Amelioration of anti-hepatotoxic effect by Lichen rangiferinus against alcohol induced liver damage in rats

Background: Reindeer lichen, Lichen rangiferinus syn. or Cladonia rangiferina (L.) F. H. Wigg. (Cladoniaceae) has been traditionally reported as a remedy to treat fever, colds, arthritis as well as convulsions,liver infections, coughs, constipation, and tuberculosis. The current study is aimed at rectification ofalcohol induced liver damage by the use of L. rangiferinus extract.Objectives: The aim of the study was to compare some biochemical markers for liver injury and hematological indices in normal untreated rats and treated rats.Material and Methods: The study was performed using male Wistar rats. Animals were categorized intofive groups, negative control group (normal diet only), treated groups (2 groups were lichen treatedalong with 10% ethanol & 1 group was only ethanol treated) and positive control group (Silymarin + 10%ethanol) of six animals in each group. Biochemical markers for liver injury and hematological indices ofall animals were measured using standard diagnostic tools. The animals were then sacrificed and liverswere sent to the pathology lab for histopathological analysis.Results: Lichen extract showed a significant restoration of altered biochemical parameters towardsnormal in both in vitro and in vivo conditions. The total phenolic and flavonoid content of the LRE wasfound to be 21.78 mg PE/mg of extract and 5.13 mg RE/mg of extract respectively. The IC50 values foratranorin and fumarprotocetraric acid were found to be 128.48 and 218.46 mg/mL respectively.Reducing power of the extract was found to be quite significant. After administration of lichen extract,endothelial cells were less injured around central vein and number of fat vacuoles was also lesser inhepatocytes.Conclusion: Conclusively, treatment with lichen extract assuages alcohol-related damage and guardshepatic tissue from alcohol-induced toxicity.© 2017 Transdisciplinary University, Bangalore and World Ayurveda Foundation. Publishing Services byElsevier B.V. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).

Cytotoxic, apoptotic and cell migration inhibitory effects of atranorin on SPC212 mesothelioma cells

Objective: To investigate the effects of atranorin, a lichen secondary metabolite, on SPC212 malignant mesothelioma cells in vitro. Methods: SPC212 malignant mesothelioma cell line was used. 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay was used to evaluate cytotoxic effects of atranorin and cisplatin at 24, 48 and 72 h. Hematoxylin-eosin staining and 4',6-diamidino-2-phenylindole, dihydrochloride staining were used for determining cell and nucleus morphology, respectively. Wound healing assay was used for investigating cell migration. The xCELLigence real-time cell analysis system was used for determining cell proliferation. Results: Atranorin at 5-450 μM decreased cell viability at 24, 48 and 72 h. IC50 values of atranorin were 300.94, 292.6 and 278.02 μM at 24, 48 and 72 h, respectively; meanwhile, the IC50 values of cisplatin were 128.00, 34.37 and 17.05 μM at 24, 48 and 72 h, respectively. Furthermore, atranorin disrupted cell and nuclear morphology with increasing concentrations. Atranorin significantly reduced cell migration by 38%, 37% and 35% at 300, 250 and 200 μM, respectively (P＜0.000). Atranorin at 160-450 μM decreased cell proliferation at 72 h (P＜0.000). Conclusions: Atranorin has cytotoxic, antiproliferative, apoptotic and cell migration inhibitory effects on SPC212 malignant mesothelioma cancer cells.

Cytotoxic, apoptotic and cell migration inhibitory effects of atranorin on SPC212 mesothelioma cells

Objective: To investigate the effects of atranorin, a lichen secondary metabolite, on SPC212 malignant mesothelioma cells in vitro. Methods: SPC212 malignant mesothelioma cell line was used. 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay was used to evaluate cytotoxic effects of atranorin and cisplatin at 24, 48 and 72 h. Hematoxylin-eosin staining and 4',6-diamidino-2-phenylindole, dihydrochloride staining were used for determining cell and nucleus morphology, respectively. Wound healing assay was used for investigating cell migration. The xCELLigence real-time cell analysis system was used for determining cell proliferation. Results: Atranorin at 5-450 μΜ decreased cell viability at 24, 48 and 72 h. IC

Evaluation of wound healing activity of atranorin, a lichen secondary metabolite, on rodents

The aim of this study was to investigate the wound healing activity of atranorin cream (Patent requested) on excision wounds. Seventy-two male rats were anesthetized and an excisional wound was performed. Then the rats were randomly assigned into three groups: untreated control group; atranorin 1 (group treated with 1% AT ointment); and atranorin 5 (group treated with 5% AT ointment). Six animals of each group were euthanized 3, 7, 14 or 21 days after surgical procedures and the wounded areas were analyzed and removed. Serial histological sections were obtained and stained by histochemical techniques (Hematoxilin-Eosin-HEand Sirius red) and immunohistochemical techniques. Topical application of atranorin reduced wound areas, induced earlier granulation tissue formation, increased cell proliferation, improved collagenization and modulated the myofibroblasts differentiation when compared to control animals. It is suggested that atranorin modulates the wound healing process. These data suggest that this formulation based on atranorin extracted from Cladina kalbii AHTI may be a new biotechnological product for wound healing clinical applications.

Anti-inflammatory and toxicity studies of atranorin extracted from Cladina kalbii Ahti in rodents

Atranorin (ATR) is the main compound from the lichen Cladina kalbii Ahti, which grows in the arid regions of northeastern Brazil. This study was conducted to evaluate the anti-inflammatory and toxicological properties of ATR. To evaluate anti-inflammatory properties, paw edema was induced by injecting 0.1 mL of carrageenan into the subplantar region of the right hind paw of rats, and leukocyte migration was induced by injection of 500 µL of carrageenan into the peritoneal cavity of mice. In addition, we determined ATR cytotoxicity in L929 cells by MTT assay and acute (5 g/kg-single dose) and subchronic (50 mg/kg-30 days) toxicity tests in Wistar rats. The results showed that ATR (100 mg/kg and 200 mg/kg) exhibited significant anti-inflammatory activity (paw edema and leukocyte migration). In the acute toxicity test, the animals showed hypoactivity and lethargy during the initial period (first 6 hours) and increase in total protein, total and indirect bilirubin, and alkaline phosphatase after 14 days in ATR-treated male rats. The subchronic toxicity test revealed increases in total protein, globulin, gamma-glutamyl transferase, alkaline phosphatase, and total and direct bilirubin in ATR-treated female rats. Histological analysis revealed no changes in the architecture and morphology of the organs. These results suggest that ATR has significant anti-inflammatory activity, with no significant acute and subchronic toxicity or cytotoxicity.

Atranorina (ATR) é o principal composto do líquen Cladina kalbii Ahti, que cresce em terras áridas do nordeste brasileiro. Este estudo foi realizado para avaliar as propriedades antiinflamatórias e toxicológicas da ATR. Para avaliar as propriedades antiinflamatórias, o edema de pata foi induzido, administrando-se 0,1 mL de carragenina na região subplantar da pata traseira direita e a migração leucocitária foi induzida pela injeção de 500 µL de carragenina no peritônio. Além disso, determinou-se a citotoxicidade da ATR, utilizando-se a linhagem celular L929, através do teste de MTT e dos testes de toxicidade aguda (5 g/kg - dose única) e subcrônica (50 mg/kg-30 dias) em ratos Wistar. Os resultados mostraram que nas doses de (100 mg/kg e 200 mg/kg) a ATR exibiu atividade antiinflamatória significativa nos ensaios de edema de pata e migração leucocitária. Nos testes de toxicidade aguda, os animais apresentaram hipoatividade e letargia no período inicial (primeiras 6 horas) e aumento das proteínas totais, bilirrubinas total e indireta e fosfatase alcalina depois de 14 dias nos machos tratados. Para o ensaio subcrônico, houve aumento das proteínas totais, gama-glutamil-transferase, fosfatase alcalina e bilirrubina total e direta nas fêmeas tratadas com ATR. Não foram encontradas alterações na arquitetura e morfologia das lâminas histológicas observadas. Esses resultados sugerem que a ATR apresenta atividade antiinflamatória significativa, sem apresentar significativa toxicidade aguda, subcrônica e citotoxicidade.