6‐Gingerol‐rich fraction from Zingiber officinale ameliorates carbendazim‐induced endocrine disruption and toxicity in testes and epididymis of rats

This study evaluated the protective effects of 6‐gingerol‐rich fraction (6‐GRF) from Zingiber officinale on carbendazim (CBZ)‐induced reproductive toxicity in rats. Adult male rats were treated with either CBZ (50 mg/kg) alone or in combination with 6‐GRF (50, 100 and 200 mg/kg) for 14 consecutive days. Gas chromatography–mass spectrometry (GCMS) analysis revealed that 6‐GRF consists of ten bioactive chemical components with 6‐gingerol being the most abundant (30.76%). Administration of 6‐GRF significantly (p < .05) prevented CBZ‐mediated increase in absolute and relative testes weights as well as restored the sperm quantity and quality in the treated rats to near control. In testes and epididymis, 6‐GRF significantly abolished CBZ‐mediated increase in oxidative damage as well as augmented antioxidant enzymes activities and glutathione level in the treated rats. Moreover, CBZ administration alone significantly decreased plasma levels of testosterone, thyrotropin, triiodothyronine and tetraiodothyronine, whereas follicle‐stimulating hormone was significantly elevated without affecting luteinising hormone and prolactin levels when compared with the control. Conversely, 6‐GRF ameliorated the disruption in the hormonal levels and restored their levels to near normalcy in CBZ‐treated rats. Collectively, 6‐GRF inhibited the adverse effects of CBZ on the antioxidant defence systems, hormonal balance and histology of the testes and epididymis in rats.     

Testosterone influences the expression and distribution of the cation‐dependent mannose‐6‐phosphate receptor in rat epididymis. Implications in the distribution of enzymes

The mammalian epididymis plays a role in sperm maturation through its secretory activity. Among the proteins secreted by the epithelium, there are significant amounts of acid hydrolases. In most cell types, the normal distribution of lysosomal enzymes is mediated by mannose‐6‐phosphate receptors (MPRs). In this study, we analysed the expression and distribution of the cation‐dependent MPR (CD‐MPR) in epididymis from control, castrated or castrated rats with testosterone replacement. It was observed that expression of CD‐MPR increased due to castration in all regions of the epididymis, which was reversed by injection of testosterone. We also measured the activity of α‐mannosidase and observed that the castration tends to increase the retention of this enzyme in the tissue, which is reversed by the hormone replacement. In corpus, this resulted in a reduced secretion of the enzyme. Immunohistochemistry showed that CD‐MPR has a supranuclear location (different from the cation‐independent MPR), most likely in principal cells, and low reactivity in other cell types. The signal in castrated animals was more intense and tended to redistribute towards the apical cytoplasm. Thus, we concluded that expression and distribution of CD‐MPR is affected by decrease of testosterone in rat epididymis, and this could change the distribution of lysosomal enzymes.     

Melatonin prevents gentamicin‐induced testicular toxicity and oxidative stress in rats

This study investigated the protective effects of melatonin (MT) against gentamicin (GM)‐induced testicular toxicity and oxidative damage in rats. GM (100 mg kg^?1) was injected intraperitoneally (i.p.) to rats for 6 days. MT (15 mg kg^?1) was administered i.p. to rats for 6 days at 1 hr after the GM treatment. GM caused a decrease in prostate and seminal vesicle weights, sperm count and sperm motility. Histopathological examination showed various morphological alterations in the testis, characterised by degeneration of spermatogonia/spermatocytes, decrease in the number of early spermatogenic cells and vacuolisation. In addition, an increased malondialdehyde concentration and decreased glutathione content and glutathione reductase, catalase and glutathione‐S‐transferase activities were found in the testis. In contrast, MT treatment significantly attenuated the testicular toxicity of GM, including decreased reproductive organ weights, sperm count, and sperm motility and increased histopathological alterations. MT also had an antioxidant benefit by decreasing the lipid peroxidative product malondialdehyde and increasing the level of the antioxidant glutathione and the activities of antioxidant enzymes in the testis. These results indicate that MT prevents testicular toxicity induced by GM in rats, presumably due to its potent antioxidant activity, and its ability to inhibit lipid peroxidation, and restore antioxidant enzyme activity.     

Tocotrienol‐rich fraction supplementation prevents foetal loss in females mated with corticosterone‐treated male Sprague–Dawley rats

This study examined whether tocotrienol supplementation to corticosterone‐treated male rats could prevent foetal loss in females upon their mating. Epididymides of adult male Sprague–Dawley (SD) rats with proven fertility were surgically separated at the testis‐caput junction. Twenty‐four hours post‐surgery, these animals received for 7 days either: tocopherol‐stripped corn oil (Control), corticosterone 25 mg/kg s.c. (CORT), CORT 25 mg/kg s.c. and tocotrienol‐rich fraction (TRF) 100 mg/kg orally (CORT + TRF) or TRF 100 mg/kg orally (TRF). On day 8, males were cohabited with proestrus females. A spermatozoa‐positive vaginal smear indicated pregnancy. Males were euthanised for analysis of testosterone and antioxidant activities. Reproductive organs were weighed. On day 8 of pregnancy, females were laparotomised to count the number of implantation sites. Pregnancy was continued until term. Number of pups delivered and their weights were determined. Data were analysed using ANOVA. Malondialdehyde levels were significantly lower in CORT + TRF group compared with CORT group. Enzymatic antioxidant activities, testosterone level and reproductive organ weights were significantly higher in CORT + TRF group compared with CORT group. Number of implantation sites and live pups delivered, and their birth weights from females mated with CORT + TRF males were significantly higher compared to CORT group. Therefore, TRF prevents foetal loss in females mated with CORT + TRF‐treated males.     

Aminoguanidine prevents testicular damage–induced‐2,3,7,8‐tetrachlorodibenzo‐p‐dioxin (TCDD) in male rats

In this study, it was aimed to determinate protective effects of aminoguanidine (AG) against reproductive toxicity caused by 2,3,7,8‐tetrachlorodibenzo‐p‐dioxin (TCDD), an environmental contaminant. Thirty‐two rats were equally divided into four groups; the first group was kept as control and given corn oil as carrier. In second and third groups, TCDD and AG were orally administered at the dose of 2 μg kg^?1 per week and 100 mg kg^?1 per day for 45 days, respectively. In fourth group, TCDD and AG were given together at the same doses. Although TCDD significantly increased the formation of TBARS, it caused a significant decline in the levels of GSH, CAT, GPx and SOD in rats. On the other hand, AG, given together TCDD, reversed TCDD effects on TBARS SOD, GSH, GPx and CAT. In addition, sperm characteristics negatively affected and histopathological deformation occurred with TCDD exposure. However, AG treatment partly prevented these toxic effects of TCDD on spermatological parameters and histopathological changes. In conclusion, TCDD exposure induces testicular damage (oxidative stress, histopathological damage and sperm parameters), and AG treatment reversed TCDD‐induced testicular damage in rats. Thus, AG may be useful for the prevention and treatment of TCDD‐induced male infertility problems.     

Relationship of cytokine levels and clinical effect on platelet‐rich plasma‐treated lateral epicondylitis

Lateral epicondylitis (LE) is difficult to manage and can result in significant patient morbidity. Currently, the clinical use of platelet‐rich plasma (PRP) for painful tendons has received attention, but its efficacy remains controversial. This study aimed to investigate the clinical effects of PRP and its biological components. A total of 156 patients with LE were randomly divided into group 1, treated with a single injection of 2‐ml autologous PRP, and group 2, treated with a control received only physical therapy without injection. Both groups used a tennis elbow strap and performed stretching and strengthening exercises during 24 weeks’ follow‐up. Pain and functional improvements were assessed using the visual analog scale (VAS), Modified Mayo Clinic Performance Index for the elbow, and magnetic resonance imaging (MRI). White blood cell count, platelet count, and levels of platelet‐derived growth factor‐AB (PDGF‐AB), PDGF‐BB, transforming growth factor‐β (TGF‐β), vascular endothelial growth factor, epithelial growth factor, and interleukin‐1 β in PRP were measured and investigated for statistical correlation with the clinical score. At 24 weeks, all pain and functional variables, including VAS score, Mayo Clinic performance scores, and MRI grade, improved significantly in group 1 (p < 0.05). PDGF‐AB, PDGF‐BB, and TGF‐β levels were more significantly increased in PRP than in whole blood. TGF‐β level significantly correlated with Mayo Clinic performance score and MRI grade improvement. Thus, TGF‐β level in PRP is considered to play a pivotal role in tendon healing. These results may contribute to identifying the best protocol for PRP application in tendinopathies. © 2017 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 36:913–920, 2018.

     

The preventive role of wheat germ oil against sertraline‐induced testicular damage in male albino rats

Sertraline is an antidepressant medication used extensively in the therapy of depression. The present investigation was intended to estimate the actual protective role of wheat germ oil on sertraline‐caused testicular injury in albino rats. Sertraline (human therapeutic dose, 15.63 mg/kg) was orally administrated to rats for 28 successive days. Sertraline‐administered rats were concurrently supplemented with wheat germ oil (human therapeutic dose, 68.75 mg/kg) for 28 successive days. Sertraline administration induced an elevation in testicular DNA damage and acute testicular damage illustrated by the histopathological alterations including marked degeneration and necrosis of germ cells lining seminiferous tubules, as well as interstitial oedema, congestion of interstitial blood vessel. Wheat germ oil administration potentially mitigated the histopathological alterations of sertraline‐administered rats. Lipid peroxidation, oxidative stress biomarker, showed a significant elevation in testicular tissue of sertraline‐administered rats. Furthermore, glutathione content and catalase activity were decreased in testicular tissue of sertraline‐administered rats. Serum testosterone level was elevated in sertraline‐administered rats. Wheat germ oil significantly reduced lipid peroxidation of testicular tissue and improved the antioxidant defences. Finally, wheat germ oil has a preventive role against testicular damage induced by sertraline in rats probably via its potential to prevent reactive oxygen species.     

Effects of Anthocleista djalonensis root extracts on reproductive hormones and testicular marker enzymes in adult male Wistar rats

This study evaluated the effect of methanol and aqueous ethanol root extracts (200 mg/kg body weight) of Anthocleista djalonensis on sex hormone concentrations and testicular marker enzymes of adult rats after 60 days of administration followed by 60 days of treatment withdrawal. The results showed no significant changes in testosterone and follicle‐stimulating hormone (FSH) levels during the 60 days of extract treatment. Interestingly, 60 days after treatment withdrawal, there was an increase in intratesticular and serum testosterone and serum FSH in the methanol but not aqueous ethanol extract post‐treatment groups. Intratesticular 3β‐hydroxysteroid dehydrogenase (3β‐HSD) activity remained unaffected while that of 17β‐HSD increased slightly during treatment of both extracts and the increase reached a statistical significance level (p < .05) during post‐treatment. Gamma‐glutamyltranspeptidase activity in the testis of the methanol but not aqueous ethanol extract‐treated animals remained high during post‐treatment compared to initial treatment values. Phytochemical analysis of the plant extracts showed that phenol and flavonoid constituents were higher in the methanol than the aqueous ethanol extract and has higher antioxidant activity. Altogether, post‐treatment effect of the extract on the testis was more effective than treatment‐related effect and the methanol extract appears to have better and consistent effects on the investigated parameters probably due to higher antioxidant activity conferred to it by its phenolic and flavonoid contents.     

Anti‐inflammatory effect of platelet‐rich plasma on nucleus pulposus cells with response of TNF‐α and IL‐1

The purpose of this study was to investigate the anti‐inflammatory effect of platelet‐rich plasma (PRP) with collagen matrix on human nucleus pulposus (NP) cell in response to pro‐inflammatory cytokines such as tumor necrosis factor‐alpha (TNF‐α) and interleukin‐1 (IL‐1). NP cells from human disks were cultured in a monolayer and maintained in the collagen matrix prior to the addition of recombinant human IL‐1 and TNF‐α. After applying IL‐1 and TNF‐α, PRP prepared by using a commercially available platelet concentration system was added. The response was investigated using real‐time PCR for mRNA expression of type II collagen, aggrecan, matrix metalloproteinase‐3 (MMP‐3), and cyclooxygenase‐2 (COX‐2). The combination of IL‐1β and TNF‐α led to decrease of matrix synthesis gene expression such as collagen type II and aggrecan and increase of the degradation gene expression of COX‐2 and MMP‐3, compared to the control. Consecutive PRP exposure significantly recovered the down‐regulated gene expression of collagen type II and aggrecan and significantly reduced the increased MMP‐3 and COX‐2 gene expression, compared to that of control groups with pro‐inflammatory cytokines. The administration of PRP with collagen matrix markedly suppressed cytokine‐induced pro‐inflammatory degrading enzymes and mediators in the NP cell. It also rescued gene expression concerning matrix synthesis, thereby stabilizing NP cell differentiation. © 2013 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 32:551–556, 2014.     

Carnosine prevents testicular oxidative stress and advanced glycation end product formation in D‐galactose‐induced aged rats

D‐Galactose is shown to mimic natural ageing in rodents by exacerbating oxidative stress and glycation. Steroid production and having a poor antioxidant system make testis vulnerable to galactose‐induced ageing. Antioxidation and antiglycating actions of carnosine may be intriguing for prevention of testicular ageing. In this study, male Wistar rats were applied D‐galactose (300 mg/kg; subcutaneously 5 days a week) and carnosine (250 mg/kg; intraperitoneally 5 days a week) along with D‐galactose for 2 months. D‐Galactose treatment increased testicular reactive oxygen species, thiobarbituric acid reactive substances, diene conjugates, protein carbonyls, advanced oxidation products of proteins and advanced glycation end products. Carnosine was capable of repelling oxidative stress and glycation produced by D‐galactose. Johnsen's score, which describes histopathological evaluation, was also significantly improved with preserved spermatogenesis by carnosine. It appears that carnosine deters the testicular oxidative stress due to galactose‐induced ageing directly by its antioxidative and antiglycating properties.     

Strontium fructose 1, 6‐diphosphate alleviate cyclophosphamide‐induced oligozoospermia by improving antioxidant and inhibiting testicular apoptosis via FAS/FASL pathway

This study investigated the treatment effects of a new compound, strontium fructose 1, 6‐diphosphate (FDP‐Sr), in cyclophosphamide (CP)‐induced oligozoospermia. FDP‐Sr, with extra high‐energy supply, could reverse male hypogonadism in the testis. Male Wistar rats were randomly divided into three groups: control group (vehicle treated), CP group and CP + FDP‐Sr group. Both CP group and CP + FDP‐Sr groups were orally administered CP (20 mg kg^?1) consecutively for the first 7 days to establish CP‐induced testicular toxic models. Subsequently, CP group was given orally distilled water per day, whereas CP + FDP‐Sr group was received FDP‐Sr (200 mg kg^?1) for 49 days. Compared to the CP group, the FDP‐Sr group showed significantly increased levels of serum testosterone, testis relative weights and epididymal sperm counts in rats. In addition, rats treated by FDP‐Sr showed the recuperative activities of testicular marker enzymes and normalised levels of antioxidants in tissue. Testicular protection of FDP‐Sr was further demonstrated by enhancing expression of P450scc, reducing ability of FAS/FASL and generating cytoprotection in the histopathological study. FDP‐Sr appeared to possess an ability to attenuate CP‐induced reproduction toxicity via the activation of antioxidants and steroidogenesis enzymes, and alleviate oligozoospermia via inhibition of testicular apoptosis by FAS/FASL pathway.     

Legg‐Calvé‐Perthes Disease Produces Chronic Hip Synovitis and Elevation of Interleukin‐6 in the Synovial Fluid

Legg‐Calvé‐Perthes disease (LCPD) is a childhood hip disorder of ischemic osteonecrosis of the femoral head. Hip joint synovitis is a common feature of LCPD, but the nature and pathophysiology of the synovitis remain unknown. The purpose of this study was to determine the chronicity of the synovitis and the inflammatory cytokines present in the synovial fluid at an active stage of LCPD. Serial MRI was performed on 28 patients. T2‐weighted and gadolinium‐enhanced MR images were used to assess synovial effusion and synovial enhancement (hyperemia) over time. A multiple‐cytokine assay was used to determine the levels of 27 inflammatory cytokines and related factors present in the synovial fluid from 13 patients. MRI analysis showed fold increases of 5.0 ± 3.3 and 3.1 ± 2.1 in the synovial fluid volume in the affected hip compared to the unaffected hip at the initial and the last follow‐up MRI, respectively. The mean duration between the initial and the last MRI was 17.7 ± 8.3 months. The volume of enhanced synovium on the contrast MRI was increased 16.5 ± 8.5 fold and 6.3 ± 5.6 fold in the affected hip compared to the unaffected hip at the initial MRI and the last follow‐up MRI, respectively. In the synovial fluid of the affected hips, IL‐6 protein levels were significantly increased (LCPD: 509 ± 519 pg/mL, non‐LCPD: 19 ± 22 pg/mL; p = 0.0005) on the multi‐cytokine assay. Interestingly, IL‐1β and TNF‐α levels were not elevated. In the active stage of LCPD, chronic hip synovitis and significant elevation of IL‐6 are produced in the synovial fluid. Further studies are warranted to investigate the role of IL‐6 on the pathophysiology of synovitis in LCPD and how it affects bone healing. © 2015 American Society for Bone and Mineral Research     

Co‐administration of caffeine and caffeic acid alters some key enzymes linked with reproductive function in male rats

This study assessed the effects of caffeine combined with caffeic acid on some biomarkers of male reproductive function using normal albino Wistar rats. Rats were divided into four groups (n = 6) and treated for seven successive days; group 1 represents the control rats; group 2 rats were treated with 50 mg/kg body weight (BW) of caffeine only; group 3 rats were treated with 50 mg/kg BW of caffeic acid, while the rats in group 4 were cotreated with an equal combination of caffeine and caffeic acid. The results revealed significant increase in reproductive hormone, testicular and epididymal nitric oxide levels of the rats. Moreover, decreased oxidative stress in the testes and epididymides of the treated rats was evidenced by significant increase in total and nonprotein thiol levels, catalase and superoxide dismutase activities. Similarly, decreased testicular cholesterol level with concomitant elevation in testicular steroidogenic enzyme activities, glycogen and zinc levels were observed in the treated rats. No morphological changes were observed as revealed by the photomicrographs from light microscopy in treated rats. Nevertheless, the combination therapy exhibited additive/synergistic effect on these biochemical indices than when they were administered singly. This study suggests the combination therapy of caffeine and caffeic acid at the dose tested for improving male reproductive function.     

Effects of different n6/n3 ratios and supplementation with DHA and EPA on the testicular histology and lipogenesis in streptozotocin‐treated rats

The aim of this study was to investigate the possible influence of different dietary n6/n3 ratios and DHA/EPA addition on the testis histology, antioxidative status and lipogenesis of streptozotocin‐treated rats. Diabetes was induced by a single intraperitoneal injection of streptozotocin (55 mg/kg). The rats were divided into five groups: CON (n6/n3 ratio, 7), CON‐DM1 (STZ; n6/n3 ratio, 7), N3‐DM1 (STZ; n6/n3 ratio, 1), N6‐DM1 (STZ; n6/n3 ratio, 60) and DHA‐DM1 (STZ; n6/n3 ratio, 1; added DHA/EPA). Antioxidative status was improved in the DHA‐DM1 group. Seminiferous tubule diameter, testicular pathohistological scoring and total lipid content were improved in the N6‐DM1 group compared to the other streptozotocin‐treated groups. Streptozotocin treatment induced changes in testis fatty acid profile depending on n6/n3 ratio. The fatty acid profile of N6‐DM1 group was characterised by similar or increased values for n6 fatty acids compared to the CON group, while the DHA‐DM1 group had the lowest content of n6 fatty acids. The content of n3 fatty acids was increased in the N3‐DM1 and DHA‐DM1 groups. These results suggest that a n6/n3 ratio could significantly influence testicular antioxidative status, histology and lipogenesis and that these effects vary depending on the supplemented fatty acid.     

Potential pathological role of pro‐inflammatory cytokines (IL‐6, TNF‐α, and IL‐17) in xenotransplantation

The major limitation of organ transplantation is the shortage of available organs from deceased human donors which leads to the deaths of thousands of patients each year. Xenotransplantation is considered to be an effective way to resolve the problem. Immune rejection and coagulation dysfunction are two major hurdles for the successful survival of pig xenografts in primate recipients. Pro‐inflammatory cytokines, such as IL‐6, TNF‐α, and IL‐17, play important roles in many diseases and in allotransplantation. However, the pathological roles of these pro‐inflammatory cytokines in xenotransplantation remain unclear. Here, we briefly review the signaling transduction and expression regulation of IL‐6, TNF‐α, and IL‐17 and evaluate their potential pathological roles in in vitro and in vivo models of xenotransplantation. We found that IL‐6, TNF‐α, and IL‐17 were induced in most in vitro or in vivo xenotransplantation model. Blockade of these cytokines using gene modification, antibody, or inhibitor had different effects in xenotransplantation. Inhibition of IL‐6 signaling with tocilizumab decreased CRP but did not increase xenograft survival. The one possible reason is that tocilizumab can not suppress IL‐6 signaling in porcine cells or organs. Other drugs which inhibit IL‐6 signaling need to be investigated in xenotransplantation model. Inhibition of TNF‐α was beneficial for the survival of xenografts in pig‐to‐mouse, rat, or NHP models. Blockade of IL‐17 using a neutralizing antibody also increased xenograft survival in several animal models. However, the role of IL‐17 in the pig‐to‐NHP xenotransplantation model remains unclear and needs to be further investigated. Moreover, blockade of TNF‐α and IL‐6 together has got a better effect in pig‐to‐baboon kidney xenotransplantation. Blockade two or even more cytokines together might get better effect in suppressing xenograft rejection. Better understanding the role of these cytokines in xenotransplantation will be beneficial for choosing better immunosuppressive strategy or producing genetic modification pig.     

Aerobic exercise blocks interleukin‐6 levels and germ cell apoptosis in obese rats

To investigate the effect of a high‐fat diet and aerobic exercise intervention and its related mechanism on rat germ cell apoptosis. Forty male Sprague‐Dawley rats were randomly divided into control group, high‐fat diet group, control exercise group and high‐fat exercise group. Rats were fed with high‐fat diet or were given weight‐free swimming. The levels of TG, TC, HDL, LDL and IL‐6 in serum of rats were measured. The body weight, body length and inguinal fat weight were measured to calculate the Lee's index and lipid/body weight ratio. The expression of IL‐6 mRNA in inguinal fat and IL‐6R,Bcl‐2 and Bax mRNA in testis was detected by RT‐PCR. The morphological structure of testis was observed, and the Johnsen's ten‐point score was calculated by HE staining, and the germ cell apoptosis was detected by TUNEL method. We got from the experimental results: a high‐fat diet induces obesity and lipid metabolism disorder, alters testis morphological structure and increases germ cell apoptosis in rats. Aerobic exercise improves the lipid metabolism disorder and interferes with germ cell apoptosis by reducing interleukin‐6 and interleukin‐6 receptor expression.     

Platelet rich plasma promotes skeletal muscle cell migration in association with up‐regulation of FAK,paxillin, and F‐Actin formation

Platelet rich plasma (PRP) contains various cytokines and growth factors which may be beneficial to the healing process of injured muscle. The aim of this study was to investigate the effect and molecular mechanism of PRP on migration of skeletal muscle cells. Skeletal muscle cells intrinsic to Sprague–Dawley rats were treated with PRP. The cell migration was evaluated by transwell filter migration assay and electric cell‐substrate impedance sensing. The spreading of cells was evaluated microscopically. The formation of filamentous actin (F‐actin) cytoskeleton was assessed by immunofluorescence staining. The protein expressions of paxillin and focal adhesion kinase (FAK) were assessed by Western blot analysis. Transfection of paxillin small‐interfering RNA (siRNAs) to muscle cells was performed to validate the role of paxillin in PRP‐mediated promotion of cell migration. Dose‐dependently PRP promotes migration of and spreading and muscle cells. Protein expressions of paxillin and FAK were up‐regulated dose‐dependently. F‐actin formation was also enhanced by PRP treatment. Furthermore, the knockdown of paxillin expression impaired the effect of PRP to promote cell migration. It was concluded that PRP promoting migration of muscle cells is associated with up‐regulation of proteins expression of paxillin and FAK as well as increasing F‐actin formation. © 2017 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 35:2506–2512, 2017.