bluuebunny/crossref_metadata_2025_split huggingface dataset:
ANTIOXIDANT AND CHEMOPREVENTIVE EFFECT OF AZADIRACHTA INDICA ON LEAD ACETATE INDUCED HEPATOTOXICITY IN RATS
Authors: Omobowale Temidayo , Oyagbemi Ademola , AdeyeOluwa Temitayo Esther , Akinboye Olajide , Adedapo Adeolu 
Published: November 2023

Lead (Pb), a ubiquitous heavy metal is known to be injurious to all systems. Oxidative damage has been shown to be one of the principal mechanisms by which Pb causes toxicity. In this study, hepatic antioxidant status and markers of oxidative stress were assessed in experimental animals orally exposed to lead acetate (PbA) and was compared with groups treated with the methanolic extract of Azadirachta indica at 2 different doses.Seventy rats were used in this study. They were divided into 7 groups each containing 10 animals. Control group (group A) received water only. Group B, C and D were dosed 0.1% lead and was withdrawn after 6 weeks. On withdrawal, Group B received water only while groups C and D were treated with 100mg/kg and 200mg/kg of the methanolic extract of Azadirachta indica respectively for 6 weeks. Similarly, groups E, F, G were given 0.2% lead orally and was withdrawn after 6 weeks. On withdrawal, group E received water only while groups F and G were treated with 100mg/kg and 200mg/kg A. indica respectively for 6 weeks.Exposure of rats to lead acetate led to a significant (p<0.05) reduction in hepatic reduced Glutathione (GSH), Glutathione peroxidase (GPX) and Glutathione S‐ transferase. Similarly, Malondialdehyde (MDA) content increased (p<0.05) significantly in animals dosed PbA alone while there was a significant (p<0.05) reduction in rats treated with A. indica. There was significant (p<0.05) increase in nitric oxide (NO) and H2O2 concentrations in rats treated with PbA alone whereas treatment with A. indica led to significant (p<0.05) reduction in these markers of oxidative stress. Histology showed the presence of high periportal infiltration with inflammatory cells and focal necrosis in PbA treated groups but periportal infiltration was mild in Rats dosed with A. indica. In conclusion, animals exposed to PbA showed hepatotoxicity and disruption of antioxidant defence system through free radical generation and oxidative stress. Treatment of PbA exposed rats with A. indica led to significant amelioration of the hepatotoxicity induced by PbA. In conclusion, this study demonstrated the hepatoprotective effect of A. indica via antioxidant and free radical scavenging activities.Support or Funding InformationNone
Autophagy: a Novel Mechanistic Contributor to Ursodeoxycholic Acid in Alleviating Progression of Hepatic Fibrosis
Authors: Ye Hui‐Lan , Zhang Ji‐Wang , Chen Xing‐Zhou , Chen Li , Zhang Fa‐Can , Zhang Guo 
Published: November 2023

Ursodeoxycholic acid (UDCA) has been found an increasingly wide utilization in the treatment of cholestatic liver fibrosis, especially for primary biliary cholangitis (PBC) and its detailed mechanisms remain unclear. Herein, this study was aimed to determine whether autophagic signaling is involved in the therapeutic effect of UDCA for liver fibrosis. Firstly, by using immunohistochemistry and fluorescent microscopic technology, autophagic molecule LC3 was found to highly express in human liver fibrotic tissues and TGFβ1 treated LX2 hepatic stellate cells(HSCs). With qRT‐PCR, autophagy promotion with rapamycin, the autophagic agonist, resulted in dramatic increment of cellular COL1A2 mRNA level, whereas blocking autophagic signaling with 3‐MA or hydroxychloroquine (HCQ) or small interference RNAs lead to decrement of COL1A2 mRNA transcription. Secondly, western blotting, qRT‐PCR and trypan blue were conducted to analyze autophagy flux, cell proliferation and collagen production in UDCA treated HSCs. UDCA treatment was found to dose‐dependently inhibit autophagy, cell proliferation as well as COL1A2 mRNA generation in TGFβ1 activated HSCs. The presence of rapamycin was shown to substantially impair the antic‐fibrotic effects of UDCA on these cells, while autophagy suppression with siRNAs targeting Atg5 or Beclin1 achieved marginally therapeutic effect. Finally, the anti‐fibrotic effect and autophagy were analyzed in CCl4‐induced liver fibrosis of rats by H&E, masson staining, qRT‐PCR and western blotting. It was found that administration of rapamycin mildly thickened fibrous septa in rats with hepatic fibrosis. However, UDCA alone or in combined with HCQ notably restored the CCl4‐induced tissue architecture disruption and collagen aggregation following inhibitory impact on autophagic signaling hallmark as LC3 IIprotein expression and P62 degradation in these rodent models. Taken together, the present study firstly revealed that UDCA displays the anti‐fibrotic role of by protecting HSC against excessive production of collagen through autophagy inhibition and provide a new insight into the pharmacological basis of UDCA treatment for hepatic fibrosis.Support or Funding InformationNational Natural Science Foundation of China, NO.30960145, NO.81360077, NO.81172260 and Guangxi Natural Science Foundation, NO.2015GXNSFDA139022, zhangguogx@hotmail.com
Pharmacologic Treatment with a Melanocortin Analogue Protects Against Pressure‐Overload Induced Cardiac Hypertrophy
Authors: Ruohonen Saku , Rinne Petteri , Ruohonen Suvi , Röyttä Matias , Savontaus Eriika 
Published: November 2023

PurposeHypertrophic growth of the heart is a consequence of heart ability to adapt to an increased afterload and wall stress. Although left ventricular hypertrophy (LVH) may resolute with adequate therapy, this process increases the amount of irreversible cardiac fibrosis.These morphological and functional changes in the myocardium may manifests as arrhythmias and congestive heart failure (CHF).Melanocortins regulate important physiological functions such as energy balance and blood pressure, but their long‐term effects on cardiac remodeling remain unexplored.In the present study, we investigated whether melanocortins affect physiological or pathologic cardiac growth and function.MethodsFirst, we studied the effects of transgenic melanocortin overexpression (MSH‐OE) on physiological age‐related cardiac growth in mice. Second, we examined the effects of a stable melanocortin analogue (MT‐II) on pressure overload‐ induced cardiac hypertrophy. We subjected 8‐week‐old C57Bl/6N mice to pressure‐overload by transverse aortic constriction (TAC) and randomly assigned the mice to receive daily i.p. injections of either vehicle (physiological saline) or MT‐II (0.3 mg/kg) for 6 weeks starting 2 weeks after surgery.ResultsAt the age of 6 months, MSH‐OE mice (n=11) had lower heart weight and smaller myocytes with maintained cardiac function compared to their age‐matched wild‐type littermates (n=15) while no differences in heart weight were noted at the age of 3‐months, indicating that melanocortin overactivity restrains age‐related cardiac growth. Eight weeks of pressure‐overload induced significant left ventricular (LV) hypertrophy with decreased cardiac function in saline‐treated TAC mice (n=15) compared to sham‐operated control mice (n=9).This pathologic cardiac remodeling was attenuated in MT‐II‐treated TAC mice (n=9) (Figure 1. and 2.). Importantly, MT‐II‐treatment did not compromise cardiac function, but tended to maintain LV contractility of TAC mice. Using quantitative NMR scanning to analyze body composition, we demonstrate that MT‐II treatment also prevented the deterioration of body weight development associated with the hypertrophic disease state.MT‐II treatment attenuated also the expression of genes regulating cardiac remodeling related mRNAs such as NppA, NppB, MMP‐2 and Col1a2.ConclusionsAlthough the underlying mechanisms remain largely unclear, these results, for the first time, establish melanocortin signaling as an antihypertrophic regulator and suggest that melanocortin analogues may be potential therapeutic tools for the treatment of cardiac hypertrophy and heart failure.Support or Funding InformationAcademy of Finland, Finnish Cultural Foundation
Pharmaceutical formulation Flavonoids and saponins extracted from black bean (Phaseolus vulgaris L.) seed coats and their hypocholesterolemic properties for the prevention of cardiovascular diseases
Authors: Jimenez‐Ramos Angelica , Chavez‐Santoscoy Rocio Alejandra 
Published: November 2023

Beans exert hypolipidemic activity, and it has been attributed to flavonoids and saponins from seed coats, by attenuating lipogenesis and stimulating fatty acid oxidation and cholesterol biliary excretion through the differentially regulation of LXR probably mediated by AMPK phosphorylation. The aim of this study was to extract and quantify the major saponins and flavonoids contained in black bean seed coats and to formulate an appropriated pharmaceutical form for the delivery of previously mentioned bioactive compounds. Phytochemical extraction was performed with 90 % ethanol in acidified water with citric acid (1%). The resulting extract was concentrated and lyophilized. Phytochemicals were identified and quantified by HPLC‐UV‐ELSD. Design experiment was conducted to obtain a formulation with the best quality characteristics. Formulations were evaluated according to Mexican Official Standard NOM‐073‐SSA1‐2005 for Stability of drugs and medicines. Briefly, friability, disintegration, weight variation and hardness were measured in 5 replicates of each formulation. In vitro and in vitro assays will be performed to analyses the effects of formulation on key proteins involved in lipid metabolism and biliary cholesterol secretion.Support or Funding InformationThis research was supported by the Research Chair Funds for Full Time Professors (2014) of Autonomous University of Baja California (UABC), program Number 3888. The authors would like to acknowledge Consejo Nacional de Ciencia y Tecnología (CONACYT) in Mexico.
Protein S‐glutathionylation: the Redox Mechanism of Metabolic Stress‐induced Endothelial Barrier Dysfunction and Atherosclerosis
Authors: Han Jingyan , Shao Di , Watanabe Yosuke , Bachschmid Markus , Matsui Reiko 
Published: November 2023

BackgroundProtein S‐glutathionylation (Pr‐SSG), the reversible formation of mixed disulfides between protein cysteinyl thiol groups and glutathione, is the most stable and abundant oxidative modification of proteins, and emerging as a critical redox signaling mechanism in cardiovascular diseases (CVD). However, its role in atherosclerosis (AS), the major cause of CVD, is not known. This study thus aimed to explore the role of Pr‐SSG in pathogenesis of AS with specific focus of vascular endothelial barrier dysfunction, an important initiating factor in the development of AS.Methods and ResultsEmploying the biotin switch assay, a well‐developed approach to assess the level of reversible oxidation of proteins in cells and tissues, we found a marked increase in cysteine oxidation of aortic proteins in whole‐tissue lysates of aortas from ApoE−/− mice on Western Diet for 8 weeks, concomitant with significant decrease in reduced form of glutaredoxin‐1(Glrx‐1), a small cytosolic thiol‐transferase that can efficiently and specifically remove the GSH abducts and appears as a pivotal regulator of redox signaling cascades governing a plethora of cellular processes. Pr‐SSG in aortic tissue of ApoE−/− mice visualized by immuno‐fluorescence staining with specific antibody against protein‐GSH complex, was markedly increased in atherosclerotic lesions. Furthermore, an increased Pr‐SSG level was mainly observed in metabolic stressed aortic ECs in vivo and in vitro, which was diminished by up‐regulation of Glrx‐1 in Glrx TG/ApoE−/− mice and in Glrx adenovirus infected human aortic endothelial cells (HAECs). More importantly, the pro‐atherogenic hyper‐permeability of aortic endothelium induced by two‐week WD feeding or disturbed flow was prevented in Glrx TG/ApoE−/− mice without alternation of plasma lipid profiles and proinflammatory responses. Mechanistically, induction of Pr‐SSG in aortic endothelium disorganized cytoskeletal actin structure, while up‐regulation of Glrx‐1 strengthened aortic barrier integrity through activation of small RhoGTPase rac1 and consequent cortical actin formation. Up‐regulation of Glrx thus preserve aortic barrier function and alleviate atherosclerotic lesion development.ConclusionsPr‐SSG in aortic ECs is highly induced by atherogenic stimulus and plays a causative role in aortic barrier dysfunction and development of AS. S‐glutathionylation and Glrx‐1 are thus positioned as excellent target and bio‐therapeutic for cardiovascular injuries associated with oxidative stress, respectively.Support or Funding InformationThis study is supported by AHA SDG20140036 (J.H.), 1UL1TR001430 (BU CTSI), T32 HL07224, and R37 HL104017 (R.A.C.).
PARTICIPATION OF ADENYLYL CYCLASE/cAMP PATHWAY IN VASORELAXANT MECHANISM OF THE PYRANOCOUMARIN BRAYLIN IN MESENTERIC ARTERY
Authors: Albuquerque Jamile , Santos William , Jesus Rafael , Costa Rafael , Filho Otávio , Velozo Eudes , Casais e Silva Luciana , Silva Darízy 
Published: November 2023

IntroductionBraylin is a pyranocoumarin isolated from the species Zanthoxylum tingoassuiba (Rutaceae). Previous studies have shown that Braylin has a possible inhibitory action in phosphodiesterase enzymes.AimsThe aim of this study was to investigate the mechanism involved in the vasorelaxant effect of Braylin.MethodsThe experiments were performed using wistar rats (12–16 weeks/250–300g), euthanized in CO2 camera and the superior mesenteric artery were isolated and free from connective tissue and cut into rings (1–2 mm), suspended by cotton threads for isometric tension recordings in a Tyrode's solution at 37 °C, gassed with a 95% O2 and 5% CO2, under a resting tension of 0.75g. All surgical and experimental procedures were approved by the local Animal use and care ethics committee – CEUA/ICS/UFBA by the number 089/2015.ResultsThe cumulative administration of Braylin (0.1–300 μM) in phenylephrine (Phe 10μM) pre‐contracted mesenteric rings, induced a concentration‐dependent vasorelaxation (Emax = 111.5 ± 7.9%, CE50 = 4.8 (3.0 – 7.7) × 10−5, n = 17), the removal of the endothelium did not change this effect (Emax = 127.6 ± 13.1%, CE50 =1.7 (4.0 – 7.3) × 10−5, n = 16). The presence of ODQ (100 μM), an inhibitor of soluble guanylyl cyclase, also did not change the effect induced by Braylin (Emax = 117.3 ± 2.6%, CE50 = 1.3 (1.9 – 8.9) × 10−5, n = 5). On the other hand, the presence of SQ 22536, an adenylyl cyclase inhibitor, reduced the relaxant effect (Emax = 115.6 ± 2.8, CE50 = 4.3 (3.0 – 6.2) × 10−5, n = 5). The incubation of the superior mesenteric rings with KT‐5720, an inhibitor of PKA, reduced the effect of Braylin (Emax = 110.4 ± 5.3%, CE50 = 2.9 (2.0 – 4.1) × 10−5, n = 5). Pre‐treatment with KCl 20 mM, TEA (1mM), BaCl2 (30μM), or 4‐aminopiridin (1mM) decreased the pharmacological potency of the Braylin significantly.ConclusionThose effects taken together suggest that Braylin induces vasorelaxant effects in isolated superior mesenteric artery rings from normotensive involving AC/cAMP pathway. K+ channels also seem to be important in the relaxant effect, probably BKca channels.Support or Funding InformationFundação de Amparo à Pesquisado Estado da Bahia (fapesb) and Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)
The Hyaluronan Receptor RHAMM is Necessary for the Activation of Src Kinase and the Signaling Pathway for Endothelial Nitric Oxide Production
Authors: Savani Rashmin C , Liao Jie , Longoria Christopher , Wu Qian , Yuhanna Ivan S , Ullrich Victoria , Shaul Philip W , Cheong Naeun 
Published: November 2023

RHAMM (Receptor for Hyaluronan‐Mediated Motility) is necessary for endothelial cell (EC) migration in vitro and angiogenesis in vivo, but the mechanisms are incompletely understood. Nitric oxide (NO) production by endothelial nitric oxide synthase (eNOS) involves a signaling cascade through Src kinase and Akt, and results in eNOS phosphorylation. A number of diverse agonists, including VEGF, HDL, estradiol, acetylcholine and insulin, activate eNOS to produce NO. We hypothesized that RHAMM is necessary for the activation of Src kinase and eNOS to produce NO. To determine the molecular mechanism by which RHAMM regulates eNOS activation, we used cultured EC and RHAMM KO mice.RHAMM siRNA knockdown in HUVEC, BAEC or HPAEC inhibited the increased cell migration after single scratch wounding and eNOS activity observed with treatment with all EC agonists, and the exogenous NO donor NONOate rescued the migratory defect, suggesting that RHAMM is universally required for eNOS activation and NO production. RHAMM siRNA‐mediated decrease in cell migration and eNOS activity correlated with decreased Akt and eNOS phosphorylation but not Erk phosphorylation. In RHAMM KO mice, normal alveolarization and re‐endothelialization after arterial injury, processes that require NO, were delayed. RHAMM KO mice also did not respond to exogenous VEGF, but treatment with either inhaled NO (iNO) or Molsidomine normalized lung alveolarization and restored re‐endothelialization after arterial injury respectively.Since activation of Src is a proximal step in the signaling cascade, we examined the effect of RHAMM knockdown on Src activation. RHAMM siRNA treatment of EC was associated with hyperphosphorylation of the inhibitory site of Src (Y527) and an inhibition of the phosphorylation of the activation site of Src (Y416). In this process, the phosphatase Shp2 is critical in the dephosphorylation of Y527 and Src activation, and associates with VEGFR2. Interestingly, Shp2 and Src both co‐immunoprecipitated with RHAMM in vivo as well as in vitro. Compared to EC treated with scrambled siRNA, co‐immunoprecipitation of Shp2 and VEGFR2 after VEGF stimulation was significantly reduced in RHAMM siRNA‐treated EC. These data suggest that the RHAMM‐Shp2 complex facilitates Shp2 tethering to VEGFR2, which is necessary for the dephosphorylation of Y527 in Src and the downstream activation of the signaling pathway for NO production in endothelial cells. We speculate that RHAMM is a novel target for the regulation of the wide array of physiologic and pathological functions that endothelial NO regulates.Support or Funding InformationNIH: HL093535, HL079090, HL075930; Children's Medical Center Foundation; William Buchanan Chair in Pediatrics.
Endogenous tissue transglutaminase critically regulates cardiac remodeling through distinct enzymatic and non‐enzymatic actions
Authors: Shinde Arti Vijay , Dobaczewski Marcin , de Haan Judith J , Saxena Amit , Lee Kang‐Kon , Xia Ying , Chen Wei , Su Ya , Palanski Brad A , Fujikura Kana , Garcia Mario J , Melino Gerry , Frangogiannis Nikolaos G 
Published: November 2023

The multifunctional protein Tissue transglutaminase (tTG) is induced in remodeling tissues and regulates cellular phenotype and matrix metabolism through enzymatic (matrix cross‐linking) effects and non‐enzymatic actions. We investigated the role of endogenous tTG in cardiac remodeling and dissected the role of enzymatic and non‐enzymatic actions in vivo and in vitro. tTG expression was upregulated in the pressure‐overloaded mouse heart and was localized in the cardiomyocytes, interstitial cells and in the extracellular matrix. tTG −/− mice had increased mortality and enhanced ventricular dilation following pressure overload, but were protected from diastolic dysfunction. In vivo, tTG loss was associated with a hypercellular cardiac interstitium, reduced collagen cross‐linking, and with accentuated matrix metalloproteinase (MMP)2 activity in the pressure‐overloaded myocardium. In contrast, inhibition of tTG activity through administration of a specific small molecule inhibitor was protective, attenuating hypertrophy, fibrosis and dysfunction. In vitro, tTG did not modulate TGF‐β‐mediated responses in cardiac fibroblasts, but exerted antiproliferative actions and, when bound to the matrix, induced synthesis of tissue inhibitor of metalloproteinases (TIMP)‐1. Following pressure overload, endogenous tTG exerts enzymatic actions that increase ventricular stiffness promoting hypertrophy and fibrosis, and protective non‐enzymatic effects that prevent chamber dilation.Support or Funding InformationSupported by NIH grants R01 HL76246, R01 HL85440 and R01DK63158. Dr. Shinde is supported by an American Heart Association Founders’ affiliate postdoctoral grant.
Activation of SIRT1 Attenuates Klotho Deficiency‐induced Arterial Stiffness and Hypertension via Enhancing AMPKα and eNOS activity
Authors: Gao Diansa , Zuo Zhong , Ali Quaisar , Tian Jing , Lei Han , Lin Yi , Sun Zhongjie 
Published: November 2023

ObjectiveHaplodeficiency of klotho causes arterial stiffness and hypertension. W found that Klotho, an aging‐suppressor protein, was significantly decreased in serum of patients with arterial stiffness and hypertension. Deficiency of klotho induces arterial stiffness and hypertension. This study was designed to investigate the underlying mechanism.Methods & ResultsKlotho mutant heterozygous (+/−) mice and wild‐type (WT) mice were used. Notably, expression and activity of SIRT1, an important deacetylase, were significantly decreased in aortas of KL (+/−) mice which coincided with elevated pulse wave velocity (PWV) and blood pressure (BP). Treatment with SRT1720 (15mg/kg per day IP), a specific SIRT1 activator, abolished klotho deficiency‐induced arterial stiffness and hypertension in KL (+/−) mice. Interestingly, Klotho deficiency was associated with decreases in the activity of AMP‐activated protein kinase (AMPKa) and endothelial nitric oxide synthase (eNOS) in aortas which can be abolished by SRT1720. Furthermore, klotho deficiency upregulated NADPH oxidase activity and superoxide production, increased collagen expression, and decreased elastin expression in the media of aortas. These klotho deficiency‐related changes were blocked by SRT1720.ConclusionThis study provided the first evidence that Klotho deficiency‐induced arterial stiffness and hypertension via downregulation of SIRT1. Pharmacological activation of SIRT1 may be an effective therapeutic approach for arterial stiffness and hypertension.Support or Funding InformationSupported by NIH R01 HL118558, DK093403, HL105302, and HL102074.