Effects of electroacupuncture at ST36 and BL20 on the diabetic rat testis.

OBJECTIVE: We aimed to evaluate the effects of electroacupuncture (EA) at ST36 and BL20 on the testicular tissues in a rat model of diabetes and to explore the mechanisms of action. METHODS: A total of 34 male Sprague-Dawley rats were allocated to a control group (n = 10), diabetes (D) group (n = 12) or diabetes + acupuncture (DA) group (n = 12). To model diabetes, rats in groups D and DA received an intraperitoneal injection of a single dose of 35 mg/kg streptozotocin (STZ) dissolved in citrate buffer (pH = 4.5; 0.1 M) after 2 weeks of high-fat diet administration. Under xylazine/ketamine anesthesia, stainless steel needles (30 mm × 0.25 mm) were inserted bilaterally at ST36 and BL20. The needles were connected to an EA device via cables, and EA was applied for 30 min (15 Hz frequency and 0.2-1 mA intensity) twice a week for 5 weeks. RESULTS: The effects of EA at ST36 and BL20 on blood glucose levels and body weight, biochemical parameters, histopathological, morphometric and immunohistochemical findings, and terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) analysis were evaluated. A significant decrease was detected in DA versus D groups in blood glucose levels, basement membrane thickness and apoptotic cell/tubule indices. In addition, there was a significant increase in the Johnsen scores, seminiferous tubule diameters, serum levels of follicle-stimulating hormone (FSH), luteinizing hormone (LH) and testosterone, proliferation indices, and sex hormone-binding globulin (SHBG) and insulin-like peptide 3 (INSL3) immunoreactivities. CONCLUSION: EA had multiple positive effects on blood glucose homeostasis and testicular structure/function in this rat model of diabetes. EA may be effective at preventing or eliminating histopathological damage in the diabetic testis.

Investigation of the relationship between the Th17/IL-23 pathway and innate-adaptive immune system in TNBS-induced colitis in rats.

OBJECTIVES: This study was aimed at investigating immune activations of the 2,4,6-trinitrobenzene sulfonic acid (TNBS)-induced colitis model in colonic mucosa by immunohistochemical and Western blot methods. MATERIALS AND METHODS: For this purpose, 16 female Wistar albino rats were divided into two random groups of control (n=8) and colitis (n=8). The experimental colitis model was induced by intracolonic administration of TNBS (25 mg/rat). Control animals received only rectal saline for the same time. The animals were sacrificed on the 15th day after TNBS administration, and colon tissue was removed and examined morphologically. Colon samples were stained immunohistochemically with anti-CD3, anti-CD4, anti-CD5, anti-CD8, anti-CD11b, anti-CD45, anti-TNF-α, anti-IL-17, anti-IL-22 and anti-IL-23 antibodies. Additionally, the colonic tissue IL-17 and IL-22 expressions were examined by the Western blot method. RESULTS: In the experimental results, it was determined that there was a significant decrease in body weight and an increase in colon weight in the colitis group when comparing initial experiments. The colon tissue ulcerations, inflammation, crypt loss and Goblet cell loss were observed in the colitis group in microscopic examinations. The immunohistochemical positive cell numbers significantly increased in the colitis group. The immunoreactive lymphocytes in the propria, intracryptal and submucosal layers were found to be increased in the colitis group of rats. In addition, IL-17 and IL-23 expressions were increased in colitis colon mucosa found by Western blot analysis. CONCLUSION: The Th17/IL-23 pathway and IL-22 serve important roles in the pathogenesis of ulcerative colitis, and will be further examined by study.

Antioxidant and antiapoptotic effects of erdosteine in a rat model of ovarian ischemia-reperfusion injury.

OBJECTIVES: To evaluate the protective effect of erdosteine, an antiapoptotic and antioxidant agent, on torsion-detorsion evoked histopathological changes in experimental ovarian ischemia-reperfusion (IR) injury. MATERIALS AND METHODS: Eighteen female Wistar albino rats were used in control, IR, and IR+Edosteine (IR-E) groups, (n=6 in each). The IR-E group received the erdosteine for seven days before the induction of torsion/retorsion, (10 mg/kg/days). The IR and IR-E groups were exposed to right unilateral adnexal torsion for 3 hr. Three hours later, re-laparotomy was performed, and the right ovaries were surgically excised. Oxidant and antioxidants levels were determined in serum. The ovarian tissue samples were received and fixed with 10% neutral buffered formalin. The sections were stained with H&E, anti-PCNA, and TUNEL. RESULTS: The IR group were showed severe acute inflammation, polynuclear leukocytes and macrophages, stromal oedema and haemorrhage. Treatment with erdosteine in rats significantly retained degenerative changes in the ovary PCNA (+) cell numbers were significantly decreased in the IR and IR-E groups unlike the control group. However, its numbers were significantly increased in the IR-E group unlike the IR group. TUNEL (+) cell numbers were significantly increased in the IR group unlike the control and the IR-E groups. In erdosteine treated group, TUNEL (+) cells were detected significantly less than the IR group (P<0.05). CONCLUSION: In conclusion, erdosteine maybe a protective agent for ovarian damage and decreasing lipid peroxidation products and leukocytes aggregation after adnexal torsion in animals.

Evaluation of the protective effects of hesperetin against cisplatin-induced ototoxicity in a rat animal model.

OBJECTIVES: We aimed to investigate the effects of hesperetin as a flavanon both histopathologically and immunohistochemically on cochlear apoptosis in a rat model of cisplatin-induced ototoxicity (CIO). The evaluation of the effects of hesperetin on cisplatin-induced hearing loss was performed using distortion product otoacoustic emission (DPOAE). METHODS: Twenty-eight wistar albino rats were used in the current study. The rats were randomly divided into four groups with seven rats in each group. Group C was exposed to a single dose of cisplatin (12mg/kg) by intraperitoneal injection. Group CH received intraperitoneally cisplatin (12mg/kg) and hesperetin (20mg/kg). Group H was exposed to hesperetin (20mg/kg) intraperitoneally. The sham group (group S) received normal saline (6cc) intraperitoneally. The measurements of DPOAE and signal-noise ratios (SNR) were performed before the treatment and again on the first and 6 days after administration of the drugs. Rats were sacrificed and cochleae were dissected 10 days after drug administration. The cochlear tissue was assessed in all groups by histopathologic, immunohistochemical and TUNEL assay. In addition, serum oxidative stress markers and antioxidant parameters were analyzed. RESULTS: There was a significant difference between the basal value and the sixth day at frequencies 8.4, 9.6 and 9.96 for group C. We also found a significant difference between the first and sixth day at frequencies 7.2, 8.4, 9.6 and 9.96. On the 6th day, there were significant differences between C and S groups at all frequencies except 2.4. We showed a significant difference between C and H groups at frequencies 4.8, 6.0, 8.4, 9.6 and 9.96. There was also a significant difference between C and CH groups at frequencies 2.4, and 3.6. We found lower levels of oxidants and higher levels of antioxidants in CH group as compared to C group. C group had a significantly greater number of TUNEL-positive cells than did S, H and CH groups. The number of TUNEL-positive cells in CH group was higher than in S and H groups. There was a significant difference between the positive PCNA cells of CH group compared to S and H groups in spiral ganglion and stria vascularis. In addition, there were no positive PCNA cells in C group. CONCLUSIONS: Hesperetin may prevent ototoxicity by increased antioxidant enzymes and reduced oxidant parameters and protected against apoptosis resulting from a proliferation of cochlear cells in CIO.

The efficacy of tyrosine kinase inhibitor dasatinib on colonic mucosal damage in murine model of colitis.

BACKGROUND AND OBJECTIVE: Ulcerative colitis is an inflammatory condition of the colon in the gastrointestinal system. Currently, the most potent medications used for ulcerative colitis produce no response in 20-30% of cases. There is a need for more efficient and reliable medications. Tyrosine kinase inhibitors have shown efficacy in some inflammatory diseases. Although dasatinib, a tyrosine kinase inhibitor, suppresses proinflammatory cytokines in colonic tissue, there are a few cases of hemorrhagic colitis with dasatinib. There is no study investigating the effect of dasatinib on experimental colitis. We aimed to investigate the effect of dasatinib in a colitis model induced with acetic acid in our study. METHODS: In the study, 24 male Sprague-Dawley rats randomly distributed into 4 groups of 6 rats each as control, dasatinib, colitis and dasatinib+colitis groups. For colitis induction, 4% acetic acid was used. Sacrificing of the rats was performed on the seventh day. Disease activity, morphologic and histological injury, superoxide dismutase, myeloperoxidase and malondialdehyde activity, TNFα and CD3 expression were assessed in colonic tissue. RESULTS: Apart from malondialdehyde, significant difference in all parameters between the control and colitis groups was determined. Difference between the colitis and colitis+dasatinib groups was not significant in only weight loss and biochemical parameters. Though dasatinib does not fully resolve the changes in colitis, there was significant regression. CONCLUSIONS: Dasatinib decreased the inflammation in a rodent model of colitis. It may be provide this effect by the suppression of TNFα. Dasatinib may be one of the treatment options for ulcerative colitis.

Effect of Topically Applied Azithromycin on Corneal Epithelial and Endothelial Apoptosis in a Rat Model of Corneal Alkali Burn.

PURPOSE: To investigate the antiapoptotic effect of topically administered azithromycin (AZM) on corneal epithelial and endothelial cells in a rat model of corneal alkali burn. METHODS: Twenty-four Wistar albino rats were divided into 4 equal groups as pseudovehicle (group 1), control (group 2), alkali burned (group 3), and treatment (group 4) groups. Alkali injury was induced only in the right corneas of rats belonging to groups 3 and 4 using 1N NaOH. The rats in group 3 and the rats in group 4 were respectively treated either with an artificial tear gel or with 1.5% AZM eye drops for 5 days. At the fifth day of the experiment, the apoptosis in the corneal epithelium and endothelium of all rats was assessed using a terminal dUTP nick-end labeling (TUNEL) assay. In addition, tumor necrosis factor-alpha (TNF-α) density in the corneal epithelium was measured in all rats. RESULTS: The mean numbers of TUNEL+ cells in the corneal epithelium and endothelium of rats in group 3 were 117.1 ± 23.8 and 34.6.± 11.3, respectively, whereas in group 4, they were 75.8 ± 15.7 and 14.7 ± 3.5, respectively. Also the mean TNF-α densities in the corneal epithelium in group 3 and group 4 were 2.65 ± 1.3 and 1.65 ± 1.1, respectively. There was a significant decrease in the mean number of TUNEL+ cells in the corneal epithelium and endothelium and in the mean TNF-α density in the corneal epithelium of rats in group 4, when compared with group 3. CONCLUSIONS: Topically applied AZM can decrease TNF-α-induced apoptosis in corneal alkali burn.

Efficacy of diffusion-weighted magnetic resonance imaging in the evaluation of extrahepatic cholestasis-related hepatic fibrosis.

BACKGROUND/AIM: To investigate the efficacy of diffusion-weighted magnetic resonance imaging (DWI) in the diagnosis and staging of fibrosis induced by experimental bile duct ligation (BDL). MATERIALS AND METHODS: Twenty-four rats were divided randomly into four groups: control, BDL--3 days, BDL--2 weeks, and BDL--4 weeks. DWI was performed with b-values of 100 and 500 on the rats from control group at day zero, on the rats from the BDL--3 days group at the end of day 3, on the rats from the BDL--2 weeks group at the end of day 14, and on the rats from the BDL--4 weeks at the end of day 28. RESULTS: When fibrosis scores generated in all groups were evaluated together, a strong negative correlation was detected between fibrosis scores and apparent diffusion coefficient (ADC) values measured using b 100 and b 500. ADC values obtained using b 100 were found to be significantly higher compared to the fibrosis observed in both the BDL--2 weeks and BDL--4 weeks groups (P < 0.003 and P < 0.001, respectively). CONCLUSION: We think that DWI may be an alternative to liver biopsy for the diagnosis and staging of hepatic fibrosis with underlying extrahepatic cholestasis.

Spleen tyrosine kinase (Syk) inhibitor fostamatinib limits tissue damage and fibrosis in a bleomycin-induced scleroderma mouse model.

OBJECTIVES: The pathogenesis of fibrosis in scleroderma (SSc) is unknown. TGF-ß and platelet-derived growth factor are important in the development of fibrosis and tyrosine kinases are involved in these pathways. The possible antifibrotic effects of various kinase inhibitors in SSc have been studied before. Spleen tyro-sine kinase (Syk) is a protein tyrosine kinase which activates intracellular signal transduction pathways; and has been claimed to be involved in the pathogenesis of systemic autoimmune diseases. Inhibition of Syk suppresses IgE- and IgG-associated FcR signal activation in various cell types; and suppresses experimental arthritis and skin and kidney disease in lupus-prone mice. We investigated the ability of a small drug, the Syk inhibitor, fostamatinib, to protect mice from bleomycin-induced SSc. METHODS: Four study groups of BALB/c mice were included into this study: control, bleomycin (administered subcutaneously to BALB/c mice for 21 days), bleomycin and fostamatinib (mice fed with chow containing a Syk inhibitor for 21 days), and fostamatinib alone groups. Skin and lung tissue specimens were obtained and evaluated histologically. RESULTS: Treatment with fostamatinib significantly reduced skin thickness and fibrosis. Mice treated with fostamatinib also displayed less fibrosis and inflammation in the lung tissue. Following fostamatinib treatment, Syk, phospho-Syk, and TGF-ß expression decreased in both skin and lung tissues. CONCLUSIONS: The Syk inhibitor fostamatinib prevented bleomycin-induced fibrosis and inflammation in the skin and in the lung. The anti-fibrotic effect of fostamatinib is linked to reduced Syk phosphorylation and TGF-ß expression. The Syk pathway appears as a potential molecular target for therapeutic intervention in SSc.

The Syk Inhibitor Fostamatinib Decreases the Severity of Colonic Mucosal Damage in a Rodent Model of Colitis.

BACKGROUND AND AIMS: Inflammatory bowel disease is a chronic inflammatory disease of the gastrointestinal system. In some cases, current medications used for inflammatory bowel disease may not be enough for remission, creating a need for more potent and reliable medications. There is no study showing the efficacy of fostamatinib, with proven effects on some inflammatory diseases, on ulcerative colitis. In our study we planned to research the efficacy of fostamatinib, a spleen tyrosine kinase inhibitor, on acetic acid-induced colitis. METHODS: The study included 28 male Sprague-Dawley rats, randomly divided into control group, fostamatinib group, colitis group and fostamatinib + colitis group, each containing seven rats. Colitis induction was performed with 4% acetic acid. Colonic inflammation was assessed with disease activity index, macroscopic and histological damage scores, colonic myeloperoxidase, malondialdehyde and superoxide dismutase activity, and tumour necrosis factor alpha [TNFα], CD3, Syk, and phospho-Syk expression. RESULTS: There was a significant difference between the colitis and control groups in terms of all parameters. The disease activity index, macroscopic and microscopic damage scores, immunohistochemical TNFα, CD3, Syk, and phospho-Syk expression, and tissue myeloperoxidase activity were found to be significantly lower in the colitis + fostamatinib group compared with the colitis group. There was no significant difference between the two groups in terms of myeloperoxidase and malondialdehyde activity. CONCLUSIONS: Fostamatinib reduced the inflammatory damage in the experimental colitis. This effect may be due to suppression of TNFα, T-lymphocytes, and neutrophils in colonic mucosa via suppression of Syk. Fostamatinib may be an appropriate treatment alternative for ulcerative colitis. Further clinical studies are required to support this.

Protective effect of royal jelly in 2,4,6 trinitrobenzene sulfonic acid-induced colitis in rats.

OBJECTIVES: In the present study, we evaluated immunological and immunomodulatory properties of royal jelly (RJ) in 2,4,6 trinitrobenzene sulfonic acid (TNBS)-induced colitis in rats. MATERIALS AND METHODS: Eighteen adult female Wistar albino rats were divided into three groups of six animals each: a control group that received only saline solution, a TNBS-induced colitis group, and a TNBS-colitis+RJ group that received 250 mg/kg/day of RJ for seven days before the induction of colitis, following by the same treatment for an additional seven days. At the end of the experiment, cardiac blood and colon samples were obtained under deep anaesthesia from the animals in all groups. Serum interleukin-1ß (IL-1ß), tumour necrosis factor-alpha (TNF-α) and IL-10 levels were analyzed with an enzyme-linked immunosorbent assay (ELISA). Five-micrometre-thick sections were stained with haematoxylin-eosin (H&E) for microscopic evaluations. For immunohistochemical evaluations, the paraffin sections were stained with anti-CD3 (cluster of differentiation), anti-CD5, anti-CD8 and anti-CD45. RESULTS: The results showed that the oral RJ treatment inhibited proinflammatory cytokines, IL-1ß and TNF-α secretion, while increasing anti-inflammatory cytokine IL-10 production in the TNBS-induced colitis+RJ group compared with the colitis group not treated with RJ. The colitis was not as severe in the colitis+RJ group, with ulcerative damage, weight loss and inflammatory scores suggesting that impaired CD3-, CD5-, CD8- and CD45-positive T cell immune responses likely mediated the anti-inflammatory effect. CONCLUSION: The antioxidant and anti-inflammatory properties of RJ protected colon mucosa against TNBS-induced colitis in rats orally treated with RJ.

Effects of sphingosylphosphorylcholine against oxidative stress and acute lung injury induced by pulmonary contusion in rats.

BACKGROUND/PURPOSE: The goal of this study was to evaluate effects of exogenous sphingosylphosphorylcholine (SPC) administration on acute lung injury induced by pulmonary contusion in rats. METHODS: Eight animals were included in each of the following five groups: control, contusion, contusion phosphate-buffered solution (PBS), contusion SPC 2, contusion SPC 10. SPC was administered 3 days at a daily two different doses of 2 µm/ml and 10 µm/ml intraperitoneally. The severity of lung injury was determined by the neutrophil activation and histological and immunohistochemical changes in the lung. Malondialdehyde (MDA), nitric oxide (NO), superoxide dismutase (SOD), glutathione peroxidase (GPx) and glutathione (GSH) were determined to evaluate the oxidative status in the lung tissue. RESULTS: Treatment with 2 µM SPC inhibited the increase in lung MDA and NO levels significantly and also attenuated the depletion of SOD, GPx, and GSH in the lung injury induced by pulmonary contusion. These data were supported by histopathological findings. The inducible nitric oxide synthase (iNOS) positive cells and apoptotic cells in the lung tissue were observed to be reduced with the 2 µM SPC treatment. But, the 10 µM SPC treatment did not provide similar effects. CONCLUSIONS: In conclusion, these findings suggested that 2 µM SPC can attenuate lung damage in pulmonary contusion by prevention of oxidative stress, inflammatory process and apoptosis. All these findings suggest that low dose SPC may be a promising new therapeutic agent for acute lung injury.

Protective effects of royal jelly against testicular damage in streptozotocin-induced diabetic rats.

BACKGROUND/AIM: TO examine the effects of royal jelly (RJ) on testicular damage in streptozotocin (STZ)-induced diabetic rats. MATERIALS AND METHODS: Eighteen adult Wistar albino rats were used, 6 in each of the 3 treatment groups: Group A: control, Group B: STZ-induced diabetes (untreated), Group C: STZ-induced diabetes plus RJ (400 mg/kg daily for 4 weeks). Diabetes was induced by a single intraperitoneal injection of STZ (60 mg/kg). Four weeks after the onset of diabetes, testicular apoptotic cell death was examined using immunohistochemical staining for caspase-3 and Ki67 staining for localization of proliferative cells. RESULTS: Compared with the control, the body and testicular weights of the RJ-treated and untreated diabetic rats were decreased (P < 0.05). The histopathological examination showed a significant increase in degenerative changes in the seminiferous tubules and in spermatogenesis of the STZ-treated rats. In contrast, the RJ treatment group showed near-normal morphology, in addition to an increased intensity of immunohistochemical staining for Ki67-positive cells. CONCLUSION: Diabetes induced a significant increase in testicular apoptotic cell death (caspase-3-positive cells). Caspase-3-positive cells were significantly decreased in the STZ plus RJ-treated group compared with the untreated STZ-induced diabetic group (P < 0.05).

The effect of hesperetin on ischemia-reperfusion injury in rat ovary.

PURPOSE: Hesperidin (HES), a citrus fruit extract, has beneficial effects on various ischemia/reperfusion (I/R) models. We aimed to evaluate the possible positive effects of hesperetin (HPT), an active metabolite of HES, on a rat ovarian I/R model. METHODS: We divided 24 Wistar Albino rats into four groups. Group I (n = 6) was sham operated, Group II (n = 6) was the I/R group, Group III (n = 6) was the I/R + solvent group and Group IV (n = 6) was the I/R + HPT group. Three hours of ischemia and 3 h of reperfusion were performed on each rat in Groups II, III, and IV. Dimethyl sulfoxide (DMSO) was given intraperitoneally to the rats in the III. Group, and 50 mg/kg of HPT dissolved in DMSO was given intraperitoneally to the rats in the IV. Group 30 min before reperfusion. After 3 h of reperfusion, the ipsilateral ovaries of the rats were examined immunohistochemically to detect apoptosis. RESULTS: Hematoxylin and eosin (H and E) staining demonstrated less edema and hemorrhage in the group where HPT was applied. Caspase-3 and terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) staining showed significantly lower apoptosis in the group where HPT was used when compared to either the I/R or solvent group. CONCLUSIONS: To the best of our knowledge, this is the first study that shows the beneficial effects of HPT in an ovarian I/R injury. HPT improved tissue damage and apoptosis caused by I/R injury. To identify the possible positive effects of HPT in ovarian torsion of humans and use in clinical practice, more studies must be performed.

The protective effect of quercetin on IMA levels and apoptosis in experimental ovarian ischemia-reperfusion injury.

OBJECTIVE: To investigate the protective effect of quercetin (QE), an anti-inflammatory and anti-oxidant agent, on torsion-detorsion induced histopathological changes and blood IMA levels in experimental ovarian ischemia-reperfusion (IR) injury. STUDY DESIGN: Twenty-four female Wistar rats were randomly divided into four groups in this study (n=6). Group I, (sham operation); Group II, torsion-detorsion plus saline (IR); Group III, torsion-detorsion plus solvent (dimethylsulfoxide: DMSO, IR+DMSO); Group IV, torsion-detorsion plus 15 mg/kg/bw quercetin (IR+QE) injected intraperitoneally 30 min prior to detorsion. After 3h of reperfusion, the right ovaries were removed surgically. The ovary tissue samples were fixed in 10% formalin solution for histopathological and immunohistochemical examination. Blood samples were obtained at the end of the procedures for each group of animals. RESULTS: Ovarian sections in Groups II and III showed higher follicular cell degeneration, hemorrhage, vascular congestion and edema when compared with Group I. Administration of quercetin in rats significantly prevented degenerative changes in the ovary. Significantly less histopathological changes were found in Group IV compared with Groups II and III. Caspase-3 and TUNEL positive cells were detected in the ovarian surface, follicle epithelium, and stromal cells in all experimental groups, and there was a significant increase in Groups II and III compared with Group I (P<0.05). Treatment with quercetin decreased the number of caspase-3 and TUNEL positive cells. IR increased the ischemia modified albumin (IMA) levels in comparison to the sham group (1.06 ± 0.10 ABSU and 0.92 ± 0.08 ABSU, P<0.05). Quercetin administration before IR reduced the levels of IMA (0.93 ± 0.08 ABSU, P<0.05). CONCLUSION: Administration of quercetin is effective in preventing tissue damage induced by IR injury in ovaries.