Djulis (Chenopodium formosanum) and Its Bioactive Compounds Protect Human Lung Epithelial A549 Cells from Oxidative Injury Induced by Particulate Matter via Nrf2 Signaling Pathway

The protective effects of water extracts of djulis (Chenopodium formosanum) (WECF) and their bioactive compounds on particulate matter (PM)-induced oxidative injury in A549 cells via the nuclear factor-erythroid 2-related factor 2 (Nrf2) signaling were investigated. WECF at 50–300 µg/mL protected A549 cells from PM-induced cytotoxicity. The cytoprotection of WECF was associated with decreases in reactive oxygen species (ROS) generation, thiobarbituric acid reactive substances (TBARS) formation, and increases in superoxide dismutase (SOD) activity and glutathione (GSH) contents. WECF increased Nrf2 and heme oxygenase-1 (HO-1) expression in A549 cells exposed to PM. SP600125 (a JNK inhibitor) and U0126 (an ERK inhibitor) attenuated the WECF-induced Nrf2 and HO-1 expression. According to the HPLC-MS/MS analysis, rutin (2219.7 µg/g) and quercetin derivatives (2648.2 µg/g) were the most abundant bioactive compounds present in WECF. Rutin and quercetin ameliorated PM-induced oxidative stress in the cells. Collectively, the bioactive compounds present in WECF can protect A549 cells from PM-induced oxidative injury by upregulating Nrf2 and HO-1 via activation of the ERK and JUN signaling pathways.     

Copper oxide nanoparticles from Rabdosia rubescens attenuates the complete Freund’s adjuvant (CFA) induced rheumatoid arthritis in rats via suppressing the inflammatory proteins COX-2/PGE2

Rheumatoid arthritis is a common auto-immune disease that affects nearly 1% of the total population worldwide and primarily occurs in the age group of 30–50. The copper oxide nanoparticles (CuONPs) were gained extensive research interests due to their immense biological benefits. This investigation was aimed to synthesize the CuONPs from R. rubescens leaves and examining its anti-arthritic potential against CFA-stimulated arthritis in rats. The formulated CuONPs from R. rubescens was characterized by UV–vis spectroscopy, TEM, XRD, and FT-IR. Arthritis was induced to animals by injecting 0.1 ml of CFA via an intra-planter route. Bodyweight and arthritis score was measured and tabulated and inflammatory markers i.e. IL-10, IL-6, TNF-α, IL-1β, Cox-2 and PGE2 in the serum was examined via ELISA kits. The status of TBARS, antioxidant enzymes, and liver marker enzymes were examined via standard procedures. The hind limb histology was examined microscopically by H&E staining. The UV–vis, XRD, and TEM studies were proved the existence of metallic RR-CuONPs. The RR-CuONPs treated arthritic animals displayed the enhanced bodyweight, reduced paw volume, and arthritic score. RR-CuONPs treatment also improved the antioxidant enzymes and diminished the pro-inflammatory markers. Histopathological investigation proved that the RR-CuONPs were suppressed the bone joints by preventing the inflammatory cell penetration and bone damage. The findings of this investigation were evidenced by the anti-inflammatory and anti-arthritic potentials of fabricated RR-CuONPs against CFA-stimulated arthritis in rats.     

Cytoprotection against Oxidative Stress by Methylnissolin-3-O-β-d-glucopyranoside from Astragalus membranaceus Mainly via the Activation of the Nrf2/HO-1 Pathway

Astragalus membranaceus is a famous herb found among medicinal and food plants in East and Southeastern Asia. The Nrf2-ARE assay-guided separation of an extract from Jing liqueur led to the identification of a nontoxic Nrf2 activator, methylnissolin-3-O-β-d-glucopyranoside (MNG, a component of A. membranaceus). Nrf2 activation by MNG has not been reported before. Using Western Blot, RT-qPCR and imaging, we investigated the cytoprotective effect of MNG against hydrogen peroxide-induced oxidative stress. MNG induced the expression of Nrf2, HO-1 and NQO1, accelerated the translocation of Nrf2 into nuclei, and enhanced the phosphorylation of AKT. The MNG-induced expression of Nrf2, HO-1, and NQO1 were abolished by Nrf2 siRNA, while the MNG-induced expression of Nrf2 and HO-1 was abated and the AKT phosphorylation was blocked by LY294002 (a PI3K inhibitor). MNG reduced intracellular ROS generation. However, the protection of MNG against the H₂O₂ insult was reversed by Nrf2 siRNA with decreased cell viability. The enhancement of Nrf2 and HO-1 by MNG upon H₂O₂ injury was reduced by LY294002. These data showed that MNG protected EA.hy926 cells against oxidative damage through the Nrf2/HO-1 and at least partially the PI3K/Akt pathways.     

Anti-arthritic activity of Tin oxide-Chitosan-Polyethylene glycol carvacrol nanoparticles against Freund’s adjuvant induced arthritic rat model via the inhibition of cyclooxygenase‑2 and prostaglandin E2

Rheumatoid arthritis (RA) results in increased rate of mortality in millions of people worldwide. Research utilizing Tin oxide – Chitosan- Polyethylene glycol Carvacrol (SCP-CAR) nanocomposites has gained increased attention because of its multipotent properties and application in diverse fields including medicinal preparations. The aim of the investigation was to synthesize and to examine the anti-arthritic ability of SCP-CAR nanocomposites against CFA -induced RA in rats. Arthritis induction was done by injecting 0.1 ml of Complete Freund’s adjuvant (CFA) intradermally. Body weight, weight of organs, hind paw volumes and arthritis score was assessed and the levels of inflammatory modulators such as IL-6, IL-1β, IL-10, TNF-α, PGE2 and COX-2 was examined using assay kits. Lipid peroxidation status, antioxidant enzyme activities and levels of liver function enzymes were evaluated using standard procedures. Histopathological changes observed in hind limb of experimental animals were viewed under microscope using H& E staining. The SCP-CAR nanocomposites treated arthritic animals showed increased bodyweight and reduced hind paw volume, organ weight and arthritis score together with elevated antioxidants status and depleted proinflammatory cytokines. Histopathological observation also showed reduction in bone destruction and penetration of inflammatory cells following treatment with SCP-CAR nanocomposites. Thus, together the findings depict the anti-arthritic and anti-inflammatory potential of SCP-CAR nanocomposites suggesting that it could be used as potent therapeutic agent to treat animals against arthritis induced by CFA.     

6,7,4′-Trihydroxyflavanone Mitigates Methamphetamine-Induced Neurotoxicity in SH-SY5y Cells via Nrf2/heme Oxyganase-1 and PI3K/Akt/mTOR Signaling Pathways

Methamphetamine (METH) is a synthetic psychostimulant drug that has detrimental effects on the health of its users. Although it has been investigated as a cause of neurodegenerative disease due to its neurotoxicity, whether small molecules derived from natural products attenuate these side effects remains elusive. 6,7,4′-trihydroxyflavanone (THF) is a flavanone family that possesses various pharmacological activities, including anti-rheumatic, anti-ischemic, anti-inflammatory, anti-osteoclastogenic, and protective effects against METH-induced deactivation of T cells. However, little is known about whether THF protects neuronal cells from METH-induced neurotoxicity. Here, we investigated the protective effects of THF on neurotoxicity induced by METH exposure by enhancing the Nrf2/HO-1 and PI3K/Akt/mTOR signaling pathways in SH-SY5y cells. Treatment with THF did not lead to cytotoxicity, but attenuated METH-induced neurotoxicity by modulating the expression of apoptosis-related proteins, METH-induced oxidative stress, and PI3K/Akt/mTOR phosphorylation in METH-exposed SH-SY5y cells. Moreover, we found THF induced Nrf2 nuclear translocation and HO-1 expression. An inhibitor assay confirmed that the induction of HO-1 by THF attenuates METH-induced neurotoxicity. Therefore, we suggest that THF preserves neuronal cells from METH-induced neurotoxicity by upregulating HO-1 expression through the Nrf2 and PI3K/Akt/mTOR signaling pathways. Thus, THF has therapeutic potential for use in the treatment of METH-addicts suffering from neurodegenerative diseases.     

Identification of a Sesquiterpene Lactone from Arctium lappa Leaves with Antioxidant Activity in Primary Human Muscle Cells

Many pathologies affecting muscles (muscular dystrophies, sarcopenia, cachexia, renal insufficiency, obesity, diabetes type 2, etc.) are now clearly linked to mechanisms involving oxidative stress. In this context, there is a growing interest in exploring plants to find new natural antioxidants to prevent the appearance and the development of these muscle disorders. In this study, we investigated the antioxidant properties of Arctium lappa leaves in a model of primary human muscle cells exposed to H₂O₂ oxidative stress. We identified using bioassay-guided purification, onopordopicrin, a sesquiterpene lactone as the main molecule responsible for the antioxidant activity of A. lappa leaf extract. According to our findings, onopordopicrin inhibited the H₂O₂-mediated loss of muscle cell viability, by limiting the production of free radicals and abolishing DNA cellular damages. Moreover, we showed that onopordopicrin promoted the expression of the nuclear factor-erythroid-2-related factor 2 (Nrf2) downstream target protein heme oxygenase-1 (HO-1) in muscle cells. By using siRNA, we demonstrated that the inhibition of the expression of Nrf2 reduced the protective effect of onopordopicrin, indicating that the activation of the Nrf2/HO-1 signaling pathway mediates the antioxidant effect of onopordopicrin in primary human muscle cells. Therefore, our results suggest that onopordopicrin may be a potential therapeutic molecule to fight against oxidative stress in pathological specific muscle disorders.     

Anti-inflammatory,analgesic and molecular docking studies of Lanostanoic acid 3-O-α-D-glycopyranoside isolated from Helichrysum stoechas

Helichrysum stoechas has been conventionally used as herbal tea due to its anti-inflammatory, antioxidant, antimicrobial and diuretic activities. Ethanolic extract of the aerial parts of the plant (HSE) afforded a lanostane triterpenoid glycoside. The isolated compound was characterized as Lanostan-3β-olyl-26-oic acid 3-O-α-D-glycopyranoside (HS-01) with the help of UV, IR, ¹H, ¹³C NMR and MS spectroscopic techniques. HSE (at 100 and 200 mg/kg doses) and the isolated compound, HS-01 (at 10 mg/kg dose) has been investigated for anti-inflammatory and analgesic activities against chemically challenged experimental animal. Both the HSE as well as HS-01 showed a substantial decline in paw volume when compared with the relevant control groups (p < 0.01 & p < 0.001). The HSE and HS-01 also confirms a significant prolongation of the paw licking or jumping towards the Eddy’s hot plate and reduction in quantity of writhes after the introduction of acetic acid in mice (p < 0.01 & p < 0.001). In order to have a better understanding of the binding interactions of HS-01 at molecular level, docking studies were performed with various macromolecular drug targets using AutoDock 4.2 and AutoDock Vina 1.1. Both programs predicted Galectin-3 as most favorable target for HS-01 followed by iNOS, whereas TNFα and COX-2 were among less favorable. Therefore, HS-01 could be developed as suitable therapy against inflammation and associated disorders.     

Fermented Oyster Extract Attenuated Dexamethasone-Induced Muscle Atrophy by Decreasing Oxidative Stress

It is well known that oxidative stress induces muscle atrophy, which decreases with the activation of Nrf2/HO-1. Fermented oyster extracts (FO), rich in γ-aminobutyric acid (GABA) and lactate, have shown antioxidative effects. We evaluated whether FO decreased oxidative stress by upregulating Nrf2/HO-1 and whether it decreased NF-κB, leading to decreased IL-6 and TNF-α. Decreased oxidative stress led to the downregulation of Cbl-b ubiquitin ligase, which increased IGF-1 and decreased FoxO3, atrogin1, and Murf1, and eventually decreased muscle atrophy in dexamethasone (Dexa)-induced muscle atrophy animal model. For four weeks, mice were orally administered with FO, GABA, lactate, or GABA+Lactate, and then Dexa was subcutaneously injected for ten days. During Dexa injection period, FO, GABA, lactate, or GABA+Lactate were also administered, and grip strength test and muscle harvesting were performed on the day of the last Dexa injection. We compared the attenuation effect of FO with GABA, lactate, and GABA+lactate treatment. Nrf2 and HO-1 expressions were increased by Dexa but decreased by FO; SOD activity and glutathione levels were decreased by Dexa but increased by FO; NADPH oxidase activity was increased by Dexa but decreased by FO; NF-κB, IL-6, and TNF-α activities were increased by Dexa were decreased by FO; Cbl-b expression was increased by Dexa but restored by FO; IGF-1 expression was decreased by Dexa but increased by FO; FoxO3, Atrogin-1, and MuRF1 expressions were increased by Dexa but decreased by FO. The gastrocnemius thickness and weight were decreased by Dexa but increased by FO. The cross-sectional area of muscle fiber and grip strength were decreased by Dexa but increased by FO. In conclusion, FO decreased Dexa-induced oxidative stress through the upregulation of Nrf2/HO-1. Decreased oxidative stress led to decreased Cbl-b, FoxO3, atrogin1, and MuRF1, which attenuated muscle atrophy.     

Bucillamine prevents cisplatin-induced ototoxicity through induction of glutathione and antioxidant genes

Bucillamine is used for the treatment of rheumatoid arthritis. This study investigated the protective effects of bucillamine against cisplatin-induced damage in auditory cells, the organ of Corti from postnatal rats (P2) and adult Balb/C mice. Cisplatin increases the catalytic activity of caspase-3 and caspase-8 proteases and the production of free radicals, which were significantly suppressed by pretreatment with bucillamine. Bucillamine induces the intranuclear translocation of Nrf2 and thereby increases the expression of γ-glutamylcysteine synthetase (γ-GCS) and glutathione synthetase (GSS), which further induces intracellular antioxidant glutathione (GSH), heme oxygenase 1 (HO-1) and superoxide dismutase 2 (SOD2). However, knockdown studies of HO-1 and SOD2 suggest that the protective effect of bucillamine against cisplatin is independent of the enzymatic activity of HO-1 and SOD. Furthermore, pretreatment with bucillamine protects sensory hair cells on organ of Corti explants from cisplatin-induced cytotoxicity concomitantly with inhibition of caspase-3 activation. The auditory-brainstem-evoked response of cisplatin-injected mice shows marked increases in hearing threshold shifts, which was markedly suppressed by pretreatment with bucillamine in vivo. Taken together, bucillamine protects sensory hair cells from cisplatin through a scavenging effect on itself, as well as the induction of intracellular GSH.     

Factors influencing glabridin stability

Glabridin, a polyphenolic isoflavan of Glycyrrhiza glabra, has shown a variety of pharmaceutical properties. We have previously studied the isolation of glabridin using macroporous resin and found that it is partially degraded, giving a dark color. To illustrate the degradation of glabridin, the present work studied the stability of glabridin under various conditions. Licorice extract containing about 20% glabridin, obtained from G. glabra by silica gel column chromatography, was used in the stability study. Seven different factors (temperature, illumination, humidity, pH, solvent, oxygen, and oxidant) were studied and content changes were determined through HPLC analysis. Except for oxygen, all the above factors had an effect on the stability of glabridin, with illumination being the main one. Moreover, the interactions between temperature and pH, temperature and humidity, and illumination and pH can promote the degradation of glabridin. In conclusion, we suggest that a dark, dry and airtight environment provides the optimized condition for the long-term storage of glabridin.     

RP-HPLC Method for the Quantitation of Glabridin in Yashti-madhu (Glycyrrhiza glabra)

A reverse phase high performance liquid chromatographic method is developed for the quantitation of glabridin in Glycyrrhiza glabra, using C₁₈ column with acetonitrile-water containing 2% AcOH (70:30) as an eluent. Glabridin is detected by UV absorption at 280 nm after separation by the chromatographic system. Good linearity was obtained in the working range of the concentration (0.01–0.1 mg mL^?1), with correlation coefficients 0.999. Limit of detection and limit of quantitation were 0.0195 and 0.065 mg mL^?1. The method was validated under ICH guidelines. The described method can be utilized for routine analysis (assays and stability tests) of G. glabra extracts and Ayurvedic medicine based on Yashti-madhu.     

Millettia ferruginea extract attenuates cisplatin-induced alterations in kidney functioning,DNA damage,oxidative stress,and renal tissue morphology

This study aimed to investigate the beneficial role of Millettia ferruginea extract (MF) in preventing cisplatin (Cisp) induced nephrotoxicity in rats. A total of 55 metabolites were identified using LC-MS analysis. The in vivo results indicated that MF pretreatment for 4 weeks (20 mg/kg b.w.) remarkably attenuated the altered renal biomarkers by decreasing the levels of plasma creatinine, urea, and uric acid when compared to the Cisp-group. The nephroprotective capacity of MF was further strengthened by histopathological observations, where Cisp + MF treated rats showed lower number of inflammatory cells and tubular degenerative changes than the Cisp-group. The harmful effects of cisplatin on renal oxidative stress indicators (MDA, SOD, CAT, and GPx), were restored by the treatment of MF. In addition, the reduction of inflammatory markers (IL-6 and TNF-α), associated with alleviating DNA fragmentation, highlighted the preventive effect of MF in kidney tissue. Additionally, MF components presented lower binding energies when docked into the active site of TNF-α and IL-6. The present findings concluded that M. ferruginea extract exhibited nephroprotective potential, which may be attributed to its antioxidant and anti-inflammatory properties. Further work is recommended to confirm the current results, explore the involved mechanism of action, and determine the therapeutic doses and time.     

Nephroprotective effects of 4-4(hydroxyl-3 methoxyphenyl)-2-butane against sodium tellurite induced acute kidney dysfunction by attenuating oxidative stress and inflammatory cytokines in rats

The aim of this study was to elucidate the protective action mechanism of 4-4(hydroxyl-3-methoxyphenyl)-2-butane against Sodium tellurite (ST) induced nephrotoxicity in rats. ST is a hazardous substance used in metallurgical and glassware industries, but its renal toxicities have not been well established before. Rats were distributed into four groups, six rats contain in each group. Normal control group given only vehicles only, toxic group given ST 8.5 mg/kg p o, treated groups given ST and 4-(hydroxyl-3-methoxyphenyl)-2-butane(100 mg/kg bwt), and positive control given only treatment drug 4-(hydroxyl-3-methoxyphenyl)-2-butane (100 mg/kg bwt) daily for 14 days. ST administration increases an alteration in biochemical, oxidative stress, cytokines markers, and morphological changes in toxic group. When it was treated with 4-(hydroxyl-3- methoxyphenyl)-2-butane significantly (p < 0.5) restores all these changes such as biochemical markers, antioxidant, inflammatory cytokines, and histopathological improvements in treated group as compared to toxic group. No significant (p > 0.05) changes have been seen in positive control as compared to normal control. In conclusion, 4(hydroxyl-3 methoxyphenyl)-2-butane successfully defended the kidney from oxidative stress, inflammatory cytokines and necrosis against ST intoxication. Thus, significant improvements were reflected and confirms with the improvement in histopathological changes.     

Enhancement of Protective Effects of <Emphasis Type="Italic">Radix Scutellariae</Emphasis> on UVB-induced Photo Damage in Human HaCaT Keratinocytes

Radix Scutellariae (RS) has long been used in the treatment of inflammatory and allergic diseases. Its main flavonoids, baicalin (BG) and wogonoside (WG), can be hydrolyzed into their corresponding aglycones, baicalein (B) and wogonin (W). In this study, we developed a safe and effective method of transforming these glycosides using Peclyve PR. The transformation rate of BG and WG reached 98.5 and 98.1%, respectively, with 10% enzyme at 40 °C for 60 h. Furthermore, we compared the anti-photoaging activity of RS before and after enzyme treatment, as well as their respective main components, in UVB-irradiated HaCaT cells. Results found that enzyme-treated RS (ERS) appeared to be much better at preventing UVB-induced photoaging than RS. ERS significantly inhibited the upregulation of matrix metalloproteinase-1 and IL-6 caused by UVB radiation by inactivating the MAPK/AP-1 and NF-κB/IκB-α signaling pathways. ERS treatment also recovered UVB-induced reduction of procollagen type I by activating the TGF-β/Smad pathway. In addition, ERS exhibited an excellent antioxidant activity, which could increase the expression of cytoprotective antioxidants such as HO-1 and NQ-O1, by facilitating Nrf2 nuclear transfer. These findings demonstrated that the photoprotective effects of RS were significantly improved by enzyme-modified biotransformation.     

New mechanistic insights on Justicia vahlii Roth: UPLC-Q-TOF-MS and GC–MS based metabolomics,in-vivo,in-silico toxicological,antioxidant based anti-inflammatory and enzyme inhibition evaluation

Justicia vahlii Roth. (acanthaceae) is an important medicinal food plant used in pain relief and topical inflammation. The present study aimed to evaluate phytochemical composition, toxicity, anti-inflammatory, antioxidant and enzyme inhibition potential of n-butanol extract of J. vahlii (BEJv). The extract prepared through maceration was found rich in total phenolic contents (TPC) 196.08 ± 6.01 mg of Gallic acid equivalent (mg GAE/g DE) and total flavonoid contents (TFC) 59.08 ± 1.32 mg of Rutin equivalent (mg RE/g DE). The UPLC-Q-TOF-MS analysis of BEJv showed tentative identification of 87 compounds and 19 compounds were detected in GC–MS analysis. The HPLC-PDA quantification showed the presence of 14 polyphenols amongst which kaempferol (3.45 ± 0.21 µg/ mL DE) and ferulic acid (2.31 ± 1.30 µg/ mL DE) were found in highest quantity. The acute oral toxicity study revealed the safety and biocompatibility of the extract up to 3000 mg/kg in mice. There was no effect of BEJv on human normal liver cells (HL 7702) and very low cytotoxic effect on liver cancer cells (HepG2) and breast cancer cells (MCF-7). In anti-inflammatory evaluation, the BEJv treated groups showed significant inhibition (p < 0.001) of late phase carrageenan induced paw edema at 400 mg/kg and increased the levels of oxidative stress markers; catalase, superoxide dismutase (SOD) and glutathione (GSH) while decreased the inflammatory markers; interleukin-1beta (IL-β) and tumor necrosis factor alpha (TNF-α) in paw tissue of mice. BEJv displayed highest results in Ferric reducing antioxidant power (FRAP) assay 97. 21 ± 2.34 mg TE (trolox equivalent)/g DE, and highest activity 3.32 ± 0.31 mmol ACAE (acarbose equivalent)/g D.E against α-glucosidase. Docking study showed good docking score by the tested compounds against the various clinically significant enzymes. Conclusively the current study unveiled J. vahlii as novel non-toxic source with good antioxidant-mediated anti-inflammatory potential which strongly back the traditional use of the species in pain and inflammation.     

Hibiscus sabdariffa synthesized gold nanoparticles ameliorate aluminum chloride induced memory deficits through inhibition of COX-2/BACE-1 mRNA expression in rats

Alzheimer’s disease (AD) is a major health challenge worldwide, especially among the elderly. The disease is associated with cognitive and memory deficits. This study investigated the effect of Hibiscus sabdariffa synthesized-gold nanoparticles (HS-AuNPs) on AlCl₃-induced memory deficits in rats. Forty-two male Wistar rats were divided into six groups (n = 7). Group I served as control. Rats in group II - V were exposed to AlCl₃ (100 mg/kg) to induce AD. Group III - V rats were treated with 5 mg/kg donepezil, 5 and 10 mg/kg HS-AuNPs, respectively, for 14 days. Behavioral tests were carried out on the rats on day 28 and 42. At the end of animal experiment, rats were sacrificed and used for various biochemical assays and gene expression. The AD rats showed memory and learning impairment, and these conditions were ameliorated by HS-AuNPs. Significant (p < 0.05) elevation in the activities of acetylcholinesterase, monoamine oxidase and adenosine deaminase, as well as malondialdehyde levels was noted. A significant reduction in the activities of superoxide dismutase (SOD), glutathione peroxidase (GPx) and reduced glutathione (GSH) noted in AlCl₃-induced rats were ameliorated by the 5 and 10 mg/kg b.w. doses of HS-AuNPs. In addition, the increased mRNA expression of cyclooxygenase-2 (COX-2) and beta-secretase 1 (BACE-1) caused by AlCl₃ were assuaged by the HS-AuNPs treatment. Based on the activities of HS-AuNPs against AlCl₃-induced AD, HS-AuNPs could be considered a potential therapeutic agent for managing AD.