Daily Oral Administration Research Articles

In vivo Antioxidant Related Effect of Orally Administered Aqueous Extract of Saliva triloba in Human Volunteers

Saliva triloba, belongs to the Lamiaceae family, is one in all the vital medicinal plant species. This work aims to study the antioxidant-related effects of trilobite saliva in the human body through in vivo studies and the effects on liver, kidney, and heart function tests. For five days, nine healthy participants consumed 250 mL of trilobite saliva extract orally. On the fifth day, blood samples were taken one hour before and after the first dosage of water extract (samples I and II, respectively), and again one day after the last dose (ie, day 6, sample III). Before the first dosage, the first blood sample was taken (ie sample I) was used as a control for the subsequent II and III samples. Subsequent determinations were performed: serum total antioxidant status (TAS), red blood cell reduced glutathione (GSH), red blood cell superoxide dismutation (SOD) A activity, malondialdehyde (MDA), and serum-selected biochemical tests. After 5 days of oral administration of trilobite saliva extract in healthy volunteers, serum TAS, erythrocyte GSH and erythrocyte SOD activity were significantly increased, and had no influence on serum biochemical examinations of kidney, liver, heart, pancreas, etc., contrasted with zero-time management. In Conclusion, salivary clover extract has effective anti-oxidation related effects in vivo. Because these findings were obtained in healthy people without oxidative stress, it means that clover saliva will enhance the bottom line of the defense system against probable oxidative stress while having no adverse effects, decreasing or avoiding pathological diseases associated with oxidative stress

Protective effects of apigenin on methylmercury-induced behavioral/neurochemical abnormalities and neurotoxicity in rats.

Methylmercury (MeHg) is a neurotoxin that induces neurotoxicity and cell death in neurons. MeHg increases oligodendrocyte death, glial cell activation, and motor neuron demyelination in the motor cortex and spinal cord. As a result, MeHg plays an important role in developing neurocomplications similar to amyotrophic lateral sclerosis (ALS). Recent research has implicated c-JNK and p38MAPK overactivation in the pathogenesis of ALS. Apigenin (APG) is a flavonoid having anti-inflammatory, antioxidant, and c-JNK/p38MAPK inhibitory activities. The purpose of this study is to determine whether APG possesses neuroprotective effects in MeHg-induced neurotoxicity in adult rats associated with ALS-like neuropathological alterations. In the current study, the neurotoxin MeHg causes an ALS-like phenotype in Wistar rats after 21 days of oral administration at a dose of 5mg/kg. Prolonged administration of APG (40 and 80mg/kg) improved neurobehavioral parameters such as learning memory, cognition, motor coordination, and grip strength. This is mainly associated with the downregulation of c-JNK and p38MAPK signaling as well as the restoration of myelin basic protein within the brain. Furthermore, APG inhibited neuronal apoptotic markers (Bax, Bcl-2, and caspase-3), restored neurotransmitter imbalance, decreased inflammatory markers (TNF- and IL-1), and alleviated oxidative damage. As a result, the current study shows that APG has neuroprotective potential as a c-JNK and p38MAPK signaling inhibitor against MeHg-induced neurotoxicity in adult rats. Based on these promising findings, we suggested that APG could be a potential new therapeutic approach over other conventional therapeutic approaches for MeHg-induced neurotoxicity in neurobehavioral, molecular, and neurochemical abnormalities.

DIAGNOSIS AND TREATMENT OF FELINE IMMUNE-MEDIATED THROMBOCYTOPENIA

Immune-mediated thrombocytopenia is an immune-mediated thrombocytopenia (IMT) of increased platelet destruction. The purpose of this article is to describe the diagnosis and treatment of IMT in a cat, which may be helpful for the diagnosis and treatment of Thrombocytopenia. The diagnosis and treatment of IMT were analyzed by case information, clinical examination, laboratory examination, imaging examination, treatment and prognosis. The results showed that the clinical manifestations of IMT were ocular hemorrhage, massive ecchymosis in abdomen, severe decrease of platelet concentration in whole body and black stool. The diagnosis of Thrombocytopenia was made by combining clinical symptoms, blood routine examination and ultrasound examination to exclude infectious diseases and parasitic infections. After 5 days of oral administration of Microlone and Synulox, the symptoms of the cats tended to smooth, and the prognosis was good. This paper has provided case study and a clinical practical reference for the diagnosis and treatment of Feline Immune-Mediated Thrombocytopenia.

Green Synthesis of Purslane Seeds-Selenium Nanoparticles and Evaluate its Cytokines Inhibitor in Lead Acetate Induced Toxicity in Rats

Lead is a heavy metal that produces toxicity in both humans and animals due to oxidative stress. The aim of this study was to synthesize through green channel and evaluate hepatoprotective effect of purslane seeds-selenium nanoparticles (PS-Se NPs) against lead acetate-induced liver toxicity. The PS-Se NPs were made and their particle size and zeta potential were measured. In addition, the IC50 of SP-Se NPs against the HepG-2 liver cancer cell line as well as and LD50 in rats were determined. The PS-Se NPs had a mean particle size of around 54.70 ± 4.55 nm, a negative zeta potential of +17.25 and the observed shape of the nanoparticle was spherical. Also, the IC50 of PS-Se NPs against HepG-2 liver carcinoma cell line is 92.66 μg/mL and the LD50 is 1695 mg/kg b.w. Daily oral administration of PS-Se NPs at concentrations of 33.9 and 84.75 mg/kg b.w. for 30 days to rats treated with lead acetate (0.6 mg/kg b.w.) resulted in significant improvements in ALT, AST, ALP, TNF-α, Bcl-2, P53 and MDA. In contrast, oral administration of PS-Se NPs increased the levels of SOD, GPx and GSH in the liver tissue of lead acetate-treated rats. Furthermore, PS-Se NPs almost normalized these effects in histoarchitecture and ultrasound scanning examination of liver tissue. The PS-Se NPs have protective activity against lead acetate-induced liver tissue injury in rats, according to the biochemical, histological and ultrasound scanning findings.

Synthesis of CEG-AgNPs as a Promising MMPs/COX-2/Bcl-2 Signaling Pathway Modulator in BaP-Induced Lung Carcinoma

Cucurbitacins are a class of highly oxidized tetracyclic triterpenoids. It’s hydrophobic properties and poor solubility in water, polymeric micellar systems exhibited improved antitumor efficacy because of a better solubilization and targeting after local and/or systemic administration. The aim of the present work was to evaluate the anticancer activity of CEG-AgNPs against benzo[a]pyren (BaP)-induced lung carcinoma. CEG-AgNPs was prepared, characterized and evaluated for its cytotoxic activity against A549 lung carcinoma cell line. Also, the anticancer activity of CEG-AgNPs (70.25 mg/kg) against BaP-induced lung carcinoma was evaluated in vivo, using 30 adult mice for 43 days. IC50 of CEG-AgNPs against A549 lung carcinoma cell line were approximately 94.47 μg/mL. Administration of BaP (50 mg/kg b.w.) to mice induced lung carcinoma with a significant increase in lung MMP-2, MMP-9, MMP-12, MDA, IL-6 and NF-κB as well as significant decreased in lung CAT, GPx and GSH level. Also, treatment with BaP produced significant increase in lung VEGF-C, COX-2 and Bcl-2 gene expression as compared to control group. Daily oral administration of CEG-AgNPs to mice treated with BaP showed a significant protection against-induced increase in lung MMP-2, MMP-9, MMP-12, MDA, IL-6 and NF-κB levels. The treatment also resulted in a significant increase in lung CAT, GPx and GSH level. In addition, the CEG-AgNPs could inhibit lung VEGF-C, COX-2 and Bcl-2 gene expression as compared to BaP treated mice. The histological and MRI examination showed that a significant normalization has been observed through in CEG-AgNPs treated mice. The biochemical, histological and MRI results showed that CEG-AgNPs have potent anticancer activity against BaP-induced lung carcinoma through modulating multiple cellular behaviours and signaling pathways leading to the suppression of adaptive immune responses.

Edible Green Solvent for Optimized Catechins Extraction from Green Tea Leaves: Anti-Hypercholesterolemia.

Catechin polyphenols are the major bioactive ingredients in green tea with various human health benefits. Extraction of catechins from green tea (GTE) leaves at optimized standard conditions is still a challenging approach. An optimized, rapid, and economic extraction method is industrially needed. We hypothesized that certain extraction techniques in the presence of natural polymers and antioxidants might improve GTE catechin extraction yield and its biological activity. The effect of microwave (30–60 seconds irradiation in a typical kitchen microwave) assisted extraction (MAE) and ultrasonic assisted extraction (UAE) techniques were evaluated separately and in combination. To study the effect of the extraction solvent, nine edible green solvent combinations were investigated namely water, ascorbic acid, chitosan/ascorbic acid, carboxymethylcellulose /ascorbic acid, methylcellulose /ascorbic acid, chitosan/methylcellulose/ascorbic acid, methylcellulose, chitosan/acetic acid, and ethanol. The amounts of extracted catechins from green tea leaves were quantified with HPLC-UV.Data showed that the use of MAE & UAE technique was the optimal in producing a higher extraction yield of catechins. Chitosan/ascorbic acid was the optimized solvent with high extraction efficiencies of catechins. Studies in high fat diet fed animals demonstrated significant reduction of total cholesterol and LDL-C by GTE after 3 weeks of oral daily administration. In conclusion, efficient extraction, and stabilization of catechins from green tea leaves demonstrated a significant lowering of high fat diet-mediated elevation in blood cholesterol and LDL-C levels.

Оценка информативности морфологических изменений эритроцитов при гипервитаминозе А и выбор прогностических признаков его возникновения

Introduction. Morphometric methods allowing quantitative estimation of changes in a variety of parameters of studied histological objects are often used to evaluate the results of experimental research. The selection of a more limited number of parameters characterized by diagnostically significant evidence of these changes is based on their informative value. The aims of the study were to evaluate the informative value of changes in morpho-densitometric parameters of erythrocytes under hypervitaminosis A and select signs to predict its clinical occurrence. Materials and methods. We used male Wistar rats randomized into two groups of 9 animals each. Hypervitaminosis A was induced by oral daily administration of an oily solution of retinol palmitate. The animals of the control group received an oil base. The morphodensitometric parameters of erythrocytes were measured in blood smears. We compared and evaluated the informativeness of differences in both mean values and t frequency distributions of the parameters in the samples. Results. Erythrocyte morphodensitometric parameters altered with retinol palmitate administration. The values of a number of measured parameters differed significantly from those in the control even before the onset of signs of hypervitaminosis A. We identified a list of informative parameters and selected prognostic criteria for retinol palmitate overdose, namely reduced erythrocyte surface area and increased curvature of the descending torus. A more limited number of diagnostically significant signs were obtained when assessing the informative value of differences in the frequency distributions of morphodensitometric parameters. Conclusion. In morphometric studies, in addition to estimating the informativeness of differences in averaged values, that of differences in their frequency distribution can be used, thus limiting the number of parameters measured under similar experimental conditions. Keywords: hypervitaminosis A, erythrocytes, morphodensitometry, assessment of informativeness

Vitamin A intake forms resistance to hypervitaminosis A and affects the functional activity of the liver

Vitamin A intake forms resistance to hypervitaminosis A and affects the functional activity of the liver

Gastroprotective and Antioxidative Effects of the Traditional Thai Polyherbal Formula Phy-Blica-D against Ethanol-Induced Gastric Ulcers in Rats.

Phy-Blica-D is a traditional Thai polyherbal formula that has reduced oxidative stress in non-communicable diseases. However, evidence supporting the gastroprotective effects of Phy-Blica-D has not been previously reported. Therefore, this study aimed to evaluate the gastroprotective effects of Phy-Blica-D against gastric ulcers in rats and investigate the potential underlying mechanism. To estimate the possible mechanisms of action, we examined the levels of oxidative stress markers, such as reactive oxygen species (ROS) and malondialdehyde (MDA), as well as antioxidant enzymes, including catalase (CAT), superoxide dismutase (SOD), and glutathione (GSH). According to our results, rats treated with only 80% ethanol (vehicle group) exhibited significant increases in their ulcer area and ulcer index (UI). Moreover, the levels of ROS and MDA markedly increased in the vehicle group compared with the normal control group. Daily oral administration of Phy-Blica-D (500 and 1000 mg/kg) for 7 days not only significantly decreased the ulcer area and UI, but also remarkably decreased the ROS and MDA levels in gastric tissue. Gastric ulcers induced by ethanol had significantly decreased antioxidant enzyme activities (CAT and SOD) and non-enzymatic antioxidant (GSH), whereas pretreatment with Phy-Blica-D significantly improved the activities of CAT, SOD, and GSH. Moreover, after exposure to ethanol, the rats exhibited a significantly increased level of inducible nitric oxide synthase (iNOS), which was reduced after treatment with Phy-Blica-D. These findings suggest that Phy-Blica-D potentially exerts its gastroprotective effects by suppressing oxidative stress and stimulating antioxidant enzymes, which is one of the causes of destruction of cell membranes, and it is involved in the pathogenesis of acute gastric ulcers induced by ethanol.

Hypoglycemic, Hypolipidemic, Renal Protective and Antioxidant Activity of Annona muricata in Streptozotocin-Induced Diabetic Rats

Annona muricata, an herbal plant commonly used in traditional medicine to manage numerous diseases, diabetes as other diseases could be managed with herbal medicine. This study was designed to be investigated the antidiabetic, hypolipidemic, renal protective, and antioxidant effects of aqueous extracts of Annona muricata as used alone or combined with metformin in streptozotocin (STZ)-induced diabetic rats. Methods: the study was involved twenty adult Wister albino rats in four groups (five rats in each) and designated as groups, control group (1), and experimental groups (2, 3, 4). Diabetes was induced in experimental groups by 60 mg/kg intravenous streptozotocin injection. Group 2: serves as a diabetic control group, Group 3: diabetic rats treated with oral administration of 100 mg/kg of Annona muricata aqueous extract, Group 4: diabetic rats treated with combination (100 mg/kg aqueous extract of Annona muricata + 50 mg/kg metformin). The treatment continuous daily for 4 weeks to determine the levels of blood glucose and biochemical analysis. Result: aqueous extract of Annona muricata was reduced the serum glucose level effectively in streptozotocin-induced diabetic rats, by 48% and 55% after 28 consecutive days of treatment when used alone and with metformin, respectively. These compared to the preliminary values and the reduction was statistically significant compared to a diabetic control group. Daily oral administration of 100 mg/kg aqueous extract of Annona muricata for 4 weeks to streptozotocin-induced diabetic rats significantly reduced the level of total cholesterol, urea, creatinine, and MDA, whereas the reduction was non-significant in triglyceride and VLDL-cholesterol levels as compared to the non-treated diabetic group. However, the reduction is more significant in streptozotocin-induced diabetes rats that were treated with a combination of Annona muricata and metformin when compared to the diabetic control group. Conclusion: Aqueous extracts of Annona muricata have anti-diabetic action through their hypoglycemic, hypolipidemic, renal protective, and antioxidant effects in streptozotocin-induced diabetic rats. Thus, can be used alone or with anti-hyperglycemic drugs as metformin in the management of DM. The combination is preferred in severe hyperglycemic cases with more hypoglycemic effect requirements.

Erteng Tongbi Decoction ameliorates collagen-induced arthritis in mice via modulating T cell differentiation and cytokines balance

Ethnopharmacological relevanceHerbs have been commonly used for the treatment of rheumatoid arthritis (RA). It has been verified that Erteng Tongbi Decoction has good therapeutic effects on RA, while, relatively few studies on the relationship between its components and anti-rheumatoid efficacy were carried out.Aim of the study: To discuss the anti-RA effects of Erteng Tongbi Decoction on collagen-induced arthritis (CIA) in mice and the influence of T cell differentiation and cytokines balance. Materials and methodsSeparate researches on the two traditional Chinese medicines of the Erteng Tongbi Decoction were conducted. First, a murine peritoneal macrophage model was established, and then the cytokines levels and macrophage maturity were measured to select the best extraction solvent. Furthermore, ethanol extracts were partitioned successively with four kinds of solvents, and the anti-inflammatory parts were selected by the same vitro model. Subsequently, mice were arbitrarily divided into control, CIA model, positive control, effective parts alone or in combination. After 20 days of oral administration, the weight, hind paw volume, rheumatism index value, and the pathological changes were checked to assess the obvious level of arthritis. Furthermore, the levels of IL-6, TNF-α, IL-10, and IL-17A in serum and the balance of Th17/Treg and Th1/Th2 cells in spleen and mesenteric lymph nodes (MLN) was detected. Finally, the major active constituents were identified. ResultsIn vitro, the anti-inflammatory effects of ethanol extracts was much better than water extract. In addition, the effective parts of Celastrus orbiculatus Thunb. ethanol extract were petroleum ether parts and dichloromethane parts. The effective parts of Spatholobus suberectus Dunn. ethanol extracts was petroleum ether parts and ethyl acetate parts screened. In vivo, effective parts compatibility could inhibit the progression of inflammation by modulating T cell differentiation and cytokines balance. Constituent analysis revealed that effective parts contained sesquiterpenes alkaloids, phenolic acids, and flavanols. ConclusionsErteng Tongbi Decoction could notably ameliorate CIA mice by modulating T cell differentiation and cytokines balance and support its application in folk medicine.

A Traditional Chinese Medicine Plant Extract Prevents Alcohol-Induced Osteopenia.

Traditional Chinese medicine (TCM) has been practiced in the treatment of bone diseases and alcoholism. Chronic excessive alcohol use results in alcohol-induced bone diseases, including osteopenia and osteoporosis, which increases fracture risk, deficient bone repair, and osteonecrosis. This preclinical study investigated the therapeutic effects of TCM herbal extracts in animal models of chronic excessive alcohol consumption-induced osteopenia. TCM herbal extracts (Jing extracts) were prepared from nine Chinese herbal medicines, a combinative herbal formula for antifatigue and immune regulation, including Astragalus, Cistanche deserticola, Dioscorea polystachya, Lycium barbarum, Epimedium, Cinnamomum cassia, Syzygium aromaticum, Angelica sinensis, and Curculigo orchioides. In this study, Balb/c male mice were orally administrated alcohol (3.2 g/kg/day) with/without TCM herbal extracts (0.125 g/kg, 0.25 g/kg, or 0.5 g/kg) by gavage. Our results showed that after 50 days of oral administration, TCM herbal extracts prevented alcohol-induced osteopenia demonstrated by μ-CT bone morphological analysis in young adults and middle-aged/old Balb/c male mice. Biochemical analysis demonstrated that chronic alcohol consumption inhibits bone formation and has a neutral impact on bone resorption, suggesting that TCM herbal extracts (Jing extracts) mitigate the alcohol-induced abnormal bone metabolism in middle-aged/old male mice. Protocatechuic acid, a natural phenolic acid in Jing extracts, mitigates in vivo alcohol-induced decline of alkaline phosphatase (ALP) gene expression in the bone marrow of Balb/c male mice and in vitro ALP activity in pre-osteoblast MC3T3-E1 cells. Our study suggests that TCM herbal extracts prevent chronic excessive alcohol consumption-induced osteopenia in male mice, implying that traditional medicinal plants have the therapeutic potential of preventing alcohol-induced bone diseases.

Amelioration of the hepatotoxic effects of nonsteroidal drugs using vitamin C and determination of their relationship with the lipid profile

ObjectiveDespite the various clinical benefits of nonsteroidal anti-inflammatory drugs, their frequent and prolonged use has led to numerous health risks, including hepatotoxicity. Hepatotoxicity mediated by oxidative stress can affect the lipid profile. The objective was to investigate whether post-treatment with vitamin C can ameliorate the effects of diclofenac and naproxen in the livers of prepubertal rats and to highlight their relationship with lipid profile. MethodsForty prepubertal female albino rats were distributed among the control group, the diclofenac-administered group (5 mg/kg/day), and the naproxen-administered group (50 mg/kg/day). This study included two phases. In Phase 1, only five rats from each group were dissected after 21 days of oral administration to assess the hepatotoxic effects of nonsteroidal drugs. In Phase 2, five of the remaining animals in each intervention group were post-treated with 25 mg/kg/day of vitamin C for an additional 21 days. After the administration and post-treatment, serum biochemical parameters and histopathological signs were evaluated. ResultsExtreme elevation in the levels of aspartate and alanine aminotransferases was observed in the diclofenac and naproxen groups compared with those in the control (p < 0.001). In addition, the levels of high- and low-density lipoproteins were significantly impacted in these drug groups (p < 0.01, p < 0.05 respectively). Several pathological signs in the liver histology were observed in both drug groups. After post-treatment with vitamin C, noticeable amelioration of these alterations was observed. There were slightly elevation in the liver enzymes and insignificant increase and decrease in the high and low-density lipoproteins respectively. ConclusionVitamin C post-treatment ameliorated the hepatotoxicity induced by diclofenac sodium and naproxen.

Abstract P165: MRTX1719: A first-in-class MTA-cooperative PRMT5 inhibitor that selectively elicits antitumor activity in MTAP/CDKN2A deleted cancer models

Abstract Nearly all oncology therapies target proteins that are hyperactivated in cancer; however, developing precision medicines for cancers that harbor a specific tumor suppressor gene loss remains one of the most significant challenges in cancer research. Previous studies demonstrated cancer cell lines harboring homozygous deletion of the MTAP gene (MTAP del) are selectively vulnerable to shRNA-mediated PRMT5 inhibition based on the concept of synthetic lethality. MTAP is adjacent to, and co-deleted with, the most commonly deleted tumor suppressor gene, CDKN2A. MTAP is deleted in ~10% of all cancers and is enriched in NSCLC, pancreatic cancer, mesothelioma, MPNST, and several other cancers. MTAP del cells accumulate the metabolite methylthioadenosine (MTA) which binds to, and partially inhibits, PRMT5 activity; however, current clinical-stage small molecule PRMT5 inhibitors have binding modes that are mutually exclusive with MTA binding and therefore are not selective for MTAP del cancers. To develop a potential precision medicine for MTAP del cancers, compounds that selectively bind to the PRMT5/MTA complex were discovered via a fragment-based approach using SPR complemented with structural insight utilizing X-ray crystallography. Optimization of the lead series identified MRTX1719, a potent inhibitor of PRMT5 activity (8 nM IC50) and cell viability (12 nM IC50) in HCT116 MTAP del cells with greater than 50-fold selectivity compared with HCT116 MTAP WT cells. MRTX1719 possesses favorable drug-like characteristics and human predicted oral PK properties. Daily oral administration of MRTX1719 to tumor xenograft-bearing mice demonstrated dose-dependent inhibition of PRMT5-dependent symmetric dimethylarginine (SDMA) protein modification in MTAP del tumors that correlated with antitumor activity. In contrast, minimal inhibition of PRMT5-dependent SDMA and tumor growth inhibition was observed in MTAP WT tumor xenografts. Repeat dosing in mice also showed reduced effects in bone marrow compared to nonselective PRMT5 inhibitors currently in clinical trials, suggesting MRTX1719 may have an improved therapeutic index relative to these agents. MRTX1719 treatment resulted in dysregulated post-translational modification of key cancer and cell growth-related proteins, dysregulated RNA splicing, decreased proliferation and increased apoptosis in MTAP del cancer cells. Screens evaluating the effect of MRTX1719 on cell viability across panels of cancer cell lines and/or patient-derived xenograft tumor models demonstrated broad anti-tumor activity across MTAP-deleted tumor models. Select targeted therapy and chemotherapy combinations augmented the activity of MRTX1719 and in some models converged on phospho-Rb inhibition to block tumor growth. Together, these data demonstrate that MRTX1719, a selective PRMT5/MTA inhibitor, represents a promising therapeutic strategy for patients with MTAP del cancers. Citation Format: Christopher R. Smith, Lars D. Engstrom, Svitlana Kulyk, Ruth Aranda, Laura Waters, Krystal Moya, Victoria Bowcut, Allan Hebbert, David Trinh, David M. Briere, J. David Lawson, Jeff Clarine, Lisa Rahbaek, James G. Christensen, Matthew A. Marx, Peter Olson. MRTX1719: A first-in-class MTA-cooperative PRMT5 inhibitor that selectively elicits antitumor activity in MTAP/CDKN2A deleted cancer models [abstract]. In: Proceedings of the AACR-NCI-EORTC Virtual International Conference on Molecular Targets and Cancer Therapeutics; 2021 Oct 7-10. Philadelphia (PA): AACR; Mol Cancer Ther 2021;20(12 Suppl):Abstract nr P165.

Butein-Enriched Fractions of Butea monosperma (Lam.) Taub. Flower Decrease Weight Gains and Increase Energy Expenditure in High-Fat Diet-Induced Obese Mice.

Butea monosperma (Lam.) Taub. has been applied to treat inflammatory, metabolic, and infectious diseases. However, the antiobesity effects of B. monosperma (Lam.) Taub. flower (BMF) and the underlying mechanisms have not been determined. In this study, we analyzed the various extraction procedures, investigated the antiobesity effects, and identified the main chemical constituents of BMF. The BMF was subjected to acid hydrolysis in 5% H2SO4 in methanol at 50°C for 48 h and partitioned with ethyl acetate. The acid-hydrolyzed BMF ethyl acetate extracts (BMFE) strongly induced the expression of uncoupling protein 1 (Ucp1) and other thermogenic genes in C3H10T1/2 adipocytes. Daily oral administration of 70 mg/kg BMFE (BMFE70) to mice with diet-induced obesity resulted in less body weight gain, increased glucose tolerance, higher rectal temperature, and increased oxygen consumption. Qualitative and quantitative analyses along with treatments in Akt1 knockout mouse embryonic fibroblasts indicate that butein is a major active ingredient of BMFE, which stimulates Ucp1 gene expression. These data show the effects of butein-containing B. monosperma flower extract on thermogenesis and energy expenditure, further suggesting the potential role of BMFE as a functional ingredient in obesity and related metabolic diseases.

Abstract 10978: Ecklonia Stolonifera Okamura Extract Suppresses Cardiac Hypertrophy and Systolic Dysfunction Through P300-HAT Inhibition

Introduction: Heart failure is the leading cause of death in the world. Cardiomyocyte hypertrophy is observed during the development of heart failure, suggesting that its inhibition is a potential target for the prevention and treatment of heart failure. In this study, we screened a natural compound library using cultured cardiomyocytes and found that Ecklonia stolonifera Okamura extract (ESE) suppressed cardiomyocyte hypertrophy. Hypothesis: ESE, a perennial brown alga, has been reported to have various bioactive effects, such as antioxidant and anti-inflammatory activity, but its effect on heart failure is still unclear. Therefore, we investigated whether ESE has an inhibitory effect on cardiomyocyte hypertrophic response and on the progression of heart failure in post-myocardial infarction (MI) rats. Methods and Results: First, primary cultured cardiomyocytes from neonatal rats were treated with ESE and then stimulated with phenylephrine (PE) for 48 hours. ESE (1000 μg/mL) significantly suppressed PE-induced increases in cardiomyocyte surface area, hypertrophic response gene transcription, and acetylation of histone H3K9. An in vitro p300-HAT assay indicated that ESE directly inhibited p300-HAT activity (IC50: 505 μg/mL). Next, one week after the ligation of the left anterior descending artery, rats with moderate MI (left ventricular fractioning shorting (LVFS) &lt;40%) were randomly assigned to three groups: vehicle (saline) (n=9), ESE (0.3 g/kg) (n=10), or ESE (1 g/kg) (n=10). Daily oral administration was repeated for 8 weeks. After treatment, LVFS was significantly higher in the ESE (1 g/kg) group (23.3 ± 0.7%, p&lt;0.05) than in the vehicle group (16.6 ± 1.3%). ESE treatment significantly suppressed MI-induced increases both in myocardial cell diameter and in the mRNA levels of hypertrophic response genes. ESE also inhibited MI-induced perivascular fibrosis and the acetylation of histone H3K9. Conclusion: These results suggest that ESE suppresses both hypertrophic responses in cardiomyocytes and the development of heart failure by inhibiting p300-HAT activity. Further studies are needed to clarify the effectiveness of ESE for heart failure therapy.

Abstract 10912: Sarpogrelate, an Antiplatelet Agent, Suppressed Cardiac Hypertrophy and Systolic Dysfunction in a 5-ht 2a Receptor-Independent Manner

Introduction: Recently, the cost of new drug development is increasing year by year, and drug repositioning is being used as a strategy for developing new treatments by saving time and costs. We used a library of approved drugs to screen for compounds that suppress cardiomyocyte hypertrophy and identified the antiplatelet drug sarpogrelate, a selective serotonin-2A receptor antagonist, as a candidate for heart failure therapy. In this study, we investigated the effect of sarpogrelate on cultured cardiomyocyte hypertrophy and development of heart failure. Methods &amp; Results: First, primary cultured cardiomyocytes were treated with 1 μM sarpogrelate and then stimulated with various hypertrophic stimuli (30 μM phenylephrine (PE), 0.1 μM angiotensin II and 0.1 μM endothelin 1). Sarpogrelate repressed hypertrophic responses induced by each stimulus, such as myofibrillar organization and increase in cell size. Moreover, PE-induced transactivation of the hypertrophic response genes promoter was significantly inhibited by sarpogrelate. Western blotting (WB) showed that sarpogrelate significantly suppressed PE-induced the phosphorylation of ERK1/2 and GATA4. Next, C57BL/6j male mice were subjected to a transverse aortic constriction (TAC) and sham operation. One day after the operation, the mice were randomly divided into 3 groups: sarpogrelate at 1 mg/kg or 5 mg/kg, and vehicle as a control. Daily oral administration was repeated for 8 weeks. Echocardiographic analysis showed that 5 mg/kg sarpogrelate significantly prevented a TAC-induced increase in posterior left ventricular wall thickness and a decrease in fractional shortening at 8 weeks after the operation. Five mg/kg sarpogrelate also suppressed a TAC-induced increase in HW/BW ratio, myocardial cell diameter, perivascular fibrosis, and mRNA levels of ANF and BNP. Moreover, the WB analysis showed that 5 mg/kg sarpogrelate significantly suppressed TAC-induced phosphorylation of the ERK1/2 and GATA4. Conclusions: Sarpogrelate significantly suppresses cardiomyocyte hypertrophy and the development of heart failure via at least, in part, by inhibition of ERK1/2-GATA4 pathway. These findings suggest that sarpogrelate may be an effective agent for heart failure therapy.

Abstract 10984: The Novel Curcumin Formulation, ASD-Cur Suppressed Myocardial Infarction Induced Heart Failure in Rats

Introduction: Curcumin prevents the development of heart failure, demonstrating that the compound is a potential treatment for the disease. Although curcumin is known to be safe, its therapeutic efficiency is limited due to its low bioavailability. To overcome this problem, we developed ASD-Cur, an amorphous formulation of curcumin. Hypothesis: ASD-Cur have greater therapeutic potency than Theracurmin ® , a colloidal submicron dispersion of curcumin. Methods: Male SD rats were orally administrated with ASD-Cur or Theracurmin ® (10 mg/kg curcumin). The plasma levels of curcumin were measured at 0.25, 0.5, 1, 2, 4 and 6 hours after administration. Twelve healthy volunteers, who had provided written informed consent, were administrated with ASD-Cur or Theracurmin ® containing 30 mg curcumin, and plasma curcumin concentrations were determined at 0.5, 1, 2, 4 and 8 hours. Next, the rats were subjected to MI or sham surgery. One week after surgery, the MI rats were randomly assigned to 4 groups: vehicle, ASD-Cur (0.2 mg/kg curcumin), or Theracurmin ® (0.2 or 0.5 mg/kg curcumin). Daily oral administration of these compounds was repeated for 6 weeks. After echocardiographic examination, myocardial cell diameter, perivascular fibrosis, mRNA levels, and the acetylation of histone H3K9 were measured. Results: After administration, the area under the plasma concentration-time curve (AUC 0-6h ) and the maximum plasma concentration (C max ) of ASD-Cur were 3.7-fold and 9.6-fold higher than those of Theracurmin ® , respectively. The AUC 0-8h and C max of ASD-Cur in humans were 3.4-fold and 5.4-fold higher than those of Theracurmin ® , respectively. Echocardiographic analysis showed that 0.2 mg/kg ASD-Cur and 0.5 mg/kg Theracurmin ® significantly improved both MI-induced reduction in FS and left ventricular hypertrophy to the same extent. Both treatments significantly suppressed MI-induced increases in myocardial cell diameter, perivascular fibrosis, mRNA levels of hypertrophic markers and cardiac fibrosis, and the acetylation of histone H3K9 to the same extent. Conclusions: These findings indicated that ASD-Cur has greater bioavailability than Theracurmin ® , and could have greater therapeutic potency towards MI-induced heart failure at a lower dose.

Abstract 11032: Compound A, a Ginger Extract, Significantly Suppresses Pressure Overload-Induced Systolic Dysfunction in Mice

Introduction: Cardiac remodeling induced by hypertrophic stresses such as hypertension is a compensatory mechanism associated with cardiomyocyte hypertrophy and cardiac fibrosis. As cardiac remodeling eventually leads to chronic heart failure (HF), there is an urgent need for compounds that can effectively suppress both cardiomyocyte hypertrophy and cardiac fibrosis. In this study, we used a natural compound library to screen for compounds that suppress these two mechanisms with cultured cardiomyocyte and cardiac fibroblasts, and identified compound A, a ginger extract, as a candidate for HF therapy. The purpose of this study is to investigate the effect of compound A on cardiomyocyte hypertrophy and cardiac fibrosis in vitro and on the development of HF in vivo . Methods &amp; Results: First, primary cultured cardiomyocytes were treated with 1 μM of compound A and stimulated with phenylephrine (PE). The results of immunofluorestaining showed that the compound significantly suppressed a PE-induced increase in the surface area of the cells. Quantitative PCR analysis demonstrated that the compound significantly suppressed PE-induced increases in the mRNA levels of hypertrophic response genes such as ANF and BNP. Next, primary cultured cardiac fibroblasts were treated with 1 μM of compound A and then stimulated with transforming growth factor-beta (TGF-β). The results of measurement of L-proline incorporation showed that the compound significantly suppressed TGF-β-induced incorporation. Quantitative PCR and western blotting demonstrated that the compound significantly suppressed TGF-β-induced mRNA and protein levels of α-smooth muscle actin (α-SMA). Finally, C57BL/6J mice were subjected to transverse aortic constriction (TAC) surgery, and were then given a daily oral administration of 1 mg/kg of compound A for 8 weeks. Echocardiographic analysis showed that the compound prevented TAC-induced increases in posterior wall thickness and systolic dysfunction. The compound also suppressed a TAC-induced increase in HW/BW ratio. Conclusions: Compound A suppressed TAC-induced development of HF via the suppression of cardiomyocyte hypertrophy and cardiac fibrosis. These findings suggest that compound A may be an effective agent for HF therapy.

CTNI-06. TRAM-01: A PHASE 2 STUDY OF TRAMETINIB FOR PATIENTS WITH PEDIATRIC GLIOMA WITH ACTIVATION OF THE MAPK/ERK PATHWAY

Abstract BACKGROUND Pediatric low-grade gliomas (PLGG) are the most frequent brain tumors in children. It is known that the majority of PLGG have activation of the MAPK/ERK pathway. METHODS This ongoing multicenter phase II trial includes three progressing/refractory PLGG groups: NF1 patients, KIAA1549-BRAF fusion patients and patients with other activation of the MAPK/ERK pathway (excluding V600E). The primary objective was to evaluate the overall response rate based on RANO criteria after daily oral trametinib administration for 18 cycles, lasting 28 days each. Secondary objectives include the assessment of progression-free survival, tolerability of trametinib, serum levels of trametinib and quality of life evaluation during treatment. RESULTS As of February 12 2021, 50 patients have been enrolled (NF1: n=10; KIAA1549-BRAF fusion: n=31; other: n=9 including 5 patients with FGFR1 alterations). Median age is 8.8 years (range 2.4-25.5). Median follow-up is 17.5 months (range 4.7-28.5). Forty-three patients are evaluable. The overall response includes: 4 partial response (PR) (9%), 18 minor response (MR) (42%), 17 stable disease (40%), 4 progressive disease (9%). Median time to response is 5.5 months (range 2.4-13.8). Median duration of response is 6.1 months (range 0.6-26.5). Progression free survival at 12 months is 79.9% (95% CI 68.5-93.6%) and median progression free survival has not yet been reached. Treatment was discontinued for 30 patients: 16 after completing 18 cycles as planned, 5 for progressive disease, 5 for adverse events, 4 for other reasons. A total of 8 patients progressed after discontinuation of treatment including 6 patients (37.5%) that completed 18 cycles. Five of these patients had achieved minor response prior to discontinuation. CONCLUSION Trametinib is a potentially effective targeted therapy for patients with recurrent/refractory PLGG. Treatment was overall well tolerated. This ongoing trial will continue to gather data on response rate, duration of response and safety of trametinib for PLGG.