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protocatechuic acid and quercetin attenuate ETEC-caused IPEC-1 cell inflammation and injury associated with inhibition of necroptosis and pyroptosis signaling pathways

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Journal of Animal Science and Biotechnology 2023年第4期14卷 1551-1568页

作者： Kan Xiao Mohan Zhou Qingqing Lv Pengwei He Xu Qin Dan Wang Jiangchao Zhao Yulan Liu Hubei Key Laboratory of Animal Nutrition and Feed Science Wuhan Polytechnic UniversityWuhan 430023People’s Republic of China Department of Animal Science Division of AgricultureUniversity of ArkansasFayettevilleAR 72701USA

Background:Necroptosis and pyroptosis are newly identified forms of programmed cell death,which play a vital role in development of many gastrointestinal disorders.Although plant polyphenols have been reported to protect intestinal health,it is still unclear whether there is a beneficial role of plant polyphenols in modulating necroptosis and pyroptosis in intestinal porcine epithelial cell line(IPEC-1)infected with enterotoxigenic Escherichia coli(ETEC)K88.This research was conducted to explore whether plant polyphenols including protocatechuic acid(PCA)and quercetin(Que),attenuated inflammation and injury of IPEC-1 caused by ETEC K88 through regulating necroptosis and pyroptosis signaling pathways.Methods:IPEC-1 cells were treated with PCA(40μmol/L)or Que(10μmol/L)in the presence or absence of ETEC K88.Results:PCA and Que decreased ETEC K88 adhesion and endotoxin level(P<0.05)in cell supernatant.PCA and Que increased cell number(P<0.001)and decreased lactate dehydrogenases(LDH)activity(P<0.05)in cell supernatant after ETEC infection.PCA and Que improved transepithelial electrical resistance(TEER)(P<0.001)and reduced fluorescein isothiocyanate-labeled dextran(FD4)flux(P<0.001),and enhanced membrane protein abundance of occludin,claudin-1 and ZO-1(P<0.05),and rescued distribution of these tight junction proteins(P<0.05)after ETEC infection.PCA and Que also declined cell necrosis ratio(P<0.05).PCA and Que reduced mRNA abundance and concentration of tumor necrosis factor-α(TNF-α),interleukin(IL)-6 and IL-8(P<0.001),and down-regulated gene expression of toll-like receptors 4(TLR4)and its downstream signals(P<0.001)after ETEC infection.PCA and Que down-regulated protein abundance of total receptor interacting protein kinase 1(t-RIP1),phosphorylated-RIP1(p-RIP1),p-RIP1/t-RIP1,t-RIP3,p-RIP3,mixed lineage kinase domain-like protein(MLKL),p-MLKL,dynamin-related protein 1(DRP1),phosphoglycerate mutase 5(PGAM5)and high mobility group box 1(HMGB1)(P<0.05)after ETEC infection.Moreover,PCA and Que reduced protein abundance of

关键词： Cell damage ETEC K88 Intestinal inflammation Necroptosis protocatechuic acid Pyroptosis Quercetin

Beneficial effects of protocatechuic acid on diabetic retinopathy in streptozocin-induced diabetic rats

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International Journal of Ophthalmology(English edition) 2023年第6期16卷 855-862页

作者： Ling Wan Jun Li Wei-Bai Chen Guo-Qiang Wu Department of Ophthalmology Sichuan Provincial People’s HospitalUniversity of Electronic Science and Technology of ChinaChengdu 610072Sichuan ProvinceChina Department of Ophthalmology Chinese Academy of Sciences Sichuan Translational Medicine Research HospitalChengdu 610072Sichuan ProvinceChina Department of Ophthalmology the First People’s Hospital of Lanzhou CityLanzhou 730050Gansu ProvinceChina

·AIM:To determine the effects of protocatechuic acid(PCA)on streptozocin-induced diabetic retinopathy(DR)in rats.·METHODS:Wistar rats were given a 50 mg/kg intraperitoneal injection of streptozocin to induce diabetes.Animals were assigned randomly one of four groups(8 rats per group):control,diabetic,diabetic plus PCA(25 mg/kg·d),and diabetic plus PCA(50 mg/kg·d).After inducing diabetes,treatments were started one week later and continued for eight weeks.After the experiment,the rats were sacrificed,and their retinas were taken for biochemical and molecular analysis.·RESULTS:PCA administration diminished the blood glucose and glycated haemoglobin levels relative to the diabetic group.In diabetic rats,PCA lowered elevated levels of advanced glycosylated end products(AGEs)and receptor for AGEs(RAGE).In the retina of diabetic rats,PCA effectively decreased inflammatory cytokine,nuclear factor-κB,tumour necrosis factor-α,interleukin-1β,and vascular endothelial growth factor,and increased antioxidant markers glutathione,superoxide dismutase,and catalase.·CONCLUSION:The protective benefits of PCA against DR may be attributable to its suppression of the AGEs and RAGE and its antioxidant and anti-inflammatory properties.

关键词： diabetic retinopathy streptozocin protocatechuic acid oxidative stress vascular endothelial growth factor

Dietary protocatechuic acid ameliorates inflammation and up-regulates intestinal tight junction proteins by modulating gut microbiota in LPS-challenged piglets

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Journal of Animal Science and Biotechnology 2021年第1期12卷 328-339页

作者： Ruizhi Hu Ziyu He Ming Liu Jijun Tan Hongfu Zhang De-Xing Hou Jianhua He Shusong Wu Hunan Collaborative Innovation Center for Utilization of Botanical Functional Ingredients College of Animal Science and TechnologyHunan Agricultural UniversityChangsha 410128China Department of Food Science and Biotechnology Faculty of AgricultureKagoshima UniversityKagoshima 890-0065Japan Beijing China-Agri HongKe Bio-Technology Co. Ltd.Beijing 102206China State Key Laboratory of Animal Nutrition Institute of Animal SciencesChinese Academy of Agricultural SciencesBeijing 100193China.

Background: Weaning is one of the major factors that cause stress and intestinal disease in piglets. protocatechuic acid(PCA) is an active plant phenolic acid which exists in Chinese herb, Duzhong(Eucommia ulmoides Oliver), and is also considered as the main bioactive metabolite of polyphenol against oxidative stress and inflammation. This study aimed to investigate the effect of PCA on growth performance, intestinal barrier function, and gut microbiota in a weaned piglet model challenged with lipopolysaccharide(LPS).Methods: Thirty-six piglets(Pig Improvement Company line 337 × C48, 28 d of age, 8.87 kg ± 0.11 kg BW) were randomly allocated into 3 treatments and fed with a basal diet(CTL), a diet added 50 mg/kg of aureomycin(AUR), or a diet supplemented with 4000 mg/kg of PCA, respectively. The piglets were challenged with LPS(10 μg/kg BW) on d 14 and d 21 by intraperitoneal injection during the 21-d experiment. Animals(n = 6 from each group) were sacrificed after being anesthetized by sodium pentobarbital at 2 h after the last injection of LPS. The serum was collected for antioxidant indices and inflammatory cytokines analysis, the ileum was harvested for detecting mRNA and protein levels of tight junction proteins by PCR and immunohistochemical staining, and the cecum chyme was collected for intestinal flora analysis using 16 S rRNA gene sequencing.Results: Dietary supplementation of PCA or AUR significantly increased the expression of tight junction proteins including ZO-1 and claudin-1 in intestinal mucosa, and decreased the serum levels of thiobarbituric acid reactive substances(TBARS) and IL-6, as compared with CTL group. In addition, PCA also decreased the serum levels of IL-2 and TNF-α(P < 0.05). Analysis of gut microbiota indicated that PCA increased the Firmicutes/Bacteroidetes ratio(P < 0.05). Spearman's correlation analysis at the genus level revealed that PCA reduced the relative abundance of Prevotella 9, Prevotella 2, Holdemanella, and Ruminococcus torques group(P < 0.05), and increased the relati

关键词： Gut microbiota Inflammation Piglets protocatechuic acid Tight junction proteins

Caffeic acid and protocatechuic acid modulate Nrf2 and inhibit Ehrlich ascites carcinomas in mice

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Asian Pacific Journal of Tropical Biomedicine 2021年第6期11卷 244-253页

作者： Venugopal R.Bovilla Preethi G.Anantharaju Sireesh Dornadula Prashanthkumar M.Veeresh Mahadevaswamy G.Kuruburu Vidya G.Bettada Kunka Mohanram Ramkumar SubbaRao V.Madhunapantula Center of Excellence in Molecular Biology and Regenerative Medicine Department of BiochemistryJSS Medical CollegeJSS Academy of Higher Education&ResearchMysore-570015KarnatakaIndia SRM Research Institute Department of BiotechnologySchool of BioengineeringSRM Institute of Science and TechnologyKattankulathur-603203Tamil NaduIndia

Objective:To assess the nuclear factor-erythroid 2-related factor-2(Nrf2)modulatory effect of caffeic acid and protocatechuic acid and determine the anti-tumor activity of these phenolic compounds against Ehrlich ascites carcinoma growth in mice.Methods:Antioxidant activity of protocatechuic acid and caffeic acid was assessed using ferric reducing antioxidant power(FRAP)and 2,2-diphenyl-1-picrylhydrazyl(DPPH).Nrf2 activation potential of phenolic compounds was tested by quantitative realtime polymerase chain reaction,and luciferase complementation reporter assays.In vivo efficacy was tested using the Ehrlich ascites carcinoma model.Results:FRAP and DPPH radical scavenging assays showed that caffeic acid and protocatechuic acid were more potent compared with cinnamic acid and benzoic acid.Luciferase complementation reporter assays identified caffeic acid and protocatechuic acid as the activators of Nrf2.Both caffeic acid and protocatechuic acid upregulated the expression of Nrf2 target genes heme oxygenase-1(HO-1),glutamate-cysteine ligase catalytic subunit(GCLC),and glutamate-cysteine ligase modifier subunit(GCLM)and the activity of NAD(P)H:quinone oxidoreductase 1(NQO1)when tested on HCT-116 cells using a cell-based assay system at 9 h.In addition,intraperitoneal administration of caffeic acid and protocatechuic acid to Ehrlich ascites carcinoma bearing mice suppressed tumor growth and angiogenesis.Conclusions:Caffeic acid and protocatechuic acid can modulate Nrf2 and inhibit Ehrlich ascites carcinoma cells.

关键词： Ehrlich ascites carcinomas Nrf2 protocatechuic acid Caffeic acid NQO1 activity

Protective and therapeutic effect of protocatechuic acid in assessment of letrozole- induced polycystic ovary syndrome in rats

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Asian pacific Journal of Reproduction 2020年第5期9卷 230-238页

作者： Rupavath Chandrashekhar Bakshi Vasudha Jagruthi Jeripothula Nelavelli Lakshmi Bhavani Bhavani Ram Department of Botany University College of ScienceSaifabadOsmania UniversityHyderabadTelangana500004India Department of Pharmacology School of PharmacyAnurag Group of InstitutionsVenkatapurGhatkesarTelangana500088India Green Evolution Laboratories Wangapally VillageNalgondaTelangana500085India

Objective:To investigate the potential activity of protocatechuic acid in female Wistar rats with letrozole-induced polycystic ovary syndrome(PCOS).Methods:Thirty rats were divided into five groups of six each.Group 1 received 0.5%carboxy methyl cellulose orally and served as the normal control group;group 2 was treated orally with 1 mg/kg of letrozole daily for 21 days and served as the PCOS induced group;group 3 was orally administered with letrozole of 1 mg/kg for 21 days and further administered with standard drug of clomiphene citrate at a dose of 1 mg/kg body weight in 0.5%carboxy methyl cellulose per oral and served as the standard group;groups 4 and 5 were administered with letrozole of 1 mg/kg for 21 days and further treated with protocatechuic acid orally at low dose of 5 mg/kg body weight and high dose of 15 mg/kg body weight respectively for 15 days.At the end of the study period,rats were subjected for the estimation of invasive blood pressure and heart rate,biochemical estimations and antioxidant assay.In addition,ovarian histomorphology was examined.Results:The PCOS was confirmed in the letrozole induced rats with increased concentration of androgen,abnormal lipid levels,glucose,glycosylated haemoglobin and also depletion of antioxidants.After protocatechuic acid treatment,the increased levels of testosterone due to induction of PCOS were restored to normal levels.Additionally,there was a consistent decrease in luteinizing hormone and follicle stimulating hormone levels in the treatment groups,followed by decrease in the number of cysts after treatment with protocatechuic acid.Histopathological observations showed a remarkable recovery of the ovarian tissue and the presence of normalized structure of antral follicle.protocatechuic acid treatment restored all the parameters to normalcy and abolished cysts formation in ovaries of female rats.Conclusions:protocatechuic acid shows potential protective effects in letrozole-induced PCOS rats.The protective effect is comparable to that of clomiphene citra

关键词： Polycystic ovary syndrome Letrozole protocatechuic acid Fertility Ovulation

Optimization of the Ultrasonic-assisted Extraction Process for protocatechuic acid from Emilia sonchifolia DC by Response Surface Methodology

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Agricultural Biotechnology 2020年第3期9卷 58-66页

作者： Wen CHEN Xiangjun WANG Lingyan LI Zhewen ZHAI Xiaoling HUANG Wubing RAO Chenghan ZOU Tengtong LI College of science Hainan Tropic Ocean UniversitySanya 572022China College of Fisheries and life Hainan Tropic Ocean UniversitySanya 572022China College of Food Science Southwest UniversityChongqing 400715China

[Objectives] This study was conducted to optimize the extraction process of protocatechuic acid from Emilia sonchifolia DC. [Methods] The optimal extraction conditions were determined by single factor,response surface analysis and variance analysis,and the content of protocatechuic acid was determined by HPLC. [Results] The protocatechuic acid standard curve equation was: y = 1 435 x + 8 403,R^2= 0. 999 8,indicating a good linear relationship. The optimal extraction conditions were as follows: a temperature at 80 ℃,an extraction time of 1 h,a material-to-liquid ratio at 1:10 and an ultrasonic power of 600 W,and the content of protocatechuic acid extracted was 1. 93 mg/g. The method showed a RSD of 0. 41%,less than 2%,and the detection limit was 0. 0000047261 g/ml.The experimental sample X1 was the low-level 0. 1 mg/ml standard solution,which showed recovery of protocatechuic acid between 100.8% and 105.2%,with a RSD of 0. 013%;and the sample X2 was the high-level 1. 0 mg/ml standard solution,which exhibited recovery between 100. 6% and 102. 2%,with a RSD of 0.076%. Thus,the recovery was high,and the requirements of the performance index were met. [Conclusions] The detection method is stable and reliable and can produce satisfactory results.

关键词： Emilia sonchifolia DC protocatechuic acid HPLC Response surface optimization Variance analysis

protocatechuic acid inhibition of tumor nuclear factor cytotoxicity in human endothelial cells is mediated by nuclear transcription factor-kB pathway

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Protocatechuic acid inhibition of tumor nuclear factor cytot...

Free and/or glucuronidated Danshensu, protocatechuic aldehyde,protocatechuic acid and caffeic acid are potentially pharmacological components in rats after oral administration of extractum for Compound DanShen Dripping Pill

2000上海科技论坛BIO-X研讨会暨中德医学生物物理...

作者： Jf. Zhou,AK.Bosserhof and GM. ArtmannDepartment of Applied Cell Biophysics and Bioengineering,AachenUniversity of Applied Sciences,Juelich 52428,Germany. Institutes of Pathology,Aachen University Hospital,Aachen 52074,Germany.

＜正＞ In this study,protocatechuic acid（Pac）, a water-soluble component of Salvia Miltiorriza Bge, on theability of the vascular endotheliun to respond to tumor necrosis factor is inveshgated. In the presenceof a protein srpthesis inhibitor, actinomycin D, tumor necrosis factor（TNF-a）induces humanumbilical vein endothelial cells（HUVECs）to undergo apoptosis. Pretreating HUVECs with Pac for 2h,enhances survival cells for 1.5 fold, suggesting that Pac inducible cytoprotective pathway occurs forcell survive. Howevef, this cell death prevention effect of Pac is markedly blocked when a nuclearfactor kappa B（NF-kB） activation iuhibitor MG132 is added. It indicates that NF-kB may be

关键词： protocatechuic acid tumor necrosis factor nuclear factor

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Free and/or glucuronidated Danshensu, protocatechuic aldehyd...

Simultaneous determination of enthalpies of dilution and dissociation of protocatechuic acid in aqueous salt solutions

中国药理学会临床药理学专业委员会会议暨第十次全国临床药...

作者： LIU Hong-tao~(1,2) LI Wei~(1,2) HU Yong~1 YANG Ling~1 1 Laboratory of Pharmaceutical Resource Discovery,Biotechnology Division,Dalian Institute of Chemical Physics,Chinese Academy of Sciences,Dalian 116023,Liaoning,China 2 Graduate School,Chinese Academy of Sciences,Beijing 100049,China

AIM:To provide primarily pharmacokinetic profiles and reveal potentially pharmacological components after oral administration of the extractum for Compund Danshen Dripping Pill(CDDP)in rats,which was mainly com- posed of protocatechuic aldehyde(PAL),Danshensu (DSS),lithospermic acid A,lithospermic acid B,salvi- anolic acid D,rosmarinic acid,salvianolic acid B and salvianolic acid A. METHODS:After orally administration of 200 mg CDDP extractum for 1 h,the portal vein blood,the femoral artery blood,the bile and urine samples were separately collect- ed.The gastrointestinal(GI)contents and mucosa were washed with physiological saline and the washing solution was pooled.The whole GI tract was removed and homoge- nized for analysis.The constituents and concentrations of ’all above samples were determined by the LC-MS analysis combined with the β-glucuronidase and sulfatase treatment. RESULTS:The investigation after oral administration of CDDP extractum for I h in rats indicated that:(1)Most of polyphenolic acids have been degraded into caffeic acid (CA)and DSS in the GI tract.Next,CA could be easily absorbed into blood circulation and a trace of DSS was also transported into the portal vein.The remains of undegrad- ed polyphenolic acids and Danshensu were detained in the GI tract lumen.(2)Part of PAL was firstly oxidized into PAC in GI epithelial cells before absorption into the portal vein.Next,PAC and the rest PAL were further transport- ed into blood circulation.(3)PAC and PAL were distrib- uted in blood circulation in free and conjugated forms after being glucuronidated in the liver and the kidney while only free CA and DSS appeared in vivo.After that,all of these metabolites were gradually secreted into bite and urine. CONCLUSION:The results suggest that DSS,PAL, PAC,CA and their conjugated forms may be potential pharmacological components for CDDP extractum.

关键词： Compound Danshen Dripping Pill protocatechuic Aldehyde protocatechuic acid caffeic acid glucuronidatio

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Simultaneous determination of enthalpies of dilution and dis...