Intraperitoneal Injection Of Saline Research Articles

Limosilactobacillus johnsoni and Limosilactobacillus mucosae and Their Extracellular Vesicles Alleviate Gut Inflammatory Injury by Mediating Macrophage Polarization in a Lipopolysaccharide-Challenged Piglet Model

BackgroundLimosilactobacillus johnsoni (L. j) and Limosilactobacillus mucosae (L. m) can alleviate the inflammatory response. ObjectivesThis study aimed to elucidate the underlying mechanisms by which L. j- and L. m-derived extracellular vesicles (EVs) mitigate lipopolysaccharide (LPS)-induced intestinal injury. MethodsPiglets were assigned to 4 groups: oral phosphate-buffered saline inoculation for 2 wk prior to intraperitoneal injection of physiological saline or LPS, and oral L. j/L. m inoculation for 2 wk prior to intraperitoneal injection of LPS. The intestinal integrity, macrophage markers, cytokine levels, and microbiota were determined. The cytokine levels and macrophage phenotype were detected after L. j/L. m and their EVs were coincubated with macrophages. The levels of cytokines, tight junction proteins, and apoptosis were measured after intestinal epithelial cells were cocultured with macrophages. ResultsLPS challenge decreased jejunal villus length; expression levels of zonula occludens-1 (ZO-1), occludin, arginase-1 (Arg1), and interleukin (IL)-10; and number of CD163+ cells and increased the expression levels of inducible nitric oxide synthase (iNOS), IL-1β, IL-6, and tumor necrosis factor (TNF)-α compared with that in the control. L. j and L. m pretreatment rescued the aforementioned indicators compared with LPS challenge. Pretreatment of L. j and L. m and their EVs reversed the levels of IL-1β, IL-6, TNF-α, and IL-10 and the gene expression of iNOS and Arg1 in the LPS group in macrophages. Pretreatment with L. j and L. m-derived EVs increased ZO-1 and occludin mRNA expression and reduced IL-1β, caspase-3, and bax gene expression in intestinal epithelial cells of the coculture system. Enzyme-treated EVs were less effective than native EVs. ConclusionsThis study suggests that EVs secreted by L. j and L. m control inflammation by modulating macrophage polarization, thereby improving intestinal barrier function.

Analyzing the Potential Antioxidative Effects of Omega-369 in Preventing Acetaminophen-Induced Liver Damage

Background: As acetaminophen (APAP) toxicity has become more common in many countries, related cases of poisoning, whether deliberate or unintentional, have been identified as a key contributor to acute liver failure. Aime: To discover if omega-369 fatty acids could protect the liver of male mice from the effects of acetamiophen. Methods: Thirty-five albino male mice were allocated to one of five groups at random. Group 1 served as the "negative control" and received a single intraperitoneal injection (10 ml/kg) of normal saline on the eleventh day of the test following ten days of receiving liquid paraffin orally at a dose of 10 ml/kg. The liquid paraffin was given to group 2 "positive control". Group 3 received Omega 369 (50 mg/kg/80 ml). Group 4 received Omega 369 (100 mg/kg/35 ml). Group 5 received N-acetylcysteine (100 mg/kg/10 ml). The mice were given Omega-369, N-acetylcysteine, and liquid paraffin via oral gavage for 10 days. Results: Group 2 had significantly lower levels of glutathione peroxidase (GP-X) and superoxide dismutase (SOD) than group 1, but significantly greater levels of malondialdehyde (MDA). GP-X and SOD levels were significantly higher in mice given the doses of omega-369, and N-acetylcysteine prior to acetaminophen administration, whereas MDA levels were significantly lower in groups 3, 4, and 5 when compared with group 2. Conclusion: Omega-369 fatty acids, when taken orally, exhibit antioxidative effects and may reduce the risk of acetaminophen-induced liver injury.

Protective effect of nanoparticle NLRP3 inhibitor (MCC950) on cranial nerves in rats with vascular cognitive impairment

Background: Chronic cerebral hypoperfusion (CCH) is a common cause of Vascular cognitive impairment (VCI). The neuroinflammation caused by ischemia play a key role in the progression of the ailment process. The over activation of NLRP3/Caspase-1/IL-1β signaling aggravates the inflammatory response. We investigated whether NLRP3 inhibitors have a protective effect on cranial nerves in rats with VCI. Methods: Male SD rats were selected arbitrarily and categorized into the sham operation group, CCH group and CCH+MCC950 group (10 mg/kg). The rat model of CCH was formed by bilateral common carotid artery occlusion (BCCAO). The changes in rat’s cerebral blood flow were observed by laser speckle imaging after operation. The sham operation and the CCH group were given intraperitoneal (i.p) injection of normal saline, and the therapeutic group was given i.p having dose of MCC950 (10 mg/kg). The behavior of the rats was analyzed through the Y maze and open field experiments, and the hematoxylin-eosin (HE) staining technique was employed to analyze the behavior of the rats. The histological changes of the rat hippocampus were observed under the following conditions, cell apoptosis was observed by Tune staining, and the protein expressions of NLRP3, IL (interleukin)-1β, and cleaved-Caspase-1 in the hippocampus were analyzed by Western blot. Results: MCC950 treatment enhanced the working memory of CCH rats, and the rats in each group showed no motor deficit. As shown by HE staining, inhibition of NLRP3 reduced neuronal damage caused by pyroptosis. In addition, MCC950 diminishes the appearance of NLRP3, IL-1β, and cleaved-Caspase-1 in the brain tissue of CCH rats by inhibiting inflammasome formation. Conclusions: inhibitors can improve the learning and cognitive ability of rats with vascular cognitive impairment, and the mechanism might be linked to inhibiting the NLRP3/Caspase-1/IL-1β signaling and decreasing apoptosis.

Melatonin Ameliorates Cyclophosphamide-Induced Spermatogenesis Disorder by Reducing Pyroptosis

As a chemotherapeutic drug, cyclophosphamide (CP) has a negative impact on male fertility due to its reproductive toxicity. Melatonin (Mel) promotes the male reproductive system and increases testosterone synthesis. This study is aimed at exploring the molecular mechanism of Mel as a protector of male fertility against CP-induced cytotoxicity. A CP toxicity model was established in adult ICR male mice by intraperitoneal injection of 100 mg/kg CP every other day for a week. Protective effects of Mel on the testis from CP-induced damage were evaluated using four groups of ICR male mice that received intraperitoneal injections of normal saline, 100 mg/kg CP, 10 mg/kg Mel, or the same dosage of CP and Mel, respectively. Testis morphology was observed by hematoxylin and eosin (HE) staining. Sperm quality parameters were evaluated, and sperm function was studied by in vitro fertilization (IVF). Proliferation, meiosis, and pyroptosis markers were examined by western blot. Results showed that CP treatment induced testis toxicity in a time-dependent manner with the most severe damage to the testis at two weeks post CP treatment. CP-treated mice showed reduced testicular weight and impaired spermatogenesis by downregulating PCNA and SYCP3, reduced serum testosterone levels, decreased sperm counts and motility, increased seminiferous tubule vacuolization, and oxidative damage to spermatogenic cells. All these effects, apart from testicular weight, could be ameliorated by Mel administration. The IVF experiment revealed that CP treatment reduced the rates of sperm fertilization and blastocyst development, which were also enhanced by Mel. Mel-treated mice also showed increased expression of proliferation-associated protein PCNA and meiosis-associated proteins REC8, STRA8, and SYCP3, which were all reduced by CP. Furthermore, Mel inhibited the pyroptosis of spermatogenic cells by reducing GSDMD and IL18 expression. In conclusion, this study indicated that Mel might protect the testis from CP-induced DNA damage to germ cells through the alleviation of pyroptosis.

Sinapic acid alleviates 5-fluorouracil-induced nephrotoxicity in rats via Nrf2/HO-1 signalling

Fluoropyrimidine 5-fluorouracil (5-FU) is a DNA analogue broadly used in chemotherapy, though treatment-associated nephrotoxicity limits its widespread clinical use. Sinapic acid (SA) has potent antioxidant, anti-inflammatory, and anti-apoptotic effects, we investigated its protective effects against 5-FU-induced nephrotoxicity in a rat model. We designated four treatment groups each Group I (control) received five intraperitoneal saline injections (once daily) from days 17 to 21; Group II received five intraperitoneal injections of 5-FU (50 mg/kg/day) from days 17 to 21; Group III received an oral administration of SA (40 mg/kg) for 21 days and five intraperitoneal injections of 5-FU (50 mg/kg/day) from days 17 to 21; and Group IV received an oral administration of SA (40 mg/kg) for 21 days (n-six rats in each group). blood samples were collected on day 22 from each group. Animals were sacrificed and their kidneys removed, and instantly frozen. 5-FU caused oxidative stress, inflammation, and activation of the apoptotic pathway by upregulating Bax and Caspase-3 and downregulating Bcl-2. However, SA exposure reduced serum toxicity indicators, boosted antioxidant defences, and reduced kidney apoptosis, which was confirmed by histopathological analysis. Therefore, prophylactic administration of SA could inhibit 5-FU-induced renal injuries in rats via suppression of renal inflammation and oxidative stress, primarily through regulation of NF-κB and proinflammatory cytokines, inhibition of renal apoptosis, and restoration of tubular epithelial antioxidant activities and cytoprotective defences.

Tissue Inhibitor of Metalloproteinase 3 as a potential staging biomarker in hepatocellular carcinoma

Objective: To assess the potential role of tissue inhibitor of metalloproteinase 3 as a staging marker of hepatocellular carcinoma.Method: The experimental study was conducted at Faculty of Pharmacy, Kafrelsheikh University, Egypt, from December 2020 to March 2022 after approval from the Faculty of Pharmacy, Kafrelsheikh University, Egypt, and comprised male albino rats. The subjects were divided into 4 groups. The control group was administrated a single intraperitoneal injection of normal saline, while the other groups were generated post-induction of hepatocellular carcinoma. The induction was done by injecting rats intraperitoneally with a single dose of 200mg/kg diethyl nitrosamine, followed by the administration of 0.05% phenobarbital sodium in drinking water daily. Rats were euthanised at 8, 16 and 24 weeks after the injection to obtain three groups related to the 3 stages of hepatocellular carcinoma. Serum was used for measuring the alpha protein level. Liver homogenates were used for the assessmentof the hepatic tissue inhibitor of metalloproteinase 3 expression, B-cell lymphoma 2-associated X protein expression, and the hepatic content of matrix metalloproteinase -9 and cyclin D1. Data was analysed using Graph Prism 8.Results: Of the 24 rats with weight range 120-130g, 6(25%) were in each of the 4 groups. The relative protein and messenger ribonucleic acid tissue inhibitor of metalloproteinase 3 expressions were significantly decreased in the intervention groups compared to the control group, with more decline as the hepatocellular carcinoma stage increased. The matrix metalloproteinase -9 and cyclin D1 concentrations and the relative hepatic protein Ki67 expression were significantly raised in the intervention groups compared to the control group (p<0.05). The relative expression of hepatic B-cell lymphoma 2-associated X protein was markedly decreased in the intervention groupscompared to the control group (p<0.05).Conclusion: Tissue inhibitor of metalloproteinase 3 might be a promising diagnostic and staging biomarker in hepatocellular carcinoma.Keywords: Carcinoma, Hepatocellular, Neoplasms, Fetoproteins, Metalloproteinase, Cyclin, Antigen, Phenobarbital, Nitrosamines.

Effects of probiotics on pancreatic inflammation and intestinal integrity in mice with acute pancreatitis

BackgroundSevere acute pancreatitis is a potentially life-threatening disease. Despite being a common disorder, acute pancreatitis lacks a specific treatment. The present study aimed to examine the effects of probiotics on pancreatic inflammation and intestinal integrity in mice with acute pancreatitis.MethodsMale ICR mice were randomly divided into 4 groups (n = 6 per group). The control group received two intraperitoneal (i.p.) injections of normal saline as a vehicle control. The acute pancreatitis (AP) group received two i.p. injections of L-arginine 450 mg/100 g body weight. AP plus probiotics groups received L-arginine to induce acute pancreatitis as above. In the single-strain and mixed-strain groups, mice received 1 mL of Lactobacillus plantarum B7 1 × 108 CFU/mL and 1 mL of Lactobacillus rhamnosus L34 1 × 108 CFU/mL and Lactobacillus paracasei B13 1 × 108 CFU/mL by oral gavage, respectively for 6 days starting 3 days prior to the AP induction. All mice were sacrificed 72 h after L-arginine injection. Pancreatic tissue was obtained for histological evaluation and immunohistochemical studies for myeloperoxidase, whereas ileal tissue was used for immunohistochemical studies for occludin, and claudin-1. Blood samples were collected for amylase analysis.ResultsSerum amylase levels and pancreatic myeloperoxidase levels in the AP group were significantly higher than in controls and significantly decreased in probiotic groups compared with the AP group. Ileal occludin and claudin-1 levels were significantly lower in the AP group than in controls. Ileal occludin levels significantly increased, whereas ileal claudin-1 levels did not significantly change in both probiotic groups as compared with the AP group. The pancreatic histopathology showed significantly higher degree of inflammation, edema, and fat necrosis in the AP group, and these changes improved in mixed-strained probiotic groups.ConclusionsProbiotics, particularly the mixed-strain ones, attenuated AP via the reduction of inflammation and the maintenance of intestinal integrity.

The protective role of curcumin nanoparticles on the submandibular salivary gland toxicity induced by methotrexate in male rats

ObjectiveTo evaluate the protective role of nanocurcumin on the toxicity induced by methotrexate in the submandibular glands of rats. DesignTwenty- four healthy male Wistar albino rats were randomly distributed into 3 groups, 8 rats each. Group I-control: rats received a single intraperitoneal injection of saline; Group II-methotrexate (MTX): rats received methotrexate 20 mg/ kg day 1 of the experiment; Group III-methotrexate and nanocurcumin (MTX+NCU): rats received methotrexate 20 mg/ kg on day 1 of the experimental period in addition to nanocurcumin 100 mg/kg/day for 7 days. After euthanasia, the submandibular salivary glands of all rats were collected and prepared for histological, histomorphometric, and immunohistochemical examination (Caspase 3, Bcl2), in addition to transmission electron microscopy. ResultsHistological and ultrastructural assessment revealed less salivary gland damage in the nanocurcumin group in comparison to the methotrexate group, and the percentage of acinar vacuolization showed significantly lower values in the nanocurcumin group. Group III (MTX+NCU) showed lower immunoexpression of caspase 3 than group II (MTX), while Bcl2 immunoreactivity was higher in the MTX group than in the MTX+NCU group. ConclusionsOur results suggest that simultaneous administration of nanocurcumin reduces apoptosis in salivary glands subjected to methotrexate.

Effects of chronic methamphetamine exposure on rewarding behavior and neurodegeneration markers in adult mice.

Recreational and medical use of stimulants among young adults have gained popularity in the United States over the last decade and their use may increase vulnerability to brain biochemical changes and addictive behaviors. The long-term effects of chronic stimulant exposure in later adulthood have not been fully elucidated.Our study investigated whether chronic exposure to methamphetamine (METH), at a dose designed to emulate human therapeutic dosing for ADHD, would promote biochemical alterations and affect sensitivity to the rewarding effects of subsequent METH dosing.Groups of 3.5-month-old male and female C57BL/6J mice were administered non-contingent intraperitoneal injections of either saline or METH (1.4 mg/kg) twice a day for 1 month (5 days/week). METH (0.5 mg/kg)-induced conditioned place preference (CPP) was tested in mice to determine the effects of previous METH exposure on reward-related behavior. Mice were randomly assigned to Experiment I (males and females) or Experiment II (females only) in which CPP testing was respectively performed either 0.5 or 5 months after the end of METH injections, at ~5 or 10 months old respectively. The midbrain and striatum, regions involved in reward circuit, were assessed for markers associated with neurotoxicity, dopaminergic function, neuroinflammation and epigenetic changes after behavioral testing.Previous exposure to chronic METH did not have significant short-term effects on CPP response but led to a decreased CPP response in 10-month-old females. Previous exposure to METH induced some short-term changes to biochemical markers measured in a brain region and sex-dependent manner, while long-term changes were only observed with GFAP and KDM5C.In conclusion, our data suggest sex- and post-exposure duration-dependent outcomes and warrant further exploration of the long-term neurobehavioral consequences of psychostimulant use in both sexes.

Abstract 307: Targeted Activation Of The Tnfa Pathway Accelerates Atherosclerosis Regression In Mice

Aggressive lipid lowering in mice with established atherosclerosis induces plaque regression and is characterized by reduced monocyte recruitment and increased expression of the motility receptor C-C chemokine receptor 7 (CCR7) to promote macrophage egress from plaques to nearby lymph nodes. Notably, genetic deletion of receptors critical for the recruitment of inflammatory Ly6C high monocytes is necessary to facilitate plaque regression. Our group previously showed that deletion of macrophage LDL receptor-related protein 1 (LRP1) in mice with established atherosclerosis accelerates plaque regression after lipid lowering and is associated with increased inflammation and macrophage CCR7-mediated egress. We tested the hypothesis that targeted activation of inflammation following lipid lowering in atherosclerotic mice accelerates atherosclerosis regression. LC-MS/MS proteomics performed on murine macrophages revealed that the loss of LRP1 upregulated the inflammatory protein tumor necrosis factor alpha (TNFα) 1.48-fold (P<0.05). Apoe -/- mice were fed a Western diet for 12 weeks to establish atherosclerosis and then subjected to bone marrow transplantation (BMT) using wildtype ( Apoe +/+ ) mice as donors to restore plasma APOE and reduce lipids. A cohort was sacrificed two weeks after BMT and used as baseline (N=11). The remaining mice (N=12/group) received intraperitoneal injections of recombinant murine TNFα (0.075mg/kg/day) or saline for 6 weeks on chow diet. After 6 weeks on chow diet, total plasma cholesterol levels ranged 82 -120 mg/dL with no significant differences between treatment cohorts. TNFα-treated mice had elevated spleen-to-body weight ratios compared to saline treated controls (P<0.0001), confirming successful activation of the immune system. Following 6 weeks of lipid lowering, TNFα treatment reduced total plaque area and lipid content 17.0% (P<0.05) and 34.4% (P<0.05), respectively, compared to saline treated controls (P<0.05).Herein, we show that the administration of TNFα, to stimulate inflammation after lipid lowering, significantly reduces plaque size and plaque lipid content in mice with established atherosclerosis.

Metabolomics for the identification of early biomarkers of nephrotoxicity in a mouse model of cisplatin-induced acute kidney injury

Background and purposeCisplatin-induced nephrotoxicity manifests as acute kidney injury (AKI) in approximately one third of patients receiving cisplatin therapy. Current measures of AKI are inadequate in detecting AKI prior to significant renal injury, and better biomarkers are needed for early diagnosis of cisplatin-induced AKI. Experimental approachC57BL/6 and FVB/N mice were treated with a single intraperitoneal injection of cisplatin (15 mg kg−1) or saline. Plasma, urine, and kidney samples were collected prior to cisplatin injection and 24-, 48-, 72-, and 96-hours following cisplatin injection. Untargeted metabolomics was employed using liquid chromatography-mass spectrometry to identify early diagnostic biomarkers for cisplatin nephrotoxicity. Principal resultsThere was clear metabolic discrimination between saline and cisplatin-treated mice at all timepoints (day 1 to day 4). In total, 26 plasma, urine, and kidney metabolites were identified as exhibiting early alterations following cisplatin treatment. Several of the metabolites showing early alterations were associated with mitochondrial function and energetics, including intermediates of the tricarboxylic acid cycle, regulators of mitochondrial function and indicators of fatty acid β-oxidation dysfunction. Furthermore, several metabolites were derived from the gut microbiome. Major conclusionsOur results highlight the detrimental effects of cisplatin on mitochondrial function and demonstrate potential involvement of the gut microbiome in the pathophysiology of cisplatin-induced AKI. We provide a panel of metabolites to guide future clinical studies of cisplatin-induced AKI and provide insight into potential mechanisms behind cisplatin nephrotoxicity.

Melatonin alleviates ovarian function damage and oxidative stress induced by dexamethasone in the laying hens through FOXO1 signaling pathway

Oxidative stress can trigger follicular atresia, and decrease follicles quantity in each development stage, thereby alleviating reproductive activity. The induction of oxidative stress in chickens through intraperitoneal injection of dexamethasone is a reliable and stable method. Melatonin has been shown to mitigate oxidative stress in this model, but the underlying mechanism remains unclear. Therefore, this study aimed to investigate whether melatonin can recover aberrant antioxidant status induced by dexamethasone and the specific mechanism behind melatonin-dependent protection. A total of 150 healthy 40-wk-old Dawu Jinfeng laying hens with similar body weights and laying rates were randomly divided into three groups, with five replicates per group and 10 hens per replicate. The hens in the control group (NS) received intraperitoneal injections of normal saline for 30 d, the dexamethasone group (Dex+NS) received 20 mg/kg dose of dexamethasone for the first 15 d, followed by the 15 d of normal saline treatment. While in the melatonin group (Dex+Mel), dexamethasone (20 mg/kg dose) was injected intraperitoneally in the first 15 d, and melatonin (20 mg/kg/d) was injected in the last 15 d. The results showed that dexamethasone treatment significantly enhanced oxidative stress (P < 0.05), while melatonin not only inhibited the oxidative stress but also notably enhanced the antioxidant enzymes superoxide dismutase (SOD), catalase activity (CAT), glutathione peroxidase (GSH-Px), and antioxidant genes CAT, superoxide dismutase 1 (SOD1), glutathione peroxidase 3 (GPX3), and recombinant peroxiredoxin 3(PRDX3) expression (P < 0.05). Melatonin treatment also markedly reduced 8-hydroxy deoxyguanosine(8-OHdG), malondialdehyde (MDA), and reactive oxygen species (ROS) levels (P < 0.05) and apoptotic genes Caspase-3, Bim, and Bax in the follicle. In the Dex+Mel group, the Bcl-2 and SOD1 protein levels were also increased (P < 0.05). Melatonin inhibited the forkhead Box Protein O1(FOXO1) gene and its protein expression (P < 0.05). In general, this investigation revealed that melatonin might decrease oxidative stress and ROS by enhancing antioxidant enzymes and genes, activating the antiapoptotic genes, and inhibiting the FOXO1 pathway in laying hens.

Lycopene attenuates D-galactose-induced cognitive decline by enhancing mitochondrial function and improving insulin signaling in the brains of female CD-1 mice

The incidence of neurodegenerative diseases is severely increasing with ageing. Lycopene (LYC), a carotenoid pigment, has been reported to have antioxidant, anti-inflammatory and neuroprotective properties. In the present study, we aimed to investigate the ameliorative effect of LYC on D-galactose (D-gal) induced cognitive defects and the underlying mechanisms. Forty-five female CD-1 mice (2 months old) were separated into three groups to be fed with either a normal diet or a LYC diet (0.03%, w/w, mixed into normal diet). Meanwhile, the mice were treated by intraperitoneal injection of normal saline or D-gal 150 mg/kg/day for 8 weeks. The behavioral test results indicated that LYC alleviated D-gal induced cognitive impairments. LYC ameliorated brain ageing by decreasing the number of SA-β-gal- stained neurons, downregulating the protein expression of the cellular senescence associated genes P19/P21/P53, increasing the activities of the antioxidant enzymes GSH and SOD, downregulating the level of ROS, inhibiting the activation of MAPKs signaling and downregulating the levels of the inflammatory cytokines IL-1β and TNFɑ in mouse brains. LYC ameliorated synaptic dysfunction by increasing the expression of the neurotrophic factor BDNF and synaptic proteins. Moreover, LYC attenuated D-gal-induced mitochondrial morphological damage, and promoted the expression of mitochondrial functional proteins. LYC also promoted insulin signal transduction in mouse brains through the regulation of IRS-1/AKT/GSK3β signaling.

Effects of gestational inflammation on age-related cognitive decline and hippocampal Gdnf-GFRα1 levels in F1 and F2 generations of CD-1 Mice

BackgroundIt has been reported that age-associated cognitive decline (AACD) accelerated by maternal lipopolysaccharide (LPS) insult during late pregnancy can be transmitted to the second generation in a sex-specificity manner. In turn, recent studies indicated that glial cell line‐derived neurotrophic factor (GDNF) and its cognate receptor (GFRα1) are critical for normal cognitive function. Based on this evidence, we aimed to explore whether Gdnf-GFRα1 expression contributes to cognitive decline in the F1 and F2 generations of mouse dams exposed to lipopolysaccharide (LPS) during late gestation, and to evaluate also the potential interference effect of pro-inflammatory cytokines.MethodsDuring gestational days 15–17, pregnant CD-1 mice (8–10 weeks old) received a daily intraperitoneal injection of LPS (50 μg/kg) or saline (control). In utero LPS-exposed F1 generation mice were selectively mated to produce F2 generation mice. In F1 and F2 mice aged 3 and 15 months, the Morris water maze (MWM) was used to evaluated the spatial learning and memory ability, the western blotting and RT-PCR were used for analyses of hippocampal Gdnf and GFRα1 expression, and ELISA was used to analyse IL-1β, IL-6 and TNF-α levels in serum.ResultsMiddle-aged F1 offspring from LPS-treated mothers exhibited longer swimming latency and distance during the learning phase, lower percentage swimming time and distance in targe quadrant during memory phase, and lower hippocampal levels of Gdnf and GFRα1 gene products compared to age-matched controls. Similarly, the middle-aged F2 offspring from the Parents-LPS group had longer swimming latency and distance in the learning phase, and lower percentage swimming time and distance in memory phase than the F2-CON group. Moreover, the 3-month-old Parents-LPS and 15-month-old Parents- and Father-LPS groups had lower GDNF and GFRα1 protein and mRNAs levels compared to the age-matched F2-CON group. Furthermore, hippocampal levels of Gdnf and GFRα1 were correlated with impaired cognitive performance in the Morris water maze after controlling for circulating pro-inflammatory cytokine levels.ConclusionsOur findings indicate that accelerated AACD by maternal LPS exposure can be transmitted across at least two generations through declined Gdnf and GFRα1 expression, mainly via paternal linage.

How Safe Is Gadobutrol? Examining the Effect of Gadolinium Deposition on the Nervous System

This study aimed to evaluate the safety of gadobutrol, a gadolinium-based contrast agent used in medical imaging, by investigating its effect on the nervous system under physiological and inflammatory conditions. Male Sprague Dawley rats were divided randomly into four groups, including gadobutrol, saline, LPS + gadobutrol, and LPS + saline, and were given intraperitoneal injections of gadobutrol (2.5 mmol/kg) or saline for 20 days. Weekly sensorimotor and cognitive behavioral tests were performed over 4 weeks, and Gd concentration in nervous tissues was analyzed using inductively coupled plasma mass spectrometry (ICP-MS). Lactate dehydrogenase (LDH) activity was measured to evaluate cytotoxicity, and electromyography (EMG) recordings from the gastrocnemius muscle were also obtained to examine signal transmission in sciatic nerves. The results indicated that gadobutrol did not induce significant behavioral changes under normal conditions. However, when administered along with LPS, the combination led to behavioral dysfunction. ICP-MS analysis revealed a higher concentration of Gd in the cerebrum and spinal cord of gadobutrol + LPS-treated rats, while peripheral nerves showed lower concentrations. In addition, there was a significant increase in LDH activity in the hippocampus of the gadobutrol group. EMG responses to electrical stimulation of the sciatic nerve demonstrated a decreased threshold of nociceptive reflexes in the gadobutrol group. Overall, while gadobutrol may be considered safe under normal physiological conditions, the findings suggest that its safety may be compromised under inflammatory conditions.

Tissue Inhibitor Of Metalloproteinase 3 As A Potential Staging Biomarker In Hepatocellular Carcinoma.

To assessthe potential role of tissue inhibitor of metalloproteinase 3 as a staging marker of hepatocellular carcinoma. The experimental study was conducted at Faculty of Pharmacy, Kafrelsheikh University, Egypt, from December 2020 to March 2022 after approval from the Faculty of Pharmacy, Kafrelsheikh University, Egypt, and comprised male albino rats. The subjects were divided into 4 groups. The control group was administrated a single intraperitoneal injection of normal saline, while the other groups were generated post-induction of hepatocellular carcinoma. The induction was done by injecting rats intraperitoneally with a single dose of 200mg/kg diethyl nitrosamine, followed by the administration of 0.05% phenobarbital sodium in drinking water daily. Rats were euthanised at 8, 16 and 24 weeks after the injection to obtain three groups related to the 3 stages of hepatocellular carcinoma. Serum was used for measuring the alpha protein level. Liver homogenates were used for the assessment of the hepatic tissue inhibitor of metalloproteinase 3 expression, B-cell lymphoma 2-associated X protein expression, and the hepatic content of matrix metalloproteinase -9 and cyclin D1. Data was analysed using Graph Prism 8. Of the 24 rats with weight range 120-130g, 6(25%) were in each of the 4 groups. The relative protein and messenger ribonucleic acid tissue inhibitor of metalloproteinase 3 expressions were significantly decreased in the intervention groups compared to the control group, with more decline as the hepatocellular carcinoma stage increased. The matrix metalloproteinase -9 and cyclin D1 concentrations and the relative hepatic protein Ki67 expression were significantly raised in the intervention groups compared to the control group (p<0.05). The relative expression of hepatic B-cell lymphoma 2-associated X protein was markedly decreased in the intervention groups compared to the control group (p<0.05). Tissue inhibitor of metalloproteinase 3 might be a promising diagnostic and staging biomarker in hepatocellular carcinoma.

Antihyperglycemic and Anticholesterolemic Effects of Fig Leaf (Ficus glomerata Linn.) Extract in Alloxan Induced Diabetes in Wistar Albino Rats

In this present scenario,medicinal plants are becoming popular for the treatment of various diseases all over the world. Some of them are being traditionally used in diabetes for human welfare. Diabetes is a common endocrine disorder that impairs glucose metabolism resulting in severe diabetic complications including retinopathy, angiopathy, nephropathy, neuropathy and causing neurological disorder due to perturbation in utilization of glucose. In the present study, an effort has been made to assess the antidiabetic properties of Ficus glomerata leaf extract treatment in alloxan monohydrate induced diabetic albino rats. Present study refers to the medicinal plant Ficus glomerata leaf metabolites compared to insulin treatment animal model. Diabetes was induced in albino rats with the help of alloxan monohydrate (150 mg/kg) intraperitoneal saline injection. Present study was carried out to evaluate the anti-diabetic activity of Ficus glomerata aqueous leaf extract in diabetic albino rats for 4 week treatment. The treatment with aqueous leaf extract at high dose (1g/kg) significantly decreased circulating blood glucose concentration in comparison to mild dose (500mg/kg) and low dose (100mg/kg) respectively in diabetic rats. The aqueous leaf extract also improved the hematological picture, weight, biochemical profile and glucose intolerance lowering effects in diabetic animal models. In conclusion, Ficus glomerata leaf metabolites have tremendous antidiabetic potential in alloxan-induced experimental models. The results suggest the possibility that Ficus glomerata leaves metabolites to cure diabetes in herbal medicine.

CAMP-Epac1 signaling is activated in DDAVP-induced endolymphatic hydrops of guinea pigs

ObjectiveTo explore whether Cyclic Adenosine Monophosphate (cAMP)-Epac1 signaling is activated in 1-Desamino-8-D-arginine-Vasopressin-induced Endolymphatic Hydrops (DDAVP-induced EH) and to provide new insight for further in-depth study of DDAVP-induced EH. MethodsEighteen healthy, red-eyed guinea pigs (36 ears) weighing 200–350 g were randomly divided into three groups: the control group, which received intraperitoneal injection of sterile saline (same volume as that in the other two groups) for 7 consecutive days; the DDAVP-7d group, which received intraperitoneal injection of 10 mg/mL/kg DDAVP for 7 consecutive days; and the DDAVP-14d group, which received intraperitoneal injection of 10 μg/mL/kg DDAVP for 14 consecutive days. After successful modeling, all animals were sacrificed, and cochlea tissues were collected to detect the mRNA and protein expression of the exchange protein directly activated by cAMP-1 and 2 (Epac1, Epac2), and Repressor Activator Protein-1 (Rap1) by Reverse Transcription (RT)-PCR and western blotting, respectively. ResultsCompared to the control group, the relative mRNA expression of Epac1, Epac2, Rap1A, and Rap1B in the cochlea tissue of the DDAVP-7d group was significantly higher (p < 0.05), while no significant difference in Rap1 GTPase activating protein (Rap1gap) mRNA expression was found between the two groups. The relative mRNA expression of Epac1, Rap1A, Rap1B, and Rap1gap in the cochlea tissue of the DDAVP-14d group was significantly higher than that of the control group (p < 0.05), while no significant difference in Epac2 mRNA expression was found between the DDAVP-14d and control groups. Comparison between the DDAVP-14d and DDAVP-7d groups showed that the DDAVP-14d group had significantly lower Epac2 and Rap1A (p < 0.05) and higher Rap1gap (p < 0.05) mRNA expression in the cochlea tissue than that of the DDAVP-7d group, while no significant differences in Epac1 and Rap1B mRNA expression were found between the two groups. Western blotting showed that Epac1 protein expression in the cochlea tissue was the highest in the DDAVP-14d group, followed by that in the DDAVP-7d group, and was the lowest in the control group, showing significant differences between groups (p < 0.05); Rap1 protein expression in the cochlea tissue was the highest in the DDAVP-7d group, followed by the DDAVP-14d group, and was the lowest in the control group, showing significant differences between groups (p < 0.05); no significant differences in Epac2 protein expression in the cochlea tissue were found among the three groups. ConclusionDDAVP upregulated Epac1 protein expression in the guinea pig cochlea, leading to activation of the inner ear cAMP-Epac1 signaling pathway. This may be an important mechanism by which DDAVP regulates endolymphatic metabolism to induce EH and affect inner ear function. Oxford Centre for Evidence-Based Medicine 2011 Levels of EvidenceLevel 5.

Pre-Test Manipulation by Intraperitoneal Saline Injection with or without Isoflurane Pre-Treatment Does Not Influence the Outcome of Social Test in Male Mice

Preclinical studies on rodents should follow the 3R principle minimising the suffering of the animals. To do so, some researchers use inhalation anaesthetic induction even before intraperitoneal injection. However, several studies suggested that both interventions might influence the behaviour of the animals. We aimed to test whether intraperitoneal injection alone or in combination with isoflurane anaesthesia is a preferable treatment method 30 min before a social test. Male C57BL/6 mice were studied using a behavioural test battery comparing three groups (one control group and intraperitoneal saline-treated groups with or without short isoflurane inhalation). Our results confirmed that both interventions had no profound influence on the conventionally measured parameters of social tests (interest in sociability, social discrimination memory, social interaction as well as resident–intruder test) and were not acutely stressful (measured by similar ACTH levels between the groups) not even after repeated administration (similar body weight gain during the one-week observation period). Taking into consideration the possible long-term harmful effect of isoflurane inhalation, we recommend using intraperitoneal injection without it as saline injection did not violate the 3R principle inducing only mild stress.

Reproductive status of male rat offspring following exposure to methamphetamine during intrauterine life: An experimental study.

Methamphetamine abuse during pregnancy is associated with maternal and fetal adverse outcomes. Methamphetamine induces reproductive damage in adults; however, its effect has not been studied during pregnancy. To investigate the effects of methamphetamine exposure during pregnancy on the reproductive system. 15 pregnant Wistar rats were divided into 3 groups (n = 5/group), they received daily intraperitoneal injections of saline or methamphetamine (5, and 10 mg/kg) from day 10 until the end of pregnancy. One adult male offspring was selected from each dam. Subjects were euthanized, and their testis was removed. Sperm samples from cauda epididymis were analyzed for sperm concentration, morphology, and motility. Terminal deoxynucleotidyl transferase dUTP nick-end labeling assay was used to detect apoptotic cells. Levels of B-cell lymphoma 2 protein (Bcl-2) and Bcl-2 associated X-protein were measured using Western blot. Methamphetamine significantly decreased sperm concentration (5 mg vs. saline: p = 0.001, 10 mg vs. saline: p 0.001), normal sperm morphology (saline vs. 10 mg: p = 0.001), and motility (p: saline vs. 5 mg = 0.004, 5 mg vs. 10 mg = 0.011, saline vs. 10 mg 0.001) in a dose-dependent manner. There was a significantly higher number of terminal deoxynucleotidyl transferase dUTP nick-end labeling -positive cells and higher exposure. Moreover, Bcl-2 associated X-protein was increased, and Bcl-2 was decreased in these rats. The present study shows that chronic methamphetamine exposure during intrauterine period can induce apoptosis of seminiferous tubules and decrease sperm quality in adult rats. Moreover, we showed that the intrinsic apoptotic pathway is involved in this process. Further studies are required to identify the complete molecular pathway of these results.