Ridha Ben Ali

An animal model of testicular toxicity by cyclosporine: evaluation and protection

CyclosporineA (CsA) improves the survival of patients who benefited from transplantation. However, its use is generally limited by its side effects. The aim of our study was to measure, in an experimental model, the changes of the testosterone plasma levels after 21 days of CsA treatment and to explain the mechanism of this modification. After treatment, the levels of CsA, testosterone, corticosterone, transaminases were measured. The cytotoxic effect of CsA was evaluated by microscopic observation. The experimental study showed that CsA had no effect on the plasmatic levels of hepatic enzymes ‐ alanine aminotransferase, aspartate aminotransferase and gamma‐glutamyl‐transferase – because their plasma concentrations in treated rats did not differ from those of the sham group. The plasma concentration of corticosterone was not modified, the plasma level of testosterone decreased when the dose of cyclosporine was increased to 4 mg/kg/day. The photonic microscope observation showed that the number of Leydig cells was increased and the electronic microscope observation showed mitochondria alteration. The treatment by CsA and trimetazidine did not correct the alteration caused by CsA. N‐benzyl‐N’‐(2‐hydrox‐3, 4‐dimethyloxybenzyl)‐pipeazine did not protect the mitochondrial function but partially protected mitochondria structure from the deleterious effect induced by CsA. The decrease of the plasma level of testosterone induced by CsA was due to the inhibition of the mitochondrial 20–22 desmolase which blocked the formation of the testosterone precursor and the destruction of the mitochondria structure.

Prophylactic and curative effect of rosemary leaves extract in a bleomycin model of pulmonary fibrosis

Abstract Context: Pulmonary fibrosis is a devastating disease without effective treatment. Rosemary is appreciated since ancient times for its medicinal properties, while biomolecules originated from the plant have an antioxidant and antifibrotic effect. Objective: The effects of Rosmarinus officinalis L. (Lamiaceae) leaves extract (RO) on bleomycin-induced lung fibrosis were investigated. Materials and methods: Male Wistar rats were given a single dose of bleomycin (BLM, 4 mg/kg, intratracheal), while RO (75 mg/kg, intraperitoneal) was administered 3 days later and continued for 4 weeks (BLM/RO1-curative group). Alternatively, RO was administered 2 weeks before BLM and continued 15 days thereafter (BLM/RO2-prophylactic group). Antioxidant activities of RO and lung tissues were studied by standard methods. Histological staining revealed lung architecture and collagen deposition. RO was characterized for its polyphenol content and by high-performance liquid chromatography. Results: RO polyphenol content was 60.52 mg/g of dry weight, carnosic and rosmarinic acids being major components (6.886 and 2.351 mg/g). Antioxidant effect of RO (DPPH and FRAP assay) expressed as IC50 values were 2.23 μg/mL and 0.074 μg/mL, respectively. In BLM/RO1 and BLM/RO2 lung architecture was less compromised compared to BLM, which was reflected in lower fibrosis score (2.33 ± 0.33 and 1.8 ± 0.32 vs 3.7 ± 0.3). Malondialdehyde levels were attenuated (141% and 108% vs 258% of normal value). Catalase and glutathione-S-transferase activities were normalized (103% and 117% vs 59%, 85% and 69% vs 23%, respectively). Discussion and conclusion: RO has a protective effect against BLM-induced oxidative stress and lung fibrosis due to its phenolic compounds.

Rosmarinic acid potentiates carnosic acid induced apoptosis in lung fibroblasts

Pulmonary fibrosis is characterized by over-population and excessive activation of fibroblasts and myofibroblasts disrupting normal lung structure and functioning. Rosemary extract rich in carnosic acid (CA) and rosmarinic acid (RA) was reported to cure bleomycin-(BLM)-induced pulmonary fibrosis. We demonstrate that CA decreased human lung fibroblast (HLF) viability with IC50 value of 17.13±1.06 μM, while RA had no cytotoxic effect. In the presence of 50 μM of RA, dose-response for CA shifted to IC50 value of 11.70±1.46 μM, indicating synergic action. TGFβ-transformed HLF, rat lung fibroblasts and L929 cells presented similar sensitivity to CA and CA+RA (20μM+100μM, respectively) treatment. Rat alveolar epithelial cells died only under CA+RA treatment, while A549 cells were not affected. Annexin V staining and DNA quantification suggested that HLF are arrested in G0/G1 cell cycle phase and undergo apoptosis. CA caused sustained activation of phospho-Akt and phospho-p38 expression and inhibition of p21 protein.Addition of RA potentiated these effects, while RA added alone had no action.Only triple combination of inhibitors (MAPK-p38, pan-caspase, PI3K/Akt/autophagy) partially attenuated apoptosis; this suggests that cytotoxicity of CA+RA treatment has a complex mechanism involving several parallel signaling pathways. The in vivo antifibrotic effect of CA and RA was compared with that of Vitamine-E in BLM-induced fibrosis model in rats. We found comparable reduction in fibrosis score by CA, RA and CA+RA, attenuation of collagen deposition and normalization of oxidative stress markers. In conclusion, antifibrotic effect of CA+RA is due to synergistic pro-apoptotic action on lung fibroblasts and myofibroblasts.

Synthesis and evaluation of analgesic, behavioral effects and chronic toxicity of the new 3,5-diaminopyrazole and its precursor the thiocyanoacetamide

This study aimed to explore the analgesic, antioxidant, behavioral and toxicological effects of 3,5-diaminopyrazole and thiocyanoacetamide. Caffeine was used as reference drug whose effects are known after oral treatment with an efficient dose (10mg/kg/day) for 30days. The preliminary bioassays indicated that both compounds at this dose have strong antioxidant capacities and present highly analgesic effects. The behavioral study showed an activation of the rat memory by thiocyanoacetamide. This molecule caused a phobia state to open areas in the elevated plus maze and specifically agoraphobia in the open field with a lack in the development of the exploratory capacity. 3,5-Diaminopyrazole caused memory troubles in rats that forgot the pathway to the exit from the maze, and induced an anxiety state revealed by immobility in closed arms of the elevated plus maze. All these observations were compared to the treatment by the known analgesic, caffeine, which increased the state of vigilance of the rats and developed their exploratory capacity. The chronic treatment with the investigated compounds showed no sign of toxicity with the absence of effect on the body and organ weights, blood count, kidney and liver function and histology. 3,5-Diaminopyrazole and thiocyanoacetamide have potent antioxidant and analgesic activities that are higher than caffeine with a safety profile. The chronic treatment by thiocyanoacetamide activated the memory and caused an emotional state of agoraphobia, but 3,5-diaminopyrazole caused a memory impairment and an emotional state of anxiety. Thus, the present study warrants further investigations involving these novel molecules for a possible development of new strong analgesic and antioxidant drugs which have an effect on the memory capacity.

The efficacy of plant extract and bioactive compounds approaches in the treatment of pulmonary fibrosis: A systematic review

Pulmonary fibrosis (PF) is a lethal, chronic and progressive respiratory disease leading to interstitial lung damage and serious breathing problems. The pathogenic mechanism involves activation, migration, proliferation and differentiation of fibroblasts into myofibroblats inducing extracellular matrix accumulation that destroy lung parenchyma. Available antifibrotic treatment options are limited to Pirfenidone and Nintedanib that prevent deterioration without an improvement of this disease. The use of plant extracts and natural bioactive compounds for the treatment of PF has been known for more than thirty years in China. Nowadays, phytotherapy has gained a considerable attention in the treatment of PF both in vivo and in vitro using bleomycin (BLM)-induced lung inflammation, oxidative stress and pulmonary fibrosis in rats. In this review, we aimed to focus on the protective effects and the mechanisms of action of several plant extracts described by various research works for the treatment of PF.

Therapeutic effect of flaxseed oil on experimental pulmonary fibrosis induced by bleomycin in rats

Pulmonary fibrosis is a chronic progressive disorder in which excessive deposition of extracellular matrix leads to irreversible scarring to interstitial lung tissue. In this study, we search to evaluate the therapeutic effect of flaxseed oil (FO) in experimental bleomycin (BLM)-induced pulmonary fibrosis. During our study, 30 male Wistar rats (weight range, 180–220 g) were divided into three groups: the control group (W) received no treatment; the second group (C) received BLM; and the third group (T) received BLM and FO for 21 days. Metabolites present in the bronchoalveolar lavage fluid (BALF) marking the changes obtained following treatment with FO were determined, histological changes in the lungs were evaluated, fatty acids present in lungs and erythrocytes of rats groups were determined by gas chromatography, and oxidative stress and antioxidant enzyme activity in the lung tissue were also recorded. Our results displayed that rat body weight decreased while fibrosis score and inflammatory index in lung tissue were significantly increased after bleomycin instillation. Administration of bleomycin followed by FO treatment reduced bleomycin-induced weight loss, increased proline, glucose, and glycerid rates in BALF and which are characterized by their anti-inflammatory effect and confirming the histological results proved by a decrease in inflammatory index and fibrosis score. This oil also significantly reduced thiobarbitunic acid reactive substance levels in the lungs of rats and increased levels of SOD and CAT and increased fatty acids levels promoting anti-inflammatory reactions especially in erythrocytes (linoleic, arachidonic, docosapentaenoic, and dihomo-γ-linoleic acids). In conclusion, these findings indicate that FO treatment significantly attenuated the increased pulmonary damage induced by bleomycin.

Gold and Female Reproductive Organs: an Ultrastructural Study

Gold, a heavy yellow-colored metal, is usually found in nature as a metallic element or as salts. This noble metal historically had a reputation as an anti-inflammatory medicine for rheumatoid arthritis, a nervine, and a remedy for nervous disorders, as well as a potential anticancer agent. It has also been used as component in dental restorations and in implant materials. The present study was undertaken to point out histological and ultrastructural effects of gold, administered by intraperitoneal route, in pregnant female reproductive organs (ovary and uterus), in order to clarify its side effects on the reproductive function. Using the transmission electron microscopy (TEM), the ultrastructural investigations of both ultrathin ovarian and uterine sections of treated pregnant rats revealed the existence of numerous heterogeneous clusters with very electron-dense inclusions characterized by various aspects in the lysosomes of granulosa, theca interna cells, and theca externa cells. Degeneration of these tissues, like cell vacuolization, marked expansion of the endoplasmic reticulum, mitochondrial alterations, and necrotic foci, were also highlighted. Moreover, huge phagolysosomes and high numbers of eosinophils as signs of inflammation were also identified especially in endometrial and myometrial cells of gold-treated rats. The ultrastructural investigations of reproductive organ sections of control pregnant rats showed a normal ultrastructural aspect and no loaded lysosomes. These results speculated the toxicity of gold at the used dose. The observed signs of toxicity allowed concluding that the important role of lysosome in the sequestration of this element under an insoluble form in all categories of cells in the studied tissues does not seem to be efficient.

Does Rosemary leaves extract can attenuate lung injury in experimental pulmonary fibrosis in rats

Background: Idiopathic Pulmonary Fibrosis (IPF) is the most devastating form of interstitial lung disease. Currently, available therapies are ineffective and have significant adverse effects. Medicinal plants rich of bioactives compounds can be a good therapeutic alternative. Objective: This study aimed to investigate the effect of Rosemary leaves extract (RO) on Bleomycin (BLM)-induced lung fibrosis in wistar rats. Methods: Animals were randomly divided into 4 groups of 10 rats each. G 1 : Control group, G 2 : BLM group, G 3 : RO group, G 4 : BLM + RO group. BLM was administered by a single intra tracheal injection (4 mg/Kg) to G 2 and G 4 . RO was administered intraperitoneally at a dose of (75 mg / kg/day) for 4 weeks to G 3 and G 4 . The later received RO 3 days after BLM injection. At the end of the protocol, all rats were sacrificed and lungs were extracted. Fibrosis was assessed conforming fibrosis Ashcroft score. Oxidative stress was evaluated in lung homogenate by measuring lipid peroxidation levels, Thiol group content, Catalase and Glutathione-S-Transferase activities. Statistical significance was determined by ANOVA followed by Sidak9s Test . Results : Histological assessments revealed a significant reduction in fibrosis score in G4 compared to G 2 . BLM increased Malondialdehyde content and decreased Glutathione S-Transferase, catalase and Thiol group levels in G 2 compared to G 1 .Treatment by RO (G 4 ) restored significantly the activities of antioxidant enzymes and Thiol group content and decreased Malondialdehyde concentration compared to G 2 . Conclusion: This study demonstrated that RO can attenuate BLM induced lung fibrosis in wistar rats by its antioxidant and antifibrotic proprieties.