Effect of Sempervivum tectorum Extract on Some Biomarkers of Reproductive Function and Levels of Some Trace Elements in Male Rats Exposed to Aluminum.

Aluminum, a contentious trace element found in the environment, has been demonstrated to have harmful effects on both humans and animals. In contrast, Sempervivum tectorum, an evergreen plant, has been found to offer numerous beneficial effects for both humans and animals. Therefore, this study aims to assess the protective effect of S. tectorum on certain reproductive biomarkers in male rats exposed to aluminum. Thirty-five Wistar rats were randomly divided into five groups: NTC (no-treatment control) received distilled water; NC (negative control) received drinking water containing 1 mg/L aluminum sulfate (AS); E1 received 1 mg/L AS along with an 8% S. tectorum extract; PC (positive control) received only 8% S. tectorum extract; E2 received 1 mg/L AS for three months followed by 8% extract for one month. The study analyzed testosterone, LH, FSH, body weight, and the histological structure of the testis, epididymis, and prostate, as well as the levels of zinc, manganese, copper, and iron in these organs. Significant decreases in body weight, testis, and epididymis size were observed in the aluminum-exposed groups compared to the control, whereas these decreases were not significant in the S. tectorum-treated groups compared to the control. Aluminum exposure led to significant decreases in testosterone and LH levels, with FSH levels showing a nonsignificant decrease in males, which were mitigated significantly by the administration of the plant extract. Histological analysis revealed alterations in the testis, epididymis, and prostate of the AS-exposed groups, including necrosis of seminiferous tubule epithelium and Leydig cells in the testis, and basal epithelial necrosis in the epididymis and prostate. Aluminum levels increased in all organs studied, while levels of zinc, copper, iron, and manganese decreased, showing a negative and significant correlation with aluminum levels. The aqueous extract of S. tectorum demonstrated a protective effect on certain studied biomarkers in male rats affected by aluminum exposure.

The antioxidant effect of Aronia melanocarpa extract in rats oxidative stress induced by cisplatin administration.

BACKGROUND: The reactive oxygen species generated by numerous xenobiotic substances has as consequences the impairment of different organs normal function. Many plants pose antioxidant activity to counteract oxidative stress, among them being the chokeberry (Aronia melanocarpa). The purpose of present study was to determine if the use of A. melanocarpa extract can counteract the oxidative stress induced by cisplatin administration in rats. MATERIAL AND METHODS: The study was made on forty Wistar rats divided in four groups as follows: C (control): receiving i.p. 1 mL of saline solution; E1: receiving cisplatin 20 mg/kg bw, i.p.; E2: receiving cisplatin 20 mg/kg bw, i.p and A. melanocarpa berry 6 % aqueous extract as drinking water, and CB (control blank): i.p 1 mL saline solution and A. melanocarpa 6 % aqueous extract for four weeks. RESULTS: Administration of Cisplatin was followed by the increase of serum superoxide dismutase (+21.18 %, P < 0.05), catalase (+25.44 %, P < 0.001), glutathione peroxidase (+17.88 %, P < 0.05) and thiobarbituric reactive substances (+28.17 %, P < 0.01) but significantly decreased glutathione reductase (-22.35 %, P < 0.001) level comparative to control, pointing out that administration of cisplatin induced oxidative stress in rats. In groups that received A. melanocarpa extract as drinking water, we noted that the levels of the oxidative stress biomarkers tended to be restored almost to normal levels, which could be a possible good antioxidant used in condition to cisplatin use. Also, we noted a significant (P < 0.001) decrease of total antioxidant capacity in liver and kidney of rats exposed to cisplatin, recovered in those that received chokeberry. Studied trace elements important for the stress oxidative enzymes (Cu, Zn, Fe and Mn) were decreased in cisplatin exposed groups compared to control and mainly all were almost to normal level in groups receiving A. melanocarpa. CONCLUSION: A. melanocarpa extract due to its antioxidants content could offer protection against free radicals produced as a consequence of cisplatin use.

Effects of Aronia melanocarpa and Hypericum perforatum aqueous extracts on hexavalent chromium induced toxicity in rat's thyrocytes.

BACKGROUND: Hexavalent chromium known as oxidizing agent is able to form reactive oxygen species. Aronia melanocarpa and Hypericum perforatum are two plants known for their antioxidant effects. Our study aimed to establish if CrVI induces apoptosis and structural changes in thyrocytes and if its effect can be counteracted by the administration of both extracts. MATERIALS AND METHODS: Wistar rats divided in five groups: C - distilled water (DW), Cr - 75 mg/L CrVI in DW for 3 months, Cr 2 - 75 mg/L CrVI in DW for 3 months followed by 1 month DW, CrA - 3 months 75 mg/L CrVI in DW and 1 month Aronia 2.5% extract, CrH - 3 months 75 mg/L CrVI in DW and 1 month Hypericum 2.5% extract. Histological assessment and qRT-PCR for evaluation of BAX and Bcl2 protein levels performed on thyroid samples. RESULTS: The Cr and Cr2 groups were those with altered cytoarchitecture: increase in the diameter of many thyroid follicles, a decrease in their number, a decrease in the height of the follicular cells. The histological examination of the CrH group revealed almost recovery of structural architecture. The BAX gene levels were higher in the Cr and Cr2 groups indicating the apoptotic activity of chromium. In extract receiving groups the BAX gene expressions were significantly lower, but the lowest level presented the CrH group. Bcl2 gene expression levels indicate antiapoptotic activity being elevated in the Cr group, followed by CrA, Cr2, and CrH groups. The BAX/Bcl2 ratio which significantly increased in the case of the Cr and Cr2 group compared to the groups that were administered the two plant extracts. CONCLUSION: The results obtained in this study confirm that CrVI has toxic effects on thyroid endocrine cells and H. perforatum has stronger antioxidant properties against the action of hexavalent chromium in thyrocytes than A. melanocarpa.

Aronia melanocarpa Ameliorates Adrenal Cytoarchitecture Against the Hexavalent Chromium-Induced Injury.

Hexavalent chromium is a toxin that penetrates the cell, triggering reactive oxygen species (ROS) production. Aronia melanocarpa, due to its proanthocyanidins, anthocyanins, and phenolic acid contents, is a valuable antioxidant. The aim was to observe the influence of hexavalent chromium Cr(VI) on the adrenal gland, and if this impact can be recovered by the administration of A. melanocarpa. Accordingly, 36 rats were divided into six groups: control; Aronia; Cr receiving Cr(VI) in distilled water for 3 months; CrA receiving a mix of Cr(VI) and A. melanocarpa at 2.5% aqueous extract for 3 months; Cr2 receiving, for 3 months, Cr(VI) in distilled water, and next, for 1 month, only distilled water; and respectively, CrA2 receiving, for 3 months, Cr(VI) in distilled water, followed by 1 month of Aronia at 2.5% extract administration. The adrenal gland samples were examined toward histological and molecular assessment, and results were statistically analyzed (ANOVA). Hexavalent chromium induced changes in the adrenal cortex expressed by focal or diffuse hypertrophies, cytoplasmic vacuolization (due to lipidic accumulation), and cells' shape and size alteration, including necrosis. These structural alterations were carried by Bax and Bcl2 gene expression: the Bax gene expression levels, increased significantly (p < 0.001) in all experimental groups, except the Aronia group, compared with control. In the Cr2, CrA, and CrA2 groups, notable reduction of Bax gene expression (p < 0.001) was reported compared with the Cr group. Regarding the Bcl2 gene expression (p < 0.001), a significant increase was observed in the experimental groups, compared with the control. The Bcl2 expression level had a similar pattern to Bax gene, consequently trying to compensate its overexpression. Aronia administered concomitantly, or after Cr(VI), diminished structural changes and expression of the studied genes, thus reducing the Bax/Bcl2 ratio and suggesting that the active ingredients from Aronia are capable of blocking apoptotic cascade induced by the pathway of Bax and Bcl2 proteins.

Sea Buckthorn and Grape Antioxidant Effects in Hyperlipidemic Rats: Relationship with the Atorvastatin Therapy.

BACKGROUND: Medications to reduce oxidative stress are preventing cellular damage associated with hyperlipidemia. In this regard, statins (e.g., atorvastatin) act primarily by decrease in low-density lipoprotein-c but, in the last decade, hepatotoxicity, associated with liver injuries in the next months after treatments' initiation, was reported. In this case, associated phytotherapy can be a solution. PURPOSE: To investigate the antioxidant potential and response to free radicals, in the case of hyperlipidemic rats treated with atorvastatin. Sea buckthorn (Hippophae rhamnoides) and a grape extract (antioxivita) efficiency in the oxidative stress were investigated, also being ascertained the rats' organs cytoarchitecture. METHODS: Eighty-four hyperlipidemic Wistar rats were divided into seven groups and orally treated as follows: ATS, atorvastatin (20 mg/kg·bw); ATS + Hr, atorvastatin + H. rhamnoides; ATS + Aox, atorvastatin + grape extract; Hr, H. rhamnoides; and Aox, grape extract (both as 100 mg/kg·bw). HFD and Control received high fat diet and normal fodder only. After two and six months, respectively, rats were euthanized and the heart, liver, and kidneys were gathered. The tissue samples were prepared by homogenization of 0.5 g tissue, in ethanol, kept for 48 hours at 4°C-10°C and then filtered, in order to assess organs' cytoarchitecture and the TAC's values (by using cupric ion reducing antioxidant capacity (CUPRAC) assay). The test tubes were incubated, at room temperature, for 30 minutes, and then analyzed using a spectrophotometer at 450-650 nm. RESULTS: The statistics (ANOVA) revealed that sea buckthorn diminished notably (p < 0.001) the oxidative stress in the heart, liver, and kidney. After six months, the TAC's reduced levels for the heart were significant (p < 0.001) in ATS + Aox. In the case of histology, the liver's cytoarchitecture in ATS revealed abnormal cytoarchitecture. In ATS + Hr, ATS + Aox, Hr, and Aox, cell regeneration improved in different stages, especially for ATS + Hr and ATS + Aox, in comparison with HFD, which exhibited fat degeneration. Kidney's cytoarchitecture revealed cellular healing, especially in ATS + Hr and ATS + Aox.

Gingival proliferative growth - stress and cytoarchitecture related with fixed and mobile orthodontic therapy.

The fixed orthodontic measures taken induce significant stress to the gingival growth process during arch wire maneuvers of aligning and leveling. We observed, for a period of one to four years, fixed orthodontic devices in 80 human subjects. From these, we selected 44 subjects (22 women and 22 men) where the inflammatory process exhibited following the orthodontic fixed treatment, and with vacuum-formed orthodontic retainers (VFR) succeeding to fixed treatment. Samples were collected from each patient and histological and immunohistochemical (IHC) methodology was made to analyze the cytoarchitecture. Statistics were made after one-way analysis of variance (ANOVA), with the Bonferroni's correction. The IHC examination performed in the early stage revealed the presence in the inflammatory infiltrate of CD8-type T-lymphocytes, and of dendritic cells in large numbers. The examination performed in the late stage revealed the presence in the inflammatory infiltrate of CD20-type B-lymphocytes, which are mature cells capable of immunoglobulin synthesis, their activation being an important step in the maturation of the antibody response. The stress generated by arch wires in both genders was significantly higher than in the case of VFR. This observation was pointed out also by the cytohistological investigation outcome but was also based on an original scale conceived by our research team, following gingival hyperplasia evaluation. Also, with statistical significance, the comparative obtained values for men (p=0.01) and for women (p=0.001) illustrate clinical observations, allowing to affirm that, in our case, men were more stressed in bearing arch wire devices (AWD) and VFR, in comparison with women.

The effect of cisplatin administration on certain trace elements homeostasis in rats and the protective effect of silver birch (Betula pendula) sap.

BACKGROUND: A clinically active structure with known antitumor activities is cisplatin (CDDP), but this it comes with toxicity characteristics which can be faded by the beneficial effects of Silver birch (Betula pendula) sap. OBJECTIVE: We aimed to assess the cisplatin activity on: Mn, Mg, Cu, Fe and Zn homeostasis in rats and to observe the effect of birch sap. METHODS: Healthy Wistar rats (n = 10/group) were divided in four groups: Control: receiving 1 mL saline I.P. way + water; E1: cisplatin 20 mg kgbw-1, I.P.; E2: cisplatin 20 mg kgbw-1, I.P. + birch sap and Control sap group: 1 mL saline I.P. + birch sap. Blood was collected: at the trial's start and after 48 h, and blood and organs (liver, kidney and spleen) for the cytoarchitecture investigation and readings were sampled after seven days. Samples were processed in nitric acid by microwave digestion and readings were completed by flame atomic absorption spectroscopy, the outcomes being statistically analyzed by ANOVA. RESULTS: Cisplatin produced a significant imbalance in the trace elements homeostasis, the birch sap administration recovering them usual homeostasis status. Comparatively with the Control, rats exposed to cisplatin presented a not significant (p > 0.05) decrease of Zn (-26.74%) and Mg (-10.25%), a significant (p < 0.05) decrease of Cu (-27.73%) at 48 h, a highly significant (p < 0.01) decrease of Cu (-56.08%) and Fe (-85.35%) at seven days after administration and a not significant (p > 0.05) increase of Mn (+28.16%). Birch sap administration after Cisplatin was followed by restoration or nevertheless significant increase (p < 0.05) of the investigated trace elements Zn (+56.88% to 48 h/+89.94% after seven days), Mg (+26.86%/+95.74%), Cu (+23.13%/+74.56%), Fe (+39.64%/+440.11%) and Mn (+4.30%/+15.87%), suggesting them defence against cisplatin. Histology revealed the order of main altered organs after the CDDP exposure: kidney, spleen and liver. CONCLUSIONS: The study recommended the important protective role of Betula pendula sap against diverse cisplatin deleterious side-effects.

The consequences of aluminium intake on reproductive function in male rats:a three-generation study.

BACKGROUND/AIM: The effects of aluminium exposure on reproductive biomarkers in male rats were followed in a three-generation study. MATERIALS AND METHODS: Forty Wistar male rats (F0) were divided into the following groups: control (C), receiving only tap water, and three experimental (E) groups, receiving aluminium sulphate (AS) (E1: 200 ppb, E2: 400 ppb, and E3: 1000 ppb) in drinking water for a 6-month exposure period. To obtain F1, three males from each group were mated with previously unexposed females (1:2 sex ratios) that during gestation and lactation were exposed to the same AS levels as males. The F1 generation male offspring were divided as described and exposed to the same AS levels. The protocol to obtain F2 was similar to that described for F1. RESULTS: Significantly lower testosterone levels in rats exposed to AS and in generations F1 and F2 compared to the parental one, luteinising hormone (LH) fluctuations in F0 and a significant LH decrease in F2 and F3 generations, testis weight decrease, increased immobile and abnormal sperm, and histoarchitecture alterations in the testes were observed. Moreover, interval between deliveries increased. CONCLUSION: Chronic exposure to AS was significantly deleterious, producing a pronounced decrease in the sperm count and testosterone levels in all experimental groups.

Effect of Euphorbia cyparissias ointments on acanthosis.

BACKGROUND: A pharmaco-chemical investigation of the Euphorbia cyparissias plant was justified by its known multiple therapeutic valences. Numerous components from extracts and latex of Euphorbiacae were identified, revealing a large plant family with a polyvalent therapeutic activity. MATERIALS AND METHODS: The aim of the study was to assess the skin tolerance level to irritation on different testing concentrations, of Euphorbia cyparissias extracts and ointments. Study was accomplished in rats and dogs, with the identification of all possible skin injuries and histological changes, after a simple patch test methodology. RESULTS: Ointment dermatological testing on rats, proved to be bearable on epilated skin at concentrations of 1, 2 and 5%. Ointments and mother tincture with higher concentrations (10% and 20%), led to irritation and cutis damages, and this was revealed through histology. CONCLUSION: Ointment tested on dog's skin was tolerable for epilated skin to concentrations of 1, 2 and/or 5%, additional testing on human volunteers confirmed the same situation.

Cytoarchitecture of steroid dependent target tissues after testosterone administration compared to nandrolone decanoate in castrated rats in the aim of Hershberger bio test.

The objective was the cytoarchitecture evaluation of known steroid dependent target tissues after administering of testosterone, compared to action of its more active ester, nortestosterone (nandrolone decanoate) in castrated rat males in the aim of Hershberger bio test. Study was performed on 30 castrated male Wistar rats, aged between 35 and 39 days, in peripubertal period, divided into five groups. Androgen doses administration begun at the rats' age of 49 days. Animals were injected i.m., daily, for 10 consecutive days as follows: Aquatest (Balkan Pharmaceuticals Ltd., Moldova) testosterone aqueous solution: Testosterone I group (0.4 mg/animal); Testosterone II (0.8 mg/animal); (Deca-Durabolin, Balkan Pharmaceuticals); nandrolone decanoate oily solution: Nortestosterone I (1.5 mg/kg body weight); Nortestosterone II (7.5 mg/kg body weight) and Control (White sesame oil, Manicos, Romania, 0.1 mL/animal). Gonadectomy (GDX) induced modifications of target tissues wet weight accompanied by important modifications in cytoarchitecture. Changes following exogenous administration of testosterone and nortestosterone decanoate were found in: liver (granular dystrophy, mega-mitochondria, tubular intumescences), prostate (increasing of the structural elements), seminal vesicles (hyalinosis, thickening of cell walls and the hyaline presence), levator ani-bulbo-cavernosus muscle (muscle fibbers dilacerations), bulbourethral glands (muscular fibbers rarefaction by fluid accumulation) demonstrating the disruptor activity especially for overdosed nandrolone decanoate.

A comparison regarding antiproliferative action between soy total extract and genistein.

The aim of this study is to make a comparison between the action of genistein and total soy extract regarding anticancer action on two different in vivo models: phytobiological test and animal model, and to see which of the two tested samples present a greater antiproliferative effect. Soybean seeds were grounded and a solvent formed of DMSO-ethanol-water in rapport 5-70-25 v/v/v was prepared. The extraction was made using an ultrasonic bath (Falc LCD Series) for 30 minutes, 59 kHz. The solvent was evaporated with a rotary evaporator at 50°C. Genistein was acquired from Extrasynthèse (France), hydroxypropyl-γ-cyclodextrin (HPGCD) from Cyclolab Hungary, 7,12-dimethylbenz[a]anthracene (DMBA), dimethylsulfoxide (DMSO), and 12-O-tetradecanoylphorbol-13-acetate (TPA) from Sigma Aldrich, Germany. Because of the poor water solubility, genistein was prepared in a complex with hydroxypropyl-γ-cyclodextrin in a molar ratio 1:2 by kneading method and total soy extract in a mass ratio 1:4 also by kneading method. Phytobiological test indicated an inhibition index over 50% in case of solutions of concentration between 8-33% in both samples, suggesting a possible antiproliferative action at a superior level. Study on C57BL/6J mice was made on which it was induced cancer with physical agents like DMBA, and it was promoted with TPA. Mice where divided in four groups: Group A - blank group, Group B - mice who received total soy extract, Group C - mice who received genistein, Group D - untreated mice. Results on animal model show that both soy total extract and genistein inhibited the initiation and promotion of chemically-induced skin tumorigenesis, but genistein had a greater success in recovering skin lesions type experimental malignant melanoma.