Rat Blood Samples Research Articles

PIPER CHABA EXTRACTS WITH ANTIBIOFILM ACTIVITY INFLUENCE ANTIHYPERGLYCEMIC AND ANTIHYPERLIPIDEMIC RESPONSES IN DIABETIC WISTER RATS

Objectives: Piper chaba, native to South and Southeast Asia, has been traditionally used as a medicinal plant. Aim of this study was to evaluate the antihyperglycemic and antihyperlipidemic activities of P. chaba root extracts (RE) in streptozotocin (STZ)-induced diabetic rats along with its antimicrobial activity. Methods: Diabetes was induced in Wister rats through the intraperitoneal administration of STZ (50 mg/kg b.w.). Antidiabetic and antilipidemic activities of the RE (in methanol, ethanol, ethyl acetate and distilled water) were evaluated by administering oral dose (200 mg/kg b.w.) for 21 days. Metformin (12.1 mg/kg b.w.) was used as a positive control. Blood samples of rats were drawn by tail vein puncture and cardiac puncture to determine the fasting blood glucose (FBG) and serum level of total cholesterol (TC), triglycerides (TG), low-density lipoprotein (LDL), and high-density lipoprotein (HDL), respectively. Standard protocols were followed to determine the antimicrobial and antibiofilm activities against two different strains of bacteria. Results: Oral administration of P. chaba RE for 21 days resulted in a significant (p< 0.001) decrease in FBG and TC, TG, and LDL levels (p<0.001), when compared to the untreated diabetic rats. Significant (p<0.001) increase of HDL was observed when ethyl acetate and aqueous RE were administered. Out of four, two extracts showed varying antimicrobial activities, particularly against the gram-positive bacteria. Conclusion: It became evident for the first time that P. chaba extracts possess antimicrobial activities and can serve as biochemical compounds with great alternative therapeutic potential in the management of diabetes and hypercholesterolemia.

Differential expressions of miR-223, miR-424, miR-145, miR-200c, miR-139 in experimental rat chronic pancreatitis model and their relationship between oxidative stress, endoplasmic reticulum stress, and apoptosis.

Objective(s):This study aimed to research the roles of miR-139, miR-221, miR-200c, miR-145, miR-223, miR-424, and miR-377 in endoplasmic reticulum stress (ERS), oxidative stress (OS), fibrosis, and apoptosis processes in chronic pancreatitis (CP) rat model. Materials and Methods:Fourteen rats were randomized into 2 groups (Group 1, sham group (n=7) and Group 2, CP group (n=7)). TGF-beta and malondialdehyde concentrations were measured in rat blood samples. qRT-PCR was used to investigate the expression levels of 7 miRNAs in the pancreas tissues. The correlations of mRNA undergoing significant changes with inflammation (TNF-α, IL-6), ERS (Ire1-α, Perk), apoptosis (Caspase 3, Bcl-2), OS (Cat, Gpx1), and fibrosis (α-Sma) were investigated.Results:The biochemical results and histopathological scores in Group 1 were statistically significantly high compared with Group 2 (P<0.5). Expression levels of seven miRNAs (miR-200c, miR-145, miR-223, miR-424) were significantly higher, while miR-139 was significantly lower in CP. In our study, we found that miR-200c, miR-145, and miR-139 may contribute to CP progression and cellular processes based on the correlation between ERS, OS, apoptosis, and inflammation with miRNA expression levels. Conclusion:miR-200c, miR-145, miR-139, miR-223, and miR-424 play roles in the CP model. They may be used as candidate biomarkers for the CP process.

MicroRNA-16 regulates lipopolysaccharide-induced inflammatory factor expression by targeting TLR4 in normal human bronchial epithelial cells.

Acute lung injury (ALI) is mainly caused by inflammation and is associated with high mortality rates. Emerging evidence has suggested that microRNAs (miRNAs or miRs) serve a significant function in ALI. However, the fundamental mechanism underlying ALI remain to be fully elucidated. Although miR-16 has been reported to be involved in the occurrence and development of a number of diseases its association with ALI has not been previously investigated. Therefore, the present study aimed to explore the role of miR-16 in the lipopolysaccharide (LPS)-induced ALI model. The expression levels of tumor necrosis factor α (TNF-α), interleukin (IL)-1β and IL-6 were measured by ELISA in the blood samples of rats with ALI and in the normal human bronchial epithelial (NHBE) cell line. The role of miR-16 in inflammation was evaluated using gene overexpression and silencing experiments in NHBE cells by reverse transcription-quantitative PCR. In addition, the expression levels of inflammatory factors TNF-α, IL-1β and IL-6 were also determined using ELISA. The potential interaction between miR-16 and TLR4 was assessed using bioinformatics analysis by the TargetScan database and then verified in 293T cells using luciferase reporter assay. The expression of miR-16 was notably decreased in the lung tissues of rats with LPS-induced ALI compared with the PBS treated-group. Additionally, the levels of the proinflammatory cytokines TNF-α, IL-1β and IL-6 were reduced following transfection of NHBE cells with miR-16 mimics compared with those in the miR-negative control group. Western blot analysis revealed that miR-16 overexpression could downregulate TLR4 expression in NHBE cells compared with that in the miR-NC group. Luciferase reporter assay confirmed that TLR4 may be directly targeted by miR-16. The effect of miR-16 on TLR4 was rescued in NHBE cells following treatment with LPS. Overall, these aforementioned findings suggest that miR-16 may serve a protective role against LPS-mediated inflammatory responses in NHBE cells by regulating TLR4, where this mechanism may be considered to be a novel approach for treating ALI in the future.

Verbascoside alleviates renal fibrosis in unilateral ureteral obstruction rats by inhibiting macrophage infiltration.

Objective(s): To explore the effect of verbascoside on renal fibrosis in unilateral ureteral obstruction (UUO) rats.Materials and Methods:Twenty Sprague-Dawley rats were randomly distributed into sham-operated, UUO, and UUO+Verbascoside groups. After two weeks of rat model construction, urine and blood samples were collected for biochemical analysis while kidney tissues were harvested for hematoxylin and eosin (H&E), Masson’s Trichrome, and immunohistochemistry staining. Pearson coefficient was used to analyze the correlation between the two proteins. Results:Verbascoside improved UUO-induced renal dysfunction as detected by decreased serum creatinine, urea nitrogen, and urinary protein excretion rate. In UUO rats, H&E staining result revealed increased total nucleated cell number, and Masson’s Trichrome staining results showed tubular interstitial fibrosis with the deposition of collagen fibrils. Besides, expressions of fibrosis-related proteins including collagen type I (COL-I), α-smooth muscle actin (a-SMA), and tissue inhibitor of metalloproteinase 2 (TIMP2) expressed higher in the UUO group. Moreover, macrophage infiltration-related factors such as CD68+, F4/80+ cells, and suppressor of cytokine signaling-3 (SOCS3) were significantly higher in the UUO group than in sham-operated rats. However, after administration with verbascoside, the accumulation of collagen fibrils and total nucleated cell numbers were mitigated. Likewise, macrophage infiltration was extenuated and fibrosis-related proteins were down-regulated in the UUO+Verbascoside rats. Correlation analysis indicated that macrophage infiltration-related markers were related to fibrosis-related factors.Conclusion:Verbascoside could alleviate renal fibrosis in UUO rats probably through ameliorating macrophage infiltration.

Repellency of Veratraldehyde (3,4-Dimethoxy Benzaldehyde) against Mosquito Females and Tick Nymphs

Arthropod-borne infectious diseases cause many deaths and a major economic burden worldwide. Repellents play an important role in protecting people from infectious biting arthropods. The repellency of veratraldehyde, a known food additive, and the WJ-1041 formulation containing 10% veratraldehyde was tested against Aedes albopictus and Culex pipiens pallens females and Haemaphysalis longicornis nymphs using arm-in-cage, indoor or filter paper tests. Veratraldehyde exhibited repellency similar to or lower than that of n,n-diethyl-meta-toluamide (DEET) against A. albopictus, but in H. longicornis, the activity of veratraldehyde was better than that of DEET. The repellency of the 10% veratraldehyde solution was comparable to that of 20% DEET against the two mosquitoes. When comparing repellency between the WJ-1041 formulation (10% veratraldehyde) and 10% DEET against C. pipiens pallens, A. Albopictus and H. longicornis, the two showed similar repellency and complete protection time (CPT) values. However, there was a small difference depending on the tested insects. The absorption of veratraldehyde via skin was minimal, if at all. The pharmacokinetic parameters (Cmax and Tmax) of veratraldehyde in blood samples of rats were not different from those of the control group. Based on these results, veratraldehyde has high potential to be commercialized as a repellent agent against infectious disease-borne pests in the near future.

The Effect of Black Cumin (Nigella sativa) Oil Supplementation on Hematotoxicity Induced by Benzene in Gasoline Vapors from Gas Stations in Male Wistar Rats

This study aimed to prove the effect of black cumin oil supplementation through analysis of levels of CYP2E1, MDA, GSH, and hematological profiles in experimental animals, Wistar rats, which were exposed to benzene in gasoline vapors from gas stations. Sixty-four experimental Wistar rats were divided into eight groups with eight rats each, two control groups (C1 and C2), and six experimental groups (T1, T2, T3, T4, T5, and T6). Blood samples of experimental rats were taken through intracardiac with ketamine-xylazineacepromazine anesthesia on day 15 for group C1, C2, T1, T3, and T5; and day 29 for group T2, T4, and T6. The ELISA method measured the levels of CYP2E1, GSH, and MDA in this study, while the hematological profile used a hematology analyzer. This study indicates that benzene in gasoline vapors exposure caused significant hematotoxicity (P &lt;0.05). Supplementation of black cumin seed oil at a dose of 2.5 ml/kg, 5 ml/kg, and 10 ml/kg, all of which have the potential to provide a protective effect against the hematotoxicity of benzene in gasoline vapors through increased internal antioxidants (GSH). The GSH can play a role in the mechanism of vapor metabolism of substances in gasoline vapors in the body through decreased CYP2E1 levels and decreased MDA levels. Further research is still needed with a more extended treatment duration to ascertain the chronic effects of benzene hematotoxicity on the genomic pathway to assess black cumin’s role, whether in the form of oil, powder, or other forms of extraction.

Investigations on the Oral and Brain Uptake Mechanisms of D‐ and L‐eflornithine in the Rat

Eflornithine is a recommended treatment of the otherwise fatal disease late stage Gambian human African trypanosomiasis (HAT). Whereas racemic eflornithine is today administered intravenously, an oral eflornithine treatment option would increase accessibility for patients in affected areas in sub-Saharan Africa. Oral absorption of eflornithine is stereoselective when dosed as a racemic mixture with the more potent L-enantiomer disfavoured [1, 2]. In late stage Gambian HAT, the brain is the primary site of pharmacological action. Knowledge on whether stereoselective passage also occurs across the blood-brain barrier is limited. This study aimed to investigate the rate, extent and potential stereoselectivity of brain uptake of D- and L-eflornithine using in vivo microdialysis in the rat. Microdialysis probes were inserted in the third brain ventricle with simultaneous catheterizing of the jugular vein for blood sampling in Sprague-Dawley rats (n=16). Racemic eflornithine was dosed (1200 mg/kg) by oral gavage. Plasma and microdialysis samples were collected up to nine hours after dosing. Stereoselective eflornithine concentrations were determined by high performance liquid chromatography [3]. Plasma data was complemented with data from a prior study [1]. Areas under the curve (AUC) were determined using pharmacokinetic modelling with naïve pooled data analysis as implemented in Phoenix (Version 8.3, Certara, Princeton, NJ, USA). Pharmacokinetics of D- and L-eflornithine were best described by a two-compartment drug disposition model. Transit compartments were used to model the delay in oral absorption. Observed AUCs in plasma and brain dialysate for D-eflornithine were 160 min•mM and 1.0 min•mM, respectively. For L-eflornithine, AUCs were 60 min•mM and 0.38 min•mM in plasma and brain dialysate, respectively. Oral uptake of D- and L-eflornithine was, as expected, stereoselective whereas the uptake to the third ventricle in the rat brain was not stereoselective since the dialysate/plasma AUC ratio was similar for the enantiomers. Overall, results suggested a brain efflux of both D- and L-eflornithine to a similar extent. The findings will be used in future translational investigations to model the feasibility of an oral eflornithine based therapy against late stage Gambian HAT.

Serial blood sampling effects in rat embryo-fetal development studies for toxicokinetics

Serial blood sampling for toxicokinetics is generally conducted in regulatory embryo-fetal development (EFD) studies in rats. EFD studies are designed to detect the potential adverse effects of pharmaceuticals on pregnant females and their fetuses; this information is useful for understanding the relationships between systemic exposure levels and toxicity profiles. However, additional satellite pregnant females are needed for toxicokinetics because comprehensive information regarding the potential impact of serial blood sampling on pregnant females is scarce. Here, in this study, we investigated the potential impact of serial blood sampling in pregnant female rats using a typical EFD study design. Additionally, we investigated the additional endpoints (clinical pathology, organ weights, and histopathology) that were deemed likely to be sensitive to blood sampling. Results indicated that serial blood sampling in pregnant females induced physiological adaptive changes and did not affect the general endpoints in EFD studies. Nevertheless, inclusion of satellite groups in EFD studies may be a more prudent approach considering the physiological changes in pregnant females and potential off-target effects of candidate pharmaceuticals. These results provide background information on the impact of serial blood sampling in pregnant females and will be useful to design the regulatory EFD studies.

Estimation of the net influx rate Ki and the cerebral metabolic rate of glucose MRglc using a single static [18F]FDG PET scan in rats

Since accurate quantification of 2-deoxy-2–18F-fluoro-D-glucose ([18F]FDG) positron emission tomography (PET) requires dynamic acquisition with arterial input function, more practical semi-quantitative (static) approaches are often preferred. However, static standardized uptake values (SUV) are typically biased due to large variations in body weight (BW) occurring over time in animal studies. This study aims to improve static [18F]FDG PET SUV quantification by better accounting for BW variations in rats. We performed dynamic [18F]FDG PET imaging with arterial blood sampling in rats (n = 27) with different BW (range 0.230–0.487 kg). By regressing the area under the curve of the input function divided by injected activity against BW (r2=0.697), we determined a conversion factor f(BW) to be multiplied with SUV and SUVglc to obtain ratSUV and ratSUVglc, providing an improved estimate of the net influx rate Ki (r = 0.758, p<0.0001) and cerebral metabolic rate of glucose MRglc (r = 0.906, p<0.0001), respectively. In conclusion, the proposed ratSUV and ratSUVglc provide a proxy for the Ki and MRglc based on a single static [18F]FDG PET SUV measurement improving clinical significance and translation of rodent studies. Given a defined strain, sex, age, diet, and weight range, this method is applicable for future experiments by converting SUV with the derived f(BW).

Determination of AB-FUBINACA and 5F-NPB-22 in rats exposed to "Bonsai" via inhalation and analysis of seized product.

Synthetic cannabinoids (SCs) are the most rapidly growing class of recreational designer drugs. Illicit drug manufacturers began to produce herbal smoking materials under a variety of brands names, e.g. "Spice, K2, Bonsai, Yucatan Fire". They were appeared on the European market in 2008. In this study, types of SCs in the herbal product sold as "Bonsai" in Turkey were determined and the identification of these substances in biological samples collected from rats depending on the inhalation of different amounts of plant material were aimed. To determine the SC species in the content of the plant product, analysis was performed via gas chromatography-mass spectrometry. Liquid-liquid extraction methods were utilized for blood and organ samples, while solid-phase extraction with β-glucuronidase enzyme treatment was applied for urine sample preparation. The relationship between the amount of burned plant and the amount of SCs accumulated in the blood, urine and organ samples of rats exposed to the plant product by inhalation was examined by liquid chromatography-tandem mass spectrometry. AB-FUBINACA and 5F-NPB-22 were detected in the herbal product. A significant correlation was found between the amount of herbal product inhaled and the prevalence of SCs, especially in lung tissues while no SCs were detected in the blood and urine samples of rats. There is currently no study on biological samples of individuals exposed to herbal products containing SCs by inhalation. Regarding the findings obtained in this study, the overall increase in the amounts of herbal product inhaled was demonstrated to pose a potential risk to humans.

Effect of Acupuncture on Ovary Morphology and Function in DHEA-Induced Polycystic Ovary Syndrome Model Rats.

To investigate the effects of acupuncture on ovary morphology and function in dehydroepiandrosterone (DHEA)-induced polycystic ovary syndrome (PCOS) model rats. A total of 40 adult female Wistar rats were randomly allocated to 4 groups by a random number table, including control, model, metformin and acupuncture groups, 10 rats in each group. PCOS rat model was developed by injecting with DHEA (6 mg/100 g body weight) in 0.2 mL of oil subcutaneously. Electrical stimulation (2 Hz, 3 mA) was applied to Guanyuan (CV 4), Zigong (EX-CA1) and Qihai (CV 6) acupoints for 30 min daily in the acupuncture group, and metformin (200 mg/kg) was given to rats in the metformin group, both once per day for 21 consecutive days, and rats in the normal group was fed with normal saline and fed regularly. After 21 days of administration, the rat blood samples were collected for detecting the reproductive hormonal levels [luteinizing hormone (LH), follicle stimulating hormone (FSH), estradiol (E2), progesterone (P), testosterone (T)] and inflammatory factors (visfatin, IL-6) analysis. Ovary tissue was used for histopathological analysis. Compared with the model group, rats in the acupuncture and metformin groups were significantly lower in weight gain, FSH, LH and T levels, and E2 and P levels significantly increased (alll P<0.05). Meanwhile, LH and FSH levels were significantly decreased, and P, T and E2 levels significantly increased in the acupuncture group, compared with the metformin group (P<0.05). Compared with the model group, IL-6 and visfatin levels were significantly decreased in the acupuncture and metformin groups (P<0.05). There were no significant differences in IL-6 and visfatin levels between the acupuncture and metformin groups (P>0.05). Ovarian diameter in the acupuncture and metformin groups were smaller than the model group (P<0.05). However, there were no significant differences in ovarian diameters between the acupuncture and metformin groups (P>0.05). Acupuncture might improve ovary morphology and its function in DHEA-induced PCOS model rats.

The Antioxidant and Hepatoprotective Potential of Temulawak (Curcuma xanthorrhiza Roxb) Ethanol Extract in Paracetamol-induced Rats

Paracetamol is widely used as analgesic and antipyretic drugs. Paracetamol can increase Radical Oxygen Species (ROS) if used exceeded the administered dose that might lead to hepatic and gastric damage. The purpose of this research is to investigate the antioxidant and hepatoprotective potential of Temulawak Ethanol Extract in Paracetamol-induced rats. This research was conducted using completely randomized design. The samples were 30 white rats, divided equally into 6 groups. The rats of group 1 were fed normally with regular feed and aquadest for 10 days (control). The rats of group 2 were fed using aquadest and temulawak extract with the dosage of 540mg/200g rats body weight for 10 days. The group 3 of rats were fed using aquadest and paracetamol with the dosage of 2g/kg rats body weight on the 7th day (positive control). The group 4,5, and 6 were fed using Temulawak ethanol extract, 270mg/200g, 540mg/200g, and 1080mg/200g rats body weight, respectively, for 10 days and were given 2g/kg rats body weight of paracetamol on 7th day. The rats blood samples were taken after 10 days treatment. The MDA, ALT and AST in rats blood serum were measured. The data were analyzed using ANOVA and LSD test. The treatment of 270mg/200g, 540mg/200g, and 1080mg/200g rats body weight Temulawak ethanol extract showed specific results of lowered ALT, ASP and MDA levels in paracetamol induced rats. It can be concluded from this research that the Temulawak ethanol extract is cosidered potential as antioxidant and hepatoprotector.

Indices of oxidative stress under intranasal administration of deicing reagent solutions in rats

Introduction. The use of road deicing reagents (RDR) is an effective measure to reduce winter traumatism and requires enhancement of methods for evaluation of deicers safety. The aim of investigation: to study markers of oxidative stress in rat blood samples during intranasal (i/n) administration of RDR solutions, as a model of intake under natural conditions, using liquid commercial RDR (22% CaCl2; 6% NaCl). Material and methods. Male Wistar rats (10 rats per group) were daily injected into the nasal cavity with 100 μl of RDR solutions in concentrations (C) 0; 0.75; 7.5 and 75 ml per liter of tap water. 5 and 28 days after the start of the experiment, the content of malondialdehyde (MDA), GSH, the activity of SOD, catalase, glutathione peroxidase (GPO) and glutathione reductase (GR) in the hemolysates were determined. Logarithmic transformation x=lg(C+0.01)+2 was used for regression analysis of dose - effect relations. Results. 5 days after the start of the experiment, adaptive dose-dependent changes in activities of SOD (R = -0.504; p=0.001); GR (R = 0.548; p&lt;0,001) and catalase (R=0.725; p&lt;0,001) were revealed, and after 28 days these effects were replaced by dose-dependent increase in MDA content (R=0.617; p&lt;0,001) and GPO activity (R=0.326; p=0.04). The revealed effects, apparently, are due to the presence of additional RDR components (such as detergents, corrosion inhibitors, etc.), since significant differences with corresponding control groups were found also during administration of 0.75 ml RDR per liter (CNa+ 200 times lower than in saline solution; CCa2+ equivalent to its serum content). In particular, sharp increase in catalase activity after 5 days may be indirect evidence of anticorrosive formates metabolism (commonly used anti-corrosive additive) in the conditions of their entry bypassing the portal vein. Conclusion. I/n administration of the studied RDR solutions (0.75-75 ml/L) gave distinct dose-dependent signs of compensated (5 days) and decompensated (28 days) oxidative stress, presumably due to the presence of additional components besides of basic chlorides.

Short Chain (≤C4) Esterification Increases Bioavailability of Rosmarinic Acid and Its Potency to Inhibit Vascular Smooth Muscle Cell Proliferation

Rosmarinic acid is a natural phenolic acid and active compound found in many culinary plants, such as rosemary, mint, basil and perilla. Aiming to improve the pharmacokinetic profile of rosmarinic acid and its activity on vascular smooth muscle cell proliferation, we generated a series of rosmarinic acid esters with increasing alkyl chain length ranging from C1 to C12. UHPLC-MS/MS analysis of rat blood samples revealed the highest increase in bioavailability of rosmarinic acid, up to 10.52%, after oral administration of its butyl ester, compared to only 1.57% after rosmarinic acid had been administered in its original form. When added to vascular smooth muscle cells in vitro, all rosmarinic acid esters were taken up, remained esterified and inhibited vascular smooth muscle cell proliferation with IC50 values declining as the length of alkyl chains increased up to C4, with an IC50 of 2.84 µM for rosmarinic acid butyl ester, as evident in a resazurin assay. Vascular smooth muscle cells were arrested in the G0/G1 phase of the cell cycle and the retinoblastoma protein phosphorylation was blocked. Esterification with longer alkyl chains did not improve absorption and resulted in cytotoxicity in in vitro settings. In this study, we proved that esterification with proper length of alkyl chains (C1–C4) is a promising way to improve in vivo bioavailability of rosmarinic acid in rats and in vitro biological activity in rat vascular smooth muscle cells.

Anemoside B4 ameliorates TNBS-induced colitis through S100A9/MAPK/NF-\u03baB signaling pathway

BackgroundDespite the increased morbidity of ulcerative colitis (UC) in the developing countries, available treatments remain unsatisfactory. Therefore, it is urgent to discover more effective therapeutic strategies. Pulsatilla chinensis was widely used for the treatment of inflamed intestinal diseases including UC for thousands of years in China. Anemoside B4, the most abundant triterpenoid saponin isolated from P. chinensis, exerts anti-inflammatory and antioxidant effects and may be the most active compounds, which is responsible for the therapeutic effects. However, the mechanism how anemoside B4 executes its biological functions is still elusive.MethodsHere, we used the 2, 4, 6-trinitrobenzene sulfonic acid (TNBS)-induced colitis rat model to evaluate the therapeutic effect of anemoside B4. Blood samples of colitis rats were collected for hematology analysis. The inflammation-associated factors were investigated by enzyme-linked immunosorbent assay (ELISA). Cell proliferation and apoptosis was determined with EdU cell proliferation assay and TUNEL assay. The proteins regulated by anemoside B4 were identified by label-free quantitative proteomics. The significantly down-regulated proteins were verified by Western blotting analysis. mRNA expression was analyzed by quantitative real-time RT-PCR.ResultsThe results showed that anemoside B4 ameliorated TNBS-induced colitis symptoms, including tissue damage, inflammatory cell infiltration, and pro-inflammatory cytokine production, apoptosis and slowed proliferation in colon. Quantitative proteomic analyses discovered that 56 proteins were significantly altered by anemoside B4 in the TNBS-induced rats. These proteins mainly clustered in tricarboxylic acid (TCA) cycle and respiratory electron transport chain. Among the altered proteins, S100A9 is one of the most significantly down-regulated proteins and associated with NF-κB and MAPK signaling pathways in the pathogenesis of UC. Further experiments revealed that anemoside B4 suppressed the expression of S100A9 and its downstream genes including TLR4 and NF-κB in colon. In vitro, anemoside B4 could inhibit the NF-κB signaling pathway induced by recombinant S100A9 protein in human intestinal epithelial Caco-2 cells. Moreover, anemoside B4 inhibits neutrophils recruitment and activation in colon induced by TNBS.ConclusionsOur results demonstrate that anemoside B4 prevents TNBS-induced colitis by inhibiting the NF-κB signaling pathway through deactivating S100A9, suggesting that anemoside B4 is a promising therapeutic candidate for colitis.

Rehmannia glutinosa polysaccharide increases the expression of erythropoietin and vascular endothelial growth factor in rats with chronic renal failure by activating hypoxia-inducible factor-2α

Background: To investigate the effect of Rehmannia glutinosa polysaccharide (RGP) in rats with chronic renal failure. Materials and Methods: In this study, we established a rat model of adenine-induced chronic renal failure. Hemoglobin (Hb), red blood cells (RBCs), total protein (TP), serum albumin (ALB), blood urea nitrogen (BUN), cystatin C (Cys C), and serum creatinine anhydride (SCA) were detected in the blood samples of rats. Hematoxylin and eosin staining was used to observe the pathological changes. Immunohistochemical was used to detect the protein expression of hypoxia-inducible factor (HIF)-2α, erythropoietin (EPO), and vascular endothelial growth factor (VEGF) and calculate the value of microvessel density (MVD) in renal tissues. M1001 and GPRRP + PT2385 groups were added to further validate the mechanism of RGP. Results: In the case of rats with chronic renal failure, GPRRP and EPO contributed to the repairment of renal tissue. The level of Hb, TP, and ALB, as well as RBC counts was significantly increased, and the renal function indexes, including BUN, SCA, and Cys C, were significantly decreased, and HIF-2α, EPO, VEGF, and MVD were significantly increased. The effects observed in M1001 group was similar to those in GPRRP group, but compared with GPRRP + PT2385 group, the biochemical indicators and renal function indexes were significantly decreased, which means HIF-2α inhibitor antagonized the protective effect of RGP. Conclusion: RGP played an essential role in repairing the renal injury in rats with chronic renal failure. The therapeutic mechanism of action may be related to the activation of HIF-2α, thereby increasing the expression of EPO and VEGF and then reducing renal anemia and renal tissue damage.

In vitro effects of temperature on red blood cell deformability and membrane stability in human and various vertebrate species.

The effects of temperature on micro-rheological variables have not been completely revealed yet. To investigate micro-rheological effects of heat treatment in human, rat, dog, and porcine blood samples. Red blood cell (RBC) - buffer suspensions were prepared and immersed in a 37, 40, and 43°C heat-controlled water bath for 10 minutes. Deformability, as well as mechanical stability of RBCs were measured in ektacytometer. These tests were also examined in whole blood samples at various temperatures, gradually between 37 and 45°C in the ektacytometer. RBC deformability significantly worsened in the samples treated at 40 and 43°C, more expressed in human, porcine, rat, and in smaller degree in canine samples. The way of heating (incubation vs. ektacytometer temperation) and the composition of the sample (RBC-PBS suspension or whole blood) resulted in the different magnitude of RBC deformability deterioration. Heating affected RBC membrane (mechanical) stability, showing controversial alterations. Significant changes occur in RBC deformability by increasing temperature, showing inter-species differences. The magnitude of alterations is depending on the way of heating and the composition of the sample. The results may contribute to better understanding the micro-rheological deterioration in hyperthermia or fever.

Siklofosfamidle İndüklenmiş Ratlarda Serbest Radikaller ve Antioksidanlar Üzerine Naringinin Protektif Etkileri

Bu calismada siklofosfamid (CYP) ile uygulanan ratlarda kan ve serum orneklerinde Naringin (NA)’in antioksidanlar ve serbest radikaller uzerine protektif etkilerinin arastirilmasi amaclandi. Arastirmamizda yaklasik 200-250 g agirliginda, 40 adet erkek eriskin Sprague Dawley rat kullanildi ve her bir grupta 8 ratin bulundugu 5 grup olusturuldu. Kontrol grubuna 10 gun boyunca intra gastrik (i.g.) serum fizyolojik (1 ml) verildi. CYP grubuna 10 gun boyunca i.g. serum fizyolojik verildi ve 10. gun tek doz intra peritonal (i.p.) CYP (200 mg/kg) enjekte edildi. NA50+CYP ve NA100+CYP gruplarina 10 gun boyunca sirasiyla serum fizyolojikte cozdurulmus 50 ve 100 mg/kg dozunda i.g. NA uygulandi ve NA’nin son enjeksiyondan sonra tek doz CYP (200 mg/kg, i.p.) enjekte edildi. NA100 grubuna ise 10 gun boyunca 100 mg/kg dozunda i.g. NA uygulandi. Deneyin 12. gununde ratlar tiopental sodyum (20 mg/kg) ile anestezi altina alinarak intra kardiyak kan ornekleri alindi ve dekapite edildiler. Ratlardan elde edilen kan ve serum orneklerinde Total Antioksidan Durumu (TAS), Total Oksidan Durumu (TOS), Oksidatif stres indeksi (OSI) ve 8-Hydroxydeoxyguanosine (8-OHdG) parametreleri analiz edildi. CYP’nin ratlarda oksidatif stresi artirdigi, antioksidanlari baskiladigi ve DNA hasarina yol actigi tespit edildi. NA’nin farkli dozlarinin, CYP’nin sebep oldugu oksidatif stresi baskiladigi, antioksidan miktarini artirdigi ve DNA hasarini onledigi belirlendi.

Protein unbound pharmacokinetics of ambroxol in the blood and brains of rats and the interaction of ambroxol with Polygala tenuifolia by multiple microdialysis

Ethnopharmacological relevanceAmbroxol elevates glucocerebrosidase (GCase) activity and reduces nigrostriatal alpha-synuclein burden to better ameliorate motor function in Parkinson's disease (PD). Polygala tenuifolia is a potential alternative botanical medicine for the treatment of many nonmotor symptoms of PD commonly used in Taiwanese patients. Co-administration of these two medicines pose potential herb-drug interaction. Aim of the studyOur hypothesis is that ambroxol and P. tenuifolia may potentially possess herbal drug synergetic effects in the blood and brain. Materials and methodsTo investigate this hypothesis, a multiple microdialysis system coupled with validated ultra-high performance liquid chromatography−tandem mass spectrometry (UPLC−MS/MS) method was developed for rat blood and brain samples. Experimental rats were divided into three groups: low-dose and high-dose ambroxol alone (10 mg/kg, i.v. and 30 mg/kg, i.v., respectively) and ambroxol (10 mg/kg, i.v.) pretreated with P. tenuifolia extract (1 g/kg, p.o. for 5 consecutive days). ResultsAmbroxol easily penetrated into the brain and reached a maximum concentration in the striatum at approximately 60 min after low- and high-dose treatment. The area under the concentration curve (AUC) ratio increased proportionally at the doses of 10 and 30 mg/kg, which suggested a linear pharmacokinetic manner of ambroxol. The brain penetration of ambroxol was approximately 30–34%, which was defined as the ambroxol AUC blood-to-brain distribution ratio (AUCbrain/AUCblood). The P. tenuifolia extract did not significantly alter the pharmacokinetics of ambroxol in the blood and brain of rats. ConclusionThe present study suggests that it is safety without pharmacokinetic interactions for this dosing regimen to use P. tenuifolia extract and ambroxol together.

Histological and Physiological Study of the Effects of Exogenous Melatonin on Liver of Female Rats Exposed to Formalin in Drinking Water

Aim: The presented study was established to evaluate the impact of formalin (FA) on the blood levels ofsuperoxide dismutase (SOD), lipid profile, total serum protein and serum albumin in female rats. Materialsand methods: The animals were assigned randomly into groups of six; control group, and treated other fivegroups as follow, first group exposed to formalin at 200ppm (F200 group), the second exposed to formalinat 400ppm (F400 group), third group exposed to formalin at 200ppm plus melatonin at 10mg/kg (F200Mgroup), fourth group exposed to formalin at 400ppm plus melatonin at 10mg/kg (F400M group), and thefifth exposed to melatonin at 10mg/kg (M group). Samples of blood from the rats were collected at day one,day and day 30 of the experimental period that was continued for 30 days. Results: Using immune-basedtechniques, the SOD activity, it has been here-resulted in significant (p?0.05) after-formalin-exposuredecreases in the blood samples of female rats from F200 and F400 groups, especially, when comparedwith the control group. The total cholesterol levels revealed significant (p?0.05) increases in the blood ofthe tested animals of the F200 and F400 groups. These increases affected the rates of LDL, VLDL, andtriglycerides in the blood of the studied rat-females of both F200 and F400 groups when compared withthe control animals. Interestingly, low levels of HDL were noticed in the blood of the animals of the F200and F400 when comparisons with control animals. The concentrations of the LDL, HDL, and triglyceridesin the serum of the formalin-exposed females showed significant (p?0.05) low levels when compared withcontrol animals. For day-30 collection, albumin concentrations revealed significant (p?0.05) elevation in theblood of the F200 and F400 animals.. Histological examination revealed that the liver of female rats exposedto 200ppm or 400 ppm of formalin for 30 days have been adversely affected. Whereas, the histologicalexamination of the liver of animals treated with melatonin and formalin were less affected due to formalinexposure. Conclusions: These results demonstrate the formalin high toxic effects on the hepatic tissues canbe overcome by the use of exogenous melatonin.