Gastric Gavage Research Articles

Acute study and the absorption, distribution, and metabolomic profile of the natural compound propyl-propane-thiosulfonate from allium in rats

Propyl-propane thiosulfonate (PTSO) is an organosulfur compound found in Allium spp., commonly used in animal nutrition and various agri-food applications. Extensive studies have demonstrated the safety of PTSO for feed use, including investigations into genotoxicity, mutagenicity, and subchronic toxicity in rats. However no kinetic or metabolic study has been previously performed. This study aimed to conduct an in vivo toxicokinetic assessment of PTSO in rats. For this purpose, eighteen Sprague Dawley rats received an oral dose of 175 mg/kg via gastric gavage. Plasma and tissue samples (brain, stomach, liver, lung, kidney, spleen, and testicle) were collected at specific intervals (10 min, 20 min, 30 min, 1h, 2h, 4h, 6h, 8h, and 24h) and analyzed using Gas Chromatography-High-Resolution Mass Spectrometry, Pyrolysis Gas Chromatography -High-Resolution Mass Spectrometry, and Ultra-High Performance Liquid Chromatography-Tandem Mass Spectrometry to detect PTSO and its metabolites. PTSO was not detected in plasma or solid tissues throughout the sampling period; however, phase I and phase II metabolites were identified in both matrices. The toxicokinetic profile of s-propyl mercaptocysteine (CSSP), a significant metabolite, exhibited a well-correlated model. In the stomach, CSSP reached peak concentrations of 495.63 ± 6.53 ng/mL, while lower concentrations of 123.59 ± 8.35 ng/mL were observed in plasma. Furthermore, CSSP demonstrated high water solubility and rapid excretion, with a plasma half-life of 0.66 ± 0.05 h. Overall, these findings substantiate the safety profile of PTSO for specific agri-food applications under the conditions investigated.

Canagliflozin exerts anti-inflammatory and antioxidant effects in the heart and skin tissues: Biochemical and histopathological assessment in a model of accelerated aging induced by D-galactose in mice

Canagliflozin was assessed for its anti-inflammatory and antioxidant effects as an anti-aging drug in animal models. 50 Swiss albino male mice were divided into five groups; all groups received their intervention using gastric gavage; group 1 received normal saline for 14 weeks; group 2 received induction by D-galactose 200 mg/kg/day for seven weeks; and group 3 to 5 received the same induction for seven weeks, followed by another seven weeks of investigated drugs; group 3 received Vitamin C (100 mg/kg/day); group 4 received canagliflozin (3 mg/kg/day); and group 5 received canagliflozin (1 mg/kg/day). At the end of 14 weeks, all animals were euthanasiad, and heart and skin tissue were harvested for further analysis. Canagliflozin at both 1 and 3 mg/kg was successful in reducing the levels of inflammatory mediators (TNF-alpha and IL6), reducing levels of MDA, increasing the levels of SOD, and increasing the levels of collagen-1 and elastin in skin tissue. Additionally, 3 mg/kg Canagliflozin showed a better effect compared to 1 mg/kg regarding its effect on IL6, SOD, and elastin. Histopathologically, treatment with both doses of Canagliflozin attenuates abnormalities induced by D-galactose (bizarre, irregular, and hyperchromatic nuclei). Canagliflozin exhibits potent antioxidant and anti-inflammatory effects in living organisms, effectively prevents cardiac and skin damage generated by D-galactose, and possibly reduces aging.

Effect of sacubitril/valsartan and dapagliflozin on athletic performance; can the popular cardiac medications of recent years be used as doping agents?

Abstract Background/Introduction Recent guidelines have highlighted the positive impacts of sacubitril/valsartan and dapagliflozin on heart failure management [1]. Given their potential misuse as doping agents in professional sports, this study evaluates their effects on athletic performance. Purpose To investigate the effects of sacubitril/valsartan and dapagliflozin on athletic performance and cardiac functions in a rodent model. Methods The study had a randomized double-blind design. 24 Sprague-Dawley male rats aged 4 months, divided into control, sacubitril/valsartan, and dapagliflozin groups. The rats were weighed at the start of the experiment. The medications (dapagliflozin 1,5 mg/kg/1 ml/once a day, sacubitril/valsartan 60 mg/kg/1 ml/once a day to their respective groups, and normal saline for control group) were administered via gastric gavage for 30 days. The rats were weighed, examined with transthoracic echocardiography and put on rotarod at the start of the experiment, and on 15th and 30th days. The rats were put on forced swimming tests for 20 days. Exhaustion, loss of coordinated movements and failure to surface for 3 seconds after submersion in the water were defined as indications for ending the test as per the guidelines regarding rat exercise testing [2]. M-mode and Doppler measurements, forced swimming test durations, rotarod durations and weight were recorded on 0th, 15th and 30th days. Results Initial performance metrics, including echocardiography, weight, and rotarod tests, showed no significant differences among the groups at baseline. The distributions in data were non-normal. Over the course of the study, median swimming times significantly differed from the 9th session, with dapagliflozin-treated rats exhibiting a substantial increase in endurance. Specifically, dapagliflozin group's swimming times increased to a median of 3580 seconds by the 20th session, compared to 2382 seconds in the control group and 2591 seconds in the sacubitril/valsartan group, indicating a pronounced improvement in exercise capacity (P<0.0001)(Figure-1). Echocardiographic assessments revealed enhanced cardiac output and ejection fraction, particularly in the sacubitril/valsartan group, suggesting increased cardiac efficiency and potential athletic performance enhancement. Conclusion(s) The study demonstrates a significant improvement in athletic performance with dapagliflozin compared to sacubitril/valsartan. These findings suggest a potential for misuse in sports doping, emphasizing the need for stringent regulation and monitoring within competitive athletics.Figure-1

Comparative effects of ranolazine and trimetazidine on cardiac structure and exercise capacity in a healthy rat model: a new doping agent?

Abstract Background/Introduction Anti-anginal drugs that have been shown to improve myocardial energy use include ranolazine and trimetazidine [1]. An inquiry is warranted into their ability to impact cardiac functions and exercise capacity. Purpose This study aims to evaluate the effects of ranolazine; and trimetazidine as a positive control group on cardiac structure and exercise capacity in a rodent model in comparison with the control group. Methods In a randomized, controlled trial, 24 Sprague-Dawley rats were divided into three groups: control, ranolazine, and trimetazidine. Rats were given daily doses of ranolazine (50 mg/kg/1 ml) [2], trimetazidine (10 mg/kg/1 ml) [3], or saline (control) via gastric gavage for 30 days. At the beginning of the trial, as well as on the 15th and 30th days, the rats were weighed, placed on rotarod, and assessed using transthoracic echocardiography (Figure 1). The rats underwent 21 days of forced swimming testing. According to the guidelines for rat exercise testing [4], signs of exhaustion, lack of coordinated movements, and failure to surface for three seconds following submersion in the water were designated as grounds for terminating the test. Weight, rotarod durations, M-mode and Doppler measurements, and forced swimming test durations were recorded on the 0th, 15th, and 30th days. Results No discernible differences were found between the groups in the initial evaluations. By using echocardiography, the left ventricular masses expanded in all three, indicating the emergence of an athlete's heart. In comparison to the control group, the ranolazine and trimetazidine groups showed considerable increases in their ability to exercise. In particular, the swimming times of the trimetazidine group and the ranolazine group improved to a mean of 1889 seconds by the 21st session, 1700 seconds in the trimetazidine group, and 1548 seconds in the control group (Figure 2), respectively, indicating enhanced cardiac performance. Conclusion(s) Ranolazine exhibits significant potential to enhance exercise capacity and cardiac efficiency in rats, surpassing the effects observed with trimetazidine. Trimetazidine is already registered as a doping agent in WADA [5]. These findings highlight the potential use of ranolazine as a performance enhancing agent in healthy subjects for a malicious purpose of creating unfair advantages in professional sports. Further studies in human subjects are needed.Figure 1Figure 2

Failure of colonization following gut microbiota transfer exacerbates DSS-induced colitis.

To study the impact of differing specific pathogen-free gut microbiomes (GMs) on a murine model of inflammatory bowel disease, selected GMs were transferred using embryo transfer (ET), cross-fostering (CF), and co-housing (CH). Prior work showed that the GM transfer method and the microbial composition of donor and recipient GMs can influence microbial colonization and disease phenotypes in dextran sodium sulfate-induced colitis. When a low richness GM was transferred to a recipient with a high richness GM via CH, the donor GM failed to successfully colonize, and a more severe disease phenotype resulted when compared to ET or CF, where colonization was successful. By comparing CH and gastric gavage for fecal material transfer, we isolated the microbial component of this effect and determined that differences in disease severity and survival were associated with microbial factors rather than the transfer method itself. Mice receiving a low richness GM via CH and gastric gavage exhibited greater disease severity and higher expression of pro-inflammatory immune mediators compared to those receiving a high richness GM. This study provides valuable insights into the role of GM composition and colonization in disease modulation.

Carvacrol improves cognitive dysfunction by decreasing amyloid-β accumulation and regulating neuroinflammation in ovariectomized renovascular hypertensive rats.

Hypertension contributes to both the development and progression of brain damage and cognitive dysfunction in the postmenopausal period in women. Carvacrol (CAR), which can easily cross the blood-brain barrier, exhibits neuroprotective properties due to its antioxidant, anti-inflammatory, and anti-apoptotic effects. In the present study, we have examined the effect of CAR treatment on learning-memory impairment in a post-menopausal hypertensive rat model that was induced by ovariectomy following two-kidney, one-clip renovascular hypertension surgery. From the third week after the establishment of renovascular hypertension in ovariectomized rats, CAR (40 mg/kg) was administered once daily for consecutive 7 weeks by gastric gavage. Systolic blood pressure was estimated by the tail-cuff method once a week. At the end of the study, cognitive functions were evaluated with behavioral tests and also neurochemical changes were measured in serum, cortex, and hippocampus by ELISA test. Blood pressure was decreased with CAR treatment in hypertensive rats. Serum estrogen levels decreased in ovariectomized rats and did not change with CAR treatment. CAR demonstrated beneficial effects on learning and memory tests as determined by increased recognition index, the number of platforms crossed, and time spent in the target quadrant. Due to CAR treatment, there was a marked reduction in the hippocampal and cortex amyloid-β, osteopontin, interleukin-6 and tumor necrosis factor-alpha levels, and acetylcholinesterase activity, while an increment in neprilysin and interleukin-10 levels was found. In conclusion, since CAR suppressed amyloid-β deposition and neuroinflammation in ovariectomized-hypertensive rats, it is thought that it may be protective against memory disorders in postmenopausal hypertensive women.

Evaluation of antioxidant prophylactic effects of parsley (Petroselinum crispum) on sepsis following cecal ligation and puncture

Objective: Sepsis causes the release of free oxygen radicals that disrupt membrane integrity, and damage to Mitochondria due to the production of free oxygen radicals and oxidation leads to exacerbation of sepsis, Cecal ligation and puncture (CLP) in rats mimics the characteristics and course of clinical sepsis (Hubbard et al., 2005). Methods: We evaluated the antioxidant effects of Petroselinum crispum (Pc) in a cecal ligation and puncture (CLP)-induced rat sepsis model, in a rat model of sepsis caused by cecal ligation and puncture (CLP). Wistar albino rats were separated into four groups of eight groups: a sham group with incised and sutured abdomens; a Pc extract (PcE) group, which was given 2 g/kg parsley extract for 14 days by gastric gavage; a CLP group, which was subjected to sepsis caused by CLP; and a PcE + CLP group, which was given parsley extract for 14 days. PcE was given for 14 days, after which sepsis was induced via the CLP procedure. The groups were compared in terms of hemogram, biochemical and histological characteristics. Results: The administration of PcE before CLP-induced sepsis increases neutrophil counts, PLTs and TASs, which decrease with sepsis, and decreases biochemical changes (BUN, AST, ALT, LDH, TOS, and OSI), which increase with sepsis, to protect against sepsis. Compared with that in the CLP group, the severity of intestinal infiltration was significantly lower in the PcE + CLP group; however, the degree of epithelial damage in the PcE + CLP group was similar to that in the CLP group. In the PcE + CLP group, the crypt and villus lengths were greater, and the decrease in Paneth cell degranulation intensity was greater than that in the CLP group. Conclusion: Additionally, the morphology of the cells in the PcE + CLP group was similar to that in the sham group. PcE has potential as a prophylactic agent for sepsis.

The potential protective effect of propolis on diabetic nephropathy induced by streptozotocin in adult albino rats

ABSTRACT Diabetes mellitus is a common metabolic disorder. It is associated with serious life-threatening complications if not properly managed. The current study aimed at investigating the possible protective role of propolis on streptozotocin-induced diabetic nephropathy. A diabetic rat model was induced by a single intraperitoneal injection of 55 mg/kg streptozotocin. After 4 days, the diabetic rats received oral propolis (300 mg/kg/day) via gastric gavage for 28 days. Biochemical, histopathological and ultrastructural evaluations were performed. The results showed that: streptozotocin-induced diabetes was associated with a marked decrease in the serum high-density lipoproteins and antioxidant enzymes. However, a significant elevation in the levels of serum creatinine, blood urea nitrogen, uric acid, cholesterol, triglycerides and low-density lipoproteins was detected. Furthermore, streptozotocin treatment induced histopathological alterations of the renal cortex; in the form of distorted glomerular capillaries, widened Bowman’s space and signs of epithelial tubular degeneration. Ultra-structurally, thickening and irregularity of the glomerular basement membrane and podocytes foot processes effacement were observed. The tubular epithelial cells showed swollen vacuolated mitochondria, scarce basal infoldings and loss of microvilli. Conversely, propolis partially restored the normal lipid profile, antioxidant biomarkers and renal cortical morphology. Propolis exhibited a sort of renoprotection through hypoglycemic, anti-hyperlipidemic and antioxidant effects.

Nobiletin restores HFD-induced enteric nerve injury by regulating enteric glial activation and the GDNF/AKT/FOXO3a/P21 pathway

BackgroundTo explore whether nobiletin has a protective effect on high-fat diet (HFD)-induced enteric nerve injury and its underlying mechanism.MethodsAn obesity model was induced by a HFD. Nobiletin (100 mg/kg and 200 mg/kg) and vehicle were administered by gastric gavage for 4 weeks. Lee’s index, body weight, OGTT and intestinal propulsion assays were performed before sacrifice. After sampling, lipids were detected using Bodipy 493/503; lipid peroxidation was detected using MDA and SOD kits and the expression of PGP 9.5, Trem2, GFAP, β-tubulin 3, Bax, Bcl2, Nestin, P75 NTR, SOX10 and EDU was detected using immunofluorescence. The GDNF, p-AKT, AKT, p-FOXO3a, FOXO3a and P21 proteins were detected using western blotting. The relative mRNA expression levels of NOS2 were detected via qPCR. Primary enteric neural stem cells (ENSCs) were cultured. After ENSCs were treated with palmitic acid (PA) and nobiletin, CCK-8 and caspase-3/7 activity assays were performed to evaluate proliferation and apoptosis.ResultsHFD consumption caused colon lipid accumulation and peroxidation, induced enteric nerve damage and caused intestinal motor dysfunction. However, nobiletin reduced lipid accumulation and peroxidation in the colon; promoted Trem2, β-tubulin 3, Nestin, P75NTR, SOX10 and Bcl2 expression; inhibited Bax and GFAP expression; reduced NOS2 mRNA transcription; and regulated the GDNF/AKT/FOXO3a/P21 pathway. Nobiletin also promoted PA-induced impairment of ENSCs.ConclusionsNobiletin restored HFD-induced enteric nerve injury, which may be associated with inhibiting enteric nerve apoptosis, promoting enteric nerve survival and regulating the GDNF/AKT/FOXO3a/P21 pathway.

Hepatotoxic Effects of an Oral Amiodarone in White Albino Rats: Sub-Acute Biochemical, and Histopathological Assessments

Amiodarone, potent antidysrhythmic, widely used drug that has been associated with hepatic toxicity in long-term or excessive use. In the presented study twelve rats were allocated into two groups and given daily doses via gastric gavage orally for two weeks as follows; The first group served as a control normal group, whereas the second got amiodarone at a dosage of 300mg/kg/day orally. Liver tissues were processed for light microscopy, and blood samples were examined for serum transaminases (alanine aminotransferase (ALT) and aspartate aminotransferase (AST)) and phospholipase a2 enzyme (Pla2) (which are thought to be an indicator of phospholipidosis and lipid buildup). Amiodarone was shown to produce hepatic histological abnormalities such as blood vessel congestion, leucocytic infiltration, liver degeneration, and stages of steatosis and necrosis of hepatocytes. The biochemical assessments were revealed that there was an elevation in amiodarone group's ALT and AST levels as a result of hepatic necrosis leading to leakage of enzymes content, while serum Pla2 was lowered significantly. Also, histopathology indicates stages of steatosis (Lipid accumulation) in hepatocytes, which may lead to farther damage in the late stages of hepatotoxicity.

The Hepatoprotective Effect of Glycyrrhiza glabra Roots Extract Against Amiodarone Induced Hepatotoxicity in Male Rats

Amiodarone (AMD), a powerful antidysrhythmic medication, can cause hepatotoxicity when used long-term or in high doses. It affects both mitochondrial and lysosomal function because of its lipophilic nature and accumulation in tissues. It also has direct production of ROS. Glycyrrhiza glabra is one of the medicinal plants that have been commonly used for thousands of years. It possesses anti-ulcer, anti-inflammatory and antioxidant properties. In this study, the antihepatotoxic effect of Gg was evaluated against AMD liver toxicity in rats model, thirty rats were taken and divided evenly into five groups given daily doses by gastric gavage oral tube for 3 weeks as follows: the 1st group acted as a control group given D.W., and the 2nd group received AMD at a dose of 300mg/kg for 2 weeks. 3rd, 4th, and 5th groups were pretreated with Gg in doses of 200, 400, and 800 mg, respectively, for one week, then along with AMD for 2 weeks. Hepatic damage, revealed by histology and the increased activities of serum aspartate aminotransferase (AST), alanine aminotransferase (ALT), and serum phospholipase A2 (PLA2) was decreased in the AMD group. Gg extract significantly reduced the elevated AST and ALT levels caused by AMD intoxication, rendering PLA2 to its normal level. It also enhanced liver superoxide dismutase (SOD), glutathione (GSH), and malondialdehyde (MDA) levels. Gg extract markedly reverses the increased serum TNF levels induced by AMD; therefore, it may be considered a good hepatoprotective agent.

An analysis combining proteomics and transcriptomics revealed a regulation target of sea cucumber autolysis

Sea cucumber is a valuable seafood product and autolysis is the main concern for the aquaculture industry. This study employed proteomics and transcriptomics to investigate the autolysis mechanism of sea cucumbers. The fresh sea cucumber was exposed to UV light to induce autolysis. The body wall samples were cut off to analyze by proteomics and transcriptomics. The angiotensin-converting enzyme (ACE) inhibitor of teprotide and the activator of imatinib were gastric gavage to live sea cucumbers, respectively, to identify the regulation target. Autolysis occurrence was evaluated by appearance, soluble peptide, and hydroxyproline content. Four gene-protein pairs were ACE, AJAP10923, Heme-binding protein 2-like, and Ficolin-2-like. Only the ACE protein and gene changed synchronously and a significant down-regulation of ACE occurred in the autolysis sea cucumbers. Teprotide led to a 1.58-fold increase in the TCA-soluble protein content and a 1.57-fold increase in hydroxyproline content. No significant differences were observed between imatinib-treated sea cucumbers and fresh ones regarding TCA-soluble protein content or hydroxyproline levels (P > 0.05). ACE inhibitor accelerated the autolysis of sea cucumber, but ACE activator inhibited the autolysis. Therefore, ACE can serve as a regulatory target for autolysis in sea cucumbers.

Anti-Inflammatory, Wound Healing, and Anti-Diabetic Effects of Pure Active Compounds Present in the Ryudai Gold Variety of Curcuma longa.

Turmeric (Curcuma longa) contains curcumin, demethoxycurcumin (DMC), and bisdemethoxycurcumin (BDMC). Nevertheless, curcumin is the most researched active ingredient for its numerous pharmacological effects. We investigated the impact of these curcuminoids found in Ryudai gold, an approved cultivar of Curcuma longa, on wound healing, inflammation, and diabetes. Sub-planter injections of carrageenan induced acute paw inflammation in rats. The wound-healing ability of 1% curcuminoids was examined by making a 6 mm round wound on the shaved dorsum of the mice with a biopsy punch. A single intraperitoneal injection of streptozotocin (50 mg/kg) was used to induce diabetes in mice. Curcuminoids at a dose rate of 100 mg/kg body weight were used with feed and as a gastric gavage to treat diabetes and inflammation in experimental animals. Paw thickness was measured at 1, 3, and 6 h following carrageenan injection. After three hours, mean paw volume was 58% in carrageenan-injected mice, which was 35%, 37%, and 31% in the curcumin, DMC, and BDMC groups, respectively. Histopathology of the paw tissue demonstrated severe infiltration of inflammatory cells and thickening of the dermis, which were remarkably improved by the curcuminoids. The wound-healing abilities were significantly higher in the curcumin- (95.0%), DMC- (93.17%), and BDMC-treated (89.0%) groups, in comparison to that of the control (65.09%) group at day nine. There were no significant differences in wound-healing activity among the groups treated with 1% curcuminoids throughout the study. Streptozotocin-induced diabetes was characterized by an increased blood glucose (552.2 mg/dL) and decreased body weight (31.2 g), compared to that of the control rats (145.6 mg/dL and 46.8 g blood glucose and body weight, respectively). It also caused an increase in serum alanine aminotransferase (ALT; 44.2 U/L) and aspartate aminotransferase (AST; 55.8 U/L) compared to that of the control group (18.6 U/L and 20.1 U/L, respectively). Histopathological examination of the liver showed that diabetes caused hepatic cellular necrosis, congestion of the central vein, and parenchymatous degeneration. However, all three curcuminoids significantly decreased blood glucose levels, ALT, and AST and improved the histopathological score of the liver. These results evidenced that not only curcumin but also DMC and BDMC have potent anti-inflammatory, wound healing, and anti-diabetic efficacy, and the Ryudai gold variety of turmeric could be used as a functional food supplement.

Protective Feature of Anzer Propolis in Contrast-Induced Nephropathy in Rats

Aim: The study aims to ascertain whether Anzer Propolis, a natural antioxidant compound, can protect the kidneys from iopromide. Material and Method: This study was done in 2021. Four groups of 32 adult male Wistar albino rats, weighing between 250 and 300 g, were formed. Control (C) group contained eight rats. Intraperitoneal ethanol was administered to the Ethanol (E) group (n=8), 8 g/kg intraperitoneal iopromide to the contrast-induced nephropathy (CIN) group (n=8), and a 100 mg/kg dose of Anzer Propolis on the date of application to the Anzer Propolis group (AP) (n=8), as well as the prior day, by gastric gavage and then 8 g/kg iopromide was administered intraperitoneally. Termination was carried out at the 48th hour. Histopathological examination was performed for CIN in the right and left kidneys. Results: Following has been examined as a part of the research: serum urea, creatinine, IL6, interleukin (IL)-1β, total oxidant status (TOS), tumor necrosis factor (TNF-α), total antioxidant status (TAS), myeloperoxidase (MPO). TAS had a statistically significant increase in AP, in comparison to CIN, C and E groups (p

In Vivo Tissue Distribution of Microplastics and the Systemic Metabolic Changes After Gastrointestinal Exposure in Mice

Marcus M. Garcia, Aaron S. Romero, Seth D. Merkley, Jewel L. Meyer-Hagen, Charles Forbes, Matthew J. Campen, Julie G. In, and Eliseo F. Castillo Microplastics (MP) are slated to be the next environmental threat to humans. It is estimated humans consume approximately 5 grams of MP per week. To date, the systemic impact of MP ingestion on metabolic pathways in systemic organs have not been studied utilizing environmentally relevant doses and mixtures. To investigate the impact of ingested MPs on target metabolic pathways, mice were exposed to either polystyrene (5 μm) microspheres or a mixture of MPs consisting of polystyrene (5 μm), polyethylene (1-4 μm), and the biodegradability and biocompatible plastic, poly-(lactic-co-glycolic acid) (5 μm). Exposures were performed twice a week for four weeks at a dose of either 0, 2, or 4 mg/week via oral gastric gavage. Our findings demonstrate that, in mice, ingested MPs can pass through the gut barrier and accumulate in distant tissues. Additionally, we report on the metabolic changes that occur in the colon, liver and brain which show differential responses that are dependent on dose and type of MPs exposure. This study provides critical insight into the pervasive issues of plastic pollution and highlights dose-dependent and polymer type specific metabolic changes in the colon, liver and brain caused by MPs exposure. NIH/NIEHS 1R01 ES032037-01A1. This is the full abstract presented at the American Physiology Summit 2024 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.

Mechanism of action of Taohong Siwu decoction in the alleviation of primary dysmenorrhea.

As one of the most common gynecological disorders, PD significantly impacts the quality of life for women. TSD, a well-known traditional Chinese medical prescription, has gained popularity for its use in treating gynecological cold coagulation and blood stasis syndromes such as PD. However, the lack of comprehensive data hinders our understanding of its molecular mechanism. The objective of the present study is to investigate the therapeutic effects of TSD on PD and elucidate its plausible mechanism. HPLC was employed to confirm the presence of the principal metabolites of TSD. The rat model of PD was induced by OT exposure following IWM and EB pretreatment, and subsequently treated with TSD via gastric gavage. The effects and potential mechanisms of TSD on PD rats were explored, encompassing general behavior, morphological alterations in the uterus and ovaries, biochemical indicators in the uterus and serum, and levels of proteins related to the PI3K/AKT signaling pathway. Gallic acid, hydroxysafflower yellow A, albiflorin, paeoniflorin, and ferulic acid were determined to be the primary active metabolites of TSD. The pharmacological studies yielded results indicating the successful establishment of the PD model in rats. Additionally, TSD demonstrated its ability to protect PD rats by ameliorating general behavior, mitigating pathological damage to uterine and ovarian tissues, and modulating the expression levels of correlated factors (PGE2, PGF2α, Ca2+, TXB2, IL-6, TNF-α, NO, and COX-2) as well as p-PI3K/PI3K and p-AKT/AKT proteins. TSD exhibited protective effects against PD in rats through its interaction with multiple targets including P13K/AKT signaling pathway, indicating that TSD holds therapeutic potential for PD treatment and providing evidence supporting the rational utilization of TSD.

Investigation of the effects of cilostazol on the myocardial ischemia-reperfusion injury of rats.

<b><i>Background</i></b>. Myocardial ischemia, occurring as a consequence of imbalance between oxygen supply and demand, causes a rapid metabolic and structural<br /> impairment within the tissue. After a period of ischemia, sudden onset of reperfusion causes a transition to aerobic metabolism within living cells. Afterwards, emerging substrates initiate a chain of reactions leading to tissue injury. This situation is called “ischemia reperfusion injury”. Despite all technical advancements in anesthesia, myocardial protection and cardiac surgical techniques, we still face the clinical reflections of ischemia reperfusion (IR) injury.<br /> <b><i>Materials and methods</i></b>. The protective effect of cilostasole on IR injury in an animal model of experimental myocardial ischemia and reperfusion was investigated. In this regional myocardial ischemia model, male Wistar-Albino rats were used as subjects and they were allocated into three groups; ischemia (n=8), sham (n=8), and cilostazole (n=8). LAD was occluded for 45 minutes, and then reperfused for three hours. Rats received Cilostazole 20 mg/kg/d by gastric gavage once daily. During IR hemodynamic parameters were recorded. Serum analysis for CK-MB and Troponin T were analysed at 180th minute of ischemia. Ischemic zone was measured by dying with Evans Blue and infarct area was measured by dying with triphenyltetrazolium chloride.<br /> <b><i>Results.</i></b> Before the onset of LAD occlusion, as well as at 25th, 60th and 120th minutes of occlusion, all groups were similar in terms of blood pressure and pulse rate.<br /> The total area, affected area and necrotic area were calculated by using formulas; affected area ratio= affected area/total area X 100, necrotic area ratio = necrotic area/total affected area X 100, necrotic area and affected area ratio = necrotic area /affected area X 100.  Affected area and total area ratio was significantly higher in IR group, compared with cilostazole group (t=8.965; p<0.001). Similarly, necrotic area and total area ratio was higher in IR group, compared with cilostazole group (t=8.965; p<0.001). The necrotic area and affected area ratios were similar in IR and cilostazole groups (t=0.245; p=0.810). CK-MB level differences were not statistically significant between two groups (Z=0.382; p=0.721).<br /> Troponin levels were similar between IR and cilostazole groups and the difference was not statistically significant (Z=0.630; p=0.574). Pathological specimens of the heart were scanned for myocytolysis, PMNL and hemorrhage.  The difference between mean value of MDA enzyme levels were statistically significant (p<0.001) between all groups.  MDA enzyme levels, from higher to lower was IR, cilostazole and Sham group.  SOD levels (F=5.910; p=0.009) were significantly lower in Sham group when compared with IR group (p=0.008). The differences between Sham and cilostazole groups and IR and cilostazole gropus were not statistically significant (p=0.008). According to planimetric values and enzyme levels, cilostazole was found to be effective in reducing the ischemic zone, without effecting the necrotic zone in cardiac ischemia reperfusion damage. Therefore cilostazole has protective effects agains ischemia reperfusion damage.<br /> <br /> <b>Conclusion</b>. This study explored how cilostazol affects myocardial ischemia-reperfusion injury in rats, finding that cilostazol administration during reperfusion may protect against such injury. Through various analyses, we observed positive outcomes associated with cilostazol treatment, suggesting its potential in reducing myocardial damage. Further research is needed to understand the underlying mechanisms and optimize therapeutic strategies, but our findings highlight cilostazol's promise in improving clinical outcomes in cardiac interventions.

In Vivo Tissue Distribution of Polystyrene or Mixed Polymer Microspheres and Metabolomic Analysis after Oral Exposure in Mice.

Global plastic use has consistently increased over the past century with several different types of plastics now being produced. Much of these plastics end up in oceans or landfills leading to a substantial accumulation of plastics in the environment. Plastic debris slowly degrades into microplastics (MPs) that can ultimately be inhaled or ingested by both animals and humans. A growing body of evidence indicates that MPs can cross the gut barrier and enter into the lymphatic and systemic circulation leading to accumulation in tissues such as the lungs, liver, kidney, and brain. The impacts of mixed MPs exposure on tissue function through metabolism remains largely unexplored. This study aims to investigate the impacts of polymer microspheres on tissue metabolism in mice by assessing the microspheres ability to translocate across the gut barrier and enter into systemic circulation. Specifically, we wanted to examine microsphere accumulation in different organ systems, identify concentration-dependent metabolic changes, and evaluate the effects of mixed microsphere exposures on health outcomes. To investigate the impact of ingested microspheres on target metabolic pathways, mice were exposed to either polystyrene () microspheres or a mixture of polymer microspheres consisting of polystyrene (), polyethylene (), and the biodegradability and biocompatible plastic, poly-(lactic-co-glycolic acid) (). Exposures were performed twice a week for 4 weeks at a concentration of either 0, 2, or via oral gastric gavage. Tissues were collected to examine microsphere ingress and changes in metabolites. In mice that ingested microspheres, we detected polystyrene microspheres in distant tissues including the brain, liver, and kidney. Additionally, we report on the metabolic differences that occurred in the colon, liver, and brain, which showed differential responses that were dependent on concentration and type of microsphere exposure. This study uses a mouse model to provide critical insight into the potential health implications of the pervasive issue of plastic pollution. These findings demonstrate that orally consumed polystyrene or mixed polymer microspheres can accumulate in tissues such as the brain, liver, and kidney. Furthermore, this study highlights concentration-dependent and polymer type-specific metabolic changes in the colon, liver, and brain after plastic microsphere exposure. These results underline the mobility within and between biological tissues of MPs after exposure and emphasize the importance of understanding their metabolic impact. https://doi.org/10.1289/EHP13435.

Abstract 691: Preclinical evaluation of valproic acid in combination with radiation in orthotopic models of glioblastoma

Abstract Background: Valproic acid (VPA) is an FDA-approved anti-seizure medication that has been demonstrated to have an anti-tumor and radiosensitization effect on multiple cancer cells including aggressive brain cancer, glioblastoma (GBM). VPA is thought to exert its anti-tumor effects in part through inhibition of histone deacetylases (HDACs), upregulation of apoptosis and autophagy, and disruption of DNA repair mechanisms. Overall survival of patients treated with VPA in combination with TMZ and radiation therapy (RT) was significantly improved compared to historical controls (mOS 29.6 months). Hence, to establish a preclinical benchmark, we investigated the effectiveness of VPA alone and in combination with the radiotherapy in patient-derived xenograft (PDX) models of GBM in vitro and in vivo. Methods: Anti-proliferative effect of VPA in combination with radiation was assessed across multiple GBM cells lines (long-term cultures and PDX lines). Athymic mice were implanted with PDX GBM cells orthotopically and were randomized into four cohorts: vehicle, VPA, IR (6 gy), VPA + IR (6 gy). Animals were treated with 300mg/kg/day of VPA alone or in combination with 6Gy of radiation. Treatments were performed either via intraperitoneal route for 3 consecutive days or via a gastric gavage 5 days on/2 days off for the duration of the study. For pharmacodynamics (PD), animals were sacrificed after 3 days of drug treatment at 2hr, 24hr and 4 days post last dose and processed for immunohistochemical (IHC) analysis for acetylated H3K9/14, KI67, pH2AX, and cleaved caspase 3. Results: VPA treatment demonstrated a dose-dependent decrease in GBM cell viability and enhancement of radiation sensitivity. Pharmacodynamic analysis of tumor-bearing mice demonstrated a significant increase in acetylated H3K9/14 levels upon treatment with combination of VPA and irradiation. Elevation of acetylated H3K9/14 levels was present 2 hours after the administration of the last dose of VPA and it persisted for 4 days. Treatment of mice with irradiation upregulated levels of the apoptosis marker, cleaved caspase 3, which was further enhanced upon VPA and irradiation combination treatment. No reduction in tumor volume or survival benefit was seen in VPA alone or combination cohorts. Conclusion: Consistent with previous reports, treatment of PDX GBM line with VPA reduced cell viability in vitro, increased histone acetylation, and cell death. However, these changes were not sufficient to provide survival benefit in the intracranial model and warrants further investigation into the genetic requirements for radiation sensitization by VPA. Citation Format: Mariya Stavnichuk, Zorana Opachich, Dylan Huttenlocher, James McNamara, Jennifer Molloy, Xu He, Sonam Patel, Nader Sanai, An-Chi Tien, Shwetal Mehta. Preclinical evaluation of valproic acid in combination with radiation in orthotopic models of glioblastoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 691.

Early detection of anthracycline-induced cardiotoxicity using [68Ga]Ga-FAPI-04 imaging.

Anthracycline-induced cardiotoxicity (AIC), whose major manifestation is diffuse myocardial fibrosis, is an important clinical problem in cancer therapy. Therefore, early identification and treatment are clinically important. This study aims to explore the feasibility of using 68Ga-labelled fibroblast activation protein (FAP) inhibitor ([68Ga]Ga-FAPI) positron emission tomography/computed tomography (PET/CT) for the early identification of the fibrotic process and guidance of antifibrosis therapy in AIC. An AIC rat model was induced by the intravascular administration of doxorubicin (DOX) once per week for 1, 2, 3 and 6weeks (2.5mg/kg/injection, groups 1-4), whereas intravascular saline was administered to control rats. Experimental and control groups (n = 4) underwent [68Ga]Ga-FAPI PET/CT following disease induction. Groups 5 and 6 received DOX injections for 3 and 6weeks, treated with angiotensin-converting enzyme (ACE) inhibitor starting at 3weeks, treated with enalapril (20mg/kg, gastric gavage) daily and underwent echocardiography and [68Ga]Ga-FAPI PET/CT at 3weeks after treatment. Rat hearts were subjected to haematoxylin and eosin staining, FAP immunohistochemistry, Sirius red staining and Masson's trichrome staining to investigate the pathological changes and deposition of collagen fibres. Rat blood was sampled weekly for the enzyme-linked immunosorbent assay of various markers of myocardial injury, such as plasma cardiac troponin I, B-type natriuretic peptide and angiotensin II. [68Ga]Ga-FAPI-04 uptake by the heart was significantly higher in the cardiotoxicity group than in the control group at weeks 3 (SUVmax: 1.21 ± 0.23 vs 0.67 ± 0.01, P < 0.05) and 6 (SUVmax: 1.48 ± 0.28 vs 0.67 ± 0.08, P < 0.001), whereas left ventricle ejection fraction (LVEF) did not significantly differ between normal and AIC rats at week 3. FAP+ expression began to increase starting at week 3, before irreversible fibrotic changes were detected, until week 6. After 3weeks of enalapril treatment, [68Ga]Ga-FAPI-04 accumulation decreased in groups 5 and 6 (SUVmax decreased from 1.21 ± 0.23 to 0.77 ± 0.08 and 1.48 ± 0.28 to 1.09 ± 1.06, P < 0.05). Cardiac function was preserved (LVEF was 75.7% ± 7.38% in group 3 vs 74.5% ± 2.45% in group 5, P > 0.05) and improved (LVEF increased from 51.6% ± 9.03% in group 4 to 65.2% ± 4.27% in group 6, P < 0.05), and myocardial fibrosis attenuated (from 6.5% ± 1.2% in group 4 to 4.31% ± 0.37% in group 6, P < 0.01). [68Ga]Ga-FAPI PET/CT can be used for the early detection of active myocardial fibrosis in AIC and the evaluation of the efficacy of therapeutic interventions. Early treatment guided by [68Ga]Ga-FAPI PET/CT may reduce anthracycline-induced myocardial injury and improve heart function.