Metabolomics and anti-inflammatory activity of Commiphora madagascariensis jacq. leaves extract using in vitro and in vivo models.

C. madagascariensis, an unexplored species of Burseraceae is used by local population for the management of inflammation and throat pain. The disease alleviation by this plant could be due to the presence of rich repository of active compounds with various pharmacological importances. In this study, therefore, the profiling of metabolites and isolation of active compounds of C. madagascariensis was performed. Furthermore, the ethanol, ethyl acetate extracts and a selected active compound was subjected for in vitro and in vivo anti-inflammatory activities. Metabolomic analysis identified and quantified 116 metabolites from leaves, young stem and gum-resins of C. madagascariensis (Burseraceae) followed by multivariate PCA analysis. NMR, GC-MS and HPLC were used to analyze primary and secondary metabolites. Subsequently, five main isolated compounds were identified as trimethoxy tetrahydrobenzo dioxolo isochromene (TTDI), butyl phenol, butyl propionate phenol, germacrone and ß-elemenone. Amongst them, TTDI was found to be a novel compound. Hence, a process was developed to obtain the enriched fraction of TTDI in ethanol and ethyl acetate extracts of leaves. Furthermore, TTDI and extracts were subjected for their in vitro anti-inflammatory activity in LPS sensitized murine splenocytes. The results showed that TTDI and both extracts significantly suppressed the levels of pro-inflammatorycytokines (TNF-α, IFN-γ). Interestingly, the suppression of pro-inflammatory cytokines was evenmore significant by the similar concentration of TTDI when compared with colchicine. However, the level of anti-inflammatory cytokine (IL-10) was found to be unchanged. Additionally, in vivo anti-inflammatory study revealed a significant reduction in carrageenan induced paw edema by TTDI and both the extracts. In the docking study, TTDI was more active than colchicine with strong binding affinity to COX-2, PLA2, and 5ß reductase. Our results highlighted that the presence of metabolites with medicinal and nutraceutical importance in C. madagascariensis, could provide opportunities for the development of a new plant-based therapeutics for inflammation.

Cinnamomum verum-derived bioactives-functionalized gold nanoparticles for prevention of obesity through gut microbiota reshaping.

Existing drugs have limited success in managing obesity in human due to their low efficacy and severe side-effects. Surface-modified gold nanoparticles have now received considerable attention of researchers for efficient biomedical applications owing to their superior uptake by cells, biocompatibility, hydrophilicity and non-immunogenicity. Here we prepared Cinnamomum verum derived bioactives-functionalized gold nanoparticles (Au@P-NPs) and assessed their impact on obesity and related immune-metabolic complications in high-fat diet (HFD)-induced obese mice using metabolic experiments along with 16S RNA gene-based gut microbial profiling and faecal microbiota transplantation (FMT). Au@P-NPs treatment prevented weight gain, decreased fat deposition, reduced metabolic inflammation and endotoxaemia in HFD-fed mice. Au@P-NPs-treated group exhibited better glucose tolerance and insulin sensitivity than HFD-fed control mice, and got completely protected against hepatic steatosis. These impacts were related to increased energy expenditure and enhanced Ucp1 expression in the brown adipose tissues of Au@P-NPs-administered animals, which strongly linked with the mRNA expression of the membrane bile acid receptor TGR5. Treatment of HFD-fed animals with Au@P-NPs altered plasma bile acid profile, and increased Akkermansia muciniphila and decreased Lactobacillus populations in the faeces. Au@P-NPs-treated animals revealed altered plasma bile acid profile, and increased Akkermansia muciniphila and decreased Lactobacillus populations in the faeces. FMT experiments showed lesser weight gain and greater energy expenditure in the mice fed with faecal suspension from Au@P-NPs-treated animals than that from HFD-fed mice. These results clearly establish that gold nanoparticles functionalized with bioactive compounds of C. verum have high potential to be an anti-obesity drug.

Nanocurcumin Potently Inhibits SARS-CoV-2 Spike Protein-Induced Cytokine Storm by Deactivation of MAPK/NF-κB Signaling in Epithelial Cells.

Interleukin-mediated deep cytokine storm, an aggressive inflammatory response to SARS-CoV-2 virus infection in COVID-19 patients, is correlated directly with lung injury, multi-organ failure, and poor prognosis of severe COVID-19 patients. Curcumin (CUR), a phenolic antioxidant compound obtained from turmeric (Curcuma longa L.), is well-known for its strong anti-inflammatory activity. However, its in vivo efficacy is constrained due to poor bioavailability. Herein, we report that CUR-encapsulated polysaccharide nanoparticles (CUR-PS-NPs) potently inhibit the release of cytokines, chemokines, and growth factors associated with damage of SARS-CoV-2 spike protein (CoV2-SP)-stimulated liver Huh7.5 and lung A549 epithelial cells. Treatment with CUR-PS-NPs effectively attenuated the interaction of ACE2 and CoV2-SP. The effects of CUR-PS-NPs were linked to reduced NF-κB/MAPK signaling which in turn decreased CoV2-SP-mediated phosphorylation of p38 MAPK, p42/44 MAPK, and p65/NF-κB as well as nuclear p65/NF-κB expression. The findings of the study strongly indicate that organic NPs of CUR can be used to control hyper-inflammatory responses and prevent lung and liver injuries associated with CoV2-SP-mediated cytokine storm.

ZnO/Curcumin Nanocomposites for Enhanced Inhibition of Pseudomonas aeruginosa Virulence via LasR-RhlR Quorum Sensing Systems.

The indiscriminate and excessive use of antibiotics has ultimately led to the emergence of bacterial resistant mutants or superbugs. These superbugs are difficult to control with conventional antibiotics. Disabling quorum sensing (QS), a population-density-dependent cell-to-cell communication process used by bacteria to coordinate the expression of virulence genes and biofilm formation, with dietary phytochemicals is emerging as a non-antibiotic strategy to inhibit bacterial pathogenicity. Although curcumin is an anti-QS agent and its delivery to cells has been a challenge due to poor bioavailability, ZnO/curcumin nanocomposites (ZnC-NCs) were fabricated with enhanced delivery of curcumin inside the bacterial superbug Pseudomonas aeruginosa PAO1 for effective inhibition of its QS and biofilm formation. Sustained release of curcumin from ZnC-NCs was observed where 51% curcumin at pH 7.2 and 83% curcumin at pH 5.5 were released within 48 h. ZnC-NCs also decreased the production of virulence factors and biofilm formation without affecting planktonic cell growth. Both LasR and RhlR QS systems were inhibited by ZnC-NCs. ZnC-NCs were also capable of protecting both mice as well as lung epithelial cells from killing by PAO1. The superoxide anions (O2·-) were also found as key players in suppressing PAO1 QS systems by ZnC-NCs. Overall, ZnC-NCs enhanced curcumin bioavailability for effective inhibition of QS signaling in P. aeruginosa via LasR-RhlR suppression and O2·- generation.

Gastroprotective effect of standardized leaf extract from Careya arborea on experimental gastric ulcers in rats.

CONTEXT: The leaf of Careya arborea Roxb. (Lecthidaceae) has been advocated in Ayurveda for the treatment of various disorders, including ulcers, healing of wounds and several skin diseases. OBJECTIVE: The 70% ethanol (EtOH) extract of C. arborea leaves (CALE) was investigated for its gastroprotective effect in different gastric ulcer models. MATERIALS AND METHODS: CALE (100, 200, and 400 mg/kg body weight) was administered orally, twice daily for 5 d, for preventing aspirin (ASP)-, EtOH-, pylorus ligation (PL)-, and cold restraint stress (CRS)-induced ulcer in rats. The status of the antioxidant enzymes in CRS-induced ulcers, H(+)K(+)ATPase activity, gastric wall mucous in EtOH-induced ulcer, and gastric secretion parameters were estimated in the PL-induced ulcer model. RESULTS: CALE exhibited significant (p < 0.01) dose-dependent inhibition of ulcer index in ASP 12.90-51.61%, EtOH 11.97-40.35%, PL 28.63-63.92%, and CRS 38.30-66.37%, respectively. A significant (p < 0.001) decrease occurred in the level of H(+)K(+)ATPase, volume of gastric juice, and acid output. Simultaneously, the level of gastric wall mucus was increased significantly (p < 0.05). The antioxidant enzyme levels of LPO and SOD were decreased with concomitant increase in catalase activity in CRS-induced ulcers. High-performance thin-layer chromatography (HPTLC) showed the presence of quercetin, ellagic acid, and gallic acid (0.31%, 0.24%, and 0.71% w/w, respectively) in CALE. CONCLUSIONS: Our results show that C. arborea possesses significant gastro-protective activity, probably due to its free radical scavenging activity, and validate the folklore claim.

Protective effect of standardized extract of Cleome viscosa against experimentally induced gastric lesions in the rat.

CONTEXT: Cleome viscosa Linn. (Capparidaceae) is used traditionally in the Indian system of medicine as a carminative, anthelmintic, and diuretic, and used for healing wounds, ulcers and diarrhea. OBJECTIVE: A 70% ethanol (EtOH) extract of the aerial parts of Cleome viscosa extract (CVE) was investigated for gastroprotective activity in different gastric ulcer models in order to validate ethnobotanical claims regarding the plant use in ulcers. MATERIALS AND METHODS: CVE (100, 200 and 400 mg/kg body weight) was administered orally, twice daily for 5 d, for prevention from EtOH, pylorus ligation (PL) and cold restraint stress (CRS)-induced ulcers in rats. Estimation of H(+)K(+)ATPase activity and gastric wall mucous were performed in EtOH-induced ulcer, antioxidant enzyme activities in supernatant mitochondrial fraction of CRS-induced ulcer, and gastric secretion parameters were estimated in PL-induced ulcer model. RESULTS: CVE showed significant (p < 0.01) dose-dependent inhibition of lesion index in EtOH 15.93-42.30%, PL 26.34-59.28% and CRS 22.58-54.03%, respectively. CVE prevents the oxidative damage of gastric mucosa by blocking lipid peroxidation and by a significant (p < 0.001) decrease in superoxide dismutase, and an increase in catalase activity. A significant (p < 0.01) decrease occurred in the level of H(+)K(+)ATPase, volume of gastric juice and total acidity. Simultaneously, the level of gastric wall mucus and pH were increased significantly (p < 0.05). High performance thin layer chromatography analysis showed the presence of quercetin and gallic acid (0.3% and 0.25% w/w, respectively) in CVE. CONCLUSIONS: Results of our study showed that C. viscosa possesses significant gastroprotective activity, probably due to free radical scavenging activity, and validates the folklore claim.

Evaluation of chemopreventive effect of Fumaria indica against N-nitrosodiethylamine and CCl4-induced hepatocellular carcinoma in Wistar rats.

OBJECTIVE: To investigation the chemopreventive potential of Fumaria indica (F. indica) extract (FIE) on N-nitrosodiethylamine and CCl(4)-induced hepatocarcinogenesis in Wistar rats. METHODS: The experimental animals were divided into six groups (n=6). Hepatocellular carcinoma was induced by single intraperitoneal injection of N-nitrosodiethylamine (NDEA) in normal saline at a dose of 200 mg/kg body weight followed by weekly subcutaneous injections of CCl(4)(3 mL/kg/week) for 6 weeks, as the promoter of carcinogenic effect. After administration of the carcinogen, 200 and 400 mg/kg of FIE were administered orally once a day throughout the study. At the end of 20 weeks, the body weight, liver weight and relative liver weight were measured. The percentage of nodule incidence and liver cancer markers such as aspartate transaminase (AST), alanine transaminase (ALT), alkaline phosphatase (ALP), γ-glutamyl transferase (γ-GT), total bilirubin level (TBL), α-feto protein (AFP) and carcinoembryonic antigen were estimated along with histopathological investigation in experimental groups of rats. RESULTS: Obtained results demonstrated that the cotreatment with FIE significantly prevented the decrease of the body weight and also increased in relative liver weight caused by NDEA. The treatment with FIE significantly reduced the nodule incidence and nodule multiplicity in the rats after NDEA administration. The levels of liver cancer markers such as AST, ALT, ALP, γ-glutamyl transferase, TBL, AFP and carcinoembryonic antigen were substantially increased by NDEA treatment. However, FIE treatment significantly reduced the liver injury and restored the entire liver cancer markers. Histological observations of liver tissues too correlated with the biochemical observations. CONCLUSIONS: These finding powerfully supports that F. indica exert chemopreventive effect by suppressing the tumor burden and restoring the activities of hepatic cancer marker enzymes on NDEA and CCl(4)-induced hepatocarcinogenesis in Wistar rats.

Gastroprotective effect of standardized leaf extract from Argyreia speciosa on experimental gastric ulcers in rats.

ETHNOPHARMACOLOGICAL RELEVANCE: Argyreia speciosa (L.f), Sweet (Family Convolvulaceae) is used traditionally in Indian System of Medicine as aphrodisiac, rejuvenating agent, intellect promoting agent, brain tonic and in the therapy of hepatomegaly, diabetes and chronic ulcer. AIM OF THE STUDY: To study the gastroprotective effect of standardized butanol fraction of Argyreia speciosa leaf (ASE). MATERIALS AND METHODS: The butanol fraction of Argyreia speciosa leaf (ASE; 50, 100 and 200mg/kg body weight) was administered orally, twice daily for 5 days for prevention from Aspirin (ASP)-, ethanol (EtOH)-, cold-restraint stress (CRS) - and pylorus ligation (PL)-induced ulcers. Estimation of antioxidant enzymes activity was carried out in CRS-induced ulcer model, and various gastric secretion parameters like volume of gastric juice, acid output, and pH value were estimated in PL-induced ulcer model. RESULT: ASE showed dose-dependent ulcer protective effect in ASP 23.64-58.76% (p<0.01 to p<0.001), EtOH 15.45-58.45% (p<0.001), CRS 19.39-78.36% (p<0.001) and PL 19.67-69.04% (p<0.05 to p<0.01), respectively. The percentage of protection by standard drug ranitidine was 77.77-84.32% (p<0.01 to p<0.001) in various gastric ulcer models. The gastric wall mucus was significantly (p<0.001) enhanced by ASE and is regarded as the first line of defence against EtOH-induced gastric ulcers showing cytoprotective property. ASE showed a marginal decrease in volume, acid pepsin concentration and acid pepsin output. However, ASE reduced the ulcer index with significant decrease in LPO level (p<0.001), and SOD level (p<0.01 to p<0.001) as compared with CRS-induced group. A gradual and significant increase in CAT values were observed at 100 and 200mg/kg dose levels (p<0.01 to p<0.001). CONCLUSIONS: The results of our study revealed that Argyreia speciosa possess significant dose dependent gastroprotective activity, probably due to its free radical scavenging activity.

In vitro and in vivo antioxidant properties and DNA damage protective activity of green fruit of Ficus glomerata.

This study evaluated the antioxidant properties of Ficus glomerata fruits using in vitro and in vivo assay. In order to find in vitro antioxidant properties, extract/fractions from F. glomerata were studied for TPC, AOA, RP, DPPH*, O2*-, *OH scavenging activities and LPO. Among all the extract/fractions, FEF has shown potent antioxidant activity and was also found effective in protecting oxidative DNA damage. The in vivo evaluation of oxidative stress (LPO) and antioxidant defenses (concentration of GSH, as well as CAT and SOD activities) were measured in CCl4 induced toxic rats. FEF was found to inhibit the toxicity as seen from the decreased LPO and increased GSH, SOD and CAT levels. FEF has higher phenolic content and showed the presence of gallic, chlorogenic and ellagic acid. Based on these results, it is concluded that F. glomerata protects tissues from oxidative stress and these effects are probably related to the antioxidant properties.

In vitro and in vivo antioxidant properties of different fractions of Moringa oleifera leaves.

The antioxidant potency of different fractions of Moringa oleifera leaves were investigated by employing various established in vitro systems, such as beta-Carotene bleaching, reducing power, DPPH/superoxide/hydroxyl radical scavenging, ferrous ion chelation and lipid peroxidation. On the basis of in vitro antioxidant properties polyphenolic fraction of M. oleifera leaves (MOEF) was chosen as the potent fraction and used for the DNA nicking and in vivo antioxidant properties. MOEF shows concentration dependent protection of oxidative DNA damage induced by HO() and also found to inhibit the toxicity produced by CCl(4) administration as seen from the decreased lipid peroxides (LPO) and increased glutathione (GSH) levels. Among the antioxidant enzymes superoxide dismutase (SOD) and catalase (CAT) levels were restored to almost normal levels compared to CCl(4) intoxicated rats. The HPLC analysis indicated the presence of phenolic acids (gallic, chlorogenic, ellagic and ferulic acid) and flavonoids (kaempferol, quercetin and rutin). Thus, it may be concluded that the MOEF possess high phenolic content and potent antioxidant properties, which may be mediated through direct trapping of the free radicals and also through metal chelation.

Hepatoprotective effects of rubiadin, a major constituent of Rubia cordifolia Linn.

The hepatoprotective effects of rubiadin, a major constituent isolated from Rubia cordifolia Linn., were evaluated against carbon tetrachloride (CCl4)-induced hepatic damage in rats. Rubiadin at a dose of 50, 100 and 200 mg/kg was administered orally once daily for 14 days. The substantially elevated serum enzymatic activities of serum glutamic oxaloacetic transaminase (SGOT), serum glutamate pyruvate transaminase (SGPT), serum alkaline phosphatase (SALP) and gamma-glutamyltransferase (gamma-GT) due to carbon tetrachloride treatment were dose dependently restored towards normalization. Meanwhile, the decreased activities of glutathione S-transferase and glutathione reductase were also restored towards normalization. In addition, rubiadin also significantly prevented the elevation of hepatic malondialdehyde formation and depletion of reduced glutathione content in the liver of CCl4 intoxicated rats in a dose dependent manner. Silymarin used as standard reference also exhibited significant hepatoprotective activity on post treatment against carbon tetrachloride induced hepatotoxicity in rats. The biochemical observations were supplemented with histopathological examination of rat liver sections. The results of this study strongly indicate that rubiadin has a potent hepatoprotective action against carbon tetrachloride induced hepatic damage in rats.