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Indian medicinal plants are effective in the treatment and management of COVID-19

BIOCELL 2023年第4期47卷 677-695页

作者： SUBHASH CHANDRA SANTWANA PALAI EDINARDO FAGNER FERREIRA-MATIAS IVO CAVALCANTE PITA-NETO CÍCERO LUCAS GOMES-RAMALHO EDLANE MARTINS DE ANDRADE RAY SILVA DE ALMEIDA MARCELLO IRITI henrique douglas melo-coutinho Department of Pharmaceutical Chemistry SGRR UniversityUttarakhand248001India Department of Pharmaceutical Chemistry HNB Garhwal UniversitySrinagar246174India Department of Veterinary Pharmacology&Toxicology College of Veterinary Science and Animal HusbandryBhubaneswar751003India CECAPE—College of Dentistry Juazeiro do Norte63050-203Brazil Department of Biological Chemistry Regional University of CaririCrato63105-000Brazil Department of Biomedical Surgical and Dental SciencesUniversitàdegli Studi di MilanoMilan20133Italy National Interuniversity Consortium of Materials Science and Technology Florence50121Italy

Indian medicinal plants are referred to as the“nectar of life”owing to their phytochemicals and bioactive complexes that are beneficial in treating diseases.Coronavirus disease 2019(COVID-19)is a global health issue without any proper medication.The indigenous plants of India can be exploited to control the precise signs of SARS-CoV-2.The Ministry of AYUSH(Ayurveda,Yoga and Naturopathy,Unani,Siddha,and Homeopathy)has advised routine usage of medicinal plants for COVID-19.Medicinal plants like Zingiber officinalis,Azadirachta indica,Ocimum sanctum,Nigella sativa,Withania somnifera,Curcuma longa,Piper nigrum,Allium sativum,Tinospora cordifolia,etc.have immunity-boosting,antiviral,antibacterial,antioxidant and anti-inflammatory actions that can suppress and treat symptoms of COVID-19.In vitro,in vivo as well as in silico validation,these phytochemicals can help us to manage and treat COVID-19 disease.This integration of traditional knowledge in the prophylaxis of corona infection and current skills validating it for the development of precise and powerful therapeutic approaches will more efficiently resolve different clinical aspects of COVID-19.The review focuses on both traditional and emergent methods to prevent and treat COVID-19 with various Indian medicinal plants along with their phytochemicals.

关键词： Antiviral Corona virus Diagnosis Immunity Herbs

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Molecular evolution,virology and spatial distribution of HCV genotypes in Pakistan:A meta-analysis

Infectious Medicine

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Infectious Medicine 2023年第4期2卷 324-333页

作者： Arslan Habib Nadiya Habib Khalid Mahmood Anjum Riffat Iqbal Zeeshan Ashraf Muhammad Usman Taj Muhammad Asim Kanwal Javid Faezeh Idoon Saeid Dashti Cassio Rocha Medeiros Ana Pavla Almeida Diniz Gurgel henrique douglas melo coutinho Laboratory of Molecular Immunology School of Life SciencesFudan UniversityShanghai 200433China Department of Zoology University of GujratGujratPunjab 50700Pakistan Department of Zoology University of Veterinary and Animal SciencesLahore 54000Pakistan Department of Zoology Government College UniversityLahore 54000Pakistan Department of Fisheries and Aquaculture University of Veterinary and Animal SciencesLahore 54000Pakistan Department of Zoology University of NarowalNarowal 51600Pakistan Department of Geography Government College UniversityLahore 54000Pakistan Department of Surgical Technology Ferdows School of Allied Medicine and Public HealthBirjand University of Medical SciencesBirjand 97178Iran Ferdows School of Paramedical and Health Birjand University of Medical SciencesBirjand 97178Iran CECAPE College Juazeiro do Norte 63024-015Brazil Department of Engineering and Environment Federal University of ParaibaJoão Pessoa 58051-900Brazil Laboratory of Microbiology and Molecular Biology(LMBM) Department of Biological ChemistryRegional University of CaririCrato 63105-000Brazil

Background:Hepatitis C,caused by the Hepatitis C Virus(HCV),is the second most common form of viral hepatitis.The geographical distribution of HCV genotypes can be quite complex,making it challenging to ascertain the most prevalent genotype in a specific area.Methods:To address this,a review was conducted to determine the prevalence of HCV genotypes across various provinces and as a whole in Pakistan.The scientific literature regarding the prevalence,distribution,genotyping,and epidemiology of HCV was gathered from published articles spanning the years 1996-2020.Results:Genotype 1 accounted for 5.1%of the patients,with its predominant subtype being 1a at 4.38%.The frequencies of its other subtypes,1b and 1c,were observed to be 1.0%and 0.31%respectively.Genotype 2 had a frequency of 2.66%,with the most widely distributed subtype being 2a at 2.11%of the patients.Its other subtypes,2b and 2c,had frequencies of 0.17%and 0.36%respectively.The most prevalent genotype among all isolates was 3(65.35%),with the most frequent subtype being 3a(55.15%),followed by 3b(7.18%).The prevalence of genotypes 4,5,and 6 were scarce in Pakistan,with frequencies of 0.97%,0.08%,and 0.32%respectively.The prevalence of untypeable and mixed genotypes was 21.34%and 3.53%respectively.Estimating genotypes proves to be a productive method in assisting with the duration and selection of antiviral treatment.Different HCV genotypes can exhibit variations in their response to specific antiviral treatments.Different genotypes may have distinct natural histories,including variations in disease progression and severity.Some genotypes may lead to more rapid liver damage,while others progress more slowly.Conclusions:This information can guide screening and testing strategies,helping to identify individuals at higher risk of developing severe complications.Studying the distribution of HCV genotypes in a population can provide valuable insights into the transmission dynamics of the virus.

关键词： Genotypes HCV Molecular evolution Pakistan Spatial distribution

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Virulence Factors and Biofilm Formation in Vancomycin Resistant Enterococcus faecalis and Enterococcus faecium Isolates in Brazil

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Advances in Microbiology 2023年第6期13卷 299-314页

作者： Bárbara de Azevedo Ramos Marília Manta Manta Sivoneide Maria da Silva Rafael Artur Queiroz Cavalcanti de Sá Natália Lira de Souza henrique douglas melo coutinho Márcia Vanusa da Silva Túlio Diego da Silva Maria Tereza dos Santos Correia Maria Betânia melo de Oliveira Universidade Federal de Pernambuco Pernambuco Brazil Universidade Regional do Cariri Ceará Brazil Instituto de Tecnologia de Pernambuco Pernambuco Brazil

In this work, we evaluated biofilm formation of Vancomycin Resistant of E. faecalis and E. faecium (VRE) in different culture media and adhesion substrate, as well as cellular hydrophobicity and presence of virulence genes. For this, 35 isolates were collected from a public hospital in Recife, Pernambuco, Brazil and identified by the Matrix-Assisted Laser Desorption Ionization - Time-of-flight - Mass Spectrometry (MALDI-TOF-MS) technique. Biofilm formation was analyzed by the Crystal Violet (CV) method and fluorescence microscopy, cellular hydrophobicity by hydrocarbon interaction and the presence of gelE, esp and asa1 genes by Polymerase Chain Reaction (PCR). 12 isolates were identified as E. faecalis and 23 as E. faecium. Most were obtained in Coronary Units (40.0%) and Intensive Care Unit (31.4%). E. faecium isolates were more resistant to the antibiotics tested than E. faecalis;however, E. faecalis stood out as a biofilm producer. Regarding the presence and gene frequency, it was observed that gelE (54.3%) and esp (54.3%) were the most prevalent, followed by asa1 (22.9%). When comparing the gene frequency, it was observed that gelE and esp were predominant (48.6% for both species), while asa1 was more frequent in E. faecalis (20.0%). The data presented here are worrying, because they reveal the virulence potential of isolates VRE, which contributes to the dissemination and persistence of these pathogens in the hospital environment.

关键词： Biofilm Cellular Hydrophobicity Gram-Positive Hospital Environment Virulence Genes

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Polyphenolic composition, antibacterial, modulator and neuroprotective activity of Tarenaya spinosa(Jacq.) Raf.(Cleomaceae)

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Asian Pacific Journal of Tropical Biomedicine 2019年第1期9卷 12-17页

作者： Francisca Samara Muniz dos Santos Jose Weverton Almeida Bezerra Jean Paul Kamdem Aline Augusti Boligon Marli Matiko Anraku Ana Raquel Pereira da Silva Kleber Ribeiro Fidelis Nadghia Figueiredo Leite Antonio Ivanildo Pinho henrique douglas melo coutinho Jose Edilson Goncalves dos Santos Microscopy Laboratory Regional University of Cariri (URCA) Program Post graduate in Plant Biology (UFPE) Phytochemical Laboratory Department of Industrial Pharmacy Federal University of Santa Maria (UFSM) Microbiology and Molecular Biology Laboratory Regional University of Cariri (URCA)

Objective: To evaluate the antibacterial activity and neuroprotective capacity of the ethanolic and aqueous extracts of Tarenaya spinosa(T. spinosa) as well as to determine and quantify some of its polyphenols by high performance liquid chromatography with diode-array detection(HPLC-DAD). Methods: The bacterial Escherichia coli, Staphylococcus aureus and Pseudomonas aeruginosa strains, grown in Heart Agar Infusion, were tested. The drugs gentamicin, norfloxacin and imipenem were used to evaluate the modulating or antagonistic capacity of the T. spinosa extracts. The extract was analysed by HPLC-DAD to determine the main phenolic compounds. For the cell viability tests, individual heads of the Nauphoeta cinerea arthropod model were removed, homogenized in Trifluoromethyl ketone and centrifuged afterwards. Subsequently, 20 μL of NaN O2 were added to the biological material, except in the control group, to evaluate the protection capacity of the extracts. The homogenate of the insect heads was incubated for 2 h in tubes containing tetrazolium bromide. Results: HPLC-DAD demonstrated that the ethanolic extract of T. spinosa presented caffeic acid as the major compound. The ethanolic extract also showed neuroprotective effects at concentrations ≥ 10 μg/mL, while aqueous extract was shown to have a protective effect only at the concentration of 100 μg/mL. The aqueous extract demonstrated a clinically relevant antibacterial activity against the Staphylococcus aureus multidrug resistant strain-MDR, with MIC 512 μg/m L. However, when the extracts were associated with gentamicin and imipenem, a synergism was detected against Staphylococcus aureus and Escherichia coli MDR strains. Conclusions: Although it does not present an antibacterial action, the extracts of T. spinosa can be used in the pharmaceutical industries since its extracts show modulating action of drugs. Besides, these natural products have neuroprotective capacity.

关键词： Tarenaya spinosa Cleome spinose Nauphoeta cinerea Synergism Antibiotic resistance

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LC–MS characterization, anti-kinetoplastide and cytotoxic activities of natural products from Eugenia jambolana Lam. and Eugenia uniflora

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Asian Pacific Journal of Tropical Biomedicine 2017年第9期7卷 836-841页

作者： Celestina Elba Sobral de Souza Ana Raquel Pereira da Silva Janaina Esmeraldo Rocha Maria Celeste Vega Gomez Míriam Rolóm Cathia Coronel José Galberto Martins da Costa Manoel L.C.Netto Larissa A.Rolim henrique douglas melo coutinho Laboratório de Microbiologia e Biologia Molecular Universidade Regional do Cariri Center for the Development of Scientific Research Moisés Bertoni Foundation/Díaz Gill Laboratories Laboratório de Pesquisa de Produtos Naturais Universidade Regional do Cariri Centro Universitário Dr.Leao Sampaio–UNILEAO Universidade Federal do Vale do Sao Francisco–UNIVASF

Objective: To evaluate the trypanocidal, leishmanicidal and cytotoxic activity of Eugenia jambolana(E. jambolana) and Eugenia uniflora(E. uniflora) extracts and fractions.Methods: The products were characterized by LC–MS. Antiparasitic assays were performed and cytotoxicity was evaluated in fibroblastos. In vitro assays were performed using spectrophotometric evaluation. All assays were performed in thrice.Results: The results showed that the extracts and the tannic fraction from E. jambolana inhibited 100% of the epimastigote lines. The ethanolic extract was the most efficient in all concentrations tested against the three parasite strains. In the cytotoxicity assay the flavonoid fraction showed low toxicity. All E. uniflora samples showed cytotoxicity at the highest concentration tested, but the extract showed no toxic effect on the fibroblasts at the lowest concentration. The flavonoid and tannic fractions were more efficient against Leishmania braziliensis promastigotes compared to the extract. However, the extracts and the tannic fraction were more effective against Leishmania infantum strains. The effect on epimastigote cells was observed at all concentrations tested, with all E. uniflora samples. However, the samples were more effective at the highest concentration, where there was inhibition in 100% of the Trypanosoma cruzi strains.Conclusions: The species E. jambolana and E. uniflora presented antiparasitic activity against all tested parasite strains, indicating that these species can serve as an alternative therapy as they were efficient in the tests performed. The E. uniflora extract and the E. jambolana flavonoid fraction presented a low cytotoxicity, opening the floor for new biological studies.

关键词： Antiparasitic effect Cytotoxicity Eugenia jambolana Eugenia uniflora

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HPLC-DAD analysis and antifungal effect of Hyptis martiusii Benth(Lamiaceae) against Candida strains

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Asian Pacific Journal of Tropical Biomedicine 2019年第3期9卷 123-128页

作者： Janaína Esmeraldo Rocha henrique douglas melo coutinho Cícero Roberto Nascimento Saraiva Antonia Thassya Lucas dos Santos Ant?nio Judson Targino Machado Jo?o Tavares Calixto Júnior Irwin Rose Alencar Menezes José Galberto Martins da Costa Jaime Ribeiro-Filho Aracelio Viana Colares Laboratory of Microbiology and Biology Molecular Department of Biological Chemistry Regional University of Cariri Leao Sampaio University Center Department of Biological Sciences Regional University of Cariri Laboratory of Pharmacology and Molecular Chemistry Department of Biological Chemistry Regional University of Cariri Laboratory of Natural Product Research Department of Biological Chemistry Regional University of Cariri

Objective: To evaluate the anti Candida activity of Hyptis martiusii decoction and its major compound, caffeic acid alone or in the presence of fluconazole, as well as their cytotoxic effect. Methods: The decoction was characterized using high performance liquid chromatography coupled with diode array detector. For the antifungal activity, the minimum inhibitory concentration(MIC) and the potential effect of the decoction with the fluconazole were evaluated by microdilution method using 96-well microtiter trays. The osmotic fragility test was performed using erythrocytes under saline stress. All tests were performed in triplicate. Results: The chemical characterization of the decoction was performed by high performance liquid chromatography and revealed the presence of seven compounds, including caffeic acid as major constituent. The antifungal tests demonstrated that both decoction(DHm) and caffeic acid obtained from Hyptis martiusii presented MIC and MFC ≥4096 μg/mL against Candida albicans and Candida tropicalis strains. However, in the presence of fluconazole, DHm and caffeic acid presented IC_(50) of 2.60 and 2.53 μg/mL respectively, demonstrating significant synergistic effects against Candida strains. The modulator activity of DHm might be due to the presence of caffeic acid. Moreover, DHm and caffeic acid did not cause significant hemolytic effects, indicating that they present low cytotoxicity. Conclusions: These data indicate that DHm potentiates the activity of the fluconazole, without enhancement of the toxicity, encouraging further toxicological, pharmacological and phytochemical studies to provide consistent evidence of the potential of this plant to be used in drug development.

关键词： Hyptis martiusii Caffeic acid Candida albicans Candida tropicalis Antifungal properties

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Antibacterial enhancement of antibiotic activity by Enterolobium contortisiliquum(Vell.)Morong

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Asian Pacific Journal of Tropical Biomedicine 2017年第10期7卷 945-949页

作者： Zildene de Sousa Silveira Nair Silva Macêdo Thiago Sampaio de Freitas Ana Raquel Pereira da Silva Joycy Francely Sampaio dos Santos Maria Flaviana Bezerra Morais-Braga José Galberto Martins da Costa Raimundo Nonato Pereira Teixeira Jean Paul Kamdem henrique douglas melo coutinho Francisco Assis Bezerra da Cunha Laboratorio de Bioprospeccao do Semioarido Universidade Regional do CaririCratoBrazil Laboratorio de Microbiologia e Biologia Molecular Universidade Regional do CaririCratoBrazil Laboratorio de Pesquisas de Produtos Naturais Universidade Regional do CaririCratoBrazil Departamento de Ciencias biologicas Universidade Regional do Cariri-URCACratoCEP:63105-000Brazil Programa de Pos-graduacao em Bioprospeccao Molecular Universidade Regional do Cariri–URCACratoBrazil

Objective: To identify the main chemical classes of compounds from aqueous extract of Enterolobium contortisiliquum(E. contortisiliquum) seed bark and to evaluate its antibacterial activity, as well as its potential to increase the activity of antibiotics against strains of Staphylococcus aureus, Pseudomonas aeruginosa and Escherichia coli.Methods: Different classes of compounds in the aqueous extract of E. contortisiliquum were evaluated based on the visual changes in the coloration and the formation of precipitate after the addition of specific reagents. The antibacterial activity of the extract and its potential to increase of antibiotic activity of antibiotics drugs, gentamicin and norfloxacin was determined by using the microdilution method.Results: Our results demonstrated that the following secondary metabolites were presented in E. contortisiliquum seed bark: flavones, flavonols, xanthones, flavononols,chalcones, aurones, flavones and catechins. The extract itself had very low antibacterial activity against all bacterial strains tested(MIC ≥ 1024 μg/m L), but there was an increase in the antibiotic activity of gentamicin and norfloxacin when combined in the subinhibitory concentration(i.e., MIC/8).Conclusions: Our data suggests that E. contortisiliquum seed bark may be an alternative source for new drugs with the potential to increase antibiotic activity against different strains of bacteria.

关键词： Antibiotic activity Enhancement Synergism Chemical compounds Bioactivity

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Antibacterial activity and inhibition against Staphylococcus aureus NorA efflux pump by ferulic acid and its esterified derivatives

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Asian Pacific Journal of Tropical Biomedicine 2021年第9期11卷 405-413页

作者： Patrícia Gonçalves Pinheiro Gilvandete Maria Pinheiro Santiago Francisco Erivaldo Freitas da Silva Ana Carolina Justino de Araújo Cícera Rejane Tavares de Oliveira Priscilla Ramos Freitas Janaína Esmeraldo Rocha JoséBezerra de Araújo Neto Maria Milene Costa da Silva Saulo Relison Tintino Irwin Rose Alencar de Menezes henrique douglas melo coutinho JoséGalberto Martins da Costa Departamento de Química Biológica Programa de Pos-Graduação em Química BiológicaUniversidade Regional do Cariri–URCACrato–CEBrazil Programa de Pós-Graduação em Química Centro de CiênciasUniversidade Federal do Ceará–UFCFortaleza–CEBrazil

Objective:To evaluate the inhibitory activity of ferulic acid and four of its esterified derivatives(methyl,ethyl,propyl,and butyl)against resistance mechanisms in Staphylococcus aureus strains.Methods:Ferulic acid derivatives were obtained by esterification with methanol,ethanol,propanol,and butanol,and then characterized by hydrogen and carbon-13 nuclear magnetic resonance analysis.The minimum inhibitory concentrations(MIC)of ferulic acid and its esterified derivatives,ethidium bromide,and norfloxacin were obtained using the microdilution test,while the efflux pump inhibition test was conducted by examining reduction in the MICs of norfloxacin and ethidium bromide.Molecular docking was also carried out using the Schrodinger Suite 2015 molecular modeling software.A three-dimensional model of NorA efflux pump was generated using I-TASSER.The best scoring model was used as a receptor for ligand-receptor docking.Results:The methyl and butyl ester derivatives did not demonstrate significant antimicrobial activity.However,a significant synergic effect was evidenced when norfloxacin was combined with the ethyl and propyl esterified derivatives.The docking study demonstrated favorable energy of interaction between ferulate derivatives and NorA,and amino acid residues TYR57,TYR58,and LEU255 were present commonly in stabilizing all complexes.The PCA analysis corroborated the docking hypothesis that the lipophilic character and hydrogen bond interactions were the most relevant characteristics involved with NorA inhibitors.The pharmacokinetic parameters of ferulic acid derivatives showed good ADMET properties,demonstrating that they can be easily absorbed and have no effect or inhibit the cytochrome P450 enzyme complex,revealing their potential as drug candidates.Conclusions:This study provides strong evidence that the molecular basis for this activity is potentially due to the NorA efflux pump.

关键词： Ferulic acid Esterified derivatives Efflux pump Staphylococcus aureus Resistance mechanisms

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Chemical composition, antiparasitic and cytotoxic activities of aqueous extracts of Ziziphus joazeiro Mart

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Asian Pacific Journal of Tropical Biomedicine 2019年第5期9卷 222-226页

作者： Jacqueline Cosmo ANDrade Ana Raquel Pereira da Silva Ant?nia Thassya Lucas dos Santos Maria Audilene Freitas Yedda Maria Lobo Soares de Matos Maria Flaviana Bezerra Morais Braga Camila Fonseca Bezerra Maria Isabeli Pereira Gon?alo Maria Celeste Vega Gomez Míriam Rolóm Cathia Coronel Paulo Riceli Vasconcelos Ribeiro Edy Sousa de Brito henrique douglas melo coutinho Microbiology and Molecular Biology Laboratory Biological Chemistry Department Regional University of Cariri-URCA Bioassays Laboratory-LABIO Federal University of Cariri-UFCA Brejo Santo University campus (CE) Center for the Development of Scientific Research Moisés Bertoni Foundation/Díaz Gill Laboratories Natural Products Chemistry Multiuser Laboratory LMQPN Embrapa Tropical Agroindustry

Objective: To compare the in vitro antiparasitic activity of aqueous extracts from Ziziphus joazeiro leaves and stem bark against Trypanosoma cruzi, Leishmania braziliensis, and Leishmania infantum, as well as to evaluate its cytotoxicity in mammalian cells, in addition to identifying the chemical composition of the extracts. Methods: Ziziphus joazeiro leaf and stem bark aqueous extracts were prepared by cold extraction maceration and subjected to ultra-efficient liquid chromatography coupled to a quadrupole/time of flight system. The susceptibility assays used Trypanosoma cruzi CLB5 strains and promastigote forms of Leishmania braziliensis and Leishmania infantum for antiparasitic activity of the extracts. Moreover, mammalian fibroblasts NCTC clone 929 were used for cytotoxicity analysis. Results: Terpenoid compounds, flavonoids and phenolic acid were identified in extracts. The stem bark aqueous extracts presented more significant results in terms of antiparasitic activity compared with the leaf aqueous extracts, especially against Leishmania braziliensis and Leishmania infantum promastigote forms with an IC50 < 500 μg/mL. The cytotoxicity evaluation showed moderate toxicity of the stem bark aqueous extracts, which is relevant information for the rational use of this plant part since it is widely used by the population. Conclusions: These preliminary results may contribute to the formulation of new therapeutic agents against this group of neglected diseases, so further investigations are required to delineate the mechanisms of action mainly of the aqueous extract of stem bark of Ziziphus joazeiro.

关键词： Antiepimastigote Antipromastigote UPLC-MS-ESI-QTOF

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Study of the roles of caspase-3 and nuclear factor kappa B in myenteric neurons in a P2X7 receptor knockout mouse model of ulcerative colitis

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World Journal of Gastroenterology 2023年第22期29卷 3440-3468页

作者： henrique Inhauser Riceti Magalhães Felipe Alexandre Machado Roberta Figueiroa Souza Marcos Antônio Ferreira Caetano Vanessa Ribeiro Figliuolo Robson coutinho-Silva Patricia Castelucci Department of Surgery School of Veterinary Medicine and Animal SciencesUniversity of São PauloSão Paulo 05508-270Brazil Department of Anatomy University of São PauloSão Paulo 05508-000Brazil Biophysics Institute Carlos Chagas Filho Federal University of Rio de JaneiroRio de Janeiro 21941-902Brazil

BACKGROUND The literature indicates that the enteric nervous system is affected in inflammatory bowel diseases(IBDs)and that the P2X7 receptor triggers neuronal death.However,the mechanism by which enteric neurons are lost in IBDs is unknown.AIM To study the role of the caspase-3 and nuclear factor kappa B(NF-κB)pathways in myenteric neurons in a P2X7 receptor knockout(KO)mouse model of IBDs.METHODS Forty male wild-type(WT)C57BL/6 and P2X7 receptor KO mice were euthanized 24 h or 4 d after colitis induction by 2,4,6-trinitrobenzene sulfonic acid(colitis group).Mice in the sham groups were injected with vehicle.The mice were divided into eight groups(n=5):The WT sham 24 h and 4 d groups,the WT colitis 24 h and 4 d groups,the KO sham 24 h and 4 d groups,and the KO colitis 24 h and 4 d groups.The disease activity index(DAI)was analyzed,the distal colon was collected for immunohistochemistry analyses,and immunofluorescence was performed to identify neurons immunoreactive(ir)for calretinin,P2X7 receptor,cleaved caspase-3,total caspase-3,phospho-NF-κB,and total NF-κB.We analyzed the number of calretinin-ir and P2X7 receptor-ir neurons per ganglion,the neuronal profile area(μm^(2)),and corrected total cell fluorescence(CTCF).RESULTS Cells double labeled for calretinin and P2X7 receptor,cleaved caspase-3,total caspase-3,phospho-NF-κB,or total NF-κB were observed in the WT colitis 24 h and 4 d groups.The number of calretinin-ir neurons per ganglion was decreased in the WT colitis 24 h and 4 d groups compared to the WT sham 24 h and 4 d groups,respectively(2.10±0.13 vs 3.33±0.17,P<0.001;2.92±0.12 vs 3.70±0.11,P<0.05),but was not significantly different between the KO groups.The calretinin-ir neuronal profile area was increased in the WT colitis 24 h group compared to the WT sham 24 h group(312.60±7.85 vs 278.41±6.65,P<0.05),and the nuclear profile area was decreased in the WT colitis 4 d group compared to the WT sham 4 d group(104.63±2.49 vs 117.41±1.14,P<0.01).The number of P2X7 receptor-ir neurons per ganglion was decrea

关键词： Cell death Enteric nervous system Gastroenterology Inflammatory bowel diseases P2X7 receptor Purinergic signaling

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