Unraveling the Mechanism of Platelet Aggregation Suppression by Monoterpenoids

Platelet aggregation causes various diseases and therefore challenges the development of novel antiaggregatory drugs. In this study, we report the possible mechanism of platelet aggregation suppression by newly synthesized myrtenol-derived monoterpenoids carrying different heteroatoms (sulphur, oxygen, or nitrogen). Despite all tested compounds suppressed the platelet aggregation in vitro, the most significant effect was observed for the S-containing compounds. The molecular docking confirmed the putative interaction of all tested compounds with the platelet’s P2Y12 receptor suggesting that the anti-aggregation properties of monoterpenoids are implemented by blocking the P2Y12 function. The calculated binding force depended on heteroatom in monoterpenoids and significantly decreased with the exchanging of the sulphur atom with oxygen or nitrogen. On the other hand, in NMR studies on dodecyl phosphocholine (DPC) as a membrane model, only S-containing compound was found to be bound with DPC micelles surface. Meanwhile, no stable complexes between DPC micelles with either O- or N-containing compounds were observed. The binding of S-containing compound with cellular membrane reinforces the mechanical properties of the latter, thereby preventing its destabilization and subsequent clot formation on the phospholipid surface. Taken together, our data demonstrate that S-containing myrtenol-derived monoterpenoid suppresses the platelet aggregation in vitro via both membrane stabilization and blocking the P2Y12 receptor and, thus, appears as a promising agent for hemostasis control.

Preliminary Phytochemical Analysis: In-Vitro Comparative Evaluation of Anti-arthritic and Anti-inflammatory Potential of Some Traditionally Used Medicinal Plants

Background: Colchicum autumnale, Strychnous nux-vomica and Aloe barbadensis are the medicinal plants clinically utilized for the management of rhuematic disorders. Purpose: The present work was focused to evaluate the in-vitro anti-arthritic and anti-inflammatory activities of Colchicum ( Colchicum autumnale), Nux-vomica ( Strychnous nux-vomica), and Aloe-vera ( Aloe barbadensis). Research Design: Primarily, the aqueous-ethanolic extracts of these medicinal plants were phytochemically screened followed by Fourier Transform Infrared (FTIR) analysis. Anti-arthritic activity by protein denaturation method and anti-inflammatory activity by human red blood cell (HRBC) membrane stabilization method at the concentration of 125, 250, and 500 µg/mL along with standard were performed. Results: Phytochemical screening revealed that alkaloids, saponins, terpenoids, phenols, and anthraquinones were found in all the extracts, and organic acids, amine group, aromatic or aliphatic compounds, esters and halogens, and phenolics were identified by FTIR. Protein denaturation method revealed that colchicum, nux-vomica, and aloe-vera showed maximum 98.5%, 99.6%, and 72.3% of inhibition at 500 µg/mL compared with that of standard drug, that is, Diclofenac sodium. Membrane stabilization method showed that colchicum, nux-vomica, and aloe-vera showed maximum 40.20%, 35.67%, and 40.1% protection at 500 µg/mL when compared with standard drug. Conclusion: It is concluded from the current study that extracts of colchicum, nux-vomica, and aloe-vera showed more potent effect and thus can be used as alternative options for the management of inflammatory and arthritic ailments.

Phytochemical, In vitro Anti-inflammatory and Antimicrobial Potential of Hugonia mystax L.

Hugonia mystax L., (Linaceae), is commonly distributed in the thorny scrubs and tropical dry evergreen forests of Tamil Nadu, which has been valued for centuries in traditional system of medicine for the treatment of various ailments. In the present study was an attempt to investigate the phytochemical nature and anti-inflammatory, antimicrobial potential by adopting suitable methods. Phytochemical analysis of Hugonia mystax L., plant extracts revealed the presence of various biochemical compounds such as alkaloids, flavonoids, glycosides, triterpenoids and saponins etc. Since triterpenoids and flavonoids have remarkable anti-inflammatory activity, so our present work aims at evaluating in vitro anti inflammatory activity of Hugonia mystax L., by HRBC membrane stabilization method. The inhibition of hypotonicity induced HRBC membrane lysis was taken as a measure of the anti-inflammatory activity. The percentage of membrane stabilization for ethanolic extracts and Diclofenac sodium were done at different concentrations. The maximum membrane stabilization of Hugonia mystax L., extracts was found to be 94.97 % at a dose of 2000 μg/ml. Therefore, our studies support the isolation and the use of active constituents from Hugonia mystax L., in treating inflammations.

Identifikasi Antiinflamasi Partisi Metanol, n-Heksan dan Ekstrak Etanol Daun Kemangi (Ocimum Americanum L.) Secara In Vitro

Basil plant (Ocimum americanum) is efficacious as an anti-inflammatory and analgesic activity. According to research by Sarma and Babu, 2011, Verma and Kothiyal, 2012 showed basil activity as an antioxidant, antimicrobial, anti-diabetic, anthelmintic, antifungal, insecticide, anti-inflammatory, analgesic, and lowering total cholesterol and LDL-C levels. The purpose of this study was to determine the stabilization activity of red blood cell membranes on methanol partitioning, n-hexane partitioning and ethanol extract of basil leaves in vitro. This study used the erythrocyte membrane stabilization method from the induction of a hypotonic solution with samples of methanol partitioning, n-hexane partitioning and ethanol extract to be compared with a positive control, namely Na diclofenac. By analyzing the data using UV-Vis spectrophotometry test. These results were supported by the ANOVA statistical test which stated that there was a difference in each treatment and continued with the Tukey test which stated that there was no difference between 100 ppm diclofenac sodium and 400 ppm ethanol extract.Keywords: Extract, Basil (Ocimum americanum) Leaf, In Vitro. AbstrakTumbuhan Kemangi (Ocimum americanum) berkhasiat sebagai aktivitas sebagai anti-inflamasi dan analgesik. Menurut penelitian Sarma dan Babu, 2011.,Verma dan Kothiyal, 2012 menunjukkan aktivitas kemangi sebagai antioksidan, antimikroba, anti diabetes, antihelmintik, antifungi, insektisida, antiinflamasi, analgesic, dan menurunkan kadar total kolesterol dan LDL-C. Tujuan dari penelitian ini yaitu untuk mengetahui aktivitas stabilisasi membran sel darah merah pada partisi metanol, partisi n-heksan dan ekstrak etanol daun kemangi secara in vitro. Penelitian ini menggunakan metode stabilisasi membran eritrosit dari induksi larutan hipotonik dengan sampel partisi metanol, partisi n-heksan dan ekstrak etanol yang akan dibandingkan dengan kontrol positif yaitu Na diklofenak. Dengan analisis data menggunakan uji spektrofotometri UV-Vis. Hasil ini didukung dengan uji statistik ANOVA yang menyatakan terdapat perbedaan pada setiap perlakuan dan dilanjutkan uji tukey yang menyatakan tidak ada perbedaan pada natrium diklofenak 100 ppm dengan ekstrak etanol konsentrasi 400 ppm.Kata Kunci : Ekstrak, Daun Kemangi (Ocimum americanum), In Vitro.

Isolation and Evaluation of Anti-inflammatory activity of Epigallocatechin from Senegalia rugata along with PUFAs

Senegalia rugata (Lam.) Britton & Rose, Synonym: Acacia concinna (Wild.) DC., Family: Fabaceae is one of the ayurvedic medicinal plant and commonly known as shikakai. The pods of S. rugata are normally used for cleansing of hair naturally due to the presence of higher content of saponins. In this study, we have isolated six compounds consisting of epigallocatechin (monomeric proanthocyanidin) from ethanol extract of S. rugata and a mixture of methyl esters of five polyunsaturated fatty acids (PUFA): methyl oleate, glyceryl trilinoleate, methyl linoleate methyl eicosenoate and methyl vernolate from petroleum ether extract of S rugata. The structures of the six compounds were elucidated using 1HNMR, 13CNMR and IR spectral studies. Epigallocatechin has shown significant in vitro anti-inflammatory property in a dose-dependent manner using the HRBC membrane stabilization method.

In vitro studies on anti-inflammatory, antioxidant and antihyperglycemic activities of potential probiotic Pediococcus acidilactici NCDC 252

Probiotics are live microbes which positively influence the health when consumed in adequate amount. Lactic acid bacteria (LAB) are commonly used probiotics and are generally found in yogurt and fermented foods. They provide barrier for pathogens by secreting peptides and other metabolites. Pediococcus acidilactici NCDC 252 is a LAB of dairy origin with probiotic attributes. NCDC 252 was studied for in vitro anti-inflammatory, antioxidant and antihyperglycemic activities. Anti-inflammatory activity was studied by human red blood cell (HRBC) membrane stabilization method, protein (albumin) denaturation inhibitory activity and heat induced haemolysis. Antioxidant activity was evaluated by α, α-diphenyl-β- picrylhydrazyl (DPPH) radical and hydrogen peroxide (H2O2) scavenging assays. Alpha amylase inhibition assay was performed to examine antihyperglycemic effect. NCDC 252 exhibited potent anti-inflammatory but moderate antioxidant and antihyperglycemic activities as compared to control. NCDC 252 exhibited 65%, 70% and 49% membrane stabilization, protein denaturation and heat induced activity respectively. Scavenging effect was 45 % and 60% in H2O2 and DPPH assays respectively. Alpha amylase inhibition was 48 %. These results suggest therapeutic potential of NCDC 252 and open new avenues to treat disorders related to free radical generation such as inflammation and diabetes mellitus after in vivo evaluation of NCDC 252 to confirm its efficacy in animals.

Gloeothece sp.—Exploiting a New Source of Antioxidant, Anti-Inflammatory, and Antitumor Agents

Bioactive lipidic compounds of microalgae, such as polyunsaturated fatty acids (PUFA) and carotenoids, can avoid or treat oxidation-associated conditions and diseases like inflammation or cancer. This study aimed to assess the bioactive potential of lipidic extracts obtained from Gloeothece sp.–using Generally Recognized as Safe (GRAS) solvents like ethanol, acetone, hexane:isopropanol (3:2) (HI) and ethyl lactate. The bioactive potential of extracts was assessed in terms of antioxidant (ABTS•+, DPPH•, •NO and O2•assays), anti-inflammatory (HRBC membrane stabilization and Cox-2 screening assay), and antitumor capacity (death by TUNEL, and anti-proliferative by BrdU incorporation assay in AGS cancer cells); while its composition was characterized in terms of carotenoids and fatty acids, by HPLC-DAD and GC-FID methods, respectively. Results revealed a chemopreventive potential of the HI extract owing to its ability to: (I) scavenge -NO• radical (IC50, 1258 ± 0.353 µg·mL−1); (II) inhibit 50% of COX-2 expression at 130.2 ± 7.4 µg·mL−1; (III) protect 61.6 ± 9.2% of lysosomes from heat damage, and (IV) induce AGS cell death by 4.2-fold and avoid its proliferation up to 40% in a concentration of 23.2 ± 1.9 µg·mL−1. Hence, Gloeothece sp. extracts, namely HI, were revealed to have the potential to be used for nutraceutical purposes.

STUDY OF ANTI-INFLAMMATORY POTENTIAL OF WHOLE CELL EXRACTS OF CYANOBACTERIA BY MEMBRANE STABILIZATION & TRYPSIN INHIBITION ASSAY

Inflammation is a complex mechanism in response to any infection, injury, or irritation. The prolonged inflammation leads to tissue damage and loss of function result in various disease conditions like osteoarthritis, and autoimmune diseases like lupus, rheumatoid arthritis, asthma and Crohn’s disease. Anti-inflammatory drugs are used to control inflammatory response and prevent tissue damage .Cyanobacteria are rich sources of phytochemicals like phenolics, flavonoids are known to have anti-inflammatory activity. Because of ease of cultivation and faster growth rate than plants, they are preferred candidates over plants. This study focuses on screening of cyanobacterial isolates for their anti-inflammatory activity. The human erythrocytes (HRBC) membrane stabilization assay indicated the potential of whole cell extracts of cyanobacteria to stabilize lysosomal membrane and thereby prevent tissue damage by lysosomal chemokines and enzymes. The trypsin inhibition assay is an indicator for potential to inhibit proteinases and decelerate tissue damage. The whole cell extracts of 10 cyanobacterial isolates of different genus namely Weistellopsis, Pseudophormidium, Oscillatoria, Nostoc, Phormidium, Chlorella, Hapalosiphon under study showed 85%-94% membrane stabilization in hypotonicity induced hemolysis and 92% to 97% membrane stabilization in heat induced hemolysis. The test extracts also showed 48% to 52% inhibition of trypsin. Thus, the isolates under study have application as anti-inflammatory agent.

Metabolic Adjustment of Glycine max (L.) Merril in the Presence of Nitrate and Bradyrhizobium japonicum

Reactive oxygen species are generated during the processes of photosynthesis and nitrate reduction, which can compromise the integrity of biomolecules and membranes. During the vegetative phase of Fabaceae species, around half of translocated carbohydrate is used for nodule growth, while the other half returns to the aerial part with nitrogen incorporated. These sugars may be yet involved with membrane stabilization, signaling, and activation of important genetic pathways for plant development. Thus, the aim was to study the adjustments of the photosynthetic and antioxidant systems and the accumulation of carbohydrates and biomass in Glycine–Bradyrhizobium cultivated with nitrate (NO3−). Four treatments were evaluated in completely randomized blocks. Glycine–Bradyrhizobium was grown with 1.7 mM of NO3− (GB: 1.7 mM NO3−) and without NO3− (GB: 0 mM NO3−), and Glycine was grown with 1.7 mM of NO3− (G: 1.7 mM NO3−) and without NO3− (G: 0 mM NO3−). Glycine–Bradyrhizobium symbiosis contributes to photosynthetic metabolism and total sugars, reduces the action of antioxidant enzymes, and minimizes the use of nitrate in soybean cultivation.; Glycine–Bradyrhizobium with nitrate provided greater plant dry mass in the vegetative phase, along with increased enzymatic activity and reduced nodule mass.