects of CDGSH iron sulfur domain 2 up-regulation and the activation of the nuclear-factor E2-related factor 2/heme oxygenase 1 pathway.Our data indicate that the up-regulation of CDGSH iron sulfur domain 2 can attenuate cerebral ischemia/reperfusion injury,thus providing theoretical support from the perspectives of cytology and experimental zoology for the use of this protein as a therapeutic target in patients with cerebral ischemia/reperfusion injury.

关键词： cerebral ischemia/reperfusion injury CDGSH iron sulfur domain 2 ferroptosis glutathione peroxidase 4 heme oxygenase 1 HT22 nuclear-factor E2-related factor 2 oxygen-glucose deprivation/reoxygenation injury stroke transferrin receptor 1

Demethylenetetrahydroberberine protects dopaminergic neurons in a mouse model of Parkinson's disease

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Chinese Journal of Natural Medicines 2022年第2期20卷 111-119页

作者： WEN Jing ZHANG Yuan-Qiang LIU Dong-Qing YAO Xu-Tao JIANG Hua ZHANG Yu-Bin department of Biochemistry School of Life Science and TechnologyChina Pharmaceutical UniversityNanjing 211198China

Parkinson’s disease(PD)is a multifactorial disorder of the nervous system where a progressive loss of dopaminergic neurons exist.However,the pathogenesis of PD remains undefined,which becomes the main limitation for the development of clinical PD treatment.Demethylenetetrahydroberberine(DMTHB)is a novel derivative of natural product berberine.This study was aimed to explore the neuroprotective effects and pharmacological mechanism of DMTHB on Parkinson’s disease using C57BL/6 mice.A PD model of mice was induced by administration of MPTP(20 mg·kg^(-1))and probenecid(200 mg·kg^(-1))twice per week for five weeks.The mice were administered with DMTHB daily by gavage at the dose of 5 and 50 mg·kg^(-1)for one-week prophylactic treatment and five-week theraputic treatment.The therapeutic effects of DMTHB were evaluated by behavior tests(the open field,rotarod and pole tests),immunohistochemical staining of tyrosine hydroxylase(TH),Nissl staining and biochemical assays.The molecular mechanisms of DMTHB on the key biomarkers of PD pathological states were analyzed by Western blot(WB)and qRT-PCR.DMTHB treatment alleviated the behavioral disorder induced by MPTP-probenecid.Nissl staining and TH staining showed that the damage of dopaminergic neurons in the substantia nigra was remarkably suppressed by DMTHB treatment.Western blot results showed that the ratio of Bcl-2/Bax and TH increased,but the level ofα-synuclein(α-syn)was remarkably reduced,which indicated that the apoptosis of dopaminergic neurons in mice was significantly reduced.The protein phosphorylation of p-PI3K,p-AKT and p-mTOR also increased about 2-fold,compared with the model group.Furthermore,qRT-PCR results demonstrated that the mRNA levels of pro-inflammatory cytokines,IL-1βand TNF-α,were reduced,but the level of anti-inflammatory cytokine IL-10 increased after DMTHB treatment.Finally,the cellular assay displayed that DMTHB was also a strong antioxidant to protect neuron cell line PC12 by scavenging ROS.In this study,we demonstrated DMTHB alleviates the be

关键词： Demethylenetetrahydroberberine Parkinson's disease Dopaminergic neurons Apoptosis Neuroinflammation

药用植物白芍愈伤组织中产芍药双苷的研究(英文)

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Chinese Journal of Chemical Engineering 2015年第2期23卷 451-455页

作者：胡升马莹莹江海龙冯定军俞伟代冬梅梅乐和 College of Biological and Chemical Engineering Ningbo Institute of Technology Zhejiang University department of Chemical and biochemical Engineering Zhejiang University Ningbo Liwah pharmaceutical Co. Ltd.

In order to facilitate the preparation of paeoniflorin(PF)and albiflorin(AF),two chief bioactive constituents in Paeonia lactiflora Pall(PL),induction and culture of callus from PL were studied.With a modified woody plant medium supplemented with 0.5 mg·L-16-benzylaminopurine,1.0 mg·L-1naphthylacetic acid,0.1 mg·L-1thidiazuron and 30 g·L-1sucrose,callus was induced from four kinds of explants:leaf,stems,petiole,and root.The potency to form callus varies between different explants and leaf explants exhibits the highest capacity(100%).On the other hand,root-derived callus(R-callus)produces the highest level of total amount of PF and AF,31.8 mg·g-1dry mass,which is higher than the corresponding level in the root of field cultivated PL.Furthermore,the time needed is only 40 days,remarkably shorter than the cultivation time of PL,about 4–5 years.Higher accumulation levels of PF and AF with shorter production time indicate that callus culture of PL is a promising powerful tool for production of PF and AF in the future.

关键词：愈伤组织培养芍药苷 modified level 内酯生产 plant total

Ca^(2+)-亚氨二醋酸金属离子亲和色谱法吸附内毒素(英文)

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色谱 2012年第11期30卷 1194-1202页

作者： André Moreni LOPES Jorge Sánchez ROMEU Rolando Páez MEIRELES Gabriel Marquez PERERA Rolando Perdomo MORALES Adalberto PESSOA Lourdes Zumalacárregui CáRDENAS Department of Biochemical and Pharmaceutical Technology School of Pharmaceutical SciencesUniversity of So Paulo(FCF/USP) Purification Development department Center for Genetic Engineering and Biotechnology(CIGB) Center for pharmaceuticals Research and Development(CIDEM) Higher Polytechnic Institute José Antonio Echeverría(CUJAE)

Endotoxins(also known as lipopolysaccharides(LPS)) are undesirable by-products of recombinant proteins,purified from Escherichia coli.LPS can be considered stable under a wide range of temperature and pH,making their removal one of the most difficult tasks in downstream processes during protein purification.The inherent toxicity of LPS makes their removal an important step for the application of these proteins in several biological assays and for a safe parenteral administration.Immobilized metal affinity chromatography(IMAC) enables the affinity interactions between the metal ions(immobilized on the support through the chelating compound) and the target molecules,thus enabling high-efficiency separation of the target molecules from other components present in a mixture.Affinity chromatography is applied with Ca2+-iminodiacetic acid(IDA) to remove most of the LPS contaminants from the end product(more than90%).In this study,the adsorption of LPS on an IDA-Ca2+ was investigated.The adsorption Freundlich isotherm of LPS-IDA-Ca2+ provides a theoretical basis for LPS removal.It was found that LPS is bound mainly by interactions between the phosphate group in LPS and Ca2+ ligands on the beads.The factors such as pH(4.0 or 5.5) and ionic strength(1.0 mol/L) are essential to obtain effective removal of LPS for contaminant levels between endotoxin' concentration values less than100 EU/mL and 100 000 EU/mL.This new protocol represents a substantial advantage in time,effort,and production costs.

关键词： immobilized metal affinity chromatography(IMAC) lipopolysaccharides(LPS) endotoxin removal recombinant proteins isotherms protein purification

Phase Behavior and Structural Transitions in Sodium Dodecyl Sulfonate Microemulsions

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Chinese Journal of Chemical Engineering 2002年第6期10卷 670-672页

作者：杨根生施介华李景华王普姚善泾 College of pharmaceutical Sciences Zhejiang University of Technology department of Chemical and biochemical Engineering Zhejiang University

The forming mechanism of microemulsion of sodium dodecyl sulfonate, alcohols, water and isooctane was studied, with particular emphasis on the effect of molecular weight and concentration of alcohols. Phase diagram of the four components, alcohol, sodium dodecyl sulfonate, water and isooctane, was used as a means of study, through which the microemulsion regions were deter-mined. Phase diagram of sodium dodecyl sulfonate/n-pentanol/isooctane/water system at km = 2 (km = Wn-pentanol / WSDS ) is presented. The variation of conductivities of different microemulsion samples with water was measured. From the conductivities we investigated a change in structure from water droplets in oil (W/O) at low water content to liquid crystal at intermediate water content and a struc-ture of oil droplets in water (O/W) at high water content.

关键词：正构醇十二烷基磺酸钠异辛烷水相行为微乳状液结构转变表面活性剂

Development of a surface plasmon resonance biosensor for accurate and sensitive quantitation of small molecules in blood samples

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Journal of pharmaceutical Analysis 2022年第6期12卷 929-936页

作者： Minyu Qi Diya Lv Ying Zhang Dongyao Wang Xiaofei Chen Zhenyu Zhu Zhanying Hong Yifeng Chai Hai Zhang Yan Cao department of biochemical Pharmacy Faculty of PharmacyNaval Medical UniversityShanghai200433China pharmaceutical Analysis and Testing Center Faculty of PharmacyNaval Medical UniversityShanghai200433China department of pharmaceutical Analysis Faculty of PharmacyNaval Medical UniversityShanghai200433China department of Pharmacy Shanghai First Maternity and Infant HospitalTongji University School of MedicineShanghai201204China

Therapeutic drug monitoring(TDM)has played an important role in clinical medicine for precise dosing.Currently,chromatographic technology and immunoassay detection are widely used in TDM and have met most of the needs of clinical drug therapy.However,some problems still exist in practical applications,such as complicated operation and the influence of endogenous substances.Surface plasmon resonance(SPR)has been applied to detect the concentrations of small molecules,including pesticide residues in crops and antibiotics in milk,which indicates its potential for in vivo drug detection.In this study,a new SPR-based biosensor for detecting chloramphenicol(CAP)in blood samples was developed and validated using methodological verification,including precision,accuracy,matrix effect,and extraction recovery rate,and compared with the classic ultra-performance liquid chromatographyultraviolet(UPLC-UV)method.The detection range of SPR was 0.1-50 ng/mL and the limit of detection was 0.099±0.023 ng/mL,which was lower than that of UPLC-UV.The intra-day and inter-day accuracies of SPR were 98%-114% and 110%-122%,which met the analysis requirement.The results show that the SPR biosensor is identical to UPLC-UV in the detection of CAP in rat blood samples;moreover,the SPR biosensor has better sensitivity.Therefore,the present study shows that SPR technology can be used for the detection of small molecules in the blood samples and has the potential to become a method for therapeutic drug monitoring.

关键词： Surface plasmon resonance UPLC-UV Chloramphenicol Methodological verification

Dynamic crosslinked polymeric nano-prodrugs for highly selective synergistic chemotherapy

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Asian Journal of pharmaceutical Sciences 2022年第6期17卷 880-891页

作者： Shi Wang Yining Song Jingge Ma Xinyang Chen Yuanhui Guan Hui Peng Guoqing Yan Rupei Tang Engineering Research Center for Biomedical Materials Anhui Key Laboratory of Modern BiomanufacturingSchool of Life SciencesAnhui UniversityHefei 230601China Anhui Engineering technology Research Center of biochemical pharmaceutical Bengbu Medical CollegeBengbu 233030China

To achieve highly selective synergistic chemotherapy attractive for clinical translation,the precise polymeric nano-prodrugs(PPD-NPs)were successfully constructed via the facile crosslinking reaction between p H-sensitive poly(ortho ester)s and reduction-sensitive small molecule synergistic prodrug(Pt(IV)-1).PPD-NPs endowed the defined structure and high drug loading of cisplatin and demethylcantharidin(DMC).Moreover,PPD-NPs exhibited steady long-term storage and circulation via the crosslinked structure,suitable negative potentials and low critical micelle concentration(CMC),improved selective tumour accumulation and cellular internalization via dynamic size transition and surficial amino protonation at tumoural extracellular p H,promoted efficient disintegration and drug release at tumoural intracellular p H/glutathione,and enhanced cytotoxicity via the synergistic effect between cisplatin and DMC with the feed ratio of 1:2,achieving significant tumour suppression while decreasing the side effects.Thus,the dynamic crosslinked polymeric nano-prodrugs exhibit tremendous potential for clinically targeted synergistic cancer therapy.

关键词： Nano-prodrugs Precise structure Size transition Amino protonation Synergistic chemotherapy

Screening and characterization of extracelluar L-asparaginase producing Bacillus subtilis strain hswx88,isolated from Taptapani hotspring of odisha,India

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Asian Pacific Journal of Tropical Biomedicine 2013年第12期3卷 936-941页

作者： Biswaprakash Pradhan Sashi K Dash Sabuj Sahoo pharmaceutical Biotechnology Division University Department of Pharmaceutical SciencesUtkal University department of Seed technology Orissa University of Agriculture and Technology

Objective:To screen and isolate an eco-friendly,u thermophilic and potent L-asparaginase producing bacterium,with novel immunological properties that may obviates hypersensitivity reactions.Methods:In the present study baclerial strain isolated for extracellular L-asparaginase production from hotspring,identified by morphological,biochemical and physiological tests followed by t6S rDNA technology and the L-asparaginase production ability was tested by both semi quantitative and quantitative enzymatic assay.Result:The bacterial strain was identified as Bacillus sublilis strain hswx88(GenBank Accession Number JQ237656.1).The extracellular enzyme yielding capacity isolate Bacillus subtilis strain hswx88(23.8 IU/mL)was found to be 1.7 and 14.5 limes higher than the reference organism Pectobacterium carotovorum MTCC 1428(14.2 IU/mL)and Bacillus sp.BCCS 034(1.64 IU/mL).Conclusion:The isolate is eco-friendly and useful to produce bulk quantity of extracellular,thermophilic L-asparaginase for the treatment of various tumor cases and for preparation of acrylamide free fry food preparation.

关键词： L-asparaginase Thermophilic bacteria Bacillus subtilis strain hswx88 16S rDNA

GC-MS analysis of Cocus nucifera flower extract and its effects on heterogeneous symptoms of polycystic ovarian disease in female Wistar rats

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Chinese Journal of Natural Medicines 2014年第9期12卷 677-684页

作者： V.Soumya Y.Indira Muzib P.Venkatesh K.Hariprasath department of pharmaceutical technology Sri Padmavathi Mahila Viswavidyalayam Tirupathi- 517502 Andhra Pradesh India department of pharmaceutical Chemistry Sir C.R.Reddy College of Pharmaceutical Sciences

AIM: To evaluate the effect of Cocus nucifera L. flowers in reducing the major multiple symptoms of letrozole-induced polycystic ovarian disease(PCOD) in female rats. METHOD: Female, virgin Wistar rats were treated with letrozole(1 mg/kg body wt) to induce PCOD, and after 21 days of induction rats were administered orally with 100 and 200 mg·kg-1 of Cocus nucifera flower aqueous extract, respectively. Estrus cycle and blood sugar were monitored once a week throughout the study. After scarification, various biochemical parameters, such as antioxidant status(superoxide dismutase(SOD) and glutathione reductase(GSH)) of the uterus homogenate, lipid profile(total cholesterol(TC), high density lipoprotein(HDL), low density lipoprotein(LDL), and triglycerides(TG)) of the serum were determined. Weights of the uterus and ovaries were separately monitored. The characteristics of changes in the ovary were evaluated by histopathological studies. RESULTS: GC-MS analysis of the aqueous extract showed the presence of volatile and pharmacologically active phytoconstituents. C. nucifera flower extract-treated groups showed estrus cyclicity and increased uterus weight which indicates the estrogenic effect. The improved blood sugar level, ideal lipid profile, good antioxidant status, and histopathology results revealed the recovery from poly cystic ovaries. CONCLUSION: The results indicate that C. nucifera flower is a potential medicine for the treatment of PCOD and this study supports the traditional uses of C. nucifera flower.

关键词： PCOD Cocus nucifera Poly cystic ovarian disease Estrus cyclicity Lipid profile Antioxidant Histopathology of ovary