Intraperitoneal Injection Of Normal Saline Research Articles

Luteolin inhibits pancreatitis‑induced acinar‑ductal metaplasia, proliferation and epithelial‑mesenchymal transition of acinar cells.

Luteolin, a flavone, has been demonstrated to have anti-cancer properties. In the current study, the effects of luteolin on certain carcinogenesis-associated changes induced by pancreatitis, which are significant risk factors for pancreatic cancer, were investigated. Male six-week-old C57BL6 mice used in the current study were divided into three groups; the control group, acute pancreatitis group and luteolin group. Intra-peritoneal injection of cearulein was performed in the acute pancreatitis group and luteolin group to induce acute pancreatitis whereas the luteolin group received intra-peritoneal injection of luteolin. The control group received intra-peritoneal injection of normal saline. Then, the expression of SOX9, phosphorylated (p-) STAT3, p-EGFR, cytokeratin-19, Ki67 and N-cadherin were determined by immunohistochemistry. Morphological changes of acinar cells were determined by hematoxylin and eosin staining. The mRNA expression of the epithelial-mesenchymal transition markers CDH1, CDH2, Slug, Zeb1, EpCAM, ZO1, Vimentin, Snail and Twist was determined by reverse transcription-quantitative polymerase chain reaction. It was identified that luteolin inhibits the formation of tubular complexes and ectopic expression of cytokeratin-19 and luteolin also decreased proteins of SOX9, p-STAT3 and p-EGFR. In addition, luteolin inhibits proliferation and epithelial-mesenchymal transition of acinar cells induced by acute pancreatitis. As tubular complex formation and ectopic expression of cytokeratin-19 were two prominent characters of acinar-ductal metaplasia, it was concluded that luteolin inhibits acinar-ductal metaplasia induced by pancreatitis and also inhibits pancreatitis-induced proliferation and epithelial-mesenchymal transition of acinar cells. Acinar-ductal metaplasia and proliferation have close associations with pancreatic carcinogenesis. It is suggested that luteolin has potential anti-pancreatic carcinogenesis effects and merits further investigation.

Toll-like receptor-9 is involved in the development of B cell stimulating factor-induced systemic lupus erythematosus.

The objective of the present study was to investigate the role of Toll-like receptor (TLR)-9 in B lymphocyte stimulating factor (BLyS)-induced systemic lupus erythematosus (SLE) in mice. The anti-double stranded (ds)DNA antibody titer, levels of complement proteins (C3 and C4), interleukin (IL)-10 and the disease activity [assessed by the erythrocyte sedimentation rate (ESR) and C-reactive protein (CRP) level] were measured. A total of 21 transgenic female mice (aged 8-10 weeks and weighing 30-40 g) expressing the Epstein-Barr virus membrane antigen, BLLF1, were studied. Mice were randomly divided into the control, the BLyS inhibition and the TLR-9 inhibition groups, with 7 mice in each group. Mice in the blank control group received intraperitoneal injections of normal saline, mice in the BLyS inhibition group received intraperitoneal injections of anti-BR3 monoclonal antibody (5,000 ng/day) and mice in the TLR-9 inhibition group received intraperitoneal injections of anti-human TLR-9 antibody (250 ng/day). The treatment regimens continued for 10 days, followed by the collection of peripheral venous blood. The relative levels of TLR-9 mRNA were measured by reverse transcription-quantitative polymerase chain reaction. Furthermore, the BLyS protein concentration and IL-10 levels were measured by ELISA. TLR-9 mRNA, BLyS, IL-10, anti-dsDNA antibody titer, C3, C4, ESR and CRP levels of the blank control group were significantly higher than those of the other two groups (P<0.05). The differences in comparison of these indexes between the BLyS inhibition and TLR-9 inhibition groups were not statistically significant (P>0.05), with the exception of TLR-9 mRNA and BLyS. In conclusion, the TLR-9 signaling pathway may be important for BLyS-induced SLE, and regulation of the inflammatory immune level.

Protective Effect of Moringa Oleifera Leaves Against TramadolInduced Nephrotoxicity in Mice

Tramadol, a broadly in recent years, is an effective analgesic agent for the treatment of moderate to acute pain. Its metabolites are excreted by the kidney which may cause nephrotoxicity. Moringa oleifera leaves are commonly used to provide herbal and plant-derived medicinal products especially in developing nations. The present study was carried out to determine the biochemical and histopathological changes in the kidney of tramadol-treated albino mice and to evaluate the possible protective role of Moringa oleifera leaves against tramadol-induced nephrotoxicity. Twenty adult albino mice were divided into four groups. Control group (group i) received daily intraperitoneal injection of normal saline only, group ii received oral dose of Moringa oleifera leaves extract (20 mg/kg/bw) for three weeks, group iii received daily intraperitoneal dose of tramadol (0.3 mg/kg/bw) for the same period, group iv, received daily oral dose of Moringa oleifera leaves extract, (20 mg/kg/bw) three hours before injecting intraperitoneal dose of tramadol (0.3 mg/kg/bw), for the same period. Blood samples were withdrawn at the end of the experiment for kidney function tests and specimens from the kidney were processed for histological study. No significant differences in the mean values of the kidney function tests were noticed between Moringa oleifera group and control group. However, there was highly significant increase in the mean values of serum, urea and creatinine in tramadol-treated group as compared to the control group. Although tramadol + Moringa oleifera group revealed significant difference in the mean values of urea and creatinine when compared with tramadol-treated group. So, Moringa oleifera leaves extract have been shown to attenuate the renal dysfunction, improve the renal architecture, with nearly normalization of serum urea and creatinine levels which indicate improvement of renal function. In conclusion, in the light of biochemical results and histological findings, co-administration of Moringa oleifera leaves lessened the negative effects of tramadol-induced nephrotoxicity; possibly by its antioxidant action. Further investigation of these promising protective effects of Moringa oleifera leaves against tramadol-induced renal injury may have considerable impact on developing an adjunct therapy aiming to improve the therapeutic index of some nephrotoxic drugs.

Experimental study on the glucagon-likepeptide 1 receptor agonist modulation of insulin resistance and hepatic oxidative stress in rats with diabetes mellitus combined with nonalcoholic fatty liver disease

To study the effect of glucagon-likepeptide 1(GLP-1)receptor agonist on insulin resistance and hepatic oxidative stress in rats with diabetes mellitus combined with nonalcoholic fatty liver disease. 36 male SD rats were served as the experimental animal and randomly divided into control group, model group, and GLP-1 group. The rats of control group were given routine diet with intraperitoneal injection of normal saline, those in model group were given high fat diet and intraperitoneal injection of normal saline, while GLP-1 group rats were fed with high fat diet and intraperitoneal injection of liraglutide. After 4 weeks of treatment, insulin resistance, lipid metabolism, liver injury and oxidative stress were all assessed. Serum fasting blood glucose, fasting insulin, total cholesterol, triglyceride, alanine transaminase(ALT), aspartate transaminase(AST)levels and total cholesterol, triglyceride contents in liver tissue, and as well as homeostasis model assessment for insulin resistance(HOMA-IR)levels of model group were significantly higher than those of control group, complex insulin sensitivity index(ISIcomp)level was significantly lower than that of control group; serum fasting blood glucose, fasting insulin, total cholesterol, triglyceride, ALT, AST contents and HOMA-IR levels of GLP-1 group were significantly lower than those of model group, ISIcomp level was significantly higher than that of model group; superoxide dismutase(SOD), glutathione peroxidase(GSH-Px), catalase(CAT)contents in liver tissue of model group were significantly lower, while malondialdehyde content and SOD, GSH-Px, CAT, NF-E2 related factor-2(Nrf-2), antioxidant response element(ARE), heme oxygenase-1(HO-1), quinone oxidoreductase-1(NQO-1), glutathione thiol transferase(GST)mRNA expression were significantly higher than control group; SOD, GSH-Px, CAT contents and SOD, GSH-Px, CAT, Nrf-2, ARE, HO-1, NQO-1, GST mRNA expression in the liver tissue of GLP-1 group were significantly higher, while malondialdehyde content was significantly lower than that of model group. GLP-1 receptor agonist reduces insulin resistance and liver oxidative stress injury in diabetic rats with nonalcoholic liver disease. (Chin J Endocrinol Metab, 2017, 33: 228-232) Key words: Diabetes mellitus, type 2; Nonalcoholic fatty liver disease; Glucagon like peptide -1; Insulin resistance; Oxidative stress

Effects of renal denervation on monocrotaline induced pulmonary remodeling.

Pulmonary artery hypertension (PAH) is a rapidly progressive disorder, which leads to right heart failure and even death. Overactivity of the renin-angiotensin-aldosterone system (RAAS) and sympathetic nervous system accounts for the development and progression of PAH. The role of renal denervation (RDN) in different periods of PAH has not been fully elucidated. A single intraperitoneal injection of monocrotaline (MCT, 60 mg/kg) was used to induce pulmonary remodeling in male Sprague Dawley rats (n = 40). After 24-hour of MCT administration, a subset of rats underwent RDN (RDN24h, n = 10); after 2-week of MCT injection, another ten rats received RDN treatment (RDN2w, n = 10) and the left 20 rats were divided to MCT group with sham RDN operation (MCT, n = 20). Eight rats in Control group received intraperitoneal injection of normal saline (60 mg/kg) once and sham RDN surgery. After 35 days, tissue and blood samples were collected. Histological analysis demonstrated that the collagen volume fraction of right ventricle, lung tissue and pulmonary vessel reduced significantly in RDN24h group but not in the RDN2w group, compared with MCT group. Moreover, the earlier RDN treatment significantly decreased SNS activity and blunted RAAS activation. Importantly, RDN treatment significantly improved the survival rate. In summary, earlier RDN treatment could attenuate cardio-pulmonary fibrosis and therefore might be a promising approach to prevent the development of PAH.

Effect of exendin-4 on lipid deposition in skeletal muscle of diet-induced obese mice and its underlying mechanism

Objective: To investigate the effect of exendin-4, a glucagon-like peptide-1 (GLP-1) receptor agonist, on reducing lipid deposition and improving insulin resistance in skeletal muscle and the underlying mechanisms in high-fat diet (HFD)-induced obese mice. Methods: Twelve male C57BL/6J mice were challenged with HFD for 12 weeks to induce obesity and then randomly divided into two groups: exendin-4 group (intraperitoneal injection of 24 nmol·kg-1·d-1 exendin-4 for 4 weeks) and HFD group (intraperitoneal injection of normal saline for 4 weeks), with 6 mice in each group. Additional 6 mice were also selected as control group. Body weight, fasting blood glucose were recorded. Serum triglyceride (TG), total cholesterol (TC), insulin and skeletal muscle triglyceride levels were measured by enzyme-linked immunosobent assay (ELISA). Oil red O staining was used for morphologic changes of frozen sections from skeletal muscle. The protein levels of lipid metabolic pathway mediated by AMP-activated protein kinase (AMPK) and insulin signailing pathway were determined by Western blot. Results: Compared with mice in HFD group, exendin-4 significantly decreased body weight[(37.68±1.80) vs (46.03±5.00) g, P<0.025], fasting blood glucose[(5.40±0.33) vs (7.65±1.92) mmol/L, P<0.025], serum TG[(37.78±7.14) vs (80.76±34.22) mg/dl, P<0.025], TC[(180.13±18.75) vs (217.57±22.52) mg/dl, P<0.025], insulin[(0.58±0.01) vs (1.67±1.23) ng/ml, P<0.025]and skeletal muscle TG levels[(9.84±1.08) vs (19.35±7.44) mg/g, P<0.025]of obese mice. Oil red O staining revealed that exendin-4 alleviated the accumulation of larger lipid droplets in skeletal muscle. The protein expressions of lipolysis and lipid oxidation mediated by AMPK and insulin signailing pathway were up-regulated, and the protein expressions of lipogenesis mediated by AMPK were down-regulated after intervention of exendin-4. Conclusion: Exendin-4 reduces lipid deposition and insulin resistance in skeletal muscle of HFD-induced obese mice via activating AMPK and up-regulating insulin signailing pathway.

Sympathetic Denervation Accelerates Wound Contraction but Inhibits Reepithelialization and Pericyte Proliferation in Diabetic Mice.

Previous studies focused on the effects of sympathetic denervation with 6-hydroxydopamine (6-OHDA) on nondiabetic wounds, but the effects of 6-OHDA on diabetic wounds have not been previously reported. In this study, treated mice received intraperitoneal 6-OHDA, and control mice received intraperitoneal injections of normal saline. Full-thickness wounds were established on the backs of mice. The wounds were sectioned (four mice per group) for analysis at 2, 5, 7, 10, 14, 17, and 21 days after injury. The wound areas in the control group were larger than those in the treatment group. Histological scores for epidermal and dermal regeneration were reduced in the 6-OHDA-treated group on day 21. The mast cells (MCs) in each field decreased after sympathectomy on days 17 and 21. The expression levels of norepinephrine, epidermal growth factor (EGF), interleukin-1 beta, NG2 proteoglycan, and desmin in the treatment group were less than those in the control group. In conclusion, 6-OHDA delays reepithelialization during wound healing in diabetic mice by decreasing EGF, but increases wound contraction by reducing IL-1β levels and the number of MCs. Besides, 6-OHDA led to reduced pericyte proliferation in diabetic wounds, which might explain the vascular dysfunction after sympathetic nerve loss in diabetic wounds.

Protective effects of hydrogen rich water on the intestinal ischemia/reperfusion injury due to intestinal intussusception in a rat model.

This study aimed to investigate the protective effects of hydrogen rich water on the intestinal ischemia/reperfusion (I/R) injury in a rat intestinal intussusception (II) model. Ninety Sprague-Dawley rats were randomly assigned into three groups (n = 30 per group). In sham group, rats received laparotomy, and the intestine was exposed for 15 minutes without II. In I/R + saline group and I/R + hydrogen group, rats received II after laparotomy and then intestine was relocated 8 hours later, followed by immediately intraperitoneal injection of normal saline and hydrogen rich water (HRW) (5 mL/kg), respectively. One hour later, the intestine was collected for hematoxylin-eosin staining and immunohistochemistry for apoptotic cells and 8-oxo-deoxyguanosine, and blood was harvested for detection of tumor necrosis factor-α, malondialdehyde and superoxide dismutase. Hematoxylin-eosin staining showed the intestinal mucosa was significantly damaged in I/R + saline group, which was markedly attenuated after HRW treatment. The serum tumor necrosis factor-α content increased significantly in I/R + saline group, but HRW treatment reduced serum tumor necrosis factor-α content as compared to I/R + saline group (P < 0.05). Serum malondialdehyde content and 8-oxo-deoxyguanosine positive cells in the intestine increased dramatically after II, but HRW significantly reduced them in I/R+hydrogen group (P < 0.05). In addition, superoxide dismutase activity reduced markedly and apoptotic cells increased in I/R + saline group as compared to sham group, but they HRW increased superoxide dismutase activity and reduced apoptotic cells significantly in I/R + hydrogen group (P < 0.05). Our results indicate hydrogen rich water is able to attenuate II induced intestinal I/R injury via inhibiting intestinal inflammation, attenuating intestinal/serum oxidative stress and reducing apoptotic intestinal cells.

Protective effects of oridonin on the sepsis in mice

This study aimed to investigate the protective effects of oridonin (ORI) on cecal ligation and puncture (CLP)-induced sepsis in mice. Male C57BL/6 mice weighing 22–30 g and aged 8–10 weeks were randomly assigned to three groups: Sham group, CLP group, or CLP plus ORI group. In the CLP group and ORI group, CLP was induced, and intraperitoneal injection of normal saline and oridonin (100 μg/kg) was conducted, respectively. The survival rate was determined within the following 7 days. The blood, liver, and lung were collected at 24 hours after injury. Hematoxylin–eosin staining of the lung, detection of lung wet-to-dry ratio, and serum cytokines (tumor necrosis factor [TNF]-α and interleukin [IL]-6), and examination of intraperitoneal and blood bacterial clearance were conducted to evaluate the therapeutic efficacy. Results showed that ORI treatment significantly reduced the lung wet-to-dry ratio, decreased serum TNF-α and IL-6, and improved liver pathology compared with the CLP group (p < 0.05). Moreover, the intraperitoneal and blood bacterial clearance increased markedly after ORI treatment (p < 0.05). The 7-day survival rate in the ORI group was also dramatically higher than in the CLP group (p < 0.05). Our findings indicate that ORI can attenuate liver and lung injuries and elevate bacterial clearance to increase the survival rate of sepsis mice.

Effects of Methane-Rich Saline on the Capability of One-Time Exhaustive Exercise in Male SD Rats.

PurposeTo explore the effects of methane-rich saline (CH4 saline) on the capability of one-time exhaustive exercise in male SD rats.MethodsThirty rats were equally divided into to three groups at random: control group (C), placebo group (P) and methane saline group (M). Rats in M group underwent intraperitoneal injection of CH4 saline, and the other two groups simultaneously underwent intraperitoneal injection of normal saline. Then, the exercise capability of rats was tested through one-time exhaustive treadmill exercise except C group. Exercise time and body weight were recorded before and after one-time exhaustive exercise. After exhaustive exercise, the blood and gastrocnemius samples were collected from all rats to detect biochemical parameters in different methods.ResultsIt was found that the treadmill running time was significantly longer in rats treated with CH4 saline. At the same time, CH4 saline reduced the elevation of LD and UN in blood caused by one-time exhaustive exercise. The low level of blood glucose induced by exhaustive exercise was also normalized by CH4 saline. Also CH4 saline lowered the level of CK in plasma. Furthermore, this research indicated that CH4 saline markedly increased the volume of T-AOC in plasma and alleviated the peak of TNF-α in both plasma and gastrocnemius. From H&E staining, CH4 saline effectively improved exercise-induced structural damage in gastrocnemius.ConclusionsCH4 saline could enhance exercise capacity in male SD rats through increase of glucose aerobic oxidation, improvement of metabolic clearance and decrease of exhaustive exercise-induced gastrocnemius injury.

Dose-dependent effect of ghrelin on gastric emptying in rats and the related mechanism of action

The aim of this study was to investigate the dose-dependent effect of ghrelin on gastric emptying in rats and the related mechanism of action. Sixty Wistar rats were randomized into control and test groups, which respectively received intraperitoneal injection of normal saline and ghrelin at different doses (0.5 nmol/kg, 1.0 nmol/kg, 1.5 nmol/kg, 2.0 nmol/kg, and 2.5 nmol/kg). After 45 minutes, all rats were gavaged with semisolid paste. The gastric emptying rate was determined 30 minutes later, and the plasma cholecystokinin level was tested by radioimmunoassay. The mean gastric emptying rate in the test groups was significantly higher than in the control group (38.24 ± 7.15% and 27.18 ± 2.37%, respectively, p < 0.05). Medium and high doses of ghrelin (1.0 nmol/kg, 1.5 nmol/kg, 2.0 nmol/kg, and 2.5 nmol/kg), but not low dose (0.5 nmol/kg), accelerated the gastric emptying. In addition, the plasma cholecystokinin level in the test groups was significantly higher than in the control group (p < 0.01). The gastric emptying rate was positively correlated with the plasma cholecystokinin level (p < 0.01). Intraperitoneal injection of ghrelin at medium and high doses significantly accelerated gastric emptying in rats.

Correlation between uncoupling protein 2 expression and myocardial mitochondrial injury in rats with sepsis induced by lipopolysaccharide

To investigate the correlation between uncoupling protein 2 (UCP2) expression and myocardial mitochondria injury in rats with sepsis induced by lipopolysaccharide (LPS). The rat model of sepsis was established through an intraperitoneal injection of LPS. Forty male Sprague-Dawley rats were randomly and equally divided into control group (an intraperitoneal injection of normal saline), sepsis 6 h group (LPS-6 h group), sepsis 12 h group (LPS-12 h group), sepsis 24 h group (LPS-24 h group), and sepsis 48 h group (LPS-48 h group). The serum and heart tissues were harvested at corresponding time points and myocardial mitochondria was extracted. The microplate reader was applied to measure creatine kinase (CK), creatine kinase-MB (CK-MB), and reactive oxygen species (ROS). Flow cytometry was applied to measure the degree of mitochondrial swelling and mitochondrial membrane potential (MMP). Western blot was used to measure the expression level of UCP2. Electron microscopy was applied to observe the morphological changes in heart tissues and myocardial mitochondria. Compared with the control group, the LPS groups had significantly increased serum levels of CK, CK-MB, and myocardial ROS, as well as a significantly increased degree of mitochondrial swelling (P<0.05), and these values reached their peaks at 24 hours after LPS injection. The LPS groups had a significant decrease in MMP (P<0.05), which reached the lowest level at 24 hours after LPS injection. Western blot showed that the LPS groups had a significant increase in the expression level of myocardial UCP2 compared with the control group (P<0.05), which reached its peak at 24 hours after LPS injection. The results of electron microscopy showed mitochondrial swelling, partial rupture of the mitochondrial membrane, and cavity formation in rats in the LPS groups. The most severe lesions occurred in the LPS-24 h group. In rats with LPS, the ROS level in the myocardial mitochondria and the degree of mitochondrial swelling were positively correlated with the expression level of UCP2 (r=0.796 and 0.893, respectively; P<0.05), while MMP was negatively correlated with the expression level of UCP2 (r=-0.903, P<0.05). In the rat model of sepsis, the myocardium and myocardial mitochondria have obvious injuries, and the expression level of UCP2 is closely correlated with mitochondrial injury. Therefore, UCP2 might play an important role in myocardial mitochondrial injury in sepsis.

Protective effect of fasudil hydrochloride against acute renal injury in septicopyemia rats.

To observe the protective effect of fasudil hydrochloride against acute renal injury in septicopyemia rats. A total of 60 Wister rats were included in the study and divided into control group (n=10), model group (n=25) and treatment group (n=25). Model group and treatment group received intraperitoneal injection of endotoxin (ET) to establish acute renal injury models while the control group only received daily intraperitoneal injection of normal saline 1mL. Five rats were taken out of model group and treatment group respectively at 1h (T1), 6h (T2), 12h (T3), 24h (T4) and 48h (T5), for intraperitoneal injection of ET 30mg/kg. Treatment group received intraperitoneal injection of fasudil hydrochloride 30mg/kg 1h before injection of ET. For three groups, 5mL blood samples were collected from postcava for determination of serum creatinine and urea nitrogen levels at different time points. Concentrations of serum tumor necrosis factor α and ET-1 were determined by using ELISA. The renal pathologic changes were observed under the microscope. Serum creatinine levels in both model group and treatment group were significantly higher than control group at T2-T5 (P<0.05) while the levels in treatment group were significantly lower than control group at T3-T5 (P<0.05). At T2-T5, blood urea nitrogen levels in model group and treatment group were significantly higher than control group (P<0.05) while the levels in treatment group were significantly lower than model group at T3-T5 (P<0.05). Concentrations of serum tumor necrosis factor α in model group and treatment group were significantly higher than control group at T1-T5 (P<0.05) while the levels in treatment group were significantly lower than model group at T1-T5 (P<0.05). Serum ET-1 concentrations in model group and treatment group were significantly higher than control group at T1-T5 (P<0.05) while the levels in treatment group at T1-T4 were significantly lower than model group (P<0.05). Rats in control group showed no swelling or hyperemia in kidney cells but normal structure and normally arranged renal tubular epithelial cells. Obvious injury was observed in model group at T3 and renal tubular epithelial cells in disorder and at swelling condition, hyperemia and angiectasis in glomerulus, degenerative opacities and vacuolar degeneration, and maximized injury were observed at T4. Injury in renal tissue in treatment group was significantly milder than model group. Fasudil hydrochloride has the significantly protective effect against acute renal injury in septicopyemia rats.

The Effects of N-acetylcystein and Epigallocatechin-3-Gallate in Ischemia-Reperfusion Injury of Rat Lungs

Background: Ischemia-reperfusion injury (IRI) is a major cause of early graft dysfunction after lung transplantation. The aim of this study was to assess the effects of N-acetylcystein (NAC) and epigallocatechin-3-gallate (EGCG) on IRI of rat lungs. Methods: Sprague-Dawley rats were divided into four groups. Sham group (n=6) did not receive IRI. Rats in the control group (n=6), NAC group (n=6), and EGCG group (n=6) were treated with an intraperitoneal injection of normal saline, NAC, and EGCG, respectively, prior to IRI. In the latter three groups, IRI was induced by clamping the left pulmonary artery, vein, and main stem bronchus for a period of 60 minutes. After ischemia, reperfusion and ventilation of the lung was allowed for a period of 180 minutes. The expression levels of inducible nitric oxide synthase (iNOS), hemeoxygenase-1 (HO-1), AMP-activated protein kinase-α (AMPK), and caveolin-1 in lung tissues were evaluated by Western blot. The pathological findings and the extent of pulmonary edema after IRI were compared among the groups. Results: The expression levels of iNOS decreased in the Sham and EGCG groups. The expression level of HO-1 was significantly higher in the EGCG group (P=0.0001). Although the expression levels of AMPK and caveolin-1 showed no differences, the extent of phosphorylation of AMPK and caveolin-1 was higher in the EGCG and NAC groups, respectively. In hematoxylin-eosin staining, the lungs in the NAC and EGCG groups showed fewer alveolar injuries and less hemorrhagic congestion compared with the control group. Conclusions: NAC and EGCG enhanced the phosphorylation of caveolin-1 and AMPK, respectively, and attenuated lung injury induced by ischemia-reperfusion.

Effect of salinomycin on metastasis and invasion of bladder cancer cell line T24.

To explore the effect of salinomycin on the metastasis and invasion of bladder cancer cell line T24 by regulating the related protein expression in the process of epithelial-mesenchymal transition (EMT), and to provide experimental basis for the treatment of urological tumors. The bladder cancer cell line T24 was cultured invitro. The rat bladder tumor model was established invivo. The rats were randomized into two groups, among which the rats in the experiment group were given intraperitoneal injection of salinomycin, while the rats in the control group were given intraperitoneal injection of normal saline. The change of tumor cells in the two groups was observed. Transwell was used to detect the cell migration and invasion abilities, Real-time PCR was used to detect the expression of mRNA, while Western-blot was utilized for the determination of the expressions of E-cadherin and vimentin proteins. The metastasis and invasion abilities of serum bladder cancer cell line T24 after salinomycin treatment in the experiment group were significantly reduced when compared with those in the control group, and the tumor metastasis lesions were decreased from an average of 1.59 to 0.6 (P<0.05). T24 cell proliferation in the experiment group was gradually decreasing. T24 cell proliferation at 48h was significantly lower than that at 12h and 24h (P<0.05). T24 cell proliferation at 24h was significantly lower than that at 12h (P<0.05). T24 cell proliferation at each timing point in the experiment group was significantly lower than that in the control group (P<0.05). The serum mRNA level and E-cadherin expression in the tumor tissues in the experiment group were significantly higher than those in the control group, while vimentin expression level was significantly lower than that in the control group (P<0.05). Salinomycin can suppress the metastasis and invasion of bladder cancer cells, of which the mechanism is probably associated with the inhibition of EMT of tumor cells.

Effects of exendin-4, a glucagon like peptide-1 receptor agonist, on neutrophil count and inflammatory cytokines in a rat model of endotoxemia

BackgroundSepsis remains a major cause of morbidity and mortality. A variety of strategies targeting modulation of the pro-inflammatory response associated with early sepsis have been reported without clinical success. GLP-1 enhances glucose-stimulated insulin secretion. In addition, it was shown to have anti-inflammatory effects. We hypothesized that treatment with exendin-4, a GLP-1 receptor agonist, would attenuate inflammation and improve glucose control in a lipopolysaccharide (LPS) rat model of inflammation.MethodsTwo-month-old male Wistar rats were randomly assigned to one of the following four groups: 1) treatment: intraperitoneal (IP) injection of LPS 10 mg/kg followed by exendin-4, 30 μg/kg, 10 minutes later; 2) control-1: IP injection of LPS 10 mg/kg, followed by normal saline (NS); 3) control-2: IP NS injection followed by exendin-4; 4) sham: IP injection of NS followed by another NS injection. Glucose concentration, total white blood count with absolute neutrophil count, and pro- and anti-inflammatory cytokine concentrations were measured at 0, 3, 6, and 10 hours following LPS injection.ResultsAt 3 hours, rats injected with LPS developed neutropenia, elevated pro- and anti-inflammatory cytokines, and mild hypoglycemia. Treatment with exendin-4 significantly modulated neutropenia, and decreased pro-inflammatory cytokine concentrations (IL-1α, IL-1β, IL-6, TNFα, and IFNγ). However, exendin-4 had no effect on IL-10 concentrations. LPS injection led to mild hypoglycemia, that was not observed in rats treated with exendin-4. Sham animals exhibited no significant change from baseline in all parameters.ConclusionIn this LPS model of acute early phase inflammation, treatment with exendin-4 decreased pro-inflammatory cytokine concentrations without changing IL-10 blood levels and improved neutropenia. Following LPS injection, rats developed a tendency toward hypoglycemia that improved with exendin-4. Overall our data suggest that exogenous exendin-4 mediates anti-inflammatory effects early in this rat model of endotoxin-induced inflammation.

Protective effect of TMP on pancreas function of acute pancreatitis rats.

To explore the protective effect and mechanism of Tetramethy1Pyrazine (TMP) on the pancreas function of acute pancreatitis rats. A total of 75 SD rats were randomly divided into three groups (A, B, C) with 25 rats in each group. Group A served as sham operation group. In the groups B and C, AP model was prepared as by injecting taurocholic acid sodium. Group B was model group. After modeling, rats were administrated by intraperitoneal injection of normal saline. Group C was TMP treatment group, which was administrated by intraperitoneal injection of 0.6% TMP after modeling. The rat blood specimens in each group were collected with 1mL/100g solution after modeling of 2, 6, 12 and 24h. Levels of amylase (AMS), blood urea nitrogen (BUN), creatinine (CR), TNF-α and IL-6 were detected, and 5 rats were sacrificed. Histopathological examination was performed in he pancreatic tissue specimens of each group to observe pancreatic tissue damage. After modeling in each time point, AMS, BUN, CR, TNF-α and IL-6 in groups B and C were significantly higher than that of in group A (P<0.05). After modeling of 2h, AMS, BUN and CR in group B increased significantly and reached the peak value at 6h. After modeling of 12h, serum level of TNF-α and IL-6 were significantly lower than that of in control group, while after 24h of modeling, serum level of AMS, BUN, CR, TNF-α and IL-6 were significantly lower than that of in control group (P<0.05). The histological observation showed that pancreatic tissue in rats of group A was normal without damage lesions. Massive bleeding, necrosis and serious injury were visible in pancreatic tissue of group B. The rat pancreatic tissue was bleeding in group C with small pieces of necrotic lesions. The degree of inflammatory cell infiltration was lower than group B, and the degree of injury was significantly lower than group B. TMP can significantly decrease the serum level of TNF-α and IL-6 in AP rats, inhibits inflammatory response of AP, and has significant protective effect on pancreatic tissue and function in AP rats.

Protective Effect of Propofol on Electroconvulsive Shock Induced Learning and Memory Function Impairment Involves Up-Regulation of NMDA Receptor NR2A/2B Subunit in a Rat Model of Depression

Backgroud: The mechanism underlying the protective effect of propofol on electroconvulsive therapy (ECT) induced learning and memory function impairment remains largely unknown. The present study aimed to explore the effect of propofol on the expression of N-methyl-D-aspartic acid (NMDA) receptor subunit NR2A/2B and α-amino-3-hydroxy-5-methyl-4-isoxazole-propionic acid (AMPA) receptor subunit glutamate receptor 1 (GluR1) in the hippocampus of rat depression model undergoing electroconvulsive shock (ECS, analog of ECT in animals) in order to uncover the mechanism underlying the damaging effect of ECS on the learning and memory function, and the mechanism underlying the protective effect of propofol on the learning and memory function in ECS treatment.Methods: This study was designed as a longitudinal study. Establishment of depression model was conducted with the application of chronic unpredictable mild stress (CUMS) in Sprague-Dawley rats. All 50 rats were randomly assigned into 5 groups (N=10) : one control group ( group C, 10 healthy rats) and four groups exposed to CUMS treatments for 28 days to construct the rat model of depression (group D, P, E, PE). Rats in group C were treated with intraperitoneal injection of 8 ml/kg normal saline and sham ECS (without the administration of current). Rats in group D received intraperitoneal injection of 8 ml/kg normal saline and then sham ECS. Rats in group P received intraperitoneal injection of propofol 80 mg/kg and then sham ECS, while rats in group E were treated with intraperitoneal injection of normal saline 8 ml/kg and then ECS. Rats in group PE were treated with intraperitoneal injection of propofol 80 mg/kg and then ECS. The treatments were conducted once a day for 7 consecutive days. The sucrose preference test was performed to assess the depression behavior. Learning and memory function of rats was evaluated with Morris water maze. Western-blot assay was applied to determine the expression levels of NR2A/2B subunit and GluR1 subunit in the hippocampus.Results: Sucrose preference percentage of the rats in group E and group PE was increased compared with rats in group D. Compared with group D, rats in group E showed increased escape latency, decreased space exploration time, as well as increased expression of GluR1, and decreased NR2A/2B expression. Shorter escape latency, longer space exploration time, and higher level of NR2A/2B expression were showed in rats in group PE than in group E.Conclusions: ECS treatment up-regulated the expression of GluR1 receptor while down-regulated the expression NR2A/2B receptors in rat hippocampus. Propofol mitigating learning and memory function impairment in depression model rats undergoing ECS may via alleviating the inhibitory effect induced by ECS on the NMDA receptor NR2A/2B subunit expression. Citation: Xue-Chao Hao, Xian-Lin Zhu, Ping Li, Jing Chen, Feng Lv, Su Min. Protective effect of propofol on electroconvulsive shock induced learning and memory function impairment involves up-regulation of NMDA receptor NR2A/2B subunit in a rat model of depression. J Anesth Perioper Med 2015; 2: 66-74. doi: 10.24015/JAPM.2015.0011This is an open-access article, published by Evidence Based Communications (EBC). This work is licensed under the Creative Commons Attribution 4.0 International License, which permits unrestricted use, distribution, and reproduction in any medium or format for any lawful purpose. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

The effect of signal transduction pathway of triggering receptor-1 expressed on myeloid cells in acute lung injury induced by paraquat in rats

To investigate the transduction pathway of triggering receptor-1 expressed on myeloid cells (TREM-1) in acute lung injure induced by paraquat in rats through the activating or blocking TREM-1, to observe the effect of signal transduction pathway in the acute lung injure induced-paraquat. 80 SD rats were randomly divided into normal saline control group (n=20) , PQ poisoning group (n=20) , antibody group (n=20) , and LP17 group (n=20). poisoning group, antibody group and LP17 group were given saline diluting PQ 80 mg/kg of disposable lavage after 2 h, a single set of intraperitoneal injection of anti-TREM-1 mAb (250 g/kg) , tail intravenous LP17 group synthetic peptide (3.5 mg/kg) , poisoning group was given equal normal saline intraperitoneal injection, control group given normal saline 1 mg/kg after 2 h after lavage, given the amount of intraperitoneal injection of saline solution. The expression of NF-κB in lung tissue was determined by immunohistochemistry.The levels of TNF-a, IL-10, TREM-1, and soluble TREM-1 (sTREM-1) in lung tissue and serum were measured by ELISA. Pathology changes of lung were observed under light microscope, and lung score of pathology was compared. Administration of anti-TREM-1 mAb after PQ poisoning modeling significantly increased the NF-κB expression in lung tissue at 48 h, resulting in a large number of pro-inflammatory cytokines releasing in the lung tissue and serum and lung pathology injury score increasing.Administration of LP17 after modeling significantly down-·regulated the expressions of NF-κB and proinflammatory cytokines, while led to a slight increase of anti-inflammatory cytokines and a decline of lung pathology injury score. TREM-1 may involve in inflammatory response by promoting the generation of inflammatory factors via NF-κB pathway, thus lead to lung pathological changes.

Peritoneal lavage using chlorhexidine gluconate at the end of colon surgery reduces postoperative intra-abdominal infection in mice

BackgroundThe use of peritoneal lavage with antiseptic solutions after bowel surgery remains controversial. This study compared peritoneal lavage using chlorhexidine gluconate at low concentrations and normal saline in mice with cecal ligation and perforation. MethodsA total of 180 mice were randomized to six groups. Groups A, B, and C received one-time intraperitoneal injections of normal saline, chlorhexidine gluconate 0.05%, and chlorhexidine gluconate 0.025%, respectively. Groups D, E, and F were all subject to cecal ligation and perforation, then underwent partial cecectomy and peritoneal lavage with normal saline only, chlorhexidine gluconate 0.05% followed by normal saline, and chlorhexidine gluconate 0.025% followed by normal saline, respectively. Animals were followed postoperatively then sacrificed and examined at necropsy for occurrence of intra-abdominal abscesses, adhesions, or other pathology. ResultsA total of 48 mice (26.7%) developed postoperative intra-abdominal abscesses. Group E mice that had chlorhexidine gluconate 0.05% lavage had significantly lower incidence of postoperative intra-abdominal abscesses compared with that of group D mice that had saline lavage only (P = 0.0113). There was no significant difference in occurrence of macroscopic adhesions among mice groups that had or did not have surgery. (P = 1 and P = 0.3728). Microscopic peritoneal fibrosis occurred significantly more among group E mice that had chlorhexidine gluconate 0.05% lavage compared with group D mice that had saline lavage only (P = <0.005). There was no significant difference in postoperative mortality between surgical groups (P = 0.8714). ConclusionsChlorhexidine gluconate 0.05% peritoneal lavage after partial colectomy (cecectomy) in mice reduces postoperative intra-abdominal infection without significant macroscopic adhesion formation.