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Evaluation of Antimycobacterial Activity of Higenamine Using Galleria mellonella as an In vivo Infection Model

Natural Products and Bioprospecting 2018年第1期8卷 63-69页

作者： Paul Erasto Richard Sunguruma Joan J.Munissi Victor Wiketye Charles de Konig Atallah F.Ahmed National Institute for Medical Research P.O.Box 9653Dar Es SalaamTanzania department of Chemistry University of Dar es SalaamDar Es SalaamTanzania School of Chemistry Witwatersrand UniversityJohannesburgRepublic of South Africa department of Pharmacognosy College of PharmacyKing Saud UniversityP.O.Box 2457Riyadh 11451Saudi Arabia

The Phytochemical investigation on MeOH extract on the bark of Aristolochia brasiliensis Mart.&Zucc(Aristolochi-aceae)led to the isolation of major compound(1)as light brown grainy *** compound was identified as 1-(4-hydroxybenzyl)-1,2,3,4-tetrahydroisoquinoline-6,7-diol(higenamine)on the basis of spectroscopic analysis,including 1D and 2D NMR *** compound was evaluated for its antimycobacterial activity against Mycobacterium indicus pranii(MIP),using Galleria mellonella larva as an in vivo infection *** survival of MIP infected larvae after a single dose treatment of 100 mg/kg body weight of higenamine was 80%after 24 *** the compound exhibited a dose dependent activity,as evidenced by the reduction of colony density from 10^(5) to 10^(3) CFU for test concentrations of 50,100,150 and 200 mg/kg body weight *** IC50 value for higenamine was 161.6 mg/kg body weight as calculated from a calibration *** analysis showed that,a complete inhibition of MIP in the *** could be achieved at 334 mg/kg body *** the fact that MIP has been found to be highly resistant against isoniazid(INH)in an in vitro assay model,in this study the microbe was highly susceptible to this standard anti-TB *** isolation of higenamine from the genus Aristolochia and the method used to evaluate its in vivo antimycobacterial activity in *** are herein reported for the first time.

关键词： Aristolochia brasiliensis Galleria mellonella Higenamine Isoniazid Mycobacterium indicus pranii Antimycobacterial activity

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Effect of Ursolic Acid on Breast Cancer Resistance Protein-mediated Transport of Rosuvastatin In vivo and Vitro

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Chinese Medical Sciences Journal 2015年第4期30卷 218-225页

作者： Jin-hua Wen Xiao-hua Wei Xiang-yuan Sheng de-qing Zhou Hong-wei Peng Yan-ni Lu Jian Zhou department of Pharmacy the First Affiliated Hospital of Nanchang University

Objective To evaluate whether ursolic acid can inhibit breast cancer resistance protein(BCRP)-mediated transport of rosuvastatin in vivo and in vitro. Methods Firstly, we explored the pharmacokinetics of 5-fluorouracil(5-FU, a substrate of BCRP) in rats in the presence or absence of ursolic acid. Secondly, we studied the pharmacokinetics of rosuvastatin in rats in the presence or absence of ursolic acid or Ko143(inhibitor of BCRP). Finially, the concentration-dependent transport of rosuvastatin and the inhibitory effects of ursolic acid and Ko143 were examined in Madin-Darby Canine Kidney(MDCK) Ⅱ-BCRP421CC(wild type) cells and MDCKⅡ-BCRP421AA(mutant type) cells. Results As a result, significant changes in pharmacokinetics parameters of 5-FU were observed in rats following pretreatment with ursolic acid. Both ursolic acid and Ko143 could significantly affect the pharmacokinetics of rosuvastatin. The rosuvastatin transport in the BCRP overexpressing system was increased in a concentration-dependent manner. However, there was no statistical difference in BCRP-mediated transport of rosuvastatin betweent the wild type cells and mutant cells. The same as Ko143, ursolic acid inhibited BCRP-mediated transport of rosuvastatin in vitro. Conclusion Ursolic acid appears to be a potent modulator of BCRP that affects the pharmacokinetic of rosuvastatin in vivo and inhibits the transport of rosuvastatin in vitro.

关键词： ursolic acid breast cancer resistance protein rosuvastatin transport

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Nanotechnology-assisted adipose-derived stem cell （ADSC） therapy for erectile dysfunction of cavernous nerve injury： In vivo cell tracking, optimized injection dosage, and functional evaluation

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Asian Journal of Andrology 2018年第5期20卷 442-447页

作者： Han Wu Wen-Hao Tang Lian-Ming Zhao de-Feng Liu Yu-Zhuo Yang Hai-Tao Zhang Zhe Zhang Kai Hong Hao-Cheng Lin Hui Jiang department of Urology Peking University Third Hospital Beijing 100191 China department of Reproductive Medicine Center Peking University Third Hospital Beijing 100191 China department of Andrology Peking University Third Hospital Beijing 100191 China department of Human Sperm Bank Peking University Third Hospital Beijing 100191 China

Stem cell therapy is a potentially promising option for erectile dysfunction; however, its risk of tumorigenicity is a clinical hurdle and the risk is positively related to the number of injected cells. Our previous study showed that nanotechnology improved adipose-derived stem cell （ADSC） therapy for erectile dysfunction of cavernous nerve injury （CNI） by attracting cells in the corpus cavernosum. These results indicated the possibility of using a reduced dosage of ADSCs for intracavernous injection. In this exploratory study, we used lower dosage （2 × 105 cells） of ADSCs for intracavernous injection （ICI） and the nanotechnology approach. Intracavernous pressure and mean arterial pressure were measured at day 28 to assess erectile function. The low-dose ADSC therapy group showed favorable treatment effects, and nanotechnology further improved these effects. In vivo imaging of ICI cells revealed that the fluorescein signals of NanoShuttle-bound ADSCs （NanoADSCs） were much stronger than those of ADSCs at days 0, 1, and 3. Both immunofluorescence and Western blot analysis showed a significant increase in smooth muscle, endothelium, and nerve tissue in the ADSC group compared to that in the CNI group; further improvement was achieved with assisted nanotechnology. These findings demonstrate that nanotechnology can be used to further improve the effect of small dosage of ADSCs to improve erectile function. Abundant NanoADSCs remain in the corpus cavernosum in vivo for at least 3 days. The mechanism of erectile function improvement may be related to the regeneration of the smooth muscle, endothelium, and nerve tissues.

关键词： adipose-derived stem cell (ADSC) cavernous nerve injury cell tracking erectile dysfunction

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Phytoestrogen genistein decreases contractile response of aortic artery in vitro and arterial blood pressure in vivo

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Acta Pharmacologica Sinica 2004年第3期25卷 59-64页

作者： Hong-fang LI1,Long-de WANG2,Song-yi QU department of Physiology,Lanzhou Medical College,Lanzhou 730000 2Affiliated Hospital,Gansu Chinese Medical College,Lanzhou 730020,China department of Physiology Lanzhou Medical College Lanzhou 730000 Affiliated Hospital Gansu Chinese Medical College Lanzhou 730020

AIM: To determine the mechanisms of effects of phytoestrogen genistein on the contracted rabbit aortic arteries in vitro, and observe the effect of genistein and 17-β estradiol on mean arterial pressure (MAP) in ovariectomized (OVX) rats. METHODS: (1) Strips of rabbit aortic smooth muscle were suspended in organ baths containing Kreb’s solution, and then isometric tension was measured. (2) Female mature Wistar rats underwent a bilateral ovariectomy (OVX). Sham-operated rats (SHAM) were used as controls. After administration of genistein (0.4 mg·kg-1·d-1, sc), 17-β estradiol (1 mg·kg-1·d-1, sc) or theirvehicle sesame oil for 21 d,MAP was measured. RESULTS: (1) Similar to 17-β estradiol, genistein could dose-dependently relax 40 mmol/L KCl-precontracted arterial strips. Incubation with Nω-L-nitro-arginine (L-NNA), methylene blue (MB), indomethacin, propranolol or endothelium removal did not affect relaxation induced by genistein. In calcium-free solution containing 0.01mmol/L egtazic acid (EGTA), genistein inhibited not only the first phase contraction induced by noradrenaline (NA), but also the second contraction induced by CaCl2. In addition, genistein could reduce the contractile responses of NA, KCl and CaCl2, and shift their cumulative concentration-response curves rightward. (2) MAP in OVX rats was significantlyhigher compared with that of SHAM rats. However,after chronically treatment with genistein or 17-β estradiol for 21 d the baseline MAP in OVX rats was reduced significantly. CONCLUSIONS: (1) The vasodilator effect of genistein in vitro is endothelium independent and not related to the nitric oxide, its mechanisms being probably due to inhibition of Ca2+ influx through calcium channels in a noncompetitive manner and Ca2+ release from intracellular store induced by NA. (2) Administration of genistein or 17-β estradiol can chronically decrease MAP in OVX rats.

关键词： genistein aorta smooth muscle muscle relaxation calciumchannel vascular endothelium blood pressure

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Preliminary studies of constructing a tissue-engineered lamellar corneal graft by culturing mesenchymal stem cells onto decellularized corneal matrix

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International Journal of Ophthalmology(English edition) 2021年第1期14卷 10-18页

作者： Yu-Jie Cen de-Bo You Wei Wang Yun Feng department of Ophthalmology Peking University Third HospitalBeijing 100191China Beijing Key Laboratory of Restoration of Damaged Ocular Nerve Peking University Third HospitalBeijing 100191China

AIM:To construct a competent corneal lamellar substitute in order to alleviate the shortage of human corneal ***:Rabbit mesenchymal stem cells(MSCs)were isolated from bone marrow and identified by flow cytometric,osteogenic and adipogenic *** decellularized corneal matrix(XDCM)was generated from dog *** were seeded and cultured on XDCM to construct the tissueengineered ***-transplantation biocompatibility of engineered corneal graft were tested by animal *** were divided into two groups then underwent lamellar keratoplasty(LK)with different corneal grafts:1)XDCM group(n=5):XDCM;2)XDCM-MSCs groups(n=4):tissue-engineered cornea made up with XDCM and *** ocular surface recovery procedure was observed while corneal transparency,neovascularization and epithelium defection were measured and *** vivo on focal exam was performed 3 mo ***:Rabbit MSCs were isolated and *** cytometry demonstrated isolated cells were CD90 positive and CD34,CD45 *** and adipogenic induction verified their multipotent ***-XDCM grafts were constructed and *** vivo transplantation showed the neovascularization in XDCMMSC group was much less than that in XDCM group ***-transplant 3-month confocal test showed less nerve regeneration and bigger cell-absent area in XDCM-MSC ***:This study present a novel corneal tissue-engineered graft that could reduce post-operatively neovascularization and remain transparency,meanwhile shows that co-transplantation of MSCs may help increase corneal transplantation successful rate and enlarge the source range of corneal substitute to overcome cornea donor shortage.

关键词： mesenchymal stem cells xenogenic decellularized corneal matrix acellular corneal matrix neovascularization tissue-engineered cornea

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The DNA-binding factor Ctcf critically controls gene expression in macrophages

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Cellular & Molecular Immunology 2014年第1期11卷 58-70页

作者： Tatjana Nikolic Dowty Movita Margaretha EH Lambers Claudia Ribeiro de Almeida] Paula Biesta Kim Kreefft Marjolein JW de Bruijn Ingrid Bergen Niels Galjart Andre Boonstra Rudi Hendriks department of Pulmonary Medicine Erasmus MC Rotterdam The Netherlands department of Gastroenterology and Hepatology Erasmus MCRotterdam The Netherlands department of Cell Biology and Genetics Erasmus MC Rotterdam The Netherlands Current address: Dr T Nikolic Department of Immunohematology and Blood Transfusion Leiden University Medical Center Leiden The Netherlands DrCR de Almeida Sir William Dunn School of Pathology University of Oxford Oxford UK. Both authors contributed equally

Macrophages play an important role in immunity and homeostasis. Upon pathogen recognition via specific receptors, they rapidly induce inflammatory responses. This process is tightly controlled at the transcriptional level. The DNA binding zinc-finger protein CCCTC-binding factor （Ctcf） is a crucial regulator of long-range chromatin interactions and coordinates specific communication between transcription factors and gene expression processes. In this study, the Ctcf gene was specifically deleted in myeloid cells by making use of the transgenic Cre-LoxP system. Conditional deletion of the Ctcfgene in myeloid cells induced a mild phenotype in vivo. Ctcf-deficient mice exhibited significantly reduced expression of major histocompatibility complex （MHC） class II in the liver. Ctcf-deficient macrophages demonstrated a normal surface phenotype and phagocytosis capacity. Upon Toll-like receptor （TLR） stimulation, they produced normal levels of the pro-inflammatory cytokines IL-12 and IL-6, but manifested a strongly impaired capacity to produce tumor-necrosis factor （TNF） and IL-IO, as well as to express the IL-IO family members IL-19, IL-20 and IL-24. Taken together, our data demonstrate a role of Ctcf that involves fine-tuning of macrophage function.

关键词： Ctcf IL-IO locus macrophage myeloid TLR

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Potential of Lunasin Orally-Administered in Comparison to Intraperitoneal Injection to Inhibit Colon Cancer Metastasis <i>in vivo</i>

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Journal of Cancer Therapy 2013年第6期4卷 34-43页

作者： Vermont P. Dia Elvira Gonzalez de Mejia department of Food Science and Human Nutrition University of Illinois at Urbana-Champaign Urbana USA

Lunasin is a bioactive peptide originally isolated from soybean and has demonstrated chemopreventive and anti-cancer properties against skin, colon and breast cancers. The objective of the study was to evaluate the capability of intraperitoneally and orally-administered soybean-derived peptide lunasin to inhibit KM12L4 human colon cancer cell metastasis in a mouse model. Intraperitoneal (i.p.) injection of lunasin (4 mg/kg bw/day) reduced the number of liver metastasis by 50% (P = 0.047) and the liver weight/body weight ratio by 23% (P = 0.039). Oral administration of lunasin reduced the number of liver metastasis by 56% (8 mg/kg bw/day, P = 0.293) and 94% (20 mg/kg bw/day, P = 0.247). Immunohistochemical staining of the liver-tissue section showed that lunasin at 20 mg/kg bw dose did not significantly reduce the expression of proliferating cell nuclear antigen, increased the expression of proapoptotic Bax by 2.7-fold but also increased the expression of the antiapoptotic Bcl-2 by 3.8-fold. Regarding epigenetic markers, H3K18 was not significantly affected by either oral dose while H4K8 was dose dependently increased by 2.3-fold (P = 0.001, 8 mg/kg bw) and 2.7-fold (P s of administration used leading to digestion of lunasin when given by oral gavage. In conclusion, lunasin reduced colon cancer metastasis in vivo;however, more studies are needed to determine the oral dose of lunasin and prevent colon cancer metastasis.

关键词： Lunasin Colon Cancer Metastasis Apoptosis Histones

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Gaussia Luciferase Reporter Assay for Assessment of Gene delivery Systems in vivo

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Chinese Medical Sciences Journal 2010年第2期25卷 95-99页

作者： Feng Chen Zhen Xu Jing Lu Xiang Lu Wen-li Mu Ya-jun Wang de-pei Liu Chih-chuan Liang National Laboratory of Medical Molecular Biology Institute of Basic Medica Sciences Chinese Academy of Medical Sciences ＆ Peking Union Medical College Beijing 100005 China

Objective To develop an alternative method for assessment of gene delivery systems in *** Mouse primary spleen lymphocytes were genetically modified in vitro by a retroviral vector harboring a Gaussia luciferase(Gluc) expression *** implantation of these cells into recipient mice,the expression of Gluc was detected in whole blood or plasma *** As little as 10 μL whole blood drawn from the recipient mice could guarantee prompt reading of Gluc activity with a *** the reading was found in good correlation with the number of genetically modified spleen lymphocytes implanted to the *** Gluc may be useful as an in vivo reporter for gene therapy researches,and Gluc blood assay could provide an alternative method for assessment of gene delivery systems in vivo.

关键词： gene delivery systems reporter Gaussia luciferase

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miRNA-4465靶向Wnt5A调控对胶质瘤细胞增殖、迁移和侵袭的影响

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中国药业 2024年第18期33卷 26-33页

作者：陈硕硕孙玉玺余德束汉生蚌埠医学院第二附属医院安徽蚌埠233000

目的探讨miRNA-4465(miR-4465)对胶质瘤细胞增殖、侵袭及迁移的影响及其作用机制。方法采用miR-4465模拟物(mimic)处理U118和U87细胞,检测细胞生物活性。双荧光素酶报告基因实验评价miR-4465和Wnt5A的结合情况。采用siRNA和miR-4465... 详细信息

目的探讨miRNA-4465(miR-4465)对胶质瘤细胞增殖、侵袭及迁移的影响及其作用机制。方法采用miR-4465模拟物(mimic)处理U118和U87细胞,检测细胞生物活性。双荧光素酶报告基因实验评价miR-4465和Wnt5A的结合情况。采用siRNA和miR-4465抑制剂(inhibitor)分别抑制Wnt5A和miR-4465的表达,采用CCK-8法、细胞克隆实验、伤口愈合实验和Transwell实验,检测细胞的增殖、迁移、侵袭能力。复制移植瘤裸鼠模型,评价miR-4465过表达和抑制对其肿瘤生长的影响。结果 miR-4465 mimic显著抑制了U118和U87细胞增殖能力。双荧光素酶报告基因实验结果显示miR-4465与Wnt5A存在靶向调节作用。沉默Wnt5A可降低细胞的增殖、迁移、侵袭能力,miR-4465被抑制后,细胞的增殖、迁移、侵袭能力均显著增强(P <0.05)。此外,沉默Wnt5A可逆转miR-4465抑制导致的细胞增殖、迁移、侵袭能力增强。体内实验结果显示,miR-4465过表达抑制肿瘤生长和ki67的表达,而miR-4465被抑制后,肿瘤生长和ki67的表达显著增强(P <0.05)。结论 miR-4465在胶质瘤中呈低表达,其过表达可抑制胶质瘤细胞的增殖、迁移、侵袭能力,其下游作用机制与Wnt5A通路有关。

关键词：胶质瘤 miR-4465 双荧光素酶报告基因细胞增殖迁移侵袭 Wnt5A

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Probiotic yeasts: Anti-inflammatory potential of various non-pathogenic strains in experimental colitis in mice

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World Journal of Gastroenterology 2010年第17期16卷 2134-2145页

作者： Benot Foligné Jo■lle dewulf Pascal Vandekerckove Georges Pignède Bruno Pot Institut Pasteur of Lille Lactic acid Bacteria & Mucosal ImmunityCenter for Infection and Immunity of Lille1 rue du Pr CalmetteBP 245F-59019 LilleFrance University of Lille Nord de France F-59000 LilleFrance Centre National de la Recherche Scientifique UMR 8204F-59021 LilleFrance Institut National de la Santé et de la Recherche Médicale U1019F-59019 LilleFrance Lesaffre International Direction of Research and Developpement147 rue Gabriel PériBP 602759700 Marcq-en-BaroeulFrance

AIM: To evaluate the in vitro immunomodulation capacity of various non-pathogenic yeast strains and to investigate the ability of some of these food grade yeasts to prevent experimental colitis in ***: In vitro immunomodulation was assessed by measuring cytokines [interleukin (IL)-12p70,IL-10,tumor necrosis factor and interferon γ] released by human peripheral blood mononuclear cells after 24 h stimulation with 6 live yeast strains (Saccharomyces ssp.) and with bacterial reference strains.A murine model of acute 2-4-6-trinitrobenzene sulfonic acid (TNBS)-colitis was next used to evaluate the distinct prophylactic protective capacities of three yeast strains compared with the performance of prednisolone ***: The six yeast strains all showed similar non-discriminating anti-inflammatory potential when tested on immunocompetent cells in vitro .However,although they exhibited similar colonization patterns in vivo ,some yeast strains showed significant anti-inflammatory activities in the TNBS-induced colitis model,whereas others had weaker or no preventive effect at all,as evidenced by colitis markers (body-weight loss,macroscopic and histological scores,myeloperoxidase activities and blood inflammatory markers).CONCLUSION: A careful selection of strains is required among the biodiversity of yeasts for specific clinical studies,including applications in inflammatory bowel disease and other therapeutic uses.

关键词： Yeast Probiotics Strain specificity Experimental colitis 2-4-6-trinitrobenzene sulfonic acid

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