Trigonella foenum-graecum Seeds Oil Attenuated Inflammation and Angiogenesis in vivo through Down-Regulation of TNF-α.

INTRODUCTION: Inflammation is a vital reaction of the natural immune system that protects against encroaching agents. However, uncontrolled inflammation can lead to complications. Trigonella foenumgraecum is traditionally used as an anti-inflammatory herb. OBJECTIVES: The current study was conducted to explore the antioxidant, anti-inflammatory, and antiangiogenic potentials of Trigonella foenum-graecum seeds oil. METHODS: Oil was extracted from seeds of Trigonella foenum-graecum by cold press method and labelled as TgSO. Phytochemical (GC-MS, Folin-Ciocalteu method) and metal analyses were conducted to evaluate the metalo-chemical profile of TgSO. In vitro antioxidant assays (2,2-diphenyl-1-picrylhydrazyl, 2,2'-azino-bis-3- ethylbenzothiazoline-6-sulfonic acid and ferric reducing antioxidant power) were performed to assess its antioxidant potential. In vitro antimicrobial activity was evaluated using agar disc diffusion method and the safety profile of TgSO was assessed in acute toxicological studies following OECD 425 guidelines. In vivo antiinflammatory activities of TgSO were assessed in carrageenan, serotonin, histamine, formalin, and cotton pelletinduced oedema models. Serum TNF-α, Superoxide Dismutase (SOD) and, Catalases (CAT) levels were assessed by ELISA kits. In vivo antiangiogenic activity of TgSO was screened in chick Chorioallantoic Membrane (CAM) assay. Histopathological studies using excised paws were conducted to observe the effects of TgSO treatment at the tissue level. In silico docking studies were conducted to screen the binding potentials of identified compounds with TNF-α. RESULTS: Extraction by cold press method yielded 16% of TgSO. Phytochemical analysis of TgSO through GCMS showed the presence of eugenol, dihydrocoumairn, heptadecanoic acid, tri- and tetradecanoic acid, and hexadecanoic acid, respectively. Total phenolic contents of TgSO were found to be 0.30±0.01mg/g gallic acid equivalent in Folin-Ciocalteu method. Metal analysis indicated the presence of different metals in TgSO. Findings of antioxidant models showed the moderate antioxidant potential of TgSO. Findings of antimicrobial assays showed that TgSO was active against bacterial (S. aureus, S. epidermidis) and fungal (C. albicans, and A. niger) strains. In vivo toxicity study data showed that TgSO was safe up to the dose of 5000 mg/kg. Data of oedema models showed a significant (p<0.05) reduction in oedema development in TgSO treated animals in both acute and chronic models. Histopathological evaluations of paws showed minimum tissue infiltration with inflammatory cells in TgSO-treated animals. Treatment with TgSO also significantly (p<0.05) down-regulated TNF-α in serum while levels of SOD and CAT were up-regulated. Findings of the CAM assay revealed the antiangiogenic activity of TgSO. Findings of in silico docking studies showed that identified phytoconstituents can bind with culprit cytokine (TNF-α). CONCLUSION: Data obtained from the current study conclude that TgSO has antioxidant, anti-inflammatory, and antiangiogenic effects that validate its traditional uses. Synergistic actions of different phytoconstituents are proposed to be responsible for the observed effects.

Methanolic extract of Ephedra ciliata promotes wound healing and arrests inflammatory cascade in vivo through downregulation of TNF-α.

Chronic wounds may lead to the development of various pathological conditions such as diabetic foot ulcers and pressure sores. The current study evaluated wound healing and anti-inflammatory potentials of methanolic extract of Ephedra ciliata using series of in vivo models. Methanolic extract of Ephedra ciliata was prepared by maceration (Ec.Me). Qualitative and quantitative (HPLC) phytochemical and metal analyses were conducted to explore the chemical and metal profiles of Ec.Me. Safety profile (behavioural) and, antimicrobial, antioxidant, wound healing, anti-inflammatory and anti-angiogenic potentials of Ec.Me were evaluated using well-established in vitro and in vivo models. ELISA assay was performed to estimate the effects of Ec.Me treatment on serum levels of TNF-α. HPLC analysis identified quercetin as one of the major compounds in Ec.Me. Safety study data showed that Ec.Me was safe up to the dose of 2000 mg/kg. Antimicrobial assay data showed that Ec.Me was active against bacterial (Staphylococcus aureus) as well as fungal (Candida albicans and Aspergillus niger) strains. Ec.Me showed modertate antioxidant potential in in vitro and in vivo models. Data of excision and burn wound healing models showed that Ec.Me, promoted wound closure in a dose and time-dependent manner. Treatment with 20% Ec.Me cream and heparin showed almost the same effects with no statistical differences (p > 0.05). Ec.Me also showed time-dependent anti-inflammatory activities in both acute and chronic models. In carrageenan model, treatment with 200 mg/kg of Ec.Me showed comparable anti-inflammatory effects (p > 0.05) with quercetin and indomethacin throughout the study. In cotton pellet granuloma model treatment with 200 mg/kg of Ec.Me and indomethacin inhibited granuloma formation significantly better (p < 0.05) as compared with the rest of the treatment groups. Histopathological examination of skin samples showed marked improvement in architecture with minimal infiltration of inflammatory cells. Data of in vivo angiogenesis assay showed marked improvement in vessels length, density, branching points, total segments and total nets after treatment with Ec.Me, indicating no toxic effects towards vasculature development. Significant (p < 0.05) downregulation of TNF-α was observed in serum samples of animals treated with Ec.Me. Based on data of the current study, it is concluded that quercetin-rich extract of Ephedra ciliata has wound healing and anti-inflammatory potentials via downregulation of TNF-α. Moreover, it is suggested that the antimicrobial activity of Ec.Me prevented microbial invasion, thus promoted natural wound healing mechanisms as well.

Evaluation of in vivo anti-inflammatory and anti-angiogenic attributes of methanolic extract of Launaea spinosa.

Launaea spinosa is used as an anti-inflammatory agent traditionally. This study was conducted to evaluate anti-inflammatory and anti-angiogenic activities of methanol extract of Launaea spinosa. Extraction was performed by maceration and the resultant green coloured extract was labelled as Ls.Me. Solubility analysis showed that Ls.Me was miscible with distilled water, normal saline, ethanol and methanol. Metal analysis following acid digestion method exhibited the presence of copper, magnesium, manganese, iron, zinc and calcium. Phytochemical analysis confirmed the presence of different classes of secondary metabolites in Ls.Me. HPLC analysis showed the presence of quercetin, gallic acid, caffeic acid, benzoic acid and sinapic acid in Ls.Me. Data of in vitro antioxidant assays showed moderate antioxidant potential of Ls.Me which was also confirmed by data of in vivo enzymes (SOD, CAT, and TSP) assays. Antimicrobial assays data showed that Ls.Me was active against S.aureus and S.epidermidis (bacterial) as well as C.albicans and A.niger (fungal) strains. Data of acute physio-pathological studies showed no abnormalities in Albino rats up to the dose of 2000 mg/kg of Ls.Me. Acute and chronic inflammatory models were used to evaluate anti-inflammatory effects of Ls.Me. Data of acute studies showed that Ls.Me has the potential to arrest inflammation produced by different mediators in a dose-dependent manner. 200 mg/kg of Ls.Me was found to produce significantly (p < 0.05) better anti-inflammatory effects than 100 mg/kg of Ls.Me. Ls.Me also significantly (p < 0.05) inhibited ear edema induced by xylene. Ls.Me showed profound anti-inflammatory responses in paw edema induced by formalin and also inhibited granuloma development in cotton pellet-induced granuloma model. Histopathological and biochemical investigations showed marked reduction in the number of inflammatory cells. TNF-α and IL-6 ELSIA kits were used to study effects of Ls.Me treatment on serum levels of TNF-α and IL-6. Data obtained showed significant (p < 0.05) reduction in TNF-α and IL-6 levels in serum of animals treated with Ls.Me. Data of in vivo angiogenesis assay showed that 200 µg/ml of Ls.Me significantly halted vasculature development indicating its potent anti-angiogenic potential. On the basis of findings of the current study, it is concluded that multiple phytochemicals present in Ls.Me act synergistically to produce anti-inflammatory and anti-angiogenic effects. Further studies are required to standardize the plant extract and explore its safety profile.