Tubules Of Rats Research Articles

The Effect of Eucalyptus globulus Hydroalcoholic Extract on LH, FSH and Testosterone Concentrations and Sperm Morphology.

Scientists have been paying attention to the life-giving properties of medicinal plants for many years. Among these plants is the eucalyptus plant. This plant has various compounds such as cineole and terpenes. It also contains compounds such as flavonoids, aliphatic aldehydes, sesquiterpene, quinotanen, catechins, salts, and vitamins. In the present study, the hydroalcoholic extract of Eucalyptus leaves with concentrations of 175, 350, and 700 mg/kg body weight, and spermatogenesis were studied in 40 adult Wistar rats in five groups of eight. Adult male mice received the extract at the above concentrations by gavage for 28 days. Control mice received only solvent and water, while control mice received no substance other than municipal tap water and normal food. After the last administration of the drug, the animals were weighed and anesthetized, and then blood samples were taken from their hearts. Concentrations of LH, FSH, and testosterone were measured by an ELISA kit. The results showed that body weight and testis, seminiferous tube diameter, Leydig cell diameter, epithelium thickness, number of Leydig cells, spermatogonium, spermatocytes, spermatids, sperm, and testosterone concentration increased significantly with the group. But no significant difference was observed in the concentration of FSH and LH hormones or the number of Sertoli cells. Therefore, it can be concluded that eucalyptus leaf extract may increase the proliferation of sex cells in the seminiferous tubules of rats.

Effects of Javanese Ginseng Root Extracts (Talinum triangulare W.) on Thickness of Seminiferous Tubules of Male Rats (Rattus norvegicus) Exposed by Cigarette Smoke

Highlights:1. Infertility in men can be caused by smoking, the content of free radicals in cigarettes can affect male reproductive organs.2. Javanese ginseng root (Talinum triangulare w.) extract contains ginsenoside which is a natural antioxidant to reduce free radical levels inside the body.3. Javanese ginseng root (Talinum triangulare w.) extract was able to reduce free radical levels as seen from the appearance of the seminiferous tubules of male white rats exposed to cigarette smoke. AbstractIntroduction: Cigarette smoke contains various radical chemical compounds that can increase oxidative stress, which may cause lipid peroxidation in the body and affect various different organs. This study aimed to examine the effects of Javanese ginseng root extracts (Talinum triangulare W.) on the thickness of seminiferous tubules of white male rats (Rattus norvegicus) exposed to cigarette smoke.Methods: This was an experimental study with a randomized controlled method and a post-test control group design. Post-test control group design with 15 white male rats was divided into five groups: a normal group, a control group (exposed to cigarette smoke), and three treatment groups (exposed to cigarette smoke and Javanese ginseng root extract with doses of 12.5mg, 25mg, and 50mg/200grBW, respectively) for 30 days. The seminiferous tubule thickness was observed using H&E staining under a light microscope with 200x magnification in five fields view.Results: One-way ANOVA showed that Javanese ginseng root extracts significantly affected seminiferous tubule thickness (p = 0.000). With a dose of 12.5mg/200mgBW, Javanese ginseng root extract started to show a significant effect (p = 0.000) on the thickness of the epithelium of the seminiferous tubules. The group exposed to cigarette smoke had an average thickness of seminiferous tubules of 7.92 mm, the normal group had an average thickness of 9.13 mm, and treatment groups with doses 12.5mg, 25mg, and 50mg/200gBW had an average thickness of 11.85 mm, 12.7mm, and 14.69 mm, respectively.Conclusion: Javanese ginseng root extract increased seminiferous tubule thickness in white male rats exposed to cigarette smoke.

Ntrk1 promotes mesangial cell proliferation and inflammation in rat glomerulonephritis model by activating the STAT3 and p38/ERK MAPK signaling pathways

BackgroundMesangial proliferative glomerulonephritis (MsPGN) accounts for a main cause of chronic kidney disease (CKD), chronic renal failure and uremia. This paper aimed to examine the effect of Ntrk1 on MsPGN development, so as to identify a novel therapeutic target for MsPGN.MethodsThe MsPGN rat model was constructed by single injection of Thy1.1 monoclonal antibody via the tail vein. Additionally, the Ntrk1 knockdown rat model was established by injection of Ntrk1-RNAi lentivirus via the tail vein. Periodic acid-schiff staining and immunohistochemistry (IHC) were performed on kidney tissues. Moreover, the rat urinary protein was detected. Mesangial cells were transfected and treated with p38 inhibitor (SB202190) and ERK inhibitor (PD98059). Meanwhile, the viability and proliferation of mesangial cells were analyzed by cell counting kit-8 (CCK-8) and 5-Ethynyl-2′-deoxyuridine assays. Gene expression was detected by quantitative real-time polymerase chain reaction (qRT-PCR) and Western-blot (WB) assays.ResultsThe proliferation of mesangial cells was enhanced in glomerulus and Ki67 expression was up-regulated in renal tubule of MsPGN rats. The urine protein level increased in MsPGN rats. Pro-inflammatory factors and Ntrk1 expression were up-regulated in glomerulus of MsPGN rats. Ntrk1 up-regulation promoted the viability, proliferation, expression of pro-inflammatory factors and activation of the STAT3, p38 and ERK signaling pathways in mesangial cells. Ntrk1 knockdown reduced mesangial cell proliferation, urine protein, pro-inflammatory factors, activation of STAT3, p38 and ERK signaling pathways in glomerulus, and decreased Ki67 expression in renal tubule of MsPGN rats. Treatment with SB202190 and PD98059 reversed the effect of Ntrk1 on promoting the viability, proliferation and inflammatory response of mesangial cells.ConclusionNtrk1 promoted mesangial cell proliferation and inflammation in MsPGN rats by activating the STAT3 and p38/ERK MAPK signaling pathways.

Effect of bioactive glasses used as dentin desensitizers on the dentin-pulp complex in rats.

Bioactive glasses have been recommended for the occlusion of dentinal tubules in treating cervical dentin hypersensitivity. This study evaluates an in vivo model of dentin exposure, and tests the efficacy of bioglass treatments. Thirty male Wistar rats received gingival recession surgery on the upper left first molar. The treatments were applied over the surface of the exposed dentin every 4 days for 28 days. The groups were as follows: Naive; Gingival recession; Cavity varnish; Biosilicate®; Strontium bioglass; and Potassium bioglass. Changes in the dentin-pulp complex, and the presence of substance P, were evaluated through hematoxylin-eosin and immunohistochemical staining. The groups had similar results. Teeth with exposed dentinal tubules in rats showed a typical pattern in the dentin-pulp complex and immunotracing for substance P. The materials did not cause pulp damage. The effects of gingival recession and open dentinal tubules on pulp tissue require further clarification.

Treatment of Male Albino Rats with 5-fluorouracil and/or Cyclophosphamide Caused Serious Alterations in Sexual Hormones, Histological Structure of the Testis, and Semen Quality

Background: The therapeutic effect of Cyclophosphamide (CPA) is thus attributed to phosphoramide mustard and acrolein that leads to the formation of high level of reactive oxygen species (ROS), which results in decreased antioxidant activity. Excessive production of ROS could also culminate in oxidative stress. Objectives: The aim of this study is to evaluate the effect of sub lethal dose of the cyclophosphamide, 5-FU combination of 5-FU and CPA on testicular function, and histopathological alterations in male albino rats. These functions were monitored through some selected biochemical parameters and confirmed by DNA, RNA concentration and histopathological examination. Materials and Methods: Twenty-eight male adult rats were grouped randomly into four groups (n=7 each group), Five for biochemical studies and 2 for histological studies. Group I (control): Rats were injected with saline intraperitoneally and at a dose of 1.0 ml/kg b.w. for 14 days. Group II cyclophosphamide (CPA): Cyclophosphamide at a dose of 10 mg/kg day by day through i.p. to rats for 14 days. Group III Fluorouracil (5-FU): 5-Fluorouracil at a dose of 10 mg/kg day by day in saline was given through i.p. to rats for 14 days. Group IV (CPA+5-FU): Rats were given CPA followed by 5-FU at a dose of 10 mg/kg per day (day by day) through i.p. to rats for 14 days. At the end of the experimental period, rats were anesthetized using light ether. Blood and testes tissue samples were taken and prepared for biochemical and histological measurements. Biochemical parameters in rat serum and tissues were evaluated. Results: Treatment of male rats with CPA, 5-FU and their combination caused a significant decrease in sperm count, motility while increased dead and abnormal sperms compared to those of control. Co-treatment of CPA and 5-FU caused a significant decrease in sperm count, sperm motility and increase in abnormal sperms compared to individual treatment of CPA and 5-FU. Levels of testosterone, luteinizing hormone (LH) and follicle stimulating hormone (FSH) were decreased in the CPA, 5-FU and their combination-treated group. Co-treatment of 5-FU and CPA synergistically reduced testicular hormones compared with individual chemotherapy. Cyclophosphamide, 5-FU and their combination decreased testis DNA and RNA concentrations compared to the control. The combination of CPA and 5-FU was more effective to DNA and RNA compared to individual administration of the chemotherapy. Epididymis of the control rats showed normal histological structure with normal sperm density. Large tubules lined by pseudostratified columnar epithelium with abundant stereocilia projection from their surface. Long and dense processes of sperm tails were observed within the Lumina. The epididymis tubules of CPA-exposed rats were smaller and the majorities of them were free from mature spermatozoa. Epithelium contained numerous numbers of sloughed cells with cell debris in Lumina. 5-FU treated rat showed atrophy as well as necrosis, the arrangement of germ cells was disordered. The number of cell layers of the seminiferous tubules was significantly reduced. Epididymis of CPA+5-FU-treared rats showed necrotic degeneration and atrophy of the germinal cells, decrease spermatid when compared to CPA. Conclusion: It could be concluded that treatment of mammals with chemotherapy is associated with the production of free radicals that lead to hazardous alterations in sexual hormones, DNA, and RNA concentration, and histopathological structure in tistes and semin quality of male albino rats. However, 5-FU and CPA combination could produce a serious alteration in these parameters. Future work should consider combined chemotherapy regimens. Toxicological studies must be performed before using drugs as combination before application. Further research is required on toxicological impacts of drugs and pollutants mixtures.

High-salt diet accelerates bone loss accompanied by activation of ion channels related to kidney and bone tissue in ovariectomized rats.

Excessive salt intake can induce a variety of diseases, such as hypertension, cardiovascular disease, kidney disease and so on，it is also one of the factors promoting bone resorption. The mechanism of osteoporosis-induced exacerbations of high salt diet is not well-defined. In this study, we used ovariectomized 6-month-old Sprague Dawley rats to construct a high bone turnover model, and then administrated with high sodium chloride diet (2.0% w/w NaCl, 8.0% w/w NaCl) for 12 weeks to observe the effect of high salt diet on bone metabolism. The results showed that high salt diet could lead to the destruction of bone microstructure, promote the excretion of urinary calcium and phosphorus and accelerate the bone turnover, as well as cause the pathologic structural abnormalities in renal tubular. At the same time, it was accompanied by the up-regulated expression of the epithelial sodium channel (ENaCα), voltage-gated chloride channels (ClC)-3 and the down-regulated expression of Na-Cl cotransporter (NCC), sodium calcium exchanger (NCX1) in femoral tissue and renal tubules. These findings confirm that high salt diet can destroy the microstructure of bone by increasing bone resorption and affect some ion channels of bone tissue and renal tubule in ovariectomized rats.

A novel decellularized nerve graft for repairing peripheral nerve long gap injury in the rat

Decellularized nerve allografts are an alternative to autograft for repairing severe nerve injuries, since they have higher availability and do not induce rejection. In this study, we have assessed the regenerative potential of a novel decellularization protocol for human and rat nerves for repairing nerve resections, compared to the gold standard autograft. A 15-mm gap in the sciatic nerve was repaired with decellularized rat allograft (DC-RA), decellularized human xenograft (DC-HX), or fresh autograft (AG). Electrophysiology tests were performed monthly to evaluate muscle reinnervation, whereas histological and immunohistochemical analyses of the grafts were evaluated at 4 months. A short-term study was also performed to compare the differences between the two decellularized grafts (DC-RA and DC-HX) in early phases of regeneration. The decellularization process eliminated cellularity while preserving the ECM and endoneurial tubules of both rat and human nerves. Higher amount of reinnervation was observed in the AG group compared to the DC-RA group, while only half of the animals of the DC-HX showed distal muscle reinnervation. The density of myelinated axons was significantly higher in AG compared to both DC grafts, being this density significantly higher in DC-RA than in DC-HX. At short term, fibroblasts repopulated the DC-RA graft, supporting regenerated axons, whereas an important fibrotic reaction was observed around DC-HX grafts. In conclusion, the decellularized allograft sustained regeneration through a long gap in the rat although at a slower rate compared to the ideal autograft, whereas regeneration was limited or even failed when using a decellularized xenograft.

Amelioration of Carboplatin-Induced Changes in Rat Renal Cortical Tubulesby Alpha-Tocopherol

Objective: To determine the effects of carboplatin and alpha-tocopherol co-administration on gross kidney features and renal cortical tubules of carboplatin-treated rats.
 Study Design: Laboratory-based experimental study.
 Place and Duration of Study: Department of Anatomy, Army Medical College/National University of Medical Sciences Rawalpindi Pakistan, in collaboration with the National Institute of Health (NIH) and Pathology Lab Pak Emirates Rawalpindi Pakistan, from Jan to Dec, 2021.
 Methodology: Thirty adults "Sprague-Dawley rats" of both genders were used and were divided into three Groups. Group-A was the Control Group. Groups B and C were given injection carboplatin 2.5 mg/kg body weight intraperitoneally on the first day. Group-C was given vitamin E at a dose of 62.7 mg/kg body weight daily via oral gavage starting from the second day of the experiment till the end of 12 weeks. Rats were euthanized at the end, kidneys were dissected, and they were preserved in 10% formalin after gross inspection. Tissue processing and staining were performed. A microscopic study was done to observe proximal and distal tubule necrosis.
 Results: Thirty adult Sprague Dawley rats, of age 10-12 weeks, and weight 250.0±50.0 grams, were included, there was statistically significant difference in the animal weight difference (p=0.001), length of right kidneys (cm) (p=0.002), width of right kidney (cm) (p= 0.001) and weight of right kidney (gm) (p=0.012) kidneys and relative tissue body weight index (RTBWI) (p = 0.048) of Control Group-A and Experimental Group B and C.
 Conclusion: Alpha-tocopherol has an ameliorative effect on gross features of rat kidneys..........

Haematological and hepatho-renal effect of cashew nut oil on male Wistar rats

The aim of this study was to evaluate the biosafety of Anacardium occidentale (cashew) nut oil using male Wistar rats as model. Cashew nut oil was extracted from cashew nuts with the aid of a Soxhlet apparatus using n-hexane as solvent. Twelve male Wistar rats were randomly assigned to four groups labeled I, II, III and IV (3 rats per group). Group I served as the control group and were administered distilled water. Group II received 1.53 mL/kg of the nut oil; group III received 4 mL/kg of the nut oil while group IV received 8.13 mL/kg of the nut oil. The administrations were via the oral route. Blood samples were collected weekly for haematological examination. At the end of four weeks study period, the rats were anaesthetized and the liver and kidneys were removed for histological examination. The haematological result showed slight deviation from the normal in packed cell volume (52%), total white blood cell count (9.38 x 103/mm3) and lymphocyte count (58.17%) in Wistar rats administered 4 to 8.13 mL/kg of the cashew nut oil, respectively yet no visible compromise of the immune system was observed. Histological analysis; however, revealed some hepatocyte morphological lesions including mild microvesicular steatosis, kupffer cell hyperplasia, shrinkage of glomeruli, glomerular distortion, necrotic renal tubules, severe congestion and atrophy of renal tubules in rats fed 8.13mL/kg of the nut oil. In conclusion, the cashew nut oil was responsible for hepatic parenchymal and matrix alteration in wistar rats administered 8.13 mL/kg of cashew nut oil.

The possible protective role of vitamin E against deferasirox-induced injury of renal cortical tubules in adult male albino rat: A histological and immunohistochemical study

The possible protective role of vitamin E against deferasirox-induced injury of renal cortical tubules in adult male albino rat: A histological and immunohistochemical study

Lycopene modulates hepatic toxicity and testicular injury induced by etoposide in male rats

Lycopene (C40H56), a carotenoid found in red colour fruits, is known as a powerful antioxidant that protects cells from damage caused by reactive oxygen species (ROS). Etoposide inhibits topoisomerase II activity and restricts the development of cancer cells, though it establishes oxidative stress. To study the effect of lycopene (Ly) against hepatotoxicity and testis injury induced by etoposide in male rats. Forty male Wister albino rats. The experiment lasted for seven consecutive weeks including one week as acclimatization time. The experiment was in a completely randomized design with a 2×2 factorial arrangement. The animals were grouped as follow: No etoposide injection and no lycopene (control), lycopene supplementation (LY), etoposide injection (ET), and rats with etoposide injection and lycopene supplement (ET+LY). At the end of the experiment, rats were sacrificed by cervical dislocation. Blood samples were harvested and analyzed for serum alkaline phosphatase (ALP), alanine aminotransferase (ALT), aspartate aminotransferase (AST), triglyceride (TG), low-density lipoprotein-Cholesterol (LDL-C), high-density lipoprotein-cholesterol (HDL-C), total cholesterol (TC), Total Protein (TP), glucose (GLU) and testosterone. The left testis was manipulated for histological examination. The result of experiment showed that rats with etoposide injection had higher ALT, AST, and ALP than the control rats. In contrast co-treated rats (ET+LY) significantly modulated the levels of the hepatic parameters. Administration of lycopene increased testosterone concentration and germinal epithelium of seminiferous tubules in testes rats. Lycopene might be a promising agent with hepatoprotective effect in restoring testis injury induced by etoposide in rats.

Changes in Expression of the Connective Tissue Growth Factor in the Kidneys of Diabetic Rats During Aging

Background: Diabetic nephropathy (DN) is the most common cause of chronic renal failure in the world. There appears to be different pathogenetic, hemodynamic and metabolic processes leading to the pathologic mechanisms in DN. We studied the expression of connective tissue growth factor (CTGF) in the kidneys of diabetic rats over time.Methods: Male Sprague-Dawley rats were injected with 55mg/kg streptozotocin (STZ) (diabetes mellitus (DM) group) or with citrate buffer (control group). The rat kidneys were harvested 2 weeks, 2 months, 6 months and 12 months following the STZ challenge, and sectioned into three different kidney structures: glomeruli, proximal convoluted tubule (PCT) and distal convoluted tubule (DCT). Sections were immunohistochemically stained to monitor the expression of CTGF. Results:Significant differences in CTGF expression were observed 2 weeks and 2 months after the STZ challenge, with higher CTGF expression in diabetic rats. Two weeks post DM onset, high CTGF expression was demonstrated in the distal tubules of the diabetic rats. The expression of CTGF in the glomeruli of diabetic rats washighest 12 months after diabetes induction, with a complete absence of CTGF expression in the control groups throughout the study.Conclusions:The major change in expression of CTGF occurs within the first 2 months of DM, particularly in the DTC, implying an early onset of pathophysiological changes in diabetic kidneys, which would normally occur with aging. These findings help to contribute to our understanding of changes associated with DN and guide towards potentially appropriate treatment modalities.Key words: diabetes mellitus (DM), nephropathy, chronic renal failure, connective tissue growth factor (CTGF)

Changes in Expression of the Vascular Endothelial Growth Factor in the Kidneys of Diabetic Rats During Aging

Background. Diabetic nephropathy is the leading cause of end-stage renal disease (ESRD) in most developed countries. A number of specific factors may contribute to the onset and progression of diabetic nephropathy. One of those specific factors is vascular endothelial growth factor (VEGF). We studied the expression of VEGF in the kidneys of diabetic rats during aging. Methods. Male Sprague-Dawley rats were injected with 55mg/kg streptozotocin (STZ) (DM group) or with citrate buffer (control group). Kidneys were collected after 2 weeks, 2 months, 6 months and 12 months, and were analyzed in three different kidney structures: glomeruli, proximal (PCT) and distal convoluted tubules (DCT). Sections were stained immunohistochemically, using VEGF.Results. Significant differences in marker expression were observed after 2 months, with higher VEGF expression in diabetic rats. Positive VEGF cells were detected exclusively in the glomeruli of diabetic rats after 2 months. After 2 weeks of DM onset, high VEGF expression was demonstrated in the distal tubules of diabetic rats, as well as a complete absence of VEGF expression in the distal tubules of the control group. Conclusions. The major change in expression of VEGF occurs after 2 months of DM onset, particularly in the DTC, implying an early onset of pathophysiological changes in diabetic kidneys, which would usually occur with aging. These findings help to contribute to our understanding of changes associated with DN and guide towards potentially appropriate treatment modalities.Key words: diabetes mellitus, kidneys, nephropathy, VEGF

A pilot dynamic analysis of formative factors of nephrolithiasis related to metabolic syndrome: evidence in a rat model

Introduction and objective To examine the dynamic changes in the formative factors of nephrolithiasis and the final micromorphological changes in an obesity-initiated metabolic syndrome (MS) rat model. Methods Forty five-week-old male Sprague–Dawley (SD) rats were randomly divided into four groups: the regular diet group (RD), high-fat diet group (HFD), regular diet with drug (ethylene glycol and ammonium chloride) group (RDD), and high-fat diet with drug group (HFDD). A dynamic assessment of MS components (body weight (BW), body length (BL), Lee’s index (LI), blood glucose (BG), total cholesterol (TC), and triglycerides (TGs)) and stone-forming factors (urinary pH, urinary calcium, and urinary oxalate acid) was carried out. In addition, the levels of oxidative stress (OS) markers (CAT, SOD, TAC, GSH-PX, and MDA) were measured, and histological analysis was carried out at the end of 16 weeks. Results MS-related parameters, such as BW, LI, BG, TC, and TG, were significantly higher in HFD-fed rats than in RD-fed rats (p < 0.001). In the HFDD group, significantly lower urinary pH, hyperoxaluria, and hypocalciuria were noted in the dynamic assessment of stone-forming factors (p < 0.001). CAT, TAC, and MDA were notably changed in the HFD-fed groups, particularly the HFDD rats. Histological analysis showed that the renal tubules of HFDD rats had the highest scores for both inflammation and renal crystallization deposition (p < 0.05). Conclusions Our results suggest that male SD rats with MS are prone to developing nephrolithiasis. Validation in an in vivo model may lead to an understanding of the underlying pathophysiological mechanisms of action of MS-related nephrolithiasis in humans. Key messages Male SD rats with metabolic syndrome are more prone to developing calcium oxalate nephrolithiasis after treatment with ethylene glycol and ammonium chloride compared to control lean rats. MS-related nephrolithiasis in rats induced by ethylene glycol and ammonium chloride is mainly related to increased hyperoxaluria and inflammation and decreased antioxidant levels. High-fat diet-fed SD rats treated with ethylene glycol and ammonium chloride are a stable and valid in vivo model for understanding the potential mechanism of action of MS-related nephrolithiasis.

Investigation of Methamphetamine Effects on the Tissue Structure of the Main Gonadal in Rats

Introduction: Methamphetamine is a stimulant of the central nervous system that is usually misused. This study aimed to determine the effects of methamphetamine on the diameter and thickness of seminiferous tubules of adult male rats’ main gonad. Method: Twenty adult male rats weighted 200 to 220 gr were used in the current study. They were divided into study and control groups, each consisting of 10 rats. The study group received 2 mg per body weight of methamphetamine in the first three days and 5 mg in the next four days in peritoneum. The control group, on the other hand, received 0.5 ml per rat of physiologic serum in the the peritoneum. Twenty-four hours after the last injection, the rats were killed, and the main gonad was taken out of the scrotum. After tissue passage and dying with hematoxylin and eosin, the main gonad was observed with an optic microscope, and the results were analyzed using an independent t-test. A value of P&lt;0.05 was considered to determine the level of significance between the two groups. Results: The findings showed that using methamphetamine significantly decreases the average diameter and thickness of the epithelium of seminiferous tubules of rats. It was also found that the weight of rats and gonads decreased, but it was not statistically significant. Conclusion: The results of this study show that methamphetamine is a poisonous compound for the main gonad of male rats due to the degeneration and atrophy of seminiferous tubules

Influence of vitamin D on the histostructure of the testis and morphometric indications of spermatogenesis of intact rats

One of the current problems is the study of the effects of vitamin D on the body, and in men its action is closely related to the pathogenesis of androgen deficiency and hypofertility. Particular attention needs to be paid to determining whether cholecalciferol (D3) has a negative effect on the gonads and spermatogenesis of intact individuals, as vitamin D therapy is used in reproductive disorders with or without vitamin D deficiency. The aim of the study was to determine the effect of vitamin D on the histological structure of gonads and morphometric parameters of spermatogenesis of adult intact male rats. The studies were performed on adult sexually active male Wistar rats. Vitamin D3 was administered orally in doses of 1000 IU, 4000 IU and 10000 IU. The solutions were made on seed oil. The control was intact rats. Vitamin D and its solvent were administered throughout the period of spermatogenesis and the time of maturation of sperm in the epididymis, after which the structural organization of the testes was determined. Gonadal samples were fixed in 10 % formalin solution, leave in alcohols of increasing strength, and embedded in paraffin. In addition to survey microscopy, morphometric evaluation of the process of spermatogenesis was performed on sections of gonads stained with hematoxylin and eosin. Micropreparations were examined using a Granum L 30 (03) light microscope, and microscopic images were taken with a Granum DSM 310 digital video camera. Photographs were processed on a Pentium 2.4 GHz computer using Toup View. Statistical processing of data results was performed in the standard software package "Statistica 6.0" using Student's t-test and using a non-parametric analogue of one-way analysis of variance – Kruskal-Wallis criterion. In rat testicular sections, seminiferous tubules were located transversely or obliquely and were oval or round in shape. The diameter of the tubules is normal, the tubular membrane, as well as the protein and vascular membranes were normal. The basal department contains the youngest cells of the germinal epithelium – spermatogonia. Cells have a pronounced functional activity. Morphometric characteristics of spermatogenesis of intact rats corresponded to the physiological norm. The introduction of the solvent throughout the period of spermatogenesis and maturation of mature sperm in the epididymis did not affect the histoarchitectonics of the testicles. The testicular lobes are filled with concentric or flattened profiles of sections of the seminal tubules, which are close enough to each other. The diameter of the tubules is normal, the intrinsic membrane of the tubules, as well as the protein and vascular membranes corresponded to those in intact animals. 3-4 generations of spermatogenic cells, which were at different stages of development, can be seen in the tubules. However, few tubules with focal destruction of the germinal epithelium and exfoliation of germ cells in the lumen of the tubules have been observed. No significant changes in the microstructure of the seminal tubules were observed after administration of vitamin D at all doses studied. Not only spermatogenesis but also spermiogenesis is clearly traced in different tubules of rats – stages of cellular transformations from spermatid to sperm. Morphometric parameters of the process of spermatogenesis of rats receiving different doses of vitamin D3 do not differ from those of intact rats. Thus, the use of vitamin D in these doses revealed the safety of its effect on the number of spermatogonia and tubules with stage 12 meiosis. When cholecaciferol was used in male rats for 68 days, the spermatogenesis index remained at the level of animals that did not receive the test substance. The use of vitamin D3 in intact adult male rats does not adversely affect the histological structure of the testes.

A comparative study of the effects of 3 testicular toxicants in cultures of seminiferous tubules of rats or of domestic cats (veterinary waste): An alternative method for reprotoxicology

In response to the EU cosmetics directive regulation and REACH legislation which encourage cell culture methods in order to reduce or replace the use of animals in toxicology studies, we settled the culture of prepubertal domestic cat seminiferous tubules in our validated BioAlter® model, usually used with prepubertal rat, called here BioAlter®-rat, by opposition to BioAlter®-cat settled here. We carried out a comparative study on the effects of 3 testicular toxicants, 1,3-dinitrobenzene at 60 μM, 2-methoxyacetic acid at 2.5 mM and carbendazim at 50 nM or 500 nM in both BioAlter®-cat and BioAlter®-rat over a 3-week culture period. Sertoli cell or each germ cell populations as well as the levels of Sertoli cell or germ cell specific mRNAs were studied. The harmful effects of the 3 toxicants on pre-meiotic, meiotic and post-meiotic cell numbers and on Sertoli or germ cell specific mRNAs were clearly observed in the two species, even if there might be some small differences in the intensity of the effects on some of the studied parameters. Hence, BioAlter®-cat might be a solution to the requirements of the EU cosmetics directive and REACH legislation for male reproductive toxicology studies.

1H‑NMR‑based metabolic profiling of rat urine to assess the toxicity‑attenuating effect of the sweat‑soaking method on Radix Wikstroemia indica

Radix Wikstroemia indica (L.) C.A. Mey. (RWI) is a toxic medicinal species primarily present in the Miao area of China. The toxicity of RWI is effectively reduced whilst maintaining the therapeutic effect when processed using the ‘sweat-soaking method’, which is a common method of Traditional Chinese Medicine preparation. However, there is a lack of scientific and medical evidence to explain the potential mechanisms by which the toxicity of RWI is reduced after preparation using this method, and the endogenous systemic metabolic effect of RWI remains uncertain. The aim of the present study was to explore the endogetnous metabolic alterations caused by RWI and to examine the possibility of reducing the toxicity of RWI using the sweat-soaking method using proton nuclear magnetic resonance (NMR) metabolomic analysis in rats. Principal Component Analysis, Partial Least Squares-Discriminant Analysis (PLS-DA) and Orthogonal PLS-DA were used to assess individual proton NMR spectra. A total of 34 metabolic products were altered after delivering raw RWI, and 32 endogenous metabolites were induced by processed RWI. The metabolic pathways that lead to a significant impact on energy and carbohydrate, amino acid, organic acids and lipid metabolism following raw and processed RWI use were identified. The mitochondria of hepatic and renal tubules of rats were injured in the raw RWI group, whereas the processed product reduced or interfered with energy substrate, carbohydrate and amino acid metabolism, whilst reducing the levels of metabolic markers of hepatotoxicity and nephrotoxicity, without causing damage to the mitochondria. Our previous study showed that the median lethal dose (LD50) value of raw RWI was 4.05 g/kg in rats after oral administration; however, the LD50 value of the processed RWI could not be measured. The maximum tolerated dose and minimum lethal dose were 20 and 30 g/kg for the processed RWI, respectively, corresponding to 109 and 164 times the clinical daily dose (0.029 g/kg). Thus, the sweat-soaking method reduced the toxicity of RWI. Moreover, after processing, the toxic component YH-10 was converted into a YH-10 + OH compound, reducing the content of the toxic YH-10 by 48%, whilst also reducing the contents of the toxic components YH-12 and YH-15 by 44 and 65%, respectively. In conclusion, the present study showed that the sweat-soaking method reduced the toxicity of RWI, as evidenced by the reduction of the levels of metabolic markers and the activity of metabolic pathways, thus providing a basis for processing of RWI for clinical use.

Evaluation of orellanine-induced toxicity from the mushroom Cortinarius orellanus and the antagonistic effect of Petroselinum crispum

Mushroom poisoning is a worldwide public health problem that may cause serious toxic consequences on renal functions. The study aimed to evaluate the acute toxicity (24 h) of orellanine (OR) from Cortinarius orellanus in rat kidney and the ameliorative effect of parsley ethanolic extract. Twelve adult male Wistar rats were used to determine intraperitoneal (ip) median lethal dose (LD50) of OR, and 32 rats were divided into 4 groups (n = 8): OR group had 500 mg OR per kg bwt; OR + parsley group had the same dose of OR and after 1 h had 500 mg/kg parsley orally; parsley group had parsley only; and control had the vehicle 0.1% DMSO. Blood and kidney samples were collected at Hour 48. The LD50 dose was 1430 mg/kg for an observation period of 24 h. There were significant reductions (p < 0.01) in the body weight, and relative kidney weight of intoxicated rats compared to parsley treated rats and to controls. Similarly, this group had significantly higher levels of creatinine (p < 0.001), uric acid and urea (p < 0.05). The antioxidant glutathione peroxidase activity was significantly reduced (p < 0.01), while Cystatin C serum levels were significantly higher (p < 0.001) in the intoxicated untreated rats compared to all groups. Histopathological examination indicated necrotic damage in glomeruli and proximal tubules of rats given OR, which was relieved by parsley extract. Overall, the study showed that parsley extract ameliorated OR-induced kidney toxicity. This could be utilized in future research on adjunct therapy for toxicity-induced renal injury.

Moringa oleifera leaf extract restored the diameter and epithelium thickness of the seminiferous tubules of rat (Rattus norvegicus) injected with gentamicin

This research aims to determine the effect of Moringa oleifera leaf extract (MLE) on the diameter and epithelium thickness of the seminiferous tubules of rats induced with gentamicin. This study used 25 male rats (Rattus norvegicus) aged 8-12 weeks with 150-200 grams body weight. Negative control group (C-) was injected with aquadest and given 0.5% Na-CMC orally, positive control group (C+) was induced with gentamicin injection of 5 mg/kg BW/day and given 0.5% Na-CMC orally, while groups T1, T2, and T3 were induced with gentamicin at the same dose and treated orally with MLE of 200, 316, 500 mg/kg BW/day. Analysis of variance showed significant differences and further analysis using Duncan Multiple Range Test showed difference in each group. The results showed that treatment with MLE of 500 mg/kg BW/day improved the diameter and epithelium thickness of the seminiferous tubules of rats injected with gentamicin. It could be concluded that MLE restored the diameter and epithelium thickness of the seminiferous tubules of rats induced with gentamicin.