Cytotoxic Effect of Ethanol Extract of Abrus precatorius Leaves on Raw 264.7 and SK-N-SH Cell lines

Aim: This study was aimed to investigate the cytotoxic effect of ethanol extract of Abrus precatorius. leaves on Raw 264.7 and SK-N-SH cell lines. Methodology: Soxhlet extraction was carried out using absolute alcohol and subsequently, the profiling of phytoconstituents of ethanol extract was performed by LC-MS analysis. Results: The results showed that the presence of anthocyanin, phenolic acid, carboxylic acid, amino acid and monoester in ethanol extract of Abrus precatorius. The phytoconstituents such as picolinic acid, N-Acetyl-DL-tryptophan, 3-Hydroxybenzoic acid, kuromanin, aflatoxin G2, monobutyl Phthalate, laurolactam, 4-Dodecylbenzenesulfonic acid, 4-Methoxycinnamic acid, caffeic acid and octyl decyl phthalate were found in ethanol extract. In addition to this, the cytotoxic effect of the ethanol extract was tested on Raw 264.7 and SK-N-SH cell lines using MTT assay. The cytotoxic study revealed that the ethanol extract of Abrus precatorius was non-toxic to Raw 264.7 cell, but it showed a toxic effect on human neuroblastoma cell line, SK-N-SH. Conclusion: In this study, it was observed that the ethanol extract of Abrus precatorius was non-toxic to Raw 264.7 cell line, but it exhibited strong inhibition on the viability of SK-N-SH cell line.

Anti-inflammatory and Anti-arthritic Activities of Glycosylated Flavonoids from Syzygium jambos in Edematogenic Agent-Induced Paw Edema in Mice

Two glycosylated flavonoids, the quercetin-3-O-β-d-xylofuranosyl-(1 → 2)-α-l-rhamnopyranoside and myricetin-3-O-β-d-xylofuranosyl-(1 → 2)-α-l-rhamnopyranoside, were isolated from the CH2Cl2/MeOH fraction of Syzygium jambos (L.) Alston, Myrtaceae. The structures of these compounds were elucidated by spectroscopic means. The cytotoxicity of the compounds was evaluated against the RAW 264.7 cell lines by the lactate dehydrogenase assay. All analyzed compounds were less cytotoxic than the positive control (actinomycin D, CC50 = 0.008 μM). The anti-inflammatory and anti-arthritic activities were evaluated by measuring inflammatory parameters in murine models. The two glycosylated flavonoids inhibited the production of tumor necrosis factor-α in RAW 264.7 cell line with IC50 of 1.68 and 1.11 μM, respectively. In addition, all flavonoids decreased the levels of tumor necrosis factor-α, C-reactive protein, and fibrinogen at a dose of 5 mg/kg in murine models. Graphical abstract

Branched chain amino Acids as in vitro and in vivo Anti-Oxidation Compounds

Background and aims: Branched chain amino acids (BCAAs) can be tightly connected to metabolism syndrome (MetS) which can be counted as a metabolic indicator in the case of insulin resistance (IR). The aim of this study was to assess the potential role of these acids under oxidative stress. Material and Methods: the in vitro antioxidant activity of BCAAs was assessed using free radical 1, 1-diphenyl-2-picryl-hydrazyl (DPPH) scavenging assays. For further check, a qRT-PCR technique was madefor detection the extent of alterations in gene expression of antioxidative enzymes (catalase and glutathione peroxidase (Gpx)) in lipopolysaccharides (LPS(-induced macrophages RAW 264.7 cell line. Additionally, BCAAs antioxidant activity was evaluated based on plasma H2O2 levels and xanthine oxidase (XO) activity in prooxidative LPS-treated mice. Results: Different concentrations of BCAAs affected on DPPH radical scavenging activity but to lesser extent than the ascorbic acid. Besides, BCAAs obviously upregulated the gene expression levels of catalases and Gpx in LPS-modulated macrophage RAW 264.7 cell line. In vivo BCAAs significantly minimized the level of plasma H2O2 as well as the activity of XO activity under oxidative stress. Conclusion: our current findings suggest that BCAAs supplementation may potentially serve as a therapeutic target for treatment of oxidative stress occurs with atherosclerosis, IR-diabetes, MetS and tumorigenesis.

Lipidated Methotrexate Microbubbles: A Promising Rheumatoid Arthritis Theranostic Medicine Manipulated via Ultrasonic Irradiation

De novo designed lipidated methotrexate was synthesized and self-assembled into microbubbles for targeted rheumatoid arthritis theranostic treatment. Controlled lipidatedmethotrexate delivery was achieved by ultrasound-targetedmicrobubble destruction technique. Methotrexate was dissociated inflammatory microenvironment of synovial cavity, owing to representive low pH and enriched leucocyte esterase. We first manipulated methotrexate controlled release with RAW 264.7 cell line in vitro and further verified with rheumatoid arthritis rabbits in vivo. Results showed that lipidated methotrexate microbubbles precisely affected infection focus and significantly enhanced rheumatoid arthritis curative effect comparing with dissociative methotrexate. This study indicates that lipidated methotrexate microbubbles might be considered as a promising rheumatoid arthritis theranostics medicine.

17β-Estradiol Attenuates LPS-Induced Macrophage Inflammation In Vitro and Sepsis-Induced Vascular Inflammation In Vivo by Upregulating miR-29a-5p Expression

Excessive release of cytokines such as IL-1β and other inflammatory mediators synthesized and secreted by macrophages is the fundamental link of uncontrolled inflammatory response in sepsis. 17β-Estradiol (E2) plays anti-inflammatory and vascular protective effects by regulating leukocyte infiltration and the expression of chemokines or cytokines induced by injury. However, the role of E2 in the inflammatory response of macrophages in sepsis and its mechanism are still not fully understood. In the present study, we show that E2 alleviates vascular inflammation in sepsis mice induced by cecal ligation puncture (CLP). E2 significantly decreases RAW 264.7 cell inflammation response by downregulating the expression of NLRP3. Furthermore, we found that miR-29a-5p was significantly downregulated in LPS-treated macrophages. Treating RAW 264.7 cells with E2 markedly upregulated the miR-29a-5p expression level. More importantly, we demonstrated that miR-29a-5p repressed NLRP3 expression by directly targeting its 3 ′ -UTR. Loss- and gain-of-function experiments revealed that transfection of the miR-29a-5p mimic abrogates LPS-induced macrophage inflammation. Moreover, depletion of miR-29a-5p by its inhibitor largely promotes LPS-induced macrophage inflammation. In summary, miR-29a-5p upregulation induced by E2 alleviated RAW 264.7 cell inflammation response by aggravating miR-29a-5p repression of NLRP3 expression. E2 exerts significant anti-inflammatory efficacy in macrophages by regulating the miR-29a-5p/NLRP3 axis. Targeting miR-29a-5p may be a novel therapeutic strategy to suppress sepsis-induced vascular inflammation.

A New Phenylpropanoid from the Roots of Solanum melongena L. and Evaluation of Anti-inflammatory Activity

Fifteen phenylpropanoids were isolated from the ethanol extract of the roots of Solanum melongena L., including a new compound, melongenapanoid A (1), together with fourteen known compounds (2-15). Their chemical structures were elucidated by 1D and 2D NMR and HR-MS data according to those values of the literatures. The fourteen known compounds (2-15) were all firstly isolated from this plant. While, the isolates were evaluated for the inhibitory activities on nitric oxide (NO) production induced by lipopolysaccharide (LPS) in RAW 264.7 cell line. Compounds 2, 4 and 5 showed moderate inhibition of NO production with IC50 values of 28.7, 24.4 and 32.6 μM, respectively.

Responses of inflammation signaling pathway by saucerneol D from elicitor-treated Saururus chinensis on pro-inflammatory responses in LPS-stimulated Raw 264.7 cell

This study confirmed the association with inflammation-related proteins, mediators, and cytokines using saucerneol D from Saururus chinensis leaf, a useful ingredient increased through elicitor treatment. To confirm the anti-inflammatory effect, saucerneol D were treated with lipopolysaccharide, which induces pro-inflammatory factors in Raw 264.7 cell. The pro-inflammatory influences were measured by dint of chemical assay and western blotting as well as ELISA. As a result, the content of saucerneol D was changed when eicitor was treated by various concentration (1.5, and 3 mg/mL) in S. chinensis leaves. In addition, the expression levels of hyaluronidase and pro-inflammatory-related factors [nitric oxide (NO), inducible NO synthase (iNOS), and cyclooxygenase-2 (COX-2)] were regulated according to the saucerneol D content in the elicitor-treated and non-treated groups. Therefore, after confirming that saucerneol D has an inhibitory effect on pro-inflammatory-related factors, saucerneol D was adjusted by concentration and compared with the control substance to verify the efficacy. Saucerneol D was adjusted to a concentration that did not toxic to macrophages through 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Saucerneol D controlled at various concentrations inhibited iNOS and COX-2 proteins. NO produced by iNOS activity, prostaglandin E2 (PGE2), an inflammatory mediator produced by COX-2 activity, and pro-inflammatory cytokines [interleukin-1β (IL-1β), IL-6, tumor necrosis factor-α (TNF-α)] were significantly suppressed. Therefore, it was confirmed that saucerneol D, an active ingredient increased by the elicitor treatment, could be used as a functional material that controls inflammatory factors.