Protective effect of saponin on sperm DNA fragmentation of mice treated with cyclophosphamide.

Cyclophosphamide (CP) is a common chemotherapy drug with the testicular damage. The aim of this study was to investigate the effect of saponin (SP) on the toxicity of CP in the male reproductive system. Following an experimental pilot study for determining SP dose, 40 male mice (32 ± 3 g) were divided into five groups (n = 8): control, sham (normal saline 0.2 ml/day), CP (15 mg/kg/week, intraperitoneally), SP (2.5 mg/kg/day, intraperitoneally) and saponin group with cyclophosphamide (SP + CP). After treatment, the left testes were removed for the measurement of malonedialdehyde (MDA) and total antioxidant capacity (TAC) levels, and sperm DNA fragmentation was assessed by SDFA kit. In the CP group, a significant decrease in motility, viability, count, normal morphology and DNA fragmentation of spermatozoa and TAC was observed, while in MDA level, a significant increase was observed compared with the control group (p < 0.05). Attenuated sperm parameters in CP group improved significantly in SP + CP group (p < 0.05). According to the findings of this study, SP was able to alter the reproductive toxicity of CP in NMRI mice and increase the antioxidant capacity of the testis.

Protection of CCl4-induced hepatic and renal damage by linalool.

The aim of the current study is to determine the protective and therapeutic effects of linalool against carbon tetrachloride (CCl4)-induced hepatoxicity and nephrotoxicity. Six-week-old male Wistar rats were divided into five groups: Control group (a regular diet); CCl4 group (1 ml/kg dissolved in olive oil, intraperitoneally at 14th day); pretreatment group (25 mg/kg linalool daily + CCl4 14thday); post-treatment group (25 mg/kg linalool 2, 6, 24, and 48 h after the injection of CCl4 at 14th day); and linalool group (25 mg/kg linalool daily, orally). All animals were sacrificed, tissue and blood samples were collected to analysis. Administration of CCl4 resulted in a marked increase in hepatic (aspartate aminotransferase, alanine transaminase, and alkaline phosphatase) and renal (blood urea nitrogen and creatinine) markers. Also, CCl4 resulted in pathological damages, a significant increase in the concentration of malondialdehyde , tumor necrosis factor-alpha, and Interleukin 6 , expression of nuclear factor kappa-light-chain-enhancer of activated B cells and a significant decrease in the levels of serum total protein, serum albumin, and antioxidants. However, in pretreatment and post treatment groups, linalool significantly inhibited CCl4- induced hepatic and nephric damages. These results demonstrate that linalool has protective and therapeutic effects in an in vitro model of CCL4-induced hepatic and nephric damage, proposing linalool as a potential therapeutic agent against chemical and drug induced hepatotoxicity and nephrotoxicity.

Treatment of cartilage defects by Low-intensity pulsed ultrasound in a sheep model.

Aim of this study was to evaluate effects of Low-intensity pulsed ultrasound on repair of articular cartilage defects. Low-intensity pulsed ultrasound (Lipus) can induce the differentiation and activation of chondrocytes. This study was designed to evaluate the effect of Lipus on articular cartilage defects in a sheep. Eight sheep were divided in to two groups. The animals received bilateraly, articular cartilage defects 4 mm in diameter and 2 mm in deep on the patellar groove and experimental groups were treated with intensity 200 mW/cm2, 20 min/day with low-intensity pulsed ultrasound for 2 month. Then both knee joints underwent surgery for remove of formed tissue sample from defects.The samples were evaluated by Quantitative real-time polymerase chain reaction (qRT-PCR), Safranin-o staining, Immunofluorescence Staining and Morphological characterization. The best and worst sample per group according to Macroscopic and micriscopic scoring were icentified. The results showed that the operated groups with-Lipus-treatment and without-Lipus treatment had considered statistically significant. Gross photography revealed that the defects in experimental groups were filled with proliferative tissue, while in control groups, a thin layer of proliferative tissue was formed in defects. qRT-PCR results showed the expression of coll2, sox9, aggrecan and Osteocalcin in experimental groups. Intense safranin-O staining show the formation cartilage tissue in ultrasound treated group, while loose safranin-o-staining were observed at the control groups. Immunofluorescence staining showed the type 2 Collagen protein expression. We suggest that low-intensity pulsed ultrasound provide the mechanistic basis force for articular cartilage repair and effective treatment modality for improving of articular cartilage defects.

The protective effect of cinnamon and ginger hydro-alcoholic extract on carbon tetrachloride-induced testicular damage in rats.

Sexual dysfunction of men is one of the most serious problems in human society. This study aimed to evaluate the protective effect of cinnamon and ginger extract on testicular damages induced by carbon tetrachloride (CCl4). Thirty-six male Wistar rats were randomly divided into 6 groups (n = 6): 1. Normal control; 2. Carbon tetrachloride (CCl4); 3. CCl4 + Cinnamon; 4. CCl4 + Ginger; 5. CCl4 + Cinnamon and Ginger; and 6. Cinnamon + Ginger. CCl4 (1 ml/kg) was injected intraperitoneally on the 14th day, and cinnamon (50 mg/kg, orally) and ginger (250 mg/kg, orally) were administered daily for 14 days. Fifty hours after the CCl4 injection, the testicles and epididymis were separated and examined as to histological alterations and oxidative stress markers. CCl4 significantly increased malondialdehyde level and decreased total antioxidant capacity when compared to the normal control group (p < .05). In addition, degenerative alterations in the testicular and epididymal tissue were observed in CCl4 group. The pre-treatment with ginger and cinnamon extract significantly improved these parameters when compared to the CCl4 group (p < .05). The results of this study indicated that co-treatment of ginger and cinnamon reduces the damages induced by CCl4 in testicular tissue by increasing antioxidant capacity and reducing lipid peroxidation.

The effect of clomiphene citrate and human chorionic gonadotropin on the expression of CatSper1, CatSper2, LHCGR, and SF1 genes, as well as the structural changes in testicular tissue of adult rats.

The purpose of this study was to investigate the effect of clomiphene citrate and human chorionic gonadotropin (HCG) on the structural changes, as well as the evaluation of the expression of cation channel sperm-associated protein 1 (CatSper1), cation channel sperm-associated protein 2 (CatSper2), luteinizing hormone/choriogonadotropin receptor (LHCGR), and steroidogenic factor 1 (SF1) genes in testicular tissue of rats. All rats divided into five groups as follows; G1 as the control group that received normal saline, G2 received olive oil, G3 received 100 IU/kg HCG, G4 received 5 mg/kg clomiphene citrate, and G5 received 5 mg/kg clomiphene citrate and 100 IU/kg HCG. At the end of the experiment period, Day 56, blood samples were taken and the serum was isolated. Then, histomorphometric analysis, hormonal assess, and real-time polymerase chain reaction to measure the expression of CatSper1, CatSper2, LHCGR, and SF1 genes were performed. The results showed that the concentrations of testosterone, follicle-stimulating hormone, and luteinizing hormone were decreased in the G4 group, whereas these parameters were increased in the G3 group. A comparison of the sperm quality indicated a significant reduction in the quality of sperm cells in the G4 group compared with other groups. The quality of sperm was significantly enhanced in the G3 and G5 groups in comparison with the G1 group. Also, our findings demonstrated that the expression of CatSper1, CatSper2, LHCGR, and SF1 genes were significantly elevated in the G3 group when compared with other experimental groups. According to the obtained results, it seems that clomiphene citrate reduces the process of spermatogenesis and the detrimental impacts of this compound would be neutralized by the administration of HCG.

Cytotoxicity of curcumin against CD44± prostate cancer cells: Roles of miR-383 and miR-708.

Objective: Cancer stem cells (CSCs) remaining in the tumor tissues after applying treatments may cause recurrence or metastasis of prostate cancer (PC). Curcumin has the promising potential to target CSCs. Here, we aim to evaluate the cytotoxic effects of curcumin on the expression of miR-383-5p and miR-708-5p and their target genes in CD44+ CSCs and CD44- non-CSCs isolated from the PC3 prostate cancer cell line. Materials and Methods: We used MTT assay to determine the optimal cytotoxic dose of curcumin on CD44± PC cells. Then, we assessed nuclear morphological changes using DAPi staining. We used Annexin V-FITC/PI to quantify apoptotic cell death. qRT-PCR was also used to detect miRNA and gene expression levels after curcumin treatment. Results: Curcumin significantly enhanced the apoptosis in both CD44- and CD44+ PC cells in a dose-dependent manner (p < 0.05). The cytotoxicity of curcumin against CD44- cells (IC50 40.30±2.32 µM) was found to be greater than that against CD44+ cells (IC50 83.31±2.91 µM). Also, curcumin promoted miR-383-5p and miR-708-5p overexpression while downregulating their target genes LDHA, PRDX3, and RAP1B, LSD1, respectively. Conclusion: Our findings indicate that curcumin, by promoting the expression of tumor suppressors, miR-383-5p and miR-708-5p, and inhibiting their target genes, induced its cytotoxicity against CD44± PC cells. We trust that curcumin could be established as a promising adjuvant therapy to current PC treatment options following more research in clinical settings.

Effect of hydroalcoholic seed extract of Nigella sativa on hepatic and pancreatic factors of Nrf2 and FGF21 in the regulation of insulin transcription factors of MafA and PDX-1 in streptozotocin-treated diabetic rats.

INTRODUCTION: Nigella sativa (N. sativa), one of the most commonly used medicinal herbs with antioxidant properties, increases blood insulin levels and lowers fasting blood sugar. Nuclear Erythroid Factor-Related Factor 2 (Nrf2) and Fibroblast Growth Factor 21 (FGF21) are two antioxidant factors that are increased by oxidative stress and hyperglycemia. The present study investigated how hydroalcoholic extract of N. sativa seed (HENS) increases blood insulin levels, taking into account changes in antioxidant factors and expression of insulin transcription factors. MATERIALS AND METHODS: Two groups of male diabetic wistar rats were treated orally with HESN at doses of 200 and 400 mg/kg-body weight for one month. Fasting blood sugar (FBS) and insulin were measured using standard kits by photometric and ELISA methods, respectively. The expression levels of the Nrf2, FGF21 and ß-Klotho genes as well as the insulin gene-stimulating transcription factors of MafA and PDX-1 were evaluated using real-time PCR. Oxidative stress was assessed by assessing serum total oxidation status (TOS), malondialdehyde (MDA), and total antioxidant capacity (TAC). RESULTS: HSEN showed a significant reducing effect on FBS and oxidative biomarkers and an increasing effect on serum insulin levels in treated diabetic rats compared to untreated diabetics (P < 0.05). The elevated levels of NRF2 and FGF21 in the liver and pancreas of the diabetic control group were significantly reduced after treatment with both HESN doses (P < 0.05). Following the ameliorative effects of HENS on pancreatic tissue and the reduction of oxidative stress, the expression level of MafA and PDX1 genes approached the level of these factors in healthy rats (P < 0.05). CONCLUSION: This study showed the therapeutic effects of HENS on diabetic pancreas by reducing oxidative stress and tissue damage, modifying the expression levels of PDX-1 and MafA genes, and regulating insulin secretion and blood glucose levels.

Impact of caffeic acid on the testicular damages in D-galactose-induced aging model in mice.

Objectives: Aging is a biological phenomenon that causes various disorders and diseases in body systems such as the reproductive system. One of the important factors in aging is oxidative stress, which facilitates the aging process through various mechanisms. The aim of this study is the investigation of effects of caffeic acid on the testicular damages in Dgalactose induced aging model in mice. Materials and Methods: Forty male mice were randomly divided into 5 groups (n=8): 1) Control, 2) Sham, 3) Aging, 4) Aging + caffeic acid, and 5) Caffeic acid. Aging was induced through daily injection of D-Galactose (300 mg/kg, intraperitoneal) for 6 weeks. Caffeic acid (60 mg/kg, intraperitoneal) was injected daily for 6 weeks. One day after the last injection mice were killed and the testicle and epididymis were removed. Then, sperm parameters, factors of oxidative stress, and histopathological changes were evaluated. Results: The results showed that aging significantly decreased the count, motility, and viability of sperm, and increased abnormal sperm and sperm DNA fragmentation in contrast to the control group (P<0.05). In addition, MDA levels increased significantly in this group, and SOD, GPx, and TAC activity decreased (P<0.05). Histological studies also showed the destruction of seminiferous tubules, and Johnson's score decreased (P<0.05). Caffeic acid administration significantly improved the above disarrays (P<0.05). Conclusion: The results showed that caffeic acid reduces the adverse effects of aging on spermatogenesis in mice by reducing oxidative stress and increasing antioxidant defenses.

Hepatoprotective Impact of Ghrelin against Cyclophosphamide-Induced Toxicity in the Male Mice.

BACKGROUND: Despite its vast spectrum of clinical usage, cyclophosphamide (CP) exerts many adverse impacts, including hepatotoxicity. Antioxidant properties of ghrelin might protect the liver from CP-induced toxicity. The current study aimed to assess the protective impacts of ghrelin on CP-induced liver toxicity. METHODS: Forty male mice were randomly divided into four groups (n=10) Group 1 as control received no intervention,group 2 received cyclophosphamide (CP) (100 mg/kg, i.p.) for five weeks and once a week. Group 3 received CP+ghrelin (CP+G), (80 µg/kg daily, i.p.) for five weeks. Group 4 received ghrelin with above-mentioned dose. At the end of the experiment, the mice were sacrificed to remove liver tissuesfor histological and biochemical examination. RESULTS: Malondialdehyde (MDA) level increased after CP treatment but ghrelin administration significantly decreased the level of MDA (P<0.05). Measurement of the total antioxidant capacity (TAC) noted a significant decrease in the CP group against the control group (P<0.05). Ghrelin treatment in the CP+G group considerably increased the TAC activity when compared to the CP group (P<0.05). Histological examinations also confirmed the hepatocyte necrosis, local bleeding and inflammation, vacuolation, and sinusoidal dilation in the CP group, ghrelin administration reduced the destructive effects of CP on the liver significantly (P<0.05). CONCLUSION: Our results reveal the hepatoprotective effect of ghrelin against CP. Therefore, ghrelin might be useful in protecting the body against the adverse impacts of injuries induced by chemotherapeutic drugs.

Protective effect of hydroalcoholic extract of Teucrium polium on diabetes-induced testicular damage and serum testosterone concentration.

BACKGROUND: Diabetes has an adverse effect on spermatogenesis by rising oxidative stress. OBJECTIVE: The aim of this study was to investigate the effect of Teucrium Polium extract administration on spermatogenesis and testicular structure in diabetic rats induced with Streptozotocin. MATERIALS AND METHODS: 32 male Wistar rats were randomly divided into four groups (n=8/each): control group, diabetic group received distilled water, and two experimental groups included diabetic rats treated with 50 and 100 mg/body weigh of Teucrium Polium extract for 6 six weeks. After six weeks, the left testis had been removed and the morphometrical study was performed. Blood samples were collected from the ophthalmic veins of the rats and plasma levels of glucose and testosterone hormone were measured afterward. RESULTS: The reduction in diameters of the seminiferous tubules and thickening of the wall of the seminiferous tubules (p=0.05) were seen in diabetic rats. Also, the degenerative changes in cells arrangement have been observed. Statistical analysis showed the use of Teucrium Polium significantly improved the above disorders in treatment group (100 mg/BW) in contrast to the non treated diabetic group (p=0.05), but no significant difference was seen between the experimental group treated with 50 mg/BW of Teucrium polium and diabetic group (p=0.08). These data also revealed that treatment of diabetic rats with 100 mg/BW of Teucrium Polium extract significantly improves the change in serum glucose (p=0.001) and testosterone (p=0.03). CONCLUSION: The results of the present study indicate that diabetes produces degenerative changes in the testis of rats and administration of Teucrium polium reduces complications resulted from diabetes.

A variation of Musculocutaneous nerve without piercing the coracobrachialis muscle while communicating to the median nerve: A case report and literature review.

INTRODUCTION: Anatomical variations of the peripheral nervous system may have not any clinical signs and symptoms. One of these variations belongs to the Musculocutaneous nerve. However, a good knowledge of nerve pathways and their variations is very important for surgeons in post-traumatic evaluations, exploratory interventions, and/or administration of neuromuscular blocks in axillary region in order to surgical therapies. PRESENTATION OF CASE: This report describes a case of variation of the musculocutaneous nerve which was observed in an old Iranian male cadaver during routine educational dissection (Fig. 1). DISCUSSION AND CONCLUSION: Anatomically, in the axilla region, the Musculocutaneous nerve is originated of the lateral cord of brachial plexus, then, by piercing the coracobrachialis muscle arrives enters to anterior compartment of the arm. But, in the present report, we observed that the Musculocutaneous nerve without piercing the coracobrachialis muscle has arrived in the left arm, then communicated to the Median nerve. To exploratory interventions of the arms for peripheral nerve repair and surgical therapies, a good knowledge of nerve pathways helps to surgeons for preventing possible mistakes during surgery.