Single Intraperitoneal Injection Research Articles

Isatin improves oligoasthenospermia caused by busulfan by regulating GSH/GPX4 axis to inhibit ferroptosis

IntroductionFerroptosis, induced by iron overload and an imbalance in redox homeostasis, promotes the generation of reactive oxygen species (ROS), leading to iron-dependent lipid peroxides (LPO) and oxidative stress. Lipid peroxidation induced by reactive oxygen species is essential for the progression of spermatogenesis. However, its imbalance can lead to reproductive system damage and oligoasthenospermia, a critical cause of oligoasthenospermia. Isatin (ISA) is a naturally occurring compound that is widely distributed in lobsters, crustaceans, shellfish and various plants. It exhibits significant antioxidant and anti-aging properties, suggesting its potential as a therapeutic agent for the treatment of oligoasthenospermia. This study aimed to investigate the effects and mechanisms of ISA on oligoasthenospermia and to elucidate the underlying molecular pathways.MethodsAll mice were divided into normal group, model group and treatment group. Both model group and treatment group received a single intraperitoneal injection of 30 mg/kg BUS to create the model of oligoasthenospermia. After 2 weeks, the treatment group received different doses of 25, 50 and 100 mg/kg ISA by gavage for 28 days, and then mice were sacrificed and tested.ResultsThe results demonstrated that ISA effectively reversed busulfan-induced reproductive system damage in mice. This included the restoration of testicular histomorphology, improvement in sperm concentration and motility, regulation of serum sex hormone levels, and normalization of various oxidative indices in testicular tissue. Furthermore, ISA successfully reversed testicular ferroptosis by restraining the translocation of nuclear factor erythroid 2-related factor 2 (NRF2) into the nucleus and improved oligoasthenospermia through the glutathione (GSH)/glutathione peroxidase 4 (GPX4) axis.DiscussionISA was found to effectively ameliorate oligoasthenospermia in mice, presenting a potential therapeutic option for patients with this condition.

GLP-1 receptor agonist liraglutide alleviates kidney injury by regulating nuclear translocation of NRF2 in diabetic nephropathy.

Diabetic nephropathy (DN) is a severe renal disorder that arises as a complication of diabetes. Liraglutide, an analogue of a glucagon-like peptide 1 (GLP-1) receptor agonist, has been shown to decrease diabetes-caused renal damage. Nevertheless, the complete understanding of the roles and mechanism remains unclear. In our study, diabetic rat models were created through a single intraperitoneal injection of streptozotocin (STZ). The level of fasting blood glucose, 24-h urine protein, serum creatinine (Scr) and blood urea nitrogen (BUN) were assessed. Periodic acid-Schiff (PAS) staining was applied to examine the pathological changes in renal tissues. Reactive oxygen species (ROS) formation was measured via dichloro-dihydro-fluorescein diacetate (DCFH-DA) probes. Western blot was conducted to examine the levels of oxidative stress-related and extracellular matrix (ECM)-associated proteins. The nuclear translocation of NRF2 was investigated through immunofluorescence and Western blot assays. We demonstrated that liraglutide attenuated DN-induced oxidative stress and ECM deposition in vitro and in vivo. Liraglutide exerted a reno-protective effect by promoting nuclear translocation of NRF2 in mesangial cells. ML385, an NRF2 inhibitor, counteracted the beneficial impact of liraglutide.

Experimental study on the treatment of diabetic cardiomyopathy in rats by using ultrasound-targeted microbubble destruction combined with coenzyme Q10 loaded long-circulating nanoliposomes

Objective: To investigate the therapeutic effects and mechanisms of Ultrasound-targeted microbubble destruction (UTMD) technology combined with CoQ10 loaded PEGylated nanoliposomes (CoQ10-PEG-lips) on diabetic cardiomyopathy (DCM) in rats. Methods: CoQ10-PEG-lips were prepared using the thin-film dispersion method combined with ultrasonic hydration, followed by quality assessment. Sixty healthy and clean male SD rats were selected, and 50 were randomly chosen using a random number table to establish a type 1 diabetes mellitus (DM) model via a single intraperitoneal injection of streptozotocin. The remaining 10 rats were assigned as the normal control group. A total of 47 rats successfully developed the DM model, and 40 were selected using the random number table method. Based on different intervention methods, these rats were then randomly divided into DM model group, CoQ10 solution group, CoQ10-PEG-Lips group, and CoQ10-PEG-Lips+UTMD group (n=10 per group). Normal control group and DM model group rats were injected with 1 ml of normal saline through caudal vein. CoQ10 solution group and CoQ10-PEG-Lips group were injected with 1 ml of CoQ10 solution or CoQ10-PEG-Lips solution containing 10 mg/kg of CoQ10 through caudal vein, respectively. Rats in the CoQ10-PEG-Lips+UTMD group were injected with 1 ml of CoQ10-PEG-Lips solution containing 10 mg/kg of CoQ10+100 mg of freeze-dried ultrasound microbubble powder through caudal vein and were given UTMD treatment at the same time, with the intervention given twice a week. After 12 weeks of intervention, cardiac function indexes [left ventricular end-systolic diameter (LVEDs), left ventricular end-diastolic diameter (LVEDd), and left ventricular ejection fraction (LVEF)]of each group were measured by ultrasonic cardiac function detection in vivo. Simultaneously, ex-vivo histopathological examination was conducted to assess myocardial cell morphology, cross-sectional area, collagen volume fraction (CVF), and apoptosis index (AI) in each group. Additionally, molecular biology techniques were employed to measure oxidative stress-related indicators and the expression of apoptosis-related pathway proteins. Results: The prepared CoQ10-PEG-lips had a well-rounded morphology, good dispersibility, and a high encapsulation efficiency of 87.45%±3.23%. After 12 weeks of intervention, the myocardial cell morphology in the CoQ10-PEG-Lips+UTMD group was intact, with orderly arrangement, closely resembling that of the normal control group. There were no statistically significant differences between the two groups in terms of LVEDs [(1.53±0.07) mm vs (1.42±0.04) mm], LVEDd [(2.93±0.15) mm vs (2.81±0.05) mm], or LVEF (80.76%±3.42% vs 84.60%±2.10%) (all P>0.05). Similarly, there were no significant differences between the two groups in myocardial cell cross-sectional area, CVF, or AI (all P>0.05). The CoQ10-PEG-Lips+UTMD group showed statistically significant differences in the above-mentioned indicators compared to the DM group, CoQ10 solution group, and CoQ10-PEG-Lips group (all P<0.05). In the CoQ10-PEG-Lips+UTMD group, the levels of myocardial superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), and the expression of the anti-apoptotic protein B-cell lymphoma-2 (Bcl-2) were significantly higher compared to the DM model group, CoQ10 solution group, and CoQ10-PEG-Lips group (all P<0.05), while malondialdehyde levels and the expression of Bcl-2-associated X protein (Bax) and caspase-3 were lower (all P<0.05). Conclusions: Using PEG-lips to encapsulate the poorly soluble drug CoQ10 in combination with UTMD technology enables targeted delivery of the drug to the myocardium, which can help reduce myocardial cell damage, fibrosis, and apoptosis caused by diabetes mellitus (DM) by inhibiting oxidative stress damage and the Bcl-2/Bax/caspase-3 apoptosis signaling pathway, ultimately improving cardiac function in DCM rats.

Hesperidin as a bioactive compound in citrus fruits reduces N-ethyl-N-nitrosourea-induced mortality and toxicity in mice: as a model for chronic lymphocytic leukemia.

The current study is aimed at determining the preventive effects of hesperidin against death, weight changes, cellular damage, and oxidative stress in mice induced by n-ethyl-n-nitrosourea as a chronic lymphocytic leukemia (CLL) model. Female mice were pretreated with hesperidin (20 mg/kg, intraperitoneally, daily for 30 days). Next, the animals received a single intraperitoneal injection of 80 mg/kg ENU on the 30th. Changes in weight and mortality were monitored for 120 days, and then the animals were sacrificed and parameters such as reactive oxygen species (ROS), mitochondrial dysfunction, lysosomal membrane integrity, oxidized/reduced glutathione (GSH/GSSG), and malondialdehyde (MDA) were analyzed in isolated lymphocytes. Hesperidin significantly increases the survival of mice up to 86% and delay in death time and prevents weight changes after exposure to ENU. Also, hesperidin improved cellular toxicity parameters such as ROS formation, MMP collapse, lysosomal membrane destabilization, and lipid peroxidation in isolated lymphocytes. These results promisingly showed that pretreatment with hesperidin increases delay in death time and reduces mortality cellular toxicities consistent with the carcinogenicity of alkylating compounds. This study confirms that the consumption of hesperidin and citrus most likely inhibits alkylating agents-induced carcinogenicity and toxicity.

Striking long-term beneficial effects of single dose psilocybin and psychedelic mushroom extract in the SAPAP3 rodent model of OCD-like excessive self-grooming.

Obsessive compulsive disorder (OCD) is a highly prevalent disorder that causes serious disability. Available treatments leave 40% or more of people with OCD significantly symptomatic. There is an urgent need for novel therapeutic approaches. Mice that carry a homozygous deletion of the SAPAP3 gene (SAPAP3 KO) manifest a phenotype of excessive self-grooming, tic-like head-body twitches and anxiety. These behaviors closely resemble pathological self-grooming behaviors observed in humans in conditions that overlap with OCD. Following a preliminary report that the tryptaminergic psychedelic, psilocybin, may reduce symptoms in patients with OCD, we undertook a randomized controlled trial of psilocybin in 50 SAPAP3 KO mice (28 male, 22 female). Mice that fulfilled inclusion criteria were randomly assigned to a single intraperitoneal injection of psilocybin (4.4 mg/kg), psychedelic mushroom extract (encompassing the same psilocybin dose) or vehicle control and were evaluated after 2, 12, and 21 days by a rater blind to treatment allocation for grooming characteristics, head-body twitches, anxiety, and other behavioral features. Mice treated with vehicle (n = 18) manifested a 118.71 ± 95.96% increase in total self-grooming (the primary outcome measure) over the 21-day observation period. In contrast, total self-grooming decreased by 14.60 ± 17.90% in mice treated with psilocybin (n = 16) and by 19.20 ± 20.05% in mice treated with psychedelic mushroom extract (n = 16) (p = 0.001 for effect of time; p = 0.0001 for time × treatment interaction). Five mice were dropped from the vehicle group because they developed skin lesions; 4 from the psilocybin group and none from the psychedelic mushroom extract group. Secondary outcome measures such as head-body twitches and anxiety all showed a significant improvement over 21 days. Notably, in mice that responded to psilocybin (n = 12) and psychedelic mushroom extract (n = 13), the beneficial effect of a single treatment persisted up to 7 weeks. Mice initially treated with vehicle and non-responsive, showed a clear and lasting therapeutic response when treated with a single dose of psilocybin or psychedelic mushroom extract and followed for a further 3 weeks. While equivalent to psilocybin in overall effect on self-grooming, psychedelic mushroom extract showed superior effects in alleviating head-body twitches and anxiety. These findings strongly justify clinical trials of psilocybin in the treatment of OCD and further studies aimed at elucidating mechanisms that underlie the long-term effects to alleviate excessive self-grooming observed in this study. Prepared with BioRender ( https://www.biorender.com/ ).

The effect of L-carnitine in reactive oxygen species reduction and apoptotic gene expression in mice after cyclophosphamide: An experimental study.

Cyclophosphamide (CP), a utilized anticancer drug, is known to cause infertility in women. However, L-carnitine (LC), an antioxidant, has been shown to offer protective benefits against infertility. This study aimed to evaluate the levels of reactive oxygen species (ROS) and apoptotic gene expression in mice treated with CP and LC. 24 NMRI female mice (6-8 wk, 30 5 gr) were divided into 4 groups: control group: received normal saline intraperitoneal (IP) injection for 10 days; CP group: received 75 mg/kg of CP as a single IP on the 10 day of the experiment; LC group: received 200 mg/kg of LC IP for 10 days; LC+CP group: received LC for 10 days and CP single IP injection on the 10 day of the experiment. After 10 days, mice were superovulated. The oviducts were then removed, and the oocytes of each group were collected for evaluating apoptotic gene expression B-cell lymphoma 2(Bcl2), Bcl2-associated X(Bax), and Caspase3 via real-time polymerase chain reaction and intracellular ROS levels by dichloro-dihydro-fluorescein diacetate fluorescence staining. Data revealed that LC in the LC+CP group significantly increased Bcl2 gene expression (p = 0.01), and decreased Bax and Caspase3 gene expression compared to the CP group (p = 0.03, p = 0.04). LC decreased the ROS level in the LC+CP group compared to the CP group (p 0.001). Findings suggest that LC can scavenge the ROS caused by CP and modulate the apoptotic pathway via downregulating the Bax and Caspase3 genes and upregulating the Bcl2 gene in oocytes of mice exposed to CP.

Oxidative Stress, Inflammation, and Altered Lymphocyte E-NTPDase Are Implicated in Acute Dyslipidemia in Rats: Protective Role of Arbutin

Background/Objectives: Dyslipidemia is frequently linked to various disorders, and its clinical relevance is now recognized. The role of inflammation and oxidative stress (OS) in dyslipidemia has been acknowledged. This study assessed the potential of arbutin (ARB) to prevent dyslipidemia and its associated OS and inflammation in rats with acute hyperlipidemia. Methods: Rats received ARB orally for 14 days and a single intraperitoneal injection of poloxamer-407 on day 15. Results: Poloxamer-407 elevated circulating cholesterol (CHOL), triglycerides (TG), very low-density lipoprotein (vLDL), and LDL, and reduced high-density lipoprotein (HDL)-C and lipoprotein lipase (LPL). ARB ameliorated the circulating lipids and LPL, and suppressed 3-hydroxy-3-methylglutaryl CoA reductase (HMGCR) in rat liver and in vitro. Fatty acid synthase (FAS) in rat liver and its in vitro activity were suppressed by ARB, which also upregulated the LDL receptor (LDL-R) and ABCA1, and had no effect on ABCG5 and ABCG8 mRNA. ARB ameliorated liver malondialdehyde and nitric oxide and enhanced antioxidants in rats with dyslipidemia. Liver NF-κB p65 and blood inflammatory cytokines were increased in dyslipidemic rats, effects that were reversed by ARB. Moreover, ARB effectively suppressed lymphocyte E-NTPDase and E-ADA activities in dyslipidemic rats. The biochemical findings were supported by in silico data showing the affinity of ARB to bind LDL-R PCSK9 binding domain, HMGCR, FAS, and E-NTPDase. Conclusions: ARB possessed anti-dyslipidemia, anti-inflammatory, and antioxidant effects mediated via the modulation of CHOL and TG synthesis, LPL, lymphocyte E-NTPDase and E-ADA, and cytokine release in rats. Thus, ARB could be an effective agent to attenuate dyslipidemia and its associated OS and inflammation, pending further studies as well as clinical trials.

Hypoglycemic and antihyperglycemic effects of Abelmoschus esculentus and Alchornea cordifolia in normal and alloxan-induced diabetic rats

BackgroundAbelmoschus esculentus and Alchornea cordifolia are commonly used in Traditional Chinese Medicine (TCM) to treat several diseases. Abelmoschus esculentus is used to treat infertility and menorrhagia, while Alchornea cordifolia is used for the treatment of venereal diseases, cough, and diarrhoea. However, very few studies assessed the antidiabetic effects of these plants. Therefore, this study aimed to investigate the hypoglycemic and antihyperglycemic effects of aqueous extract of A. esculentus fruits and A. cordifolia leaves. Material and MethodsFresh Abelmoschus esculentus fruits and the powder from the dried leaves of Alchornea cordifolia leaves were prepared by maceration in the aqueous phase (200g/100mL and 50g/100mL respectively) for 24 hours, then filtered and concentrated in an oven at 45°C. Diabete was induced to male Wistar rats by a single intraperitoneal injection of alloxan (150 mg/kg, b.w). Rats with a blood glucose level greater than or equal to 200 mg/dL were selected, divided into groups and were daily administered orally with either aqueous extracts of A. esculentus at 30 mg/kg (AEAE30) or A. cordifolia at 400 mg/kg (AEAC400) for 14 consecutive days. For comparison, acarbose (100 mg/kg), glibenclamide (5 mg/kg), and metformin (500 mg/kg) were administered orally as reference drugs. Moreover, insuline was also used as a positive control and administered intraperitoneally at a dose of 5 IU/kg. Then, blood glucose levels, oral glucose tolerance test, oral maltose tolerance club, body weight and hemoglobin were assessed. For evaluation of the aqueous extracts in the intestinal transit, imodium (2 mg/kg, p.o) and fructine (5 mg/kg, p.o) were used as a positive control to determine the spasmolytic and laxative activities, respectively. The histopathological study of the liver, kidney, pancreas, testis, epididymis, and seminal vesicle was also carried out using the hematoxyline & eosin (H&E) technique. ResultsAEAE30 and AEAC400 significantly reduced (P<0.001) fasting blood glucose (FBG) levels and significantly prevented (P<0.001) postprandial glycemia in AI-db rats following oral glucose tolerance test (OGTT) and oral maltose tolerance test (OMTT) in alloxan-induced diabetic rats (AI-db). In normoglycemic and insulin-resistant (IR) rats, AEAE30 significantly prevented the post-prandial blood glucose level during the OGTT (P<0.01) only in normoglycemic rats. At the end of treatment, AEAE30 significantly reduced the relative weight of the liver (P<0.01) and significantly increased (P<0.001) the relative weight of the testes and pancreas while AEAC400 significantly increased the relative weight of the testes compared to untreated AI-db rats. The histopathological study revealed a restoration of alloxan-induced tissue damage close to the normal control group in AI-db animals treated with plant extracts, as well as an increase in sperm density in the epididymis unlike in the untreated AI-db group. ConclusionThese findings suggested that AEAE and AEAC exhibited hypoglycemic and antihyperglycemic activities and could be therefore a useful source of antidiabetic agent.

Flaxseed extract: A potential therapeutic intervention for type-2 diabetes mellitus and cardiovascular risk reduction in Wistar rats

Introduction: Type 2 diabetes mellitus (T2DM) affects millions of people worldwide, highlighting the importance of dietary interventions. Flaxseed (Linum usitatissimum L.) has shown promise in treating chronic metabolic diseases. This study investigates the therapeutic potential of flaxseed as an antidiabetic agent and its protective effect on cardiovascular diseases. Methods: Rats were randomly divided into five groups: normal fed with drug carrier group (N), diabetes with drug carrier group (D), diabetes with metformin (M) group (D+M), and diabetes with flaxseed extract doses of 100 and 200 mg/kg (D+F1 and D+F2) groups. The diabetic rat model was established by administration of drinking water containing 25% fructose and a single intraperitoneal injection of 100 mg/kg alloxan. After a six-week intervention, the rats were sacrificed to assess the antidiabetic and cardiovascular effects. Results: The doses of 100 and 200 mg/kg flaxseed extract significantly reduced body weight and fasting glucose levels and increased insulin sensitivity index (KITT) compared to the diabetes group (P&lt;0.05). In addition, histological analysis showed a significant improvement and increase in pancreatic beta cells in the treatment group compared to the diabetes group (P&lt;0.05). Flaxseed extract doses of 100 and 200 mg/kg significantly reduced weight gain and insulin resistance, contributing to decreased arterial stiffness and atherogenic index compared to the diabetic group (P&lt;0.05). The hypoglycemic effect suggested a dose-dependent response. Conclusion: Flaxseed extract regulates blood glucose levels probably by enhancing insulin sensitivity and revitalizing damaged pancreatic beta cells. It mitigates cardiovascular risk, by reducing arterial stiffness and atherogenicity.

Ameliorative extract Syzygium cumini seed extract on lipid peroxidation and antioxidant activity in alloxan induced diabetic Wistar albino rats

Aim: This study intends to examine the protective effects of an aqueous extract of Syzygium cumini seed on lipid peroxidation and antioxidant enzyme activities in alloxan-induced diabetic Wistar albino rats. Methodology: A single intraperitoneal injection of 150 mg kg-1 b.wt of alloxan monohydrate was used to induce diabetes in rats. Syzygium cumini group rats were later administered 300 mg kg-1 of Syzygium cumini seed extract by oral gavage for 21 days. On 22nd day, the animals were given general anaesthesia, blood was drawn through the retroorbital plexus, and the kidneys were promptly removed. Lipid peroxidation levels were assessed in the blood (serum) and kidney tissues by measuring the thiobarbituric acid reactive substances, and the activities of antioxidant enzymes such as GST, GPX, SOD and CAT were examined in the blood and kidneys. Results: The acute toxicity experiments revealed that Syzygium cumini did not cause any obvious toxicity indications or mortality at a dosage of 300 mg kg-1, proving the safety of this extract with a broad therapeutic index. The results obtained showed that using aqueous Syzygium cumini seed extract for 21 days considerably (P &lt;0.05) increased the antioxidant enzyme activity for GST, GPx, SOD and CAT, while significantly (P &lt;0.01) decreasing the TBARS levels. Interpretation: Conclusively, the seed extract of Syzygium cumini might be a possible treatment for controlling hyperglycemic oxidative stress complications owing to its antioxidant properties. Key words: Albino rats, Antioxidant enzymes, Lipid peroxidation, Syzygium cumini, Seed extract

Strategy for treating MAFLD: Electroacupuncture alleviates hepatic steatosis and fibrosis by enhancing AMPK mediated glycolipid metabolism and autophagy in T2DM rats

BackgroundRecent studies have highlighted type 2 diabetes (T2DM) as a significant risk factor for the development of metabolic dysfunction-associated fatty liver disease (MAFLD). This investigation aimed to assess electroacupuncture’s (EA) impact on liver morphology and function in T2DM rats, furnishing experimental substantiation for its potential to stall MAFLD progression in T2DM.MethodsT2DM rats were induced by a high-fat diet and a single intraperitoneal injection of streptozotocin, and then randomly assigned to five groups: the T2DM group, the electroacupuncture group, the metformin group, combination group of electroacupuncture and metformin, combination group of electroacupuncture and Compound C. The control group received a standard diet alongside intraperitoneal citric acid - sodium citrate solution injections. After a 6-week intervention, the effects of each group on fasting blood glucose, lipids, liver function, morphology, lipid droplet infiltration, and fibrosis were evaluated. Techniques including Western blotting, qPCR, immunohistochemistry, and immunofluorescence were employed to gauge the expression of key molecules in AMPK-associated glycolipid metabolism, insulin signaling, autophagy, and fibrosis pathways. Additionally, transmission electron microscopy facilitated the observation of liver autophagy, lipid droplets, and fibrosis.ResultsOur studies indicated that hyperglycemia, hyperlipidemia and IR promoted lipid accumulation, pathological and functional damage, and resulting in hepatic steatosis and fibrosis. Meanwhile, EA enhanced the activation of AMPK, which in turn improved glycolipid metabolism and autophagy through promoting the expression of PPARα/CPT1A and AMPK/mTOR pathway, inhibiting the expression of SREBP1c, PGC-1α/PCK2 and TGFβ1/Smad2/3 signaling pathway, ultimately exerting its effect on ameliorating hepatic steatosis and fibrosis in T2DM rats. The above effects of EA were consistent with metformin. The combination of EA and metformin had significant advantages in increasing hepatic AMPK expression, improving liver morphology, lipid droplet infiltration, fibrosis, and reducing serum ALT levels. In addition, the ameliorating effects of EA on the progression of MAFLD in T2DM rats were partly disrupted by Compound C, an inhibitor of AMPK.ConclusionsEA upregulated hepatic AMPK expression, curtailing gluconeogenesis and lipogenesis while boosting fatty acid oxidation and autophagy levels. Consequently, it mitigated blood glucose, lipids, and insulin resistance in T2DM rats, thus impeding liver steatosis and fibrosis progression and retarding MAFLD advancement.

Characterizing the adult zebrafish model of Parkinson's disease: a systematic review of dynamic changes in behavior and physiology post-MPTP administration.

Adult zebrafish are increasingly used in Parkinson's disease (PD) research due to their well-characterized dopaminergic system. Among the toxin-based models, the neurotoxin 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) is widely utilized to induce parkinsonism in adult zebrafish. Therefore, this review presents an overview of the procedures and the dynamic changes in behavior and physiology observed in the adult zebrafish PD model following a single intraperitoneal injection of MPTP. A systematic literature search in the PubMed and Google Scholar databases was conducted to identify relevant articles. Of the 165 articles identified, 9 were included in this review. These chosen articles are original works published before March 2024, all of which utilized adult zebrafish induced with MPTP as the model for PD. Other articles were excluded based on factors such as limited relevance, utilization of zebrafish embryos or larvae instead of adults, and variations in MPTP deliveries. Studies indicated that the ideal model entails the utilization of mixed gender zebrafish aged between 4 and 6 months from the wild-type strain. The acceptable MPTP doses ranges between 20 μg/g (lowest) and 225 μg/g (highest) and doses above 292 μg/g are lethal. Furthermore, noticeable parkinsonian symptoms appear 1 day after administration and persist for more than 1 week. Mitochondrial dysfunction precedes dopaminergic neurodegeneration within this experimental regime. A single administration of MPTP effectively induces PD in adult zebrafish. This study aids in crafting the adult zebrafish PD model, outlining the progressive behavioral and physiological changes ensuing from MPTP administration.

Evaluation of the ameliorative potency of spirulina platensis against cerebellar damage induced by methotrexate in male rats: histopathological, ultrastructural, molecular, and biochemical studies

BackgroundMethotrexate (MTX), a drug utilized in cancer and rheumatoid arthritis treatment, is associated with acute and chronic neurodegenerative alterations. Spirulina platensis (SP) has several important phytochemical substances that act as free radical scavengers or natural antioxidants. The current study investigated the possible effects of the blue-green alga Spirulina platensis on cerebellar damage in male rats exposed to methotrexate. Forty (40) adult male albino rats were randomly divided into 4 groups (n = 10) and treated for one week: GI, the control group; GII was orally given 1000 mg SP/kg/daily, GIII was given a single intraperitoneal injection of MTX 75 mg/kg at the first day, and continued under the normal condition without other treatment till the end of the experiment, and GIV received both SP and MTX together with the same previous doses and duration. Neurobehavioral, histopathological, histochemical, immunohistochemical, ultrastructural, molecular, and biochemical data were recorded.ResultsMTX caused severe cerebellar degeneration in 3 cortical layers, especially the Purkinje layer. The Purkinje layer displayed a disrupted monolayer arrangement with pyknotic nuclei, a significant decrease in cell number, and shrunken cells surrounded by empty spaces. The molecular and granular layers are degenerated with elevated immunoreactions and gene expression of the glial fibrillary acidic protein (GFAP), ionized calcium-binding adapter molecule 1 (Iba-1), and neurofilament light chain antibody (NFL). Moreover, MTX significantly increased malondialdehyde (MDA) and myeloperoxidase (MPO) while decreasing the levels of reduced glutathione (GSH), serotonin, superoxide dismutase (SOD), acetylcholinesterase (ACHE), norepinephrine, and dopamine. These insults were noticeably mitigated by concomitant treatment with spirulina.ConclusionSpirulina improves neurological function by modulating the cerebellar damage elicited by MTX. This improvement may be attributed to the anti-inflammatory and antioxidant properties of spirulina.

EFFECTS OF HYDRAZONE COMPOUND DERIVED FROM 4-AMINO ANTIPYRINE AND BUTANEDIONE AND ITS Ni (II) COMPLEX ON HAEMATOLOGICAL AND DIFFERENTIAL BLOOD COUNT OF ALLOXAN INDUCED DIABETIC RATS

In this study, the effect of Schiff base compound derived from 4-Aminoantipyrine and 1-phenylbutan-1,3-dione and its Ni(II) complex, on the haematology indices of diabetic rats was investigated. A single intraperitoneal injection of Alloxan (120 mg/kg body weight), induced diabetes. The rats were treated with 200 and 400 mg/kg of body weight of 3-[2-(1,5-Dimethyl-3-oxo-2-Phenyl-2,3-Dihydro-1H-Pyrazol-4-yl)Hydrazinylidene]-1-Phenylbutanedione (HL) and its Ni(II) complex ([Ni(HL)2]Cl2) respectively for 14 days, following which they were humanely sacrificed under chloroform. A heart puncture was used to collect blood, and some of the blood samples were analyzed to evaluate white blood cell (WBC), red blood cell (RBC) counts, packed cell volume (PCV) and hemoglobin (Hb) concentration, and differential blood count profiles. Red blood cells, hemoglobin and packed cell volume concentrations in rats treated with HL and [Ni(HL)2]Cl2 in (low and high doses) increased when compared with untreated group B diabetic rats. The Eosinophils, Monocytes, and Lymphocytes were observed to be bad in the diabetic rats; this was significantly better in the treatment groups. HL and [Ni(HL)2]Cl2 have therapeutic promise as functional medicines against diabetes and complete blood count alterations linked to diabetes mellitus, as demonstrated by these studies. In the diabetic group, oxidative stress results in RBC dysfunction, platelet destruction, and tissue injury. These affect the functions of blood cells and the haemostatic parameters which may lead to various complications. As a result, these substances may be used as medication therapy for diabetes since they lower the glucose concentration without alterations in blood cells.

Protective Effects of Spermacoce hispida Against Cisplatin-Induced Nephrotoxicity: A Study on Oxidative and Nitrosative Stress

Background: Nephrotoxicity is a common and severe side effect of cisplatin, a widely used chemotherapeutic agent. The mechanism of cisplatin-induced nephrotoxicity involves oxidative stress, inflammation, and apoptosis, leading to renal damage. There is growing interest in exploring natural products with antioxidant and anti-inflammatory properties as potential protective agents against drug-induced nephrotoxicity.Objective: The present study aimed to investigate the protective effects of Spermacoce hispida against cisplatin-induced nephrotoxicity in an in vivo rat model. Methods: S. hispida was collected, and plant extracts were prepared using different solvents. The prepared extracts underwent phytochemical screening. Nephrotoxicity was induced in rats through a single intraperitoneal injection of cisplatin at a dose of 5 mg/kg. S. hispida extracts at a dose of 100 mg/kg were administered to assess their protective activity. Key parameters measured included blood urea nitrogen (BUN), serum creatinine, oxidative stress markers, proinflammatory cytokines, nitric oxide (NO) levels, and histological alterations in kidney tissue.Results: Cisplatin treatment resulted in increased levels of BUN, serum creatinine, and proinflammatory cytokines in rats, indicating nephrotoxicity. However, treatment with S. hispida extracts for 14 days significantly decreased these elevated levels. Additionally, S. hispida treatment reduced oxidative stress and NO production in cisplatin-treated rats. Histological examination revealed that cisplatin induced structural damage in kidney tissues, which was normalized by S. hispida treatment. Conclusion: The study concludes that Spermacoce hispida exhibits nephroprotective activity, likely by inhibiting oxidative stress and NO production, thereby mitigating cisplatin-induced nephrotoxicity in rats

Unraveling the RAGE-NF-κB pathway: implications for modulating inflammation in diabetic neuropathy through photobiomodulation therapy.

Diabetic peripheral neuropathy (DPN) is a primary complication observed in diabetes that severely affects quality of life. Recent evidence suggests that photobiomodulation (PBM) is a promising therapy against painful conditions and nerve damage. However, the effects of PBM on DPN remains mostly unknown. In the present study, we investigated the efficacy of PBM therapy in modulating proinflammatory cytokine expression in both central and peripheral nervous systems of rats with Streptozotocin (STZ)-induced type 1 diabetes. Male Wistar rats were allocated into control (naïve), diabetic (STZ), and treatment (STZ + PBM) groups. A single intraperitoneal (i.p.) injection of STZ (85mg/kg) was administered for the induction of diabetes. Animals were subjected to 10 treatment sessions, every other day. The results herein presented indicate that PBM treatment diminishes Receptor for Advanced Glycation End-products (RAGE) and Nuclear Factor Kappa B (NF-ϰB) expression in peripheral nervous system and suppresses TNF-α expression in central nervous system tissues. Furthermore, PBM-therapy in diabetic rats also induces increased levels of the anti-inflammatory protein IL-10 in both peripheral and central nervous system. Collectively, our findings demonstrate compelling evidence that PBM-therapy modulates cytokine dynamics and influences RAGE/NF-ϰB axis in a STZ-induced model of type 1 diabetes.

Histological assessment for investigation of dose-dependent ovarian toxicity of cyclophosphamide in the rat

BackgroundCyclophosphamide (CPA) have significant effects on ovarian follicles which lead to ovarian toxicity and impair the normal female reproductive function. This study aimed to evaluate the dose-dependent effects of CPA on rat follicle numbers. MethodsThe experimental groups consisted of rats administered a single intraperitoneal injection of CPA at doses of either 50, 75,150, or 200 mg/kg followed by daily doses of 8 mg/kg for 14 days and control group given no treatment. After the treatment period, the histological evaluation was done. ResultsPrimordial and primary follicles were affected by all doses of CPA, but differential follicle counts revealed that graaf and preantral follicles were most sensitive to CPA, followed by primary and primordial follicles. The greatest reduction in all type of studied follicles caused by CPA doses of 50 mg/kg. ConclusionDifferential follicle counts revealed that CPA-induced ovarian toxicity is exhibited in structural feature of the ovary, particularly in destruction of graaf and preantral follicles in a dose-dependent manner so that the highest decrease in all type of studied follicles caused by 50 mg/kg of CPA and is suggested as the best concentration for ovotoxicity induction. These findings give insight into ovarian response to structural disruption of folliculogenesis.

Mitigation of cisplatin-induced acute kidney injury through oral administration of FAAH Inhibitor PF-04457845.

Fatty acid amide hydrolase (FAAH) serves as the primary enzyme responsible for degrading the endocannabinoid anandamide (AEA). Inhibition of FAAH, either through pharmacological means or genetic manipulation, can effectively reduce inflammation in various organs, including the brain, colon, heart, and kidneys. Infusion of a FAAH inhibitor into the kidney medulla has been shown to induce diuretic and natriuretic effects. FAAH knockout mice have shown protection against both post-ischemia reperfusion injury and cisplatin-induced acute kidney injury (AKI), although through distinct mechanisms. The present study was based on the hypothesis that pharmacological inhibition of FAAH activity could mitigate cisplatin-induced AKI, exploring potential renoprotective mechanism. Male wild type C57BL/6 were administered an oral gavage of a FAAH inhibitor (PF-04457845, 5mg/kg) or vehicle (10% PEG200+5% Tween80+normal saline) at 72, 48, 24, and 2 hours before and 24 and 48 hours after a single intraperitoneal injection of cisplatin (Cis, 25 mg/kg). Mice were euthanatized 72 hours after cisplatin treatment. Compared to vehicle-treated mice, PF-04457845-treated mice showed a decrease of cisplatin-induced plasma creatinine, blood urea nitrogen levels, kidney injury biomarkers (NGAL and KIM-1) and renal tubular damage. The renal protection from oral gavage of PF-04457845 against cisplatin-induced nephrotoxicity was associated with an enhanced AEA tone and reduced levels of DNA damage response biomarkers p53 and p21. Our work demonstrates that PF-04457845 effectively alleviates cisplatin-induced nephrotoxicity in mice, underscoring the potential of orally targeting FAAH as a novel strategy to prevent cisplatin nephrotoxicity. Significance Statement Oral administration of FAAH inhibitor, can reduce cisplatin-induced DNA damage response, tubular damages, and kidney dysfunction. Inactivation of FAAH could be a potential strategy to prevent cisplatin-induced nephrotoxicity.

Neuroprotective Effects of Myricetin on PTZ-Induced Seizures in Mice: Evaluation of Oxidation, Neuroinflammation and Metabolism, and Apoptosis in the Hippocampus.

Epilepsy is one of the most frequently diagnosed neurological diseases, but the neurobiological basis of the disease remains poorly understood. Immunophenotyping CBA mice brain (NeuN and caspase-8) in parallel with hippocampal neurons' functional status and survival rate assessment during acute epileptic PTZ-induced seizures is of particular interest. The aims of this study were to investigate the involvement of NeuN and caspase-8 in cell cycle regulation and the death of hippocampal neurons during PTZ-induced seizures in mice and to assess the therapeutic efficacy of Myricetin in the aforementioned experimental settings. Male CBA mice (n = 340) were divided into six groups to investigate the neuroprotective and antiepileptic effects of Myricetin and Valproic Acid in the PTZ-induced seizure model. Group I (control, n = 20) received a single intraperitoneal injection of NaCl 0.9% solution. Group II (PTZ only, n = 110) received a single intraperitoneal 45 mg/kg PTZ to induce seizures. Group III (Myricetin + PTZ, n = 90) was administered Myricetin orally at 200 mg/kg for 5 days, followed by a PTZ injection. Group IV (Valproic Acid + PTZ, n = 80) received intraperitoneal Valproic Acid at 100 mg/kg for 5 days, followed by PTZ. Group V (Myricetin + NaCl, n = 20) received Myricetin and NaCl. Group VI (Valproic Acid + NaCl, n = 20) received Valproic Acid and NaCl. Seizure severity was monitored using the modified Racine scale. Behavioral assessments included sensorimotor function tests, motor coordination using the rotarod test, and cognitive function via the Morris water maze. Brain tissues were collected and analyzed for oxidative stress markers, including malondialdehyde (MDA), superoxide dismutase (SOD), and glutathione (GSH). Blood samples were analyzed for cytokine levels (IL-1β, IL-6, and TNF-α). Histological studies involved H&E and Nissl staining to evaluate general histopathology and neuronal density. Immunohistochemical analysis was conducted using antibodies against NeuN and caspase-8 to assess neuronal cell cycle regulation and apoptosis. PTZ-induced seizures caused significant oxidative stress and inflammation, leading to neuronal damage. Biochemical analyses showed elevated levels of MDA, SOD, GSH, IL-1β, IL-6, and TNF-α. Histological and immunohistochemical evaluations revealed a significant increase in caspase-8-positive neurons and a decrease in NeuN-positive neurons in the hippocampus and other brain regions, correlating with seizure severity. Myricetin and Valproic Acid treatments reduced oxidative stress markers and neuronal damage. Both treatments resulted in moderate neuronal protection, with fewer damaged neurons observed in the hippocampus, dentate gyrus, and other brain areas compared to the PTZ-only group. Summarizing, Myricetin administration showed promising neuroprotective effects. It significantly reduced oxidative stress markers, including MDA, and restored antioxidant enzyme activities (SOD and GSH), suggesting its antioxidative potential. Myricetin also effectively attenuated the elevation of pro-inflammatory cytokines IL-1β, IL-6, and TNF-α, indicating strong anti-inflammatory properties. Behavioral assessments revealed that Myricetin improved cognitive and motor functions in PTZ-treated mice, with notable reductions in seizure severity and mortality rates. Histological analyses supported these behavioral findings, with Nissl staining showing reduced neuronal damage and NeuN staining indicating better preservation of neuronal integrity in Myricetin-treated groups. Additionally, caspase-8 staining suggested a significant reduction in neuronal apoptosis.

The metabolic clock of ketamine abuse in rats by a machine learning model

Ketamine has recently become an anesthetic drug used in human and veterinary clinical medicine for illicit abuse worldwide, but the detection of illicit abuse and inference of time intervals following ketamine abuse are challenging issues in forensic toxicological investigations. Here, we developed methods to estimate time intervals since ketamine use is based on significant metabolite changes in rat serum over time after a single intraperitoneal injection of ketamine, and global metabolomics was quantified by ultra-performance liquid chromatography-quadrupole-time-of-flight mass spectrometry (UPLC-Q-TOF–MS). Thirty-five rats were treated with saline (control) or ketamine at 3 doses (30, 60, and 90 mg/kg), and the serum was collected at 21 time points (0 h to 29 d). Time-dependent rather than dose-dependent features were observed. Thirty-nine potential biomarkers were identified, including ketamine and its metabolites, lipids, serotonin and other molecules, which were used for building a random forest model to estimate time intervals up to 29 days after ketamine treatment. The accuracy of the model was 85.37% in the cross-validation set and 58.33% in the validation set. This study provides further understanding of the time-dependent changes in metabolites induced by ketamine abuse.