Single Intraperitoneal Injection Research Articles

Empagliflozin alone and in combination with metformin mitigates diabetes-associated renal complications.

Diabetes mellitus is a major public health concern, often leading to undiagnosed micro- and macrovascular complications, even in patients with controlled blood glucose levels. Recent evidence suggests that empagliflozin and metformin have renoprotective effects in addition to their hypoglycemic action. This study investigated the potential protective effect of empagliflozin and metformin on diabetic renal complications. Forty-two adult male Sprague Dawley rats were randomized into six groups: normal control, diabetic control, metformin (250 mg/kg), empagliflozin (10 mg/kg), and combination therapy groups. Type 2 diabetes was induced in rats by a single intraperitoneal injection of streptozotocin (40 mg/kg) following two weeks of 10% fructose solution in their drinking water. Blood glucose, creatinine, urea nitrogen, inflammatory markers (IL-6, TNF-α), and renal tissue caspase-3 were assessed after eight weeks. Blood glucose, urea, creatinine, serum IL-6, TNF-α, and tissue caspase-3 were significantly decreased in the treatment groups compared to the diabetic group. The histopathological findings revealed that treatment with empagliflozin and/or metformin improved the damage in the renal tissue caused by diabetes-induced nephropathy. Moreover, co-administration of empagliflozin and metformin resulted in even better outcomes. Our data revealed that empagliflozin and metformin could improve renal function and decrease inflammation and apoptosis in diabetic animals, delaying the progression of diabetic nephropathy. Combined treatment with metformin and empagliflozin proved to have an additive protective action on renal tissue.

A new mouse model of depression induced by chronic restraint stress combined with lipopolysaccharide

The aim of this study was to establish a multi-factor-induced depression model in mice within a relatively short period, specifically through the combination of intraperitoneal injection of lipopolysaccharide (LPS) and chronic restraint stress (CRS), and to evaluate the differences in depressive-like behaviors among three different strains of mice, seeking mouse strains more suitable for this combined model. The mice of each strain were randomly divided into the normal group and model group. The mice in the model group received a single intraperitoneal injection of LPS once daily (1 mg/kg/d for 7 days) and subsequent CRS for 6 h, to induce depression, while the mice in the normal group received no treatment. Behavioral tests: sucrose preference test (SPT), open field test (OFT), forced swim test (FST), and tail suspension test (TST) were conducted to assess depressive-like behaviors in the mice. Data analysis showed that there were significant intergroup differences in depressive-like behaviors in ICR and C57BL/6 J mice, while KM mice exhibited minor differences with relatively high deviation in individual behavioral score. This study indicated that the combined depression mouse model could successfully induce significant depressive-like behaviors in ICR and C57BL/6 J mice.

Bushen Huoxue recipe ameliorates ovarian function via promoting BMSCs proliferation and homing to ovaries in POI mice

BackgroundPremature ovarian insufficiency (POI) is a tricky puzzle in the field of female reproductive medicine. Bushen Huoxue recipe (BHR), a traditional Chinese medicine compound based on the combination of kidney-tonifying and blood-activating functions, has shown excellent efficacy in improving female irregular menstruation, POI, and infertility. However, the potential mechanism of BHR in POI treatment has not yet been elucidated. Bone marrow mesenchymal stem cells (BMSCs), a type of pluripotent stem cells, have received increasing attention for their significant role in improving ovarian function and restoring fertility in women with POI. PurposeThis study aimed to evaluate the therapeutic effect of BHR in POI mice and explore its potential mechanism. MethodsA POI mouse model was established with a single intraperitoneal injection of 120 mg/kg cyclophosphamide (CTX). Distilled water, BHR, or dehydroepiandrosterone was administered via gavage for 28 consecutive days. The effect of BHR on ovarian function in POI mice was evaluated by assessing the estrous cycle, ovarian morphology, follicular development, hormone levels, and angiogenesis. The proportion of BMSCs in bone marrow, peripheral blood, and ovary was analyzed via flow cytometry, and the level of molecules mediating migration and homing in ovary was measured. Cell viability assays, scratch healing assays and transwell migration assays were performed to explore the effect of BHR on BMSCs proliferation and migration in vitro, and its potential mechanism was explored. ResultsBHR significantly ameliorated estrous cycle disorders, hormone disorders, ovarian morphology, ovarian microvascular formation, and ovarian reserve in POI mice. Meanwhile, the number of BMSCs number in the bone marrow, peripheral blood, and ovary was apparently increased. Of note, BHR increased the level of hepatocyte growth factor (HGF)/cellular mesenchymal epithelial transition factor (cMET) and stromal cell-derived factor-1(SDF-1)/CXC chemokine receptor 4 (CXCR4) in the ovaries of POI mice. Moreover, BHR treatment promoted BMSCs proliferation and migration in vitro, with a significant increase in the level of proliferating cell nuclear antigen, cMET, and CXCR4. ConclusionsBHR effectively restored ovarian reserve, ovarian function, and ovarian angiogenesis in CTX-induced POI mice. In addition, BHR promoted BMSCs proliferation, migration, and homing to the ovary, which was mediated by the SDF-1/CXCR4 and HGF/cMET signaling axis. Finally, the amelioration of ovarian reserve and ovarian function in CTX-induced POI mice by BHR may be related to its promotion of endogenous BMSCs proliferation and homing.

Nephroprotective effect of Ginsenoside Rg1 in lipopolysaccharide-induced sepsis in mice through the SIRT1/NF-κB signaling.

During sepsis, the kidney is one of the most vulnerable organs. Sepsis-associated acute kidney injury (S-AKI) is hallmarked by renal inflammation, apoptosis, and oxidative injury. Ginsenoside Rg1 (Rg1) is a natural product that possesses abundant pharmacological actions and protects against many sepsis-related diseases. Nevertheless, its role and related mechanism in S-AKI remain to be determined. S-AKI was induced using lipopolysaccharide (LPS, 10 mg/kg) via a single intraperitoneal injection. Rg1 (200 mg/kg) was intraperitoneally administered for 3 consecutive days before LPS treatment. For histopathological examination, murine kidney tissues were stained with hematoxylin and eosin. Tubular injury score was calculated to evaluate kidney injury. Serum creatinine and BUN levels were measured for assessing renal dysfunction. The levels and activities of oxidative stress markers (MDA, 4-HNE, PC, GSH, SOD, and CAT) in renal tissue were measured by corresponding kits. Renal cell apoptosis was detected by TUNEL staining. The protein levels of apoptosis-related markers (Bcl-2, Bax, and Cleaved caspase-3), proinflammatory factors, SIRT1, IκBα, p-NF-κB p65, and NF-κB p65 in kidneys were determined using western blotting. Immunofluorescence staining was employed to assess p-NF-κB p65 expression in renal tissues. LPS-induced injury of kidneys and renal dysfunction in mice were ameliorated by Rg1. Rg1 also impeded LPS-evoked renal cell apoptosis in kidneys. Moreover, Rg1 attenuated LPS-triggered inflammation and oxidative stress in kidneys by inhibiting proinflammatory cytokine release, enhancing antioxidant levels and activities, and reducing lipid peroxidation. However, all these protective effects of Rg1 in LPS-induced AKI mice were reversed by EX527, an inhibitor of sirtuin 1 (SIRT1). Mechanistically, Rg1 upregulated SIRT1 protein expression, increased SIRT1 activity, and inactivated NF-κB signaling in the kidney of LPS-induced AKI mice, which was also reversed by EX527. Rg1 ameliorates LPS-induced kidney injury and suppresses renal inflammation, apoptosis, and oxidative stress in mice via regulating the SIRT1/NF-κB signaling.

Mimicking bacterial infection in male mice changes sperm small RNA profiles and multigenerationally alters offspring behavior and physiology

Paternal pre-conceptual exposures, including stress, diet, substance abuse, parasite infection, and viral immune activation via Poly I:C, have been reported to influence the brains and behavior of offspring through sperm epigenetic changes. However, the effects of paternal (F0) pre-conceptual exposure to bacterial-induced immune activation on the behavior and physiology of F1 and F2 generations remain unexplored. We examined this using C57BL/6J mice. Eight-week-old males (F0) received a single intraperitoneal injection of the bacterial mimetic lipopolysaccharide (LPS: 5 mg/kg) or 0.9 % saline (vehicle control) before mating with naïve females at four weeks post-injection. Comprehensive behavioral assessments were conducted to investigate anxiety, social behaviors, depressive-like behaviors and cognition in both the F1 and F2 generations within the age range of 8 to 14 weeks. Results demonstrated that only female offspring of LPS-exposed fathers exhibited reduced anxiety levels in the light/dark box, large open field, and novelty-suppressed feeding test. These F1 female offspring also exhibited heightened sociability in the 3-chambered social interaction test and a reduced preference for saccharin in the saccharin preference test. Additionally, the F1 male offspring of LPS-challenged males demonstrated an increased total distance traveled in the light/dark box and a longer distance covered in the light zone. They also exhibited diminished preference for social novelty in the 3-chambered social interaction test and an elevated novel arm preference index in the Y-maze. In the F2 generation, male descendants of LPS-treated fathers showed reduced latency to feed in the novelty-suppressed feeding test. Additionally, the F2 generation of LPS-challenged fathers, but not the F1 generation, displayed enhanced immune response in both sexes after an acute LPS immune challenge (5 mg/kg). Analysis of sperm small noncoding RNA profiles from LPS-treated F0 mice revealed significant changes at 4 weeks after administration of LPS. These changes included three microRNAs, eight PIWI-interacting RNAs, and two transfer RNAs, exhibiting significant upregulation (mmu-miR-146a-5p, mmu-piR-27082 and mmu-piR-29102) or downregulation (mmu-miR-5110, mmu-miR-467e-3p, mmu-piR-22583, mmu-piR-23548, mmu-piR-36341, mmu-piR-50293, mmu-piR-16583, mmu-piR-36507, Mus_musculus_tRNA-Ile-AAT-2-1 and Mus_musculus_tRNA-Tyr-GTA-1-1). Additionally, we detected 52 upregulated small noncoding RNAs (including 9 miRNAs, 41 piRNAs, and 2 tRNAs) and 7 downregulated small noncoding RNAs (3 miRNAs, 3 piRNAs, and 1 tRNA) in the sperm of F1 offspring from LPS-treated males. These findings provide compelling evidence for the involvement of epigenetic mechanisms in the modulation of brain function and immunity, and associated behavioral and immunological traits, across generations, in response to bacterial infection.

Bosutinib Stimulates Macrophage Survival, Phagocytosis, and Intracellular Killing of Bacteria.

Host-acting compounds are emerging as potential alternatives to combating antibiotic resistance. Here, we show that bosutinib, an FDA-approved chemotherapeutic for treating chronic myelogenous leukemia, does not possess any antibiotic activity but enhances macrophage responses to bacterial infection. In vitro, bosutinib stimulates murine and human macrophages to kill bacteria more effectively. In a murine wound infection with vancomycin-resistant Enterococcus faecalis, a single intraperitoneal bosutinib injection or multiple topical applications on the wound reduce the bacterial load by approximately 10-fold, which is abolished by macrophage depletion. Mechanistically, bosutinib stimulates macrophage phagocytosis of bacteria by upregulating surface expression of bacterial uptake markers Dectin-1 and CD14 and promoting actin remodeling. Bosutinib also stimulates bacterial killing by elevating the intracellular levels of reactive oxygen species. Moreover, bosutinib drives NF-κB activation, which protects infected macrophages from dying. Other Src kinase inhibitors such as DMAT and tirbanibulin also upregulate expression of bacterial uptake markers in macrophages and enhance intracellular bacterial killing. Finally, cotreatment with bosutinib and mitoxantrone, another chemotherapeutic in clinical use, results in an additive effect on bacterial clearance in vitro and in vivo. These results show that bosutinib stimulates macrophage clearance of bacterial infections through multiple mechanisms and could be used to boost the host innate immunity to combat drug-resistant bacterial infections.

Abstract LB264: Intracavitary gene therapy using miRNA-29b-encoding adeno-associated virus for peritoneal dissemination

Abstract INTRODUCTION: Peritoneal metastasis (PM) is the most critical prognostic factor in gastrointestinal cancers. Notably, the levels of microRNA (miR)-29b in peritoneal fluids have shown significant reduction in patients with PM of gastric cancer. Moreover, supplementation with miR-29b-incorporated exosomes has demonstrated the potential to suppress the formation of PM in a murine model. Nevertheless, given the technical demands associated with this method, we investigated the viability of intracavitary gene therapy as a potentially more advanced and practical approach for clinical implementation. METHODS: Using the calcium phosphate-based protocol, adeno-associated virus encoding miR-29b (AAV-miR-29b) was constructed, and its effects on mesothelial mesenchymal transition (MMT) were examined using peritoneal mesothelial cells (PMC) derived from human and murine omental tissues. PM was established in C57BL/6 mouse by intraperitoneal (IP) injection of a highly metastatic clone of gastric cancer cell, YTN16P or a pancreatic cancer cell, PAN02. In these syngeneic models, we examined the impact of IP injection of AAV-miR-29b at a single dose of 5 × 1010 vector genome (vg) on PM. RESULTS: AAV-miR-29b was effectively incorporated into PMCs, resulting in an increased level of miR-29b in extracellular vesicles produced by PMCs. Stimulation of PMCs with TGF-β1 (10ng/ml) decreased the expression of E-cadherin and increased vimentin and fibronectin expression along with enhanced migration activity. Importantly, all the changes were totally abrogated by AAV-miR-29b. Treatment of PMC with TGF-β1 notably increased the attachment of tumor cells, which was also entirely inhibited by AAV-miR-29b. Following IP administration of AAV-miR-29b in vivo, the expression level of miR-29b in peritoneal tissue and intraperitoneal exosomes increased significantly after two weeks. In both murine gastric and pancreatic cancer models, a single IP injection of AAV-miR-29b on day 0 led to a marked reduction in the number of PM nodules in the mesentery (gastric: 38.2 ± 18.7 vs. 8.0 ± 5.2, n=5, P < 0.05; pancreatic: 41.2 ± 7.5 vs. 14.4 ± 16.7, n=7, P < 0.001) and inhibited peritoneal fibrosis associated with PM. AAV-miR-29b elicits the same significant effects when administered on day3. Furthermore, when AAV-miR-29b was administered on day 7 in combination with low-dose Paclitaxel (PTX) (200 µg/mouse), a significant reduction in the number of PM nodules was observed, whereas the effect was not evident with PTX alone (35.8 ± 22.2, n=16 vs. 14.2 ± 16.0, n=12, P < 0.05). CONCLUSIONS: IP injection of AAV-miR-29b achieved robust transgene expression in PMCs and their exosomes, effectively suppressing MMT and peritoneal fibrosis leading to the inhibition of PM Intraabdominal gene therapy utilizing AAV containing tumor-suppressor miRNA could serve as a promising strategy for both the prevention and treatment of PM. Citation Format: Yuki Kaneko, Hideyuki Ohzawa, Yuki Kimura, Rei Takahashi, Misaki Matsumiya, Kohei Tamura, Yurie Futoh, Hideyo Miyato, Ryota Watano, Hiroaki Mizukami, Naohiro Sata, Joji Kitayama. Intracavitary gene therapy using miRNA-29b-encoding adeno-associated virus for peritoneal dissemination [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 2 (Late-Breaking, Clinical Trial, and Invited Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(7_Suppl):Abstract nr LB264.

Bio-analytical Estimation of Metformin Co-administered with Antidiabetic and Antihyperlipidemic Herbal Preparations by LCMS

The antidiabetic and antihyperlipidemic herbal preparations were developed in liquid and oral tablet dosage form as Saptarangi Plus Kadha (SK) and Saptarangi Tablet (ST). Streptozotocin (STZ) (45mg/kg) was injected by single intraperitoneal (i.p.) injection and diabetes was induced in Wistar rats of group II to group VII. Metformin (100mg/kg, p.o.) was co-administered orally to group IV to VII with SK and ST as a reference standard drug. There is a need of bio-analytical study and standardization of herbal preparation which is required to get approval of the regulatory agencies for the commercialization. The estimation of metformin was carried in the presence of other herbal phytoconstituents by LCMS/MS. The samples of all the groups were centrifuged and vortexed. The samples were prepared in ACN: 0.1% formic acid (70:30) and LCMS method was developed and validated. The metformin was estimated in presence of active phytoconstituents of SK and ST. SK and ST contains Salacinol and folia salaciosides E1 which was detected by LCMS at m/z 333.42 and m/z 504.63 respectively. There has been an honest effort to create and validate LCMS method for the detection of metformin in presence of phytoconstituents of SK and ST in serum.

Zingiber officinale ethanolic extract improved organs function in lipopolysaccharides-induced organ toxicity by modulating inflammation and oxidative stress in male rats

ABSTRACT Bacterial lipopolysaccharide (LPS) is the typical inflammatory and oxidative stress agent; ginger (Zingiber officinale) has long been utilized to treat inflammatory conditions. This study aimed to evaluate the possible therapeutic role of ginger extract (GE) against LPS-induced hepato-renal and testicular injury. Male rats received GE (150 mg/kg, orally, for 28 days) after the inflammation shock was provoked by a single intraperitoneal injection of LPS (5 mg/kg). Rats treated with LPS exhibited marked inflammation and oxidative stress, in addition to liver and kidney injury and disruption of male sex hormones. The treatment of LPS-injected rats with GE resulted in a remarkable return to normalcy of the oxidative stress in the rats, as evidenced by a notable decrease MDA and an amelioration of SOD and CAT activities. On the other hand, the rats administered LPS and GE had considerably decreased serum levels of TNF-α and IL-6, improvement in serum albumin, total bilirubin and ALT and AST activity, urea, creatinine, and uric acid as well as FSH and testosterone levels. This indicating improved liver, kidney and testicular functions. In conclusion, GE attenuates the inflammatory damage and oxidative stress caused by LPS, which helps to restore organ function.

Antidiabetic effect of the mixture of Cissus polyantha and Rytigynia senegalensis in high-fat diet and streptozocin-induced diabetic rats

BackgroundIn traditional Chinese medicine (TCM) practices, many plants of the Cissus and Rytigynia genera are used to treat many diseases including diabetes mellitus.The aim of this study was to evaluate the modulating effects of the mixture of the aqueous extracts of Cissus polyantha and Rytiginia senegalensis (CPRS) on some enzymes of carbohydrate metabolism, oxidative profile, and cardiovascular parameters in diabetic rats. MethodsDiabetes was induced by a diet enriched in high-fat for 30 days associated with a single intraperitoneal injection of streptozocin (35 mg/kg). Rats with a blood glucose level greater than or equal to 200 mg/dL were selected and divided into groups. Groups 1 and 2 received distilled water. Group 3 received metformin (40 mg/kg). Groups 4 and 5 received CPRS at doses of 50 and 111 mg/kg, respectively. Glycaemia, body mass, food and water intake, insulinemia, insulin resistance and insulin secretion indices, lipid profile and cardiovascular risk indices, oxidant profile, glycogen and liver enzymes were evaluated. Histology of the pancreas and heart was performed. ResultsCPRS (50 and/or 111 mg/kg) significantly (p < 0.05 to p < 0.001) reduced water and food consumption, blood glucose, HOMA-IR, coronary risk index, cholesterol, triglyceride, LDL-c, VLDL-c, and malondialdehyde levels, as well as glucose-6-phosphatase and fructose-1,6-biphosphatase activities in diabetic animals. In contrast, CPRS (50 and/or 111 mg/kg) caused an increase (p < 0.05 to p < 0.001) in body mass, insulinemia and HDL-c levels, reduced glutathione and hepatic glycogen. HOMA-β, insulin distribution, atherogenic and cardioprotective indices as well as superoxide dismutase, catalase, glucokinase, and glucose-6-phosphate dehydrogenase activities were also increased after CPRS administration. CPRS regenerated pancreatic islet β cells and decreased leukocyte infiltration in cardiomyocytes of diseased animals. ConclusionCPRS has hypoglycemic, antidyslipidemic, and antioxidant effects, thus validating its use in traditional medicine in the treatment of diabetes mellitus and its complications.

Ammoniagenic Action of Valproate without Signs of Hepatic Dysfunction in Rats: Possible Causes and Supporting Evidence.

(1) Background: Valproic acid (VPA) is one of the frequently prescribed antiepileptic drugs and is generally considered well tolerated. However, VPA neurologic adverse effects in the absence of liver failure are fairly common, suggesting that in the mechanism for the development of VPA-induced encephalopathy, much more is involved than merely the exposure to hyperammonemia (HA) caused by liver insufficiency to perform detoxification. Taking into account the importance of the relationship between an impaired brain energy metabolism and elevated ammonia production, and based on the ability of VPA to interfere with neuronal oxidative pathways, the current study intended to investigate a potential regional ammoniagenic effect of VPA on rats' brains by determining activities of the enzymes responsible for ammonia production and neutralization. (2) Methods: Rats received a single intraperitoneal injection of VPA (50, 100, 250, 500 mg/kg). Plasma, the neocortex, the cerebellum, and the hippocampus were collected at 30 min after injection. The levels of ammonia, urea, aspartate aminotransferase (AST), and alanine aminotransferase (ALT) were measured in blood plasma. The activities of glutaminase and glutamate dehydrogenase (GDH) in mitochondria and the activities of AMP deaminase (AMPD), adenosine deaminase (ADA), and glutamine synthetase (GS) in cytosolic fractions isolated from rat brain regions were measured. Ammonia, ALT, and AST values were determined in the mitochondrial and cytosolic fractions. (3) Results: Multi-dose VPA treatment did not significantly affect the plasma levels of ammonia and urea or the ALT and AST liver enzymes. Significant dose-independent increases in the accumulation of ammonia were found only in the cytosol from the cerebellum and there was a strong correlation between the ammonia level and the ADA activity in this brain structure. A significant decrease in the AMPD and AST activities was observed, while the ALT activity was unaffected. Only the highest VPA dose (500 mg/kg) was associated with significantly less activity of GS compared to the control in all studied brain structures. In the mitochondria of all studied brain structures, VPA caused a dose-independent increases in ammonia levels, a high concentration of which was strongly and positively correlated with the increased GDH and ALT activity, while glutaminase activity remained unchanged, and AST activity significantly decreased compared to the control in all studied brain structures. (4) Conclusions: This study highlights the rat brain region-specific ammoniagenic effects of VPA, which may manifest themselves in the absence of hyperammonemia. Further research should analyze how the responsiveness of the different brain regions may vary in VPA-treated animals that exhibit compromised energy metabolism, leading to increased ammoniagenesis.

Platycodin D Ameliorates Cognitive Impairment in Type 2 Diabetes Mellitus Mice via Regulating PI3K/Akt/GSK3β Signaling Pathway.

Objectives: The aim of this study was to investigate the ameliorative effect of platycodin D (PD) on cognitive dysfunction in type 2 diabetes mellitus (T2DM) and its potential molecular mechanisms of action in vivo and in vitro. Materials and methods: An animal model of cognitive impairment in T2DM was established using a single intraperitoneal injection of streptozotocin (100 mg/kg) after 8 weeks of feeding a high-fat diet to C57BL/6 mice. In vitro, immunofluorescence staining and Western blot were employed to analyze the effects of PD on glucose-induced neurotoxicity in mouse hippocampal neuronal cells (HT22). Results: PD (2.5 mg/kg) treatment for 4 weeks significantly suppressed the rise in fasting blood glucose in T2DM mice, improved insulin secretion deficiency, and reversed abnormalities in serum triglyceride, cholesterol, low-density lipoprotein, and high-density lipoprotein levels. Meanwhile, PD ameliorated choline dysfunction in T2DM mice and inhibited the production of oxidative stress and apoptosis-related proteins of the caspase family. Notably, PD dose-dependently prevents the loss of mitochondrial membrane potential, promotes phosphorylation of phosphatidylinositol 3 kinase and protein kinase B (Akt) in vitro, activates glycogen synthase kinase 3β (GSK3β) expression at the Ser9 site, and inhibits Tau protein hyperphosphorylation. Conclusions: These findings clearly indicated that PD could alleviate the neurological damage caused by T2DM, and the phosphorylation of Akt at Ser473 may be the key to its effect.

Cobalt protoporphyrin promotes heme oxygenase 1 expression and ameliorates cardiac dysfunction in long-term fasting mice

BackgroundThe association between malnutrition and cardiac dysfunction has been reported. Heme oxygenase (HO)-1 played protective roles in the animals functioning as a myocardial infarction, heart failure, or cardiomyopathy model. We hypothesized that the administration of HO-1 inducer, cobalt protoporphyrin (CoPP) reduces oxidative stress and ameliorates cardiac systolic dysfunction in long-term fasting mice. MethodsC57BL/6 J mice were classified into three groups: fed mice (fed group), 48-h fasting mice with a single intraperitoneal injection of the corresponding vehicle (fasting group), and 48-h fasting mice with a single intraperitoneal injection of 5 mg/kg CoPP (CoPP group). ResultsThe fasting group showed a significant increase in heme and 4-hydroxy-2-nonenal (4HNE) protein in the heart tissue, and reduced left ventricular ejection fraction (LVEF) when compared with the fed group. The CoPP group showed significantly increased protein levels of nuclear factor-erythroid 2-related factor 2 and HO-1, and increased mRNA expression levels of HO-1, peroxisome proliferator-activated receptor gamma coactivator 1-alpha, forkhead box protein O1, sirtuin-1, cyclooxygenase 2, and superoxide dismutase 2, and reduced levels of heme and 4HNE protein when compared with the fasting group. LVEF were significantly higher in the CoPP group than in the fasting group. ConclusionsAdministration of CoPP reduced heme accumulation and oxidative stress, and ameliorated cardiac systolic dysfunction in long-term fasting mice. This study suggests that heme accumulation may be associated with impaired cardiac function induced by long-term fasting and that HO-1 may be a key factor or therapeutic target.

Idebenone alleviates doxorubicin-induced cardiotoxicity by stabilizing FSP1 to inhibit ferroptosis

Doxorubicin (DOX)-mediated cardiotoxicity can exacerbate mortality in oncology patients, but related pharmacotherapeutic measures are relatively limited. Ferroptosis was recently identified as a major mechanism of DOX-induced cardiotoxicity. Idebenone, a novel ferroptosis inhibitor, is a well-described clinical drug widely used. However, its role and pathological mechanism in DOX-induced cardiotoxicity are still unclear. In this study, we demonstrated the effects of idebenone on DOX-induced cardiotoxicity and elucidated its underlying mechanism. A single intraperitoneal injection of DOX (15 mg/kg) was administrated to establish DOX-induced cardiotoxicity. The results showed that idebenone significantly attenuated DOX-induced cardiac dysfunction due to its ability to regulate acute DOX-induced Fe2+ and ROS overload, which resulted in ferroptosis. CESTA and BLI further revealed that idebenone's anti-ferroptosis effect was mediated by FSP1. Interestingly, idebenone increased FSP1 protein levels but did not affect Fsp1 mRNA levels in the presence of DOX. Idebenone could form stable hydrogen bonds with FSP1 protein at K355, which may influence its association with ubiquitin. The results confirmed that idebenone stabilized FSP1 protein levels by inhibiting its ubiquitination degradation. In conclusion, this study demonstrates idebenone attenuated DOX-induced cardiotoxicity by inhibiting ferroptosis via regulation of FSP1, making it a potential clinical drug for patients receiving DOX treatment.

Citrus aurantifolia (Chrism.) Swingle peel extract attenuate nephrotoxicity induced by doxorubicin

BackgroundDoxorubicin (DOX) is an anthracycline antibiotic primarily utilized in the treatment of various solid tumors, including those affecting the cervix, ovaries, breasts, uterus, lungs, as well as certain blood cancers. However, the nephrotoxicity connected to DOX therapy restricts its clinical use. In an Ancient Chinese Medicine used Lime (Citrus. aurantifolia) Peel was employed to address cramps, colic pain, cold, and as an appetite stimulant. Lime is known as "Zhi qiao" in Traditional Chinese Medicine (TCM), in addition to its nephroprotective, antioxidant and immunostimulatory properties the peel had been noted for its effectiveness in regulating vital energy, used to treat cough, abdominal distention and constipation. ObjectiveThe purpose of this research is to evaluate the potential protective effect of Citrus aurantifolia peel extract (CAPE) in alleviating the nephrotoxicity generated by doxorubicin (DOX) injection in rats. MethodsThirty Wister albino rats were randomly and equally separated into six groups (n = 5), and they were treated as follows: group I received distilled water only (baseline), toxicant groups administered with 15 mg/kg, i.p. DOX, 24 h after, groups II, III IV, V and VI were treated with distilled water (negative control), Alpha lipoic acid, 150 mg/kg (positive control) and 100, 200 and 400 mg/kg of CAPE respectively, orally for seven days. ResultsDOX caused kidney injury after a single intraperitoneal injection, as shown by substantial changes in kidney biomarkers, histological irregularities, and the reduction of antioxidant defense mechanisms (reduced glutathione, superoxide dismutase and catalase). CAPE treatment resulted in significant improvements in the amended renal biomarkers (urea, creatinine, albumin and uric acid), haematological parameters, antioxidative markers, and malondialdehyde. The present findings indicated that CAPE could reduce lipid peroxidation (LPO) and reactive oxygen species production, and observable histological improvements supported this result. ConclusionOur results point out that CAPE is a potential therapeutic alternative for the prevention of DOX-related nephrotoxicity.

Boron Facilitates Amelioration of Hepatic Injury by the Osmolyte Glycine and Resolves Injury by Improving the Tissue Redox Homeostasis

Background: Glycine is a conditional non-essential amino acid in human and other mammals. It is abundant in the liver and is known for a wide spectrum of characteristics including the antioxidant, antiinflammatory, immunomodulatory, and cryoprotective effects. The amino acid is a naturally occurring osmolyte compatible with protein surface interactions and has been reported in literature as a potent therapeutic immuno-nutrient for liver diseases such as alcoholic liver disease. Oral glycine administration protects ethanol-induced liver injury, improves serum and tissue lipid profile, and alleviates hepatic injury in various conditions. In recent years, sodium salt of boron (borax) has been reported for its beneficial effects on cellular stress, including the effects on cell survival, immunity, and tissue redox state. Incidentally both glycine and boron prevent apoptosis and promote cell survival under stress. Objective: This study investigates the beneficial effect of borax on liver protection by glycine. Methods: Briefly, liver toxicity was induced in rats by a single intraperitoneal injection of thioacetamide (400 mg/kg b. wt.). Results: Significant changes in oxidative stress and liver function test parameters, the molybdenum Fe-S flavin hydroxylase activity, nitric oxide and tissue histopathology were observed in thioacetamide treated positive control group. The changes were ameliorated both by glycine as well as borax, but the combinatorial treatment yielded a better response indicating the impact of boron supplementation on glycine mediated protection of liver injury in experimental animal model. Conclusions: The study has clinical implications as the hepatotoxicity caused by thioacetamide mimics features of hepatitis C infection in human.

Propionic acid affects the synaptic architecture of rat hippocampus and prefrontal cortex

It is well documented that propionic acid (PPA) produces behavioral, morphological, molecular and immune responses in rats that are characteristic of autism spectrum disorder in humans. However, whether PPA affects the ultrastructure and synaptic architecture of regions of autistic brain has not been adequately addressed. Earlier we show that single intraperitoneal (IP) injection of PPA (175 mg/kg) produces superficial changes in the spatial memory and learning of adolescent male Wistar rats. However, in neurons, synapses and glial cells of hippocampal CA1 area and medial prefrontal cortex transient (mainly) or enduring alterations were detected. In this study, we used electron microscopic morphometric analysis to test the effect of PPA on different structural parameters of axodendritic synapses of the hippocampus and prefrontal cortex. The animals were treated with a single IP injection of PPA (175 mg/kg). The length and width of synaptic active zone, the area of presynaptic and postsynaptic mitochondria, the distance between presynaptic mitochondria and the synapse active zone, the distance between postsynaptic mitochondria and postsynaptic density and the depth and opening diameter of neuronal porosome complex were evaluated. Our results show that synaptic mitochondria of the hippocampus and prefrontal cortex are the most vulnerable to PPA treatment: in both regions, the area of postsynaptic mitochondria were increased. In general, our results show that even small dose of PPA, which produces only superficial effects on spatial memory and learning is able to alter the synapse architecture in brain regions involved in cognition and autism pathogenesis. Therefore, the microbiome may be involved in the control of neurotransmission in these regions.

Molecular docking, characterization, ADME/toxicity prediction, and anti-ulcer activity of new quercetin derivatives on indomethacin-induced gastric ulcer in mice

Gastric ulcer (GU) is a serious upper gastrointestinal tract disorder that affects people worldwide. The drugs now available for GU treatment have a high rate of relapses and drug interactions, as well as mild to severe side effects. As a result, new natural therapeutic medications for treating GU with fewer negative side effects are desperately needed. Because of quercetin's (QCT) diverse pharmacological effects and unique structural features, we decided to semi-synthesize new QCT derivatives and test them for antiulcer activity. Docking assays were performed on the synthesized compounds to determine their affinity for TLR-4/MD-2, MyD88/TIR, and NF-κB domains, an important inflammatory pathway involved in GU development and progression. Mice were given oral famotidine (40 mg/kg/day), QCT, QCT pentamethyl (QPM), or QCT pentaacetyl (QPA) (50 mg/kg/day) for 5 days before GU induction by a single intraperitoneal injection of indomethacin (INDO; 18 mg/kg). QPM and QPA have a stronger binding affinity for TLR-4/MD-2, MyD88/TIR and NF-κB domains than QCT. In comparison, they demonstrated the greatest reduction in ulcer score and index, gastric MDA and nitric oxide (NO) contents, MyD88 and NF-κB expressions, and gastric TLR-4 immunostaining. They also enhanced the levels of GSH, CAT, COX-1, and COX-2 in the gastric mucosa, as well as HO-1 and Nrf2 expression, with histological regression in gastric mucosal lesions, with QPA-treated mice demonstrating the best GU healing. QPA is safe against all of the target organs and adverse pathways studied, with good ADME properties. However, further in vitro experiments are necessary to demonstrate the inhibitory effects of QPM and QPA on the protein targets of interest. In addition, preclinical research on its bioavailability and safety is essential before clinical management can be undertaken. Overall, the new QPA derivative could one day serve as the basis for a new class of potential antiulcer drugs.

Synergistic Interaction between Topical Application of Allicin and Quercetin Enhances Diabetic Wound Healing Via Inflammatory in Wistar Rats

Prolonged and excessive inflammatory processes at the wound site result in delayed healing of diabetic wounds. Allicin and quercetin have significantly inhibited tumor necrosis factor alpha (TNF-α) and increased levels of transforming growth factor beta (TGF-β) and vascular endothelial growth factor (VEGF) in rats. However, there is no known investigation on the combination of both compounds on TNF-α, TGF-β, and VEGF as a topical healing agent via inflammatory mechanisms. This study aims to determine the synergistic interaction between topical application of allicin and quercetin in enhancing diabetic wound healing via inflammatory in Wistar rats. A total of 45 male Wistar rats weighing 180-250 g were induced diabetes by a single intraperitoneal injection of streptozotocin (45 mg/kg bw). Diabetic rats underwent wound creation under anaesthesia and a square-shaped open excision wound was made with a full-thickness measurement of 1 cm × 1 cm on the right side of the rat's back. Rats were randomly allocated into group namely, vehicle control, allicin, quercetin, and a combination of allicin and quercetin which is applied topically once a day at a dose of 10 mg/mL for 7 days. Our finding showed that once daily topical application of allicin and quercetin, both independently and synergistically, increased levels of TGF-β and VEGF, as well as decreased levels of TNF-α of the rat diabetic wound model compared with the vehicle control group over 7th day. The synergistic effects of allicin and quercetin significantly enhanced diabetic wound healing via inflammation in rats, by suppressing inflammatory cytokines and stimulating growth factors hence offering a new window of an experimental study.

JianPiYiShen formula prevents cisplatin-induced acute kidney injury in mice by improving necroptosis through MAPK pathway

BackgroundAcute kidney injury (AKI), characterized by necroptosis and activation of MAPK pathway, causes sudden declines in renal function. To date, efficacious treatments are lacking. JianPiYiShen Formula (JPYSF) has a protective effect on the kidneys. The aim of this study is to explore the mechanism of JPYSF in cisplatin-induced AKI.MethodsMale C57/BL6J mice were divided into control group, cisplatin group and cisplatin + JPYSF group. Before establishing the model, the cisplatin + JPYSF group was administered JPYSF (18.35 g/kg/day) by gavage for 5 consecutive days. A single intraperitoneal injection of cisplatin (20 mg/kg) was used to establish AKI model. Measurement of renal function and H&E staining were performed to assess renal damage. WB, PCR, TUNEL staining and immunohistochemistry were used to detect related indicators of mitochondrial function, oxidative stress, necroptosis, inflammation and MAPK pathway. And one-way analysis of variance was used to compare group differences.ResultsCompared with the cisplatin group, JPYSF can attenuate AKI, reflected by the decrease in Scr and BUN levels, the improvement of renal tubular injury, and the downregulation of NGAL and KIM1. Cisplatin can induce mitochondrial dysfunction and oxidative stress, triggering necroptosis. In this study, JPYSF improved mitochondrial dysfunction to enhance oxidative stress, as manifested by upregulation of OPA1, PGC-1α, SOD and CAT, and downregulation of DRP1 and MFF. Then JPYSF showed a significant protective effect in necroptosis, as embodied by reduced number of TUNEL-positive cells, decreased the gene expression of RIPK3 and MLKL, as well as downregulation the proteins expression of P-RIPK1, P-RIPK3, and P-MLKL. Moreover, necroptosis can aggravate inflammation. JPYSF ameliorated inflammation by improving inflammatory and anti-inflammatory indexes, including downregulation of TNF-α, IL-6, MCP-1 and LY6G, and upregulation of IL-10. In addition, JPYSF also inhibited MAPK pathway to improve necroptosis by decreasing the expression of P-JNK and P-ERK.ConclusionOur data showed that JPYSF prevents cisplatin-induced AKI by improving necroptosis through MAPK pathway, which is related to the improvement of mitochondrial dysfunction, oxidative stress, and inflammation.