Intraperitoneal Injection Of Normal Saline Research Articles

Exogenous Ghrelin Increases Plasma Insulin Level in Diabetic Rats.

Ghrelin, a 28-amino acid peptide, is a strong growth hormone secretagogue and a regulator of food intake. In addition, ghrelin is thought to play a role in insulin secretion and in glucose homeostasis. A lot of contradictory data have been reported in the literature regarding the co-localization of ghrelin with other hormones in the islet of Langerhans, its role in insulin secretion and attenuation of type 2 diabetes mellitus. In this study, we investigate the effect of chronic ghrelin treatment on glucose, body weight and insulin level in normal and streptozotocin-induced diabetic male Wistar rats. We have also examined the distribution pattern and co-localization of ghrelin with insulin in pancreatic islet cells using immunohistochemistry and immune-electron microscopy and the ability of ghrelin to stimulate insulin release from the CRL11065 beta cell line. Control, non-diabetic groups received intraperitoneal injection of normal saline, while treated groups received intraperitoneal injection of 5 µg/kg body weight of ghrelin (amino acid chain 24–51) on a daily basis for a duration of four weeks. Our results show that the administration of ghrelin increases the number of insulin-secreting beta cells and serum insulin level in both normal and diabetic rats. We also demonstrated that ghrelin co-localizes with insulin in pancreatic islet cells and that the pattern of ghrelin distribution is altered after the onset of diabetes. Moreover, ghrelin at a dose of 10−6 M and 10−12 M increased insulin release from the CRL11065 beta cell line. In summary, ghrelin co-localizes with insulin in the secretory granules of pancreatic beta cells and enhances insulin production.

Noise-induced blood-labyrinth-barrier trauma of guinea pig and the protective effect of matrix metalloproteinase inhibitors

Objective: The research is to study the expression and distribution of matrix metalloproteinase (MMPs)-2 and -9 in the guinea pig cochlea after noise exposure, and to explore the role of MMPs in the blood-labyrinth-barrier (BLB). In addition, the role of MMPs inhibitor doxycycline in noise-induced BLB trauma was studied as well, which provides the basis for further studies and prophylaxis of noise-induced hearing loss. Methods: A total of 45 healthy adult guinea pigs were randomly divided into the control group (15 received intraperitoneal injection of 0.9% saline for 4 consecutive days), the noise-exposure group (15 exposed by 120 dB SPL white noise for 4 h per day for continuous 2 d, intraperitoneal injection of normal saline for 4 consecutive days) and the noise-exposure + doxycycline group (15 exposed by 120 dB SPL white noise exposure for 4 h per day for 2 consecutive days, and intraperitoneal injection of doxycycline 50 mg/kg/d for 4 consecutive days), respectively. Immunofluorescence staining, western blot, and real-time quantitative PCR were used to analyze the distribution and differential expression of MMP-2 and -9 in the stria vascularis of guinea pigs in comparison with the normal control group, noise only group, and noise & doxycycline treatment group. Immunofluorescence staining was used to observe the changes in tight junction (TJ) protein ZO-1 in stria vascularis in three groups and to investigate the effect of acoustic injury on TJs. And ABR tests were utilized to detect the hearing function of guinea pigs in the three groups. Intravenous Evans blue was administrated intravenously as an indicator of vascular leakage among three groups to study the changes in BLB permeability in context of acoustic injury. SPSS 22.0 was used for statistical analysis. Results: There was no significant difference in hearing function between the noise-exposure group and the noise & doxycycline group two hours after noise exposure. After seven, 14 and 28 days noise exposure, the hearing recovery of the noise & doxycycline treatment group was significantly greater than that of the noise-exposure group (P<0.05) . Immunofluorescence staining showed that there was only a small amount of MMP-2 and -9 in the stria vascular in the normal control group, and ZO-1 showed dense linear expression. While, in the noise-explore group, MMP-2 and -9 in the stria vascular was significantly elevated (P<0.05), and the configuration of ZO-1 became loose and discontinuous. However, the MMP-2 and -9 in the noise & doxycycline treatment group were not significantly different from the normal control group (P>0.05), which were significantly less than that in the noise-exposure group, and just a little break of ZO-1 was observed, however, the overall structure remained dense. The leakage of Evans blue from stria vascular capillary in the noise-exposure group was significantly increased, and the difference between the other two groups did not show any statistical significance (P>0.05). Conclusions: The damage of tight junction structure induced by MMP-2 and -9 may play an important role in BLB destruction. In addition, doxycycline can inhibit MMPs secretion, thereby, to some extent, protecting the integrity of BLB from acoustic injury, and contributing to the long-term hearing recovery.

Effect of curcumin, an active substance of turmeric, on acute hyperglycemia induced by ketamine-xylazine: role of α2-adrenergic receptor.

Objective- Medicinal plants and their active constituents are frequently used components for treating hyperglycemia. In the present study, the effect of curcumin was investigated on acute hyperglycemia and hyperinsulinemia induced by ketamine-xylazine in rats. To explore the possible mechanism, yohimbine (an α2-adrenergic receptor antagonist) was also used. Design- An experimental study. Animals– Forty-eight healthy male Wistar rats. Procedures– Rats were divided into eight groups with six rats in each group to receive intraperitoneal injection of normal saline, oral administration of curcumin (12.5, 50 and 200 mg/kg), intraperitoneal injection of yohimbine (0.5 and 2 mg/kg), and oral administration of curcumin (12.5 mg/kg) plus intraperitoneal injection of yohimbine (0.5 mg/kg). After these treatments, ketamine (100 mg/kg) and xylazine (10 mg/kg) were intraperitoneally administered to all groups. Blood glucose concentration was measured at 60 and 5 min before and at, 30, 60, 90 and 120 min after ketamine-xylazine injection. Serum insulin concentration was measured by ELISA kit at the end of experiment. Results- Ketamin-xylazine increased blood glucose and decreased serum insulin. Curcumin lowered increased blood glucose and increased decreased serum insulin. Yohimbine prevented the hyperglycemia and hypoinsulinemia induced by ketamin-xylazine produced the same results as curcumin. Low doses of curcumin and yohimbine induced documented hypoglycemic and hyperinsulinemic effects. Conclusion and Clinical Relevance- Based on the results, it is concluded that curcumin improves hyperglycemia and hypoinsulinemia induced by ketamine- xylazine and α2-adrenergic receptor may involve in this effect.

Effect of sivelestat sodium on early inflammatory reaction and pulmonary edema in rats with smoke inhalation induced lung injury

Objective To investigate the effects of sivelestat sodium on early inflammatory response and pulmonary edema in rats with smoke inhalation-induced lung injury. Methods Seventy-five Sprague-Dawley rats were randomly divided into control group, model group, low-dose group, middle-dose group and high-dose group. The smoke inhalation model was continued except for the control group. The rats were given intraperitoneal injection of normal saline, cevicell sodium 10 mg/kg, 20 mg/kg and 30 mg/kg. The general condition and lung tissue were observed after 24 hours. Results Compared with the control group, rats in each group had a certain degree of lung injury, alveolar wall thickening, alveolar interstitial hyperemia, etc. The alveolar septal thickness, lung tissue wet-dry (W/D) level, the incidence of edema and the levels of tumor necrosis factor-alpha (TNF-α), interleukin-6 (IL-6), neutrophil elastase (NE) and myeloperoxidase (MPO) in the model group were significantly higher than those of the other 4 groups. The lung function recovery was highest in the low dose group and the alveolar septal thickness was the highest among the dose group. The alveolar wall thickening incidence of W/D level and pulmonary edema in lung tissue in the low dose group were (6.76±0.40)μm, (5.18±0.13) and 40.00%, respectively, and the levels of TNF-α, IL-6, NE and MPO were (19.62±0.83)pg/L, (9.41±0.25)pg/L, (6.23±0.18)μg/L and (30.64±0.87)pg/L, significantly better than the middle dose group and high dose group (P<0.05). Conclusions Sivelestat sodium can significantly affect the smoke inhalation lung injury, and improve the recovery of liver function, which can significantly reduce the incidence of inflammation and pulmonary ede-ma in the lung, but its therapeutic effect and drug delivery. There was a significant association between doses and therapeutic effect, in the case of rats, a dose of 10 mg/kg was the best for treatment. Key words: Lung injury; Smoke inhalation injury; Sivelestat sodium; Inflammation; Pulmonary edema; Rats

Dose-Dependent Effects of Long-Term Administration of Hydrogen Sulfide on Myocardial Ischemia-Reperfusion Injury in Male Wistar Rats: Modulation of RKIP, NF-κB, and Oxidative Stress.

Decreased circulating levels of hydrogen sulfide (H2S) are associated with higher mortality following myocardial ischemia. This study aimed at determining the long-term dose-dependent effects of sodium hydrosulfide (NaSH) administration on myocardial ischemia-reperfusion (IR) injury. Male rats were divided into control and NaSH groups that were treated for 9 weeks with daily intraperitoneal injections of normal saline or NaSH (0.28, 0.56, 1.6, 2.8, and 5.6 mg/kg), respectively. At the end of the study, hearts from all rats were isolated and hemodynamic parameters were recorded during baseline and following IR. In isolated hearts, infarct size, oxidative stress indices as well as mRNA expression of H2S-, nitric oxide (NO)-producing enzymes, and inflammatory markers were measured. In heart tissue following IR, low doses of NaSH (0.28 and 0.56 mg/kg) had no effect, whereas an intermediate dose (1.6 mg/kg), improved recovery of hemodynamic parameters, decreased infarct size, and decreased oxidative stress. It also increased expression of cystathionine γ-lyase (CSE), Raf kinase inhibitor protein (RKIP), endothelial NO synthase (eNOS), and neuronal NOS (nNOS), as well as decreased expression of inducible NOS (iNOS) and nuclear factor kappa-B (NF-κB). At the high dose of 5.6 mg/kg, NaSH administration was associated with worse recovery of hemodynamic parameters and increased infarct size as well as increased oxidative stress. This dose also decreased expression of CSE, RKIP, and eNOS and increased expression of iNOS and NF-κB. In conclusion, chronic treatment with NaSH has a U-shaped concentration effect on IR injury in heart tissue. An intermediate dose was associated with higher CSE-derived H2S, lower iNOS-derived NO, lower oxidative stress, and inflammation in heart tissue following IR.

Exogenous hydrogen sulfide improves erectile dysfunction by inhibiting apoptosis of corpus cavernosum smooth muscle cells in rats with cavernous nerve injury

To explore the effects of exogenous hydrogen sulfide (H2S) on apoptosis of corpus cavernosum smooth muscle cells (CCSMCs) and erectile dysfunction (ED) in rats with bilateral cavernous nerve injury (BCNI). Twentyfour male SD rats were randomly divided into 3 groups (n=8):sham operation group, bilateral cavernous nerve injury group (BCNI group) and H2S intervention group (BCNI+NaHS group). In BCNI and BCNI+NaHS groups, BCNI was induced by clamp injury of the bilateral cavernous nerves, and the rats were subjected to daily intraperitoneal injection of normal saline and 100 μmol/kg NaHS solution for 4 weeks, respectively. After the treatment, the intracavernous pressure (ICP) and mean arterial pressure (MAP), ) of the rats were measured. Western blotting was used to detect the expressions of cystathionine β synthetase (CBS), cystathionine γ lyase (CSE), α-SMA, collagen-I, caspase-3, Bax and Bcl-2 in the penile cavernous tissue, and the expressions of CBS and CSE were also detected immunohistochemically. The ratio of cavernous smooth muscle to collagen was detected using Masson's Trichrome staining. The apoptosis level of CCSMC was detected by TUNEL + α-SMA immunofluorescence double staining. After 4 weeks of treatment, the rats in BCNI+NaHS group showed a significantly higher ICP/MAP ratio than those in BCNI group (P < 0.05). The results of Masson's Trichrome staining showed that the ratio of cavernous smooth muscle/collagen was significantly higher in BCNI + NaHS group than in BCNI group (P < 0.05). Western blotting showed a significantly higher expression of α-SMA protein but a lower expression of collagen-I protein in BCNI + NaHS group than in BCNI group (P < 0.05). TUNEL+α-SMA immunofluorescence double staining revealed a significantly lower number of apoptotic CCSMCs in BCNI+NaHS group than in BCNI group (P < 0.05). Compared with those in BCNI group, the rats in BCNI+NaHS group had significantly decreased expressions of caspase-3 and Bax proteins (P < 0.05) with significantly enhanced Bcl-2 protein expression and an increased Bcl-2/Bax ratio (P < 0.05). The expressions of CBS and CSE were significantly lower in BCNI group than in the other two groups (P < 0.05). Exogenous H2S enhance the expression of the classic apoptotic protein Bcl-2 and reduces apoptosis of CCSMC to improve the erectile function in rats with BCNI.

The new complement inhibitor CRIg/FH ameliorates lupus nephritis in lupus-prone MRL/lpr mice

BackgroundsThe aberrant activation of complement system is critically involved in lupus nephropathy. Recent study showed complement C3 inhibitor was effective in the treatment of lupus nephropathy. In this study, we investigate the effect of a novel complement C3 inhibitor, CRIg/FH, in the treatment of lupus nephropathy in MRL/lpr lupus mice.MethodsWe treated MRL/lpr female mice with a dose escalation of CRIg/FH (10, 5 and 2 mg/kg) by intraperitoneal injection twice weekly since 12 weeks age. In addition, MRL/lpr mice treated with intraperitoneal injection of normal saline or oral prednisone, along with C57BL/6 J healthy mice were maintained to serve as controls. We started 8-h urine collection weekly to screen proteinuria by measuring the levels of urine urea/creatinine. Serum samples was collected at week 16 and 20 to measure levels of urea nitrogen, creatinine, and immunological markers (C3, C4, A-ds-DNA) before the mice were sacrificed at 20 weeks age to collect kidneys for histopathological examinations.ResultsOvert skin lesions were observed in MRL/lpr mice treated with normal saline, while skin lesion was not observed in CRIg/FH treated MRL/lpr mice. There was no overt proteinuria observed in MRL/lpr mice treated with CRIg/FH. Serum creatinine and BUN levels in MRL/lpr mice was maintained in highest CRIg/FH dose (10 mg/kg twice a week) to be significantly lower than that in prednisone treated MRL/lpr mice at 20 weeks age. In addition, CRIg/FH treatment in MRL/lpr mice results in a significantly elevated serum C3 and C4 levels when compared to prednisone treatment at both 16 and 20 weeks. Furthermore, our study identified that serum level of A-ds-DNA was also significantly lower in CRIg/FH treatment than that in predisone treated MRL/lpr mice. Renal pathology confirmed that kidneys from CRIg/FH treated MRL/lpr mice suffered less from nephritis and complement disposition.ConclusionOur results showed that the complement inhibitor CRIg/FH can protect MRL/lpr mice from lupus nephropathy by preserving renal function and glomerulus complement activation. Our findings support the positive effect of complement inhibitors in the treatment of lupus nephropathy.

Fucoxanthin, a Marine Xanthophyll Isolated From Conticribra weissflogii ND-8: Preventive Anti-Inflammatory Effect in a Mouse Model of Sepsis.

Background: Fucoxanthin (FX), a xanthophyll pigment which occurs in marine brown algae with remarkable biological properties, has been proven to be safe for consumption by animals. Although FX has various pharmacological effects including anti-inflammatory, anti-tumor, anti-obesity, antioxidant, anti-diabetic, anti-malarial, and anti-lipid, in vivo protective effect against sepsis has not been reported. In this study, we aimed at evaluation the efficacy of the FX in a model of sepsis mouse. Methods: FX was successfully isolated from Conticribra weissflogii ND-8 for the first time. The FX was identified by thin-layer chromatography (TLC), high-performance liquid chromatography-mass spectrometry (HPLC-MS), and nuclear magnetic resonance (NMR). Animals were randomly divided into 9 groups, including Sham group (mouse received an intraperitoneal injection of normal saline 1.0 ml/kg), FX-treated (0.1–1.0 ml/kg), Lipopolysaccharide (LPS)-treated (20 mg/kg), FX+LPS-treated (0.1–10.0 mg/kg and 20 mg/kg, respectively), and urinastatin groups (104 U/kg). Nuclear factor (NF)-κB activation could be potential treatment for sepsis. NF-κB signaling components were determined by western-blotting. IL-6, IL-1β, TNF-α production, and NF-κB activation were evaluated by ELISA and immunofluorescent staining in vitro. Results: FX was found to decrease the expression of inflammatory cytokines including IL-6, IL-1β, and TNF-α, in a prophylactic manner in the LPS-induced sepsis mouse model. Meanwhile, FX significantly inhibits phosphorylation of the NF-κB signaling pathway induced by LPS at the cellular level and reduces the nuclear translocation of NF-κB. The IC50 for suppressing the expression of NF-κB was 11.08 ± 0.78 μM in the THP1-Lucia™ NF-κB cells. Furthermore, FX also inhibits the expression of inflammatory factors in a dose-dependent manner with the IC50 inhibition of IL-6 production was 2.19 ± 0.70 μM in Raw267.4 macrophage cells. It is likely that the molecules with the ability of targeting NF-κB activation and inflammasome assembly, such as fucoxanthin, are interesting subjects to be used for treating sepsis.

Effects of ISO-Alpha-Acids on the Hematoma Volume, Inflammation and Neurological Functions in Intracerebral Hemorrhage Rats and its Potential Mechanism

Abstract INTRODUCTION ISO-alpha-acids (IAA) was reported to be useful for the prevention of dementia as a PPAR-gamma agonist in rats, but the application of IAA in intracerebral hemorrhage (ICH) was still limited. METHODS ICH models were built in Vivo and 90 SD rats were randomized to 5 groups: ICH + IAA intraperitoneal injection (IAA), ICH + normal saline intraperitoneal injection (control), ICH without any intraperitoneal injections (vehicle), Sham, and Mock. Neurological functions of rats, hematoma volume, and brain water content were evaluated 1-d, 3-d, and 7-d postoperation. Microglia of SD rats were sub cultured in Vitro. According to different interventions, 4 groups were set, including IAA, PPAR-gamma antagonist, normal saline (control), and normal cultured (vehicle). Western blotting, immunofluorescence, flow cytometry, and RT-PCR were used to detect the expression of molecular markers, proteins and mRNAs. RESULTS In ICH rats, intraperitoneal injection of IAA was able to promote hematoma clearance, to relieve brain water content and improve neurological functions. Detected by western blot, immunofluorescence, flow cytometry, and reverse transcription polymerase chain reaction (RT-PCR), IAA was able to significantly increase the expression of CD36, CD11b, and CD206, and inhibit the expression of CD80, CD86, TLR4, NF-kappaB, TNF-alpha, and IL-1beta comparing with other groups (P &lt; .05) in Vivo and Vitro. IAA was able to significantly increase both in CD11b and CD206 double-positive anti-inflammatory type microglia and in microglia phagocytosis in the brain (P &lt; .05). CONCLUSION IAA induces microglia to M2 type, characterized by phagocytosis and anti-inflammation, via activating PPAR-gamma signaling pathway in Vivo and Vitro. In ICH rats, administration of IAA is able to facilitate intracerebral hematoma absorption, alleviate brain edema, suppress inflammatory reactions, protect neurological functions, and finally improve rat's outcome.

Effect of Dexmedetomidine on Cerebral Vasospasm and Associated Biomarkers in a Rat Subarachnoid Hemorrhage Model.

The α2 adrenergic agonist dexmedetomidine (DEX) has huge potential for protecting against cerebral vasospasm, a leading cause of death and disability after subarachnoid hemorrhage (SAH). Biomarker assays for SAH have recently emerged as tools for predicting vasospasm and outcomes. We investigated the effects of DEX on vasospasm and assessed relevant biomarkers in a rat SAH model. Male Wistar rats were randomly assigned to sham (n=10), vehicle (n=10), SAH (n=10), or SAH+ DEX (n=10) groups. The SAH and SAH+DEX groups received 0.3 mL injections of autologous blood into the cisterna magna, followed by intraperitoneal injections of normal saline or 10 μg/kg DEX. Forty-eight hours later, neurological deficits as well as the basilar artery (BA) wall thickness and cross-sectional area were measured. Cerebrospinal fluid (CSF) and blood samples were obtained to assess concentrations of interleukin (IL)-6, C-reactive protein (CRP), endothelin-1, and S100-β using enzyme-linked immunosorbent assays. The SAH and SAH+DEX groups exhibited deteriorated neurological function as well as structural and morphological BA vasospasm. The SAH+DEX group showed an improved neurological function score (ie, a 52% decrease), a 10% reduction in wall thickness, and a BA cross-sectional area enlarged by 157%. Compared with the sham group, CSF levels of IL-6 and CRP in the SAH and SAH+DEX groups, as well as serum IL-6 and CRP levels in the SAH group, were significantly elevated. The SAH+DEX group showed significantly lower CSF IL-6 levels than the SAH group. Serum and CSF levels of endothelin-1 and S100-β were similar across all groups. DEX administration reduced the severity of cerebral vasospasm and improved neurological function in SAH rats; this may be closely linked to reduced CSF IL-6 levels.

Effect of exercise on body weight and insulin sensitivity following glucose loading in chronic leptin treated rats

Leptin affects insulin secretion and action through either a central or peripheral mechanism.&nbsp; It increases glucose metabolism and energy expenditure.&nbsp; Though overweight and obese individuals have been reported to have high circulating leptin levels, these effects of leptin are not evident.&nbsp; Exercise, on the other hand, has been found to increase glucose uptake in these individuals.&nbsp; This study examined the effect of chronic leptin treatment and exercise on body weight, glucose homeostasis and insulin sensitivity in Sprague-Dawley rats.&nbsp; Eight-week old rats were treated with either intraperitoneal injection of normal saline (Control; n=8), or leptin (60 μg/kg body weight/day; Leptin; n=8), or leptin and exercise (60 μg/kg body weight/day plus running on a treadmill every other day for 30 minutes at a speed of 30 m/min with 10° inclinations; Leptin-exercise; n=8) or exercise only (running every other day for 30 minutes at a speed of 30 m/min with 10° inclinations on a treadmill; Exercise; n=8) for six weeks.&nbsp; Following six weeks of treatment, glucose challenge was performed by intravenous infusion of 100 mg/ml of glucose for 5 minutes.&nbsp; During the protocol, blood was drawn at 0, 5, 10, 15, 20, 25 and 30-min for blood glucose, serum glucose, and serum insulin levels determination.&nbsp; Data were analyzed using One Way ANOVA with post-hoc analysis and expressed as mean ± standard error of mean (SEM).&nbsp; Despite no different in body weight between the groups, leptin group had a slightly higher trend of mean body weight compared to other groups.&nbsp; Glucose clearance was delayed in the leptin group.&nbsp; This delay in glucose clearance might be associated with lower insulin level and action in leptin group.&nbsp; More importantly, exercise reversed the leptin effects by promoting glucose clearance despite a significantly lower insulin peak, indicating increase in insulin sensitivity.&nbsp; In conclusion, six weeks of daily leptin administration resulted in delayed glucose clearance, but concurrent exercise however prevented these effects of leptin by promoting glucose clearance and increasing insulin sensitivity.

Effects of prenatal exposure to chrysotile asbestos on hippocampal neurogenesis and long-term behavioral changes in adult male rat offspring

Prenatal development is a critical period of life that many environmental pollutants have been suggested to influence fetal growth. Nevertheless, there are still a few investigations into the prenatal exposure to chrysotile asbestos and its neurodevelopmental and behavioral outcome in offspring.In this study, twenty-eight pregnant Wistar rats were divided into four groups and received three-times repeated intraperitoneal injections of normal saline, chrysotile, ascorbic acid and the combination of chrysotile and ascorbic acid on gestational days 11, 14 and 17. The maternal serum levels of malondialdehyde (MDA) and prooxidant-antioxidant balance (PAB) and hippocampal MDA content in adult male offspring were measured. At postnatal day (PND) 60, elevated plus maze was performed to determine anxiety-like behavior, also depression-like behavior was examined using a forced swim test at PND 61- 62. Thereafter, the quantitative analysis of Ki-67, NeuN and GFAP positive cells in the hippocampal dentate gyrus were studied by immunostaining.Our data showed that prenatal exposure to chrysotile increased the maternal serum level of MDA and PAB as well as hippocampal MDA content in adult male offspring, also increased the depression- and anxiety-like behaviors of adult male offspring and decreased the hippocampal Ki-67+, NeuN+ and GFAP+ cells in dantate gyrus of adult male offspring. However, co-administration of ascorbic acid and chrysotile decreased hippocampal lipid peroxidation and increased the Ki-67+, NeuN+ and GFAP+ cells in adult male offspring. In summary, these results indicated that oxidative stress induced by prenatal exposure to chrysotile, lead to the long-lasting decrease of the hippocampal cell proliferation and neuronal differentiation as well as astrogliosis of adult male offspring that exhibit more depression- and anxiety-like behaviors in adulthood and co-treatment of ascorbic acid with chrysotile asbestos attenuated the changes.

Apparent Correlations Between AMPK Expression and Brain Inflammatory Response and Neurological Function Factors in Rats with Chronic Renal Failure.

To explore the correlations between AMP-activated protein kinase (AMPK) expression and brain inflammatory response and neurological function factors in rats with chronic renal failure. Chronic renal failure models in rats were established, and the healthy control group (normal group) was set. Chronic renal failure model rats were divided into model group (without any treatment), control group (intraperitoneal injection of normal saline), A-769662 group (intraperitoneal injection of AMPK specific activator), and compound C group (intraperitoneal injection of AMPK specific inhibitor). The results of HE staining showed renal tissue enlargement, and significant pathological changes. Compared with the normal group, AMPK level in peripheral blood and AMPK mRNA and protein expressions in brain tissue were significantly reduced, and AMPK pathway activation was significantly inhibited in other groups. Compared with the model group, rats in the A-769662 group had significantly decreased serum creatinine (Scr) and blood urea nitrogen (BUN) levels and γ-aminobutyric acid (γ-GABA) content, significantly increased brain-derived neurotrophic factor (BDNF) positive expressions and 5-hydroxytryptamine (5-HT) content, and decreased interleukin-1 (IL-1), tumor necrosis factor-α (TNF-α), and intercellular adhesion molecule 1 (ICAM-1) expressions (all P < 0.05), while it was just the opposite in compound C group (all P < 0.05). There is an apparent correlation between AMPK expression and brain inflammatory response in chronic renal failure rats. AMPK is expected to be an important pathway in the treatment of uremic encephalopathy.

Comparison of anti-peritoneal fibrotic effects between an mTORC1-specific blocker and a PI3K/mTOR dual-blocker

Objective: To compare the anti-peritoneal fibrotic effects between a mammalian target of rapamycin complex 1-specific blocker and a phosphatidyl-inositol 3-kinase/mammalian target of rapamycin dual-blocker.Methods: A total of 40 male Sprague-Dawley rats were randomly divided into five groups with eight animals per group. The normal group (N group) did not receive any intervention. The normal saline group (NS group) received an intraperitoneal injection of normal saline at 1 ml/100 g daily. The model group (3 W group), rapamycin (RAPA) group and BEZ235 (PI3K/mTOR dual-blocker) group all received an intraperitoneal injection of 0.1% chlorhexidine gluconate at 1 ml/100g daily. And the RAPA and BEZ235 groups also received a 0.5 mg/d RAPA or 2.5 mg/d BEZ235 gavage every day, respectively. Rats in each group were sacrificed after 3 weeks.Results: Immunohistochemistry, real-time PCR and western blotting analysis of fibrosis-related indicators (FN, Col 1, and α-SMA) confirmed that RAPA and BEZ235 significantly inhibited peritoneal fibrosis and that these two drugs had similar effects. The p-Akt, p-mTOR, p-p70S6K expression levels were significantly up-regulated in the 3 W group compared to the NS group, confirming that the mTOR pathway was significantly activated during peritoneal fibrosis. RAPA significantly inhibited the phosphorylation of mTOR and p70S6K but did not have significant effects on p-Akt upstream of mTOR. BEZ235 had significant inhibitory effects on all signaling molecules (p-Akt, p-mTOR, and p-p70S6K) in the mTOR pathway.Conclusion: RAPA did not up-regulate p-Akt in a negative feedback fashion. Both drugs effectively inhibited peritoneal fibrosis.

Effects of low-frequency repetitive transcranial magnetic stimulation on depression- and anxiety-like behaviors in epileptic rats.

Low-frequency repetitive transcranial magnetic stimulation (rTMS) has been considered as a treatment option for depression and anxiety. However, its role in epilepsy comorbid with depression and anxiety is unclear. Therefore, we evaluated whether low-frequency rTMS can alleviate depression- and anxiety-like behavior in epileptic rats. Forty-eight adult rats were allocated at random to four groups: Control, Pentylenetetrazol (PTZ), PTZ-rTMS and PTZ-Sham. The control group received intraperitoneal injections of normal saline, while the other groups received intraperitoneal injections of pentylenetetrazol (35 mg/kg/d) once a day for 15 days. Low-frequency rTMS or sham stimulation were administered to the PTZ-rTMS and PTZ-Sham group, respectively, over the two-week period. The open-field test (OFT), elevated plus-maze test (EPM) and forced swimming test (FST) were carried out before the experiment, on the 8th and 15th day to assess depression- and anxiety-like behavior in the rats. Two weeks of low-frequency rTMS treatment could not impair the increases of seizure severity in epileptic rats. However, relative to the PTZ and PTZ-Sham group, the two-week low-frequency rTMS treatment significantly reduced the immobility time in the forced swimming test and attenuated the progressive decrease in total distance traveled, frequency of rearing, velocity in the open-field test, number of entries in the open arms (%) and the time spent in the open arms (%) in the elevated plus-maze test of the PTZ-rTMS group. We proposed that low-frequency rTMS can benefit epileptic rats via amelioration of comorbid depression and anxiety, but it can not alleviate the seizure severity.

Vinpocetine Improves Oxidative Stress and Pro-Inflammatory Mediators in Acute Kidney Injury.

Background:Gentamicin-induced-acute kidney injury (AKI) is a multifaceted phenomenon which previously linked to the oxidative stress only. Vinpocetine prevents reactive free radical generation which contributed in reduction of damage. Therefore, objective of the present study was to investigate the renoprotective effect of vinpocetine on gentamicin-induced-AKI in rats.Methods:Thirty Sprague Dawley Male rat were divided into three groups. Control group (n = 10): Rats treated with distilled water + intra-peritoneal injection of normal saline 2 ml/kg/day. Gentamicin group (n = 10): Rats treated with distilled water + intra-peritoneal injection of gentamicin 100 mg/kg/day. Vinpocetine group (n = 10): Rats treated with vinpocetine + intra-peritoneal injection of gentamicin 100 mg/kg/day. Blood urea and serum creatinine were estimated by auto-analyzer. Serum malondialdehyde (MDA), superoxide dismutase (SOD), Neutrophil Gelatinase Associated Lipocalin (NGAL), kidney injury molecules (KIM-1), and Cystatin-c were measured by ELISA kit methods.Results:Vinpocetine led to significant renoprotective effect on gentamicin induced-AKI through amelioration of blood urea and serum creatinine compared with gentamicin group P < 0.01. Vinpocetine improved oxidative stress through reduction of MDA serum level and elevation of SOD significantly compared with gentamicin group P = 0.001 and P = 0.03, respectively. Indeed, vinpocetine reduced glomerular and renal tubular injury via reduction of inflammatory biomarkers including KIM-1, NGALand Cystatin-c sera levels significantly P < 0.01 compared to gentamicin group.Conclusions:Vinpocetine leads to significant attenuation of gentamicin-induced-AKI through modulation of oxidative stress and pro-inflammatory pathway.

Probucol improves erectile function via Activation of Nrf2 and coordinates the HO-1 / DDAH / PPAR-γ/ eNOS pathways in streptozotocin-induced diabetic rats

BackgroundDiabetic erectile dysfunction (DMED) is mainly attributed to oxidative stress, and Nrf2 plays an important role in cellular antioxidation and regulates NO production in the vascular endothelium. Probucol maintains endothelial function through its antioxidant activity. This study investigated the efficacy and mechanism of probucol in improving erectile function in streptozotocin-induced diabetic rats. MethodsIn our study, thirty 12-week-old Sprague-Dawley male rats were fasted for 12 h. All rats received a 1-time injection of intraperitoneal streptozotocin(60 mg/kg) or vehicle. After 72 h, STZ-treated rats (with random blood glucose concentrations consistently greater than 16.7 mmol/L) were considered diabetic. The diabetic rats were randomly assigned into 2 groups and treated with daily gavage feedings of probucol at doses of 0 and 500 mg/kg for 12 weeks. A positive control group underwent intraperitoneal injection of normal saline followed by daily gavage of saline solution. Erectile function was assessed by electrical stimulation of the cavernous nerves with real-time intracavernous pressure measurement. After euthanasia, penile tissue was investigated using immunohistochemistry, Western blot, and ELISA to assess the proteins of Nrf2/HO-1/DDAH/PPAR-γ/eNOS pathways. ResultsAfter treatment, the rats in the probucol group presented significantly improved erectile function (P < 0.05) than that of the diabetic group without probucol treatment (DM). Also, protein expression of Nrf2, DDAH, PPAR-γ, HO-1 and eNOS was significantly higher than that of the DM group (P < 0.05). CGMP concentrations and SOD concentrations of probucol-treated rats were higher than those of DM group (P < 0.05). The MDA levels and ADMA levels were significantly lower than those of DM group rats (P < 0.05). ConclusionProbucol can improve erectile function via activation of Nrf2, which coordinates the HO-1/DDAH/PPAR-γ/eNOS pathways in streptozotocin-induced diabetic rats.

Effect of glucocorticoid receptor function on the behavior of rats with attention deficit hyperactivity disorder

To investigate the ideal animal models for attention deficit hyperactivity disorder (ADHD) subtypes and the effect of glucocorticoid receptor (GR) function on the behavior of ADHD rats by comparing behavioral differences between spontaneously hypertensive rats (SHRs), Wistar Kyoto (WKY) rats, and Sprague-Dawley (SD) rats. A total of 24 male SHRs aged 21 days were randomly divided into GR agonist group, GR inhibitor group, and SHR group, with 8 rats in each group. Eight male WKY rats and 8 male SD rats, also aged 21 days, were enrolled as WKY group and SD group respectively. The GR agonist group was treated with intraperitoneal injection of dexamethasone (0.5 mg/kg daily); the GR inhibitor group was treated with intraperitoneal injection of mifepristone (RU486) (54 mg/kg daily); the SHR, WKY, and SD groups were treated with intraperitoneal injection of normal saline (0.5 mL/kg daily). The course of treatment was 14 days for all groups. The open field test and Lat maze test were used to evaluate spontaneous activity and non-selective attention. The open field test showed that before drug intervention the SHR group had significantly higher numbers of line crossings and rearings than the WKY and SD groups (P<0.05); the WKY group had a significantly higher number of line crossings than the SD group (P<0.05); the SD group had a significantly higher number of groomings than the WKY group (P<0.05). After drug intervention, the GR agonist group had significantly lower numbers of line crossings and groomings than the SHR group (P<0.05). The Lat maze test indicated that before drug intervention the SHR group had significantly higher numbers of corner crossings and rearings than the WKY and SD groups (P<0.05); the WKY group had significantly higher numbers of rearings and leanings than the SD group (P<0.05). After drug intervention, the GR agonist group had significantly lower numbers of corner crossings and rearings than the SHR group (P<0.05); the GR inhibitor group had a significantly higher number of rearings than the SHR group (P<0.05); the WKY group had significantly higher numbers of rearings and leanings than the SD group (P<0.05). SHR is an ideal animal model for mixed subtype ADHD, and further studies are needed to determine whether WKY rats can be used as an animal model for attention-deficit subtype ADHD. GR agonist can effectively improve spontaneous activity and non-selective attention in SHRs.

Role of erythropoietin in methotrexate-induced nephrotoxicity in adult male albino rats

Introduction: Methotrexate (MTX) could provoke a renal dysfunction. However, beneficial extra-hematopoietic effect of erythropoietin might guard against MTX-induced nephrotoxicity. Objectives: Determination of renoprotective erythropoietin’s role against MTX-induced nephrotoxicity through elucidating its renofunctional and renomorphological effects in adult male albino rats. Materials and Methods: The study was performed on 60 adult male Albino rats, equally divided into three groups; group 1 (control): treated with intraperitoneal injections of normal saline at a dosage of 0.5 mg/kg BW twice weekly for 9 weeks. group 2: injected with MTX hydrate intraperitoneal twice weekly at a dosage of 0.5 mg/kg BW for 9 weeks; and group 3: intraperitoneal injected with MTX hydrate in a similar dosage and duration like group 2 concomitant with subcutaneous injection of 100 IU/kg recombinant human erythropoietin once weekly for 9 weeks. At the study end, serum urea and creatinine together with albuminuria were measured, rats were sacrificed and renal sections were prepared for histopathological examination. Results: Significantly increased values of renal function analyzed substances with deteriorated histopathological renal changes were detected in the MTX-treated group compared to either the control or to the MTX and erythropoietin co-treated group. The later displayed statistically significant decreased levels of the substances accompanied by remarkably ameliorated microscopic renal changes. Additionally, insignificant statistical biochemical and morphological renal differences were noticed between the third and control groups. Conclusion: This study concluded valuable and efficient defense against MTX-induced nephrotoxicity in adult male Albino rats when co-treated with erythropoietin.

Depressant effect of Lithium on apoptosis of nerve cells of adult rats after spinal cord injury

To study whether lithium agent produces neuroprotective effect by inhibiting the nerve cell apoptosis of rats after spinal cord injury. Forty-two male SD rats weighing 200 to 250 g were randomly divided into 3 groups: blank control group(n=6) without surgery, normal saline(NS) group(n=18) with intraperitoneal injection of NS (40 mg/kg); and Lithium chloride (Licl) group (n=18) with intraperitoneal injection of Licl (40 mg/kg). After Allen method modeling, Licl group started intraperitoneal injection of Licl solution (40 mg·kg⁻¹·d⁻¹) within 15 min after operation to the second week. NS group, during the same interval, was injected with a same amount of NS. Postoperative 3, 7, 14 d, BBB scores in each group were measured;the expression of Bcl-2 and Bax protein were observed by immunohistochemisty staining;TUNEL staining was used to observe the nerve cell apoptosis. The BBB scores in blank control group were 21. Postoperative 7, 14 d, BBB scores of Licl group were higher than that of NS group(P<0.05). As for the Bcl-2 protein expression, black control group has a level of 0.081±0.003;7 d and 14 d postoperatively, the level in Licl group was 0.151±0.003, 0.163±0.003 and in NS group, 0.143±0.003, 0.154±0.002, respectively. Licl group showed significantly increased Bcl-2 protein expression(P<0.05). As for the Bax protein expression, black control group showed a level of 0.071±0.003; 7 d and 14 d postoperatively, the level in Licl group was 0.121±0.002, 0.106±0.002 and in NS group was 0.126±0.001, 0.120±0.002, respectively. The Bax protein expression is significantly inhibited in the Licl group(P<0.05). In nerve cell apoptosis by TUNEL staining, the positive cells were fewer in the black control group with apoptosis index (AI) of 1.98±0.19;while 7d and 14d postoperatively, the AI of Licl group was 13.12±0.69, 4.29±1.00 and of NS group, 18.26±0.87, 5.48±0.70, respectively. Licl group showed significant inhibition of the cell apoptosis(P<0.05). Licl can promote the Bcl-2 protein expression and inhibit the Bax proteins expression in nerve cells of rat after SCI, thereby playing a role in the inhibition of nerve cell apoptosis. This may be one of the mechanisms that Licl can promote the recovery of motor function of rats after SCI.