Unraveling Stereochemical Structure-Activity Relationships of Sesquiterpene Lactones for Inhibitory Effects on STAT3 Activation.

Sesquiterpene lactones, a class of natural compounds abundant in the Asteraceae family, have gained attention owing to their diverse biological activities, and particularly their anti-proliferative effects on human cancer cells. In this study, we systematically investigated the structure-activity relationship of ten sesquiterpene lactones with the aim of elucidating the structural determinants for the STAT3 inhibition governing their anti-proliferative effects. Our findings revealed a significant correlation between the STAT3 inhibitory activity and the anti-proliferative effects of sesquiterpene lactones in MDA-MB-231 breast cancer cell lines. Among the compounds tested, alantolactone and isoalantolactone emerged as the most potent STAT3 inhibitors, highlighting their potential as candidates for anticancer drug development. Through protein-ligand docking studies, we revealed the structural basis of STAT3 inhibition by sesquiterpene lactones, emphasizing the critical role of hydrogen-bonding interactions with key residues, including Arg609, Ser611, Glu612, and Ser613, in the SH2 domain of STAT3. Furthermore, our conformational analysis revealed the decisive role of the torsion angle within the geometry-optimized structures of sesquiterpene lactones in their STAT3 inhibitory activity (R=0.80, p<0.01). These findings not only provide preclinical evidence for sesquiterpene lactones as promising phytomedicines against diseases associated with abnormal STAT3 activation, but also highlight the importance of stereochemical aspects in their activity.

Cellular Target Proteome in Breast Cancer Cells of an Oplopane Sesquiterpenoid Isolated from Tussilago farfara.

Tussilago farfara is a traditional herbal medicine used to treat coughs, bronchitis, and asthma. Its bioactive compounds include sesquiterpenoids with anti-inflammatory, antiproliferative, neuroprotective, and other effects. Biochemical studies have highlighted the mechanisms of action, but the investigations of related molecular pathways have not specified direct molecular targets. Therefore, this study profiled cellular target proteins of a sesquiterpenoid isolated from T. farfara using quantitative chemical proteomics in MDA-MB-231 and MCF-7 human breast cancer cells. Compound 8, 7ß-(3'-ethyl-cis-crotonoyloxy)-1α-(2'-methyl butyryloxy)-3,14-dehydro-Z-notonipetranone, exhibited potent antiproliferative activity based on its α,ß-unsaturated carbonyl moiety, and its potential cellular target proteins were identified using a compound 8-based clickable probe. Among >200 identified proteins, 17 showed enrichment ratios of >3 in both cell lines, while recombinant 14-3-3 protein zeta and peroxiredoxin-1 were verified using isothermic calorimetry and their alkylation sites. Considering the interaction between the α,ß-unsaturated carbonyl moiety of compound 8 and cysteine residues of the proteins, peptides containing Cys25 and Cys94 of 14-3-3 protein zeta and Cys83 of peroxiredoxin-1 were significantly reduced by this sesquiterpene ester. Although the results did not elucidate the effects of compound 8 in breast cancer cells, identification of potential target proteins contributes to enhanced understanding of its antiproliferative and anti-inflammatory effects.

Igalan from Inula helenium (L.) suppresses the atopic dermatitis-like response in stimulated HaCaT keratinocytes via JAK/STAT3 signaling.

OBJECTIVE: This study aimed to evaluate the protective effect of igalan, a sesquiterpene lactone isolated from Inula helenium (L.), on inhibiting inflammation, regulating the epidermal differentiation gene expression, and reactive oxygen species scavenging in atopic dermatitis (AD)-like inflammatory keratinocytes. METHODS: HaCaT human keratinocytes were treated with igalan at indicated concentrations before being activated by a combination of TNF-α and IFN-γ or IL-4 representative for T-helper 1 and T-helper 2 cell cytokines, which are associated with AD pathogenesis. RESULTS: By inhibiting the NF-κB pathway as well as the STAT activation, igalan could downregulate several marker inflammatory genes in AD, such as TARC/CCL17, MDC/CCL22, and RANTES/CCL5. In contrast, igalan, acting as JAK inhibitor, could promote the mRNA expression levels of the genes FLG, LOR, KRT10, and DSC1, which encode for essential proteins responsible for keratinocyte differentiation, by inhibiting STAT3 signaling. Furthermore, igalan exerts its antioxidant effect through activating the Nrf2 pathway, triggering the expression of some enzymes that contribute to preventing intracellular ROS generation during inflammation. CONCLUSION: These findings indicate that igalan, via suppressing JAK/STAT3 signaling, could impair the production of pro-inflammatory chemokines and enhance expression levels of several genes involved in keratinocyte differentiation in AD-like stimulated keratinocytes.

Mucoprotective effects of Saikosaponin-A in 5-fluorouracil-induced intestinal mucositis in mice model.

5-Fluorouracil (5-FU)-induced intestinal mucositis (IM) is one of the most common oncological problem. It involves serious clinical consequences such as diarrhea, erythematous lesions of mucosa, and eventually development of ulcers accompanied by severe pain. The aim of the present study was to demonstrate the mucoprotective effects of Saikosaponin-A in 5-FU-induced intestinal mucositis in mice. Mucositis was induced in BALB/c mice by intraperitoneal injection of 5-FU (50 mg/kg/day) for three consecutive days and IM was assessed by both behavioral and histochemical analysis. While, Saikosaponin-A (1, 5, 10 mg/kg/day) was administered 1 h before 5-FU injection for consecutive seven days. Pre-treatment of Saikosaponin-A significantly ameliorated the severity of mucositis reflected as food intake, body weight loss, severity of diarrhea and mortality rate in a dose depended manner as compared to mice treated with 5-FU. Moreover, histopathological analysis furthered reinforced the mucoprotective potential of Saikosaponin-A against 5-FU-induced intestinal abnormalities referred as villus atrophy, mitotic crypt stem cells damage, inflammatory cells infiltration, vacuolization and edema. Furthermore, Saikosaponin-A administration strongly inhibited pro-inflammatory mediators (TNF-α, COX-2, IL-1ß and IL-6) and apoptotic markers (p-JNK, Casp-3). Saikosaponin-A pre-treatment significantly reduced the production of nitric oxide (NO) in intestinal tissue, inhibited acetic acid-induced Evans blue vascular permeability. The Siaikosaponin-A treatment markedly enhanced the anti-oxidants enzymes (Nrf2, HO-1, SOD, GSH, GST and Catalase), while decreased the oxidative stress markers i.e. Malonaldehde (MDA). Hence, these data suggest that Saikosaponin-A maybe a potential candidate for the treatment of chemotherapy-induced intestinal mucositis.

Anti-hyperalgesic properties of a flavanone derivative Poncirin in acute and chronic inflammatory pain models in mice.

BACKGROUND: Poncirin is flavanone derivative (isolated from Poncirus trifoliata) with known pharmacological activities such as anti-tumor, anti-osteoporotic, anti-inflammatory and anti-colitic. The present study aimed to explore the anti-allodynic and anti-hyperalgesic potentials of poncirin in murine models of inflammatory pain. METHODS: The analgesic potential of poncirin was evaluated in formalin-, acetic acid-, carrageenan- and Complete Freund's Adjuvant (CFA)-induced inflammatory pain models in mice. Anti-allodynic and anti-hyperalgesic activities were measured using Von Frey filaments, Randall Selitto, hotplate and cold acetone tests. The serum nitrite levels were determined using Griess reagent. The Quantitative Real-time PCR (qRT-PCR) was performed to assess the effect of poncirin on mRNA expression levels of inflammatory cytokines and anti-oxidant enzymes. RESULTS: Intraperitoneal administration of poncirin (30 mg/kg) markedly reduced the pain behavior in both acetic acid-induced visceral pain and formalin-induced tonic pain models used as preliminary screening tools. The poncirin (30 mg/kg) treatment considerably inhibited the mechanical hyperalgesia and allodynia as well as thermal hyperalgesia and cold allodynia. The qRT-PCR analysis showed noticeable inhibition of pro-inflammatory cytokines (mRNA expression levels of TNF-α, IL-1ß and IL-6) (p < 0.05) in poncirin treated group. Similarly, poncirin treatment also enhanced the mRNA expressions levels of anti-oxidant enzymes such as transcription factor such as nuclear factor (erythroid-derived 2)-like 2 (Nrf2) (p < 0.05), heme oxygenase (HO-1) (p < 0.05) and superoxide dismutase (SOD2) (p < 0.05). Chronic treatment of poncirin for 6 days did not confer any significant hepatic and renal toxicity. Furthermore, poncirin treatment did not altered the motor coordination and muscle strength in CFA-induced chronic inflammatory pain model. CONCLUSION: The present study demonstrated that poncirin treatment significantly reduced pain behaviors in all experimental models of inflammatory pain, suggesting the promising analgesic potential of poncirin in inflammatory pain conditions.

Anticancer Activity of Smallanthus sonchifolius Methanol Extract against Human Hepatocellular Carcinoma Cells.

Background: This research aimed to investigate the cytotoxicity of methanol extract of Smallanthus sonchifolius leaf (YLE) against a human hepatocellular carcinoma cell line (HepG2). This plant is currently used as a traditional herbal remedy in the treatment of liver diseases in some rural parts of Myanmar. Methods: The cytotoxic activity of the plant extract against the cancerous cell line was assessed using an MTT assay. YLE demonstrated a significant effect (IC50 = 58.2 ± 1.9 µg/mL) on anti-cancer activity, which was further investigated using various assays including an in vitro cell migration assay, a colony formation assay, cell cycle analysis, western blot analysis, and a ROS assay. The significance of the phytochemical constituents of YLE could be identified using LC/Q-TOF-MS techniques. Results: We putatively identified the active components in YLE, which were possibly melampolide-type sesquiterpenoids. YLE showed an inhibitory effect on HepG2 cell proliferation and cell migration. YLE also induced cell cycle arrest and necrosis in a dose-dependent manner. Additionally, YLE significantly suppressed ROS formation in HepG2 cells. Conclusions: These findings suggest that YLE is sufficient for application as a promising anti-liver drug in herbal medicine.

A Sesquiterpenoid from Farfarae Flos Induces Apoptosis of MDA-MB-231 Human Breast Cancer Cells through Inhibition of JAK-STAT3 Signaling.

Triple-negative breast cancers (TNBCs) are hard-to-treat breast tumors with poor prognosis, which need to be treated by chemotherapy. Signal transducer and activator of transcription 3 (STAT3) is a transcription factor involved in proliferation, metastasis, and invasion of cancer cells. Therefore, research on searching for promising compounds with metabolism that suppress phosphorylation or transcription of STAT3 in TNBC cells is important. Farfarae Flos is well known as a traditional medicine for treating inflammation. However, few studies have shown that sesquiterpenoids from Farfarae Flos have an anticancer effect. In this study, efficient separation methods and an MTT assay were conducted to isolate an anticancer compound from Farfarae Flos against TNBC MDA-MB-231 cells. Here, 7ß-(3-Ethyl-cis-crotonoyloxy)-1α-(2-methylbutyryloxy)-3,14-dehydro-Z-notonipetranone (ECN), a compound isolated from Farfarae Flos showed a potent cytotoxic effect on MDA-MB-231 cells. ECN inhibited JAK-STAT3 signaling and suppressed the expression of STAT3 target genes. In addition, ECN induced apoptosis through both extrinsic and intrinsic pathways. Furthermore, we investigated that ECN inhibited the growth of tumors by intraperitoneal administration in mice injected with MDA-MB-231 cells. Therefore, ECN can be an effective chemotherapeutic agent for breast cancer treatment.

Sesquiterpenoids from Tussilago farfara Flower Bud Extract for the Eco-Friendly Synthesis of Silver and Gold Nanoparticles Possessing Antibacterial and Anticancer Activities.

Sesquiterpenoids from the flower bud extract of Tussilago farfara were effectively utilized as a reducing agent for eco-friendly synthesis of silver and gold nanoparticles. The silver and gold nanoparticles had a characteristic surface plasmon resonance at 416 nm and 538 nm, respectively. Microscopic images revealed that both nanoparticles were spherical, and their size was measured to be 13.57 ± 3.26 nm for the silver nanoparticles and 18.20 ± 4.11 nm for the gold nanoparticles. The crystal structure was determined to be face-centered cubic by X-ray diffraction. Colloidal stability of the nanoparticle solution was retained in a full medium, which was used in the cell culture experiment. The antibacterial activity result demonstrated that the silver nanoparticles showed better activity (two- to four-fold enhancement) than the extract alone on both Gram-positive and Gram-negative bacteria. Interestingly, the highest antibacterial activity was obtained against vancomycin-resistant Enterococci Van-A type Enterococcus faecium. Cytotoxicity on cancer cell lines confirmed that gold nanoparticles were more cytotoxic than silver nanoparticles. The highest cytotoxicity was observed on human pancreas ductal adenocarcinoma cells. Therefore, both nanoparticles synthesized with the sesquiterpenoids from T. farfara flower bud extract can be applicable as drug delivery vehicles of anticancer or antibacterial agents for future nanomedicine applications.

Novel Galiellalactone Analogues Can Target STAT3 Phosphorylation and Cause Apoptosis in Triple-Negative Breast Cancer.

Aberrant activation of signal transducer and activator of transcription 3 (STAT3) has been documented in various malignancies including triple-negative breast cancers (TNBCs). The STAT3 transcription factor can regulate the different important hallmarks of tumor cells, and thus, targeting it can be a potential strategy for treating TNBC, for which only limited therapeutic options are available. In this study, we analyzed the possible effect of (-)-galiellalactone and its novel analogues, SG-1709 and SG-1721, and determined whether these agents exerted their antineoplastic effects by suppressing the STAT3 signaling pathway in TNBC cells. The two analogues, SG-1709 and SG-1721, inhibited both constitutive as well as inducible STAT3 phosphorylation at tyrosine 705 more effectively than (-)-galiellalactone, which indicates that the analogues are more potent STAT3 blockers. Moreover, SG-1721 not only inhibited nuclear translocation and DNA binding of STAT3 but also induced apoptosis, and decreased expression of diverse oncogenic proteins. Interestingly, SG-1721 also exhibited an enhanced apoptotic effect when combined with radiotherapy. Furthermore, in vivo administration of SG-1721 significantly attenuated breast xenograft tumor growth via decreasing levels of p-STAT3. Therefore, SG-1721 may be a promising candidate for further application as a pharmacological agent that can target STAT3 protein in treating TNBC.

Suppression of TRPV1 and P2Y nociceptors by honokiol isolated from Magnolia officinalis in 3rd degree burn mice by inhibiting inflammatory mediators.

Burn pain is one of the worst imaginable pain, associated with considerable morbidity and mortality worldwide. The management of pain made significant progress; however, more research is needed for burn pain. In the present study, the antinociceptive effect of honokiol extracted from Magnolia officinalis was assessed for 3 consecutive days. The third-degree burns were induced by the hot water method. The honokiol both by intraperitoneal (i.p) and intra plantar (i.pl) route and in combination with tramadol (i.p) was found to be effective in significantly reducing the mechanical allodynia, hyperalgesia, thermal hyperalgesia and paw edema. Honokiol also succeeded in reducing weight loss and spontaneous pain behavior in mice. Honokiol treatment both i.p and ipl decrease significantly the loss of total protein (3.3 and 3.4 g/dl of total protein) and albumin (2.2 and 2.6 g/dl of total albumin) respectively. It also significantly recovers the normal balance of blood electrolytes and normalizes blood profile. Effect of honokiol on cytokines and mRNA expression levels of TRPV1 and P2Y were also assessed. Honokiol significantly decreases the expression of TNF-α, IL-1ß and IL-6 and decreases expression level of TRPV1 and P2Y. Additionally, TRPV1 and P2Y proteins expression levels were also assessed by Western blot in paw skin tissue, sciatic nerve and spinal cord which were remarkably down-regulated by honokiol. Histological analysis of vehicle control and drug-treated paws were also performed through hematoxylin and eosin (H&E) staining which exhibited that honokiol significantly reduced the dermal layers distortion and inflammation associated with the burn. The antioxidant enzymes and nitric oxide (NO) were also determined through ELISA. Honokiol treatment also potentiates the expression of reduced glutathione and glutathione S-transferase, and catalase levels and reduced significantly the nitric oxide (NO) as compared to the burn-induced group. It can be concluded on the base of the results that honokiol has a significant analgesic activity through its action on cytokines and by downregulating TRPV1 and P2Y receptors. It also has a protective role against burn damage by upregulation of antioxidants.

Matrine ameliorates anxiety and depression-like behaviour by targeting hyperammonemia-induced neuroinflammation and oxidative stress in CCl4 model of liver injury.

Acute or chronic liver injury is associated with hyperammonemia which induced neuroinflammation and oxidative stress in the brain. Neuroinflammation, oxidative stress, reduced neurogenesis, and apoptosis are critical factors for the development of anxiety and depression. The present study was aimed to evaluate the anxiolytic and antidepressant properties of matrine against acute liver injury in the rodent model. Acute liver injury in mice was induced by administration of the acute hepatotoxic dose of carbon tetrachloride (CCl4) (1 ml/kg, i.p.). Pretreatment of mice with matrine (50 mg/kg i.p.) remarkably ameliorated CCl4-induced anxiety and depression-like behavior as evident from the results of open field test (OFT), elevated plus maze test (EPM), light-dark box test (LDB), forced swimming test (FST), and tail suspension test (TST). Moreover, matrine significantly inhibited CCl4-induced neuroinflammation in mice by reducing pro-inflammatory cytokines such as interleukins (IL-1ß, IL-6) and tumor necrosis factor-α (TNF-α) levels in the hippocampus (HC) and prefrontal cortex (PFC). CCl4-induced oxidative stress was reduced by matrine due to its potential to enhance the levels of reduced glutathione (GSH), catalase (CAT), glutathione-S-transferase (GST), and decreased the malondialdehyde (MDA), and nitrite level in the PFC and HC of mice brain. Matrine remarkably reduced the levels of corticosterone, ammonia, AST, ALT, and creatinine. Matrine pretreatment remarkably ameliorated CCl4-induced morphological liver injury. Acute pretreatment of matrine enhanced neurogenesis by increasing the number of GFAF (glial fibrillary acidic protein) positive astrocyte, BDNF (brain-derived neurotrophic factor), and VEGF (vascular endothelial growth factor) in the hippocampus of CCl4-treated mice. Pretreatment of matrine inhibited apoptosis and DNA damage in the hippocampus. The present data revealed that hyperammonemia produced due to liver injury induced oxidative stress, neuroinflammation, reduced neurogenesis and apoptosis in the hippocampus, thus, resulting in anxiety and depression. Taken together, the present results suggested that matrine has a significant antidepressant and anxiolytic effects through modulation of neuroinflammation, oxidative stress, reduced neurogenesis and apoptosis induced by CCl4 administration.

Igalan induces detoxifying enzymes mediated by the Nrf2 pathway in HepG2 cells.

Igalan is one of the sesquiterpene lactones found in Inula helenium L., which is used as the traditional medicine to treat inflammatory diseases. However, the pharmacological effects of igalan have not been characterized. In this study, we isolated igalan from I. helenium L. and evaluated the effects of igalan on signaling pathways and expression of target genes in HepG2 cells. Igalan activated the nuclear factor erythroid 2-related factor 2 (Nrf2) pathway by increasing the inactive form of GSK3ß, the phosphorylated form of AKT, and the nuclear accumulation of Nrf2. Thus, target genes of Nrf2 such as HO-1 and NQO1 increased in HepG2 cells. Moreover, igalan inhibited the tumor necrosis factor-α (TNF-α)-induced nuclear factor-κB activation and suppressed the expression of its target genes, including TNF-α, interleukin (IL)-6, and IL-8 in HepG2 cells. Our results indicate the potential of igalan as an activator of cellular defense mechanisms and a detoxifying agent.

Effect of 25-methoxy hispidol A isolated from Poncirus trifoliate against bacteria-induced anxiety and depression by targeting neuroinflammation, oxidative stress and apoptosis in mice.

Neuroinflammation, oxidative stress and apoptosis are implicated in the pathogenesis of neuropsychiatric diseases like anxiety and depression. 25-Methoxyhispidol A (25-MHA) is a triterpenoid isolated from the immature fruit of Poncirus trifoliate. Recently, its crude extracts have been shown to exhibit anti-bacterial and anti-inflammatory activities. The current study investigated the effect of 25-MHA (1, 5 and 10 mg/kg) against bacteria-induced anxiety and depression-like behaviors in mice. Mice were challenged intraperitoneally (i.p.) with LPS (0.83 mg/kg), S. aureus and E. coli after 30 min of 25-MHA treatment. 25-MHA (10 mg/kg) significantly mitigated the anxiety-like behavior as indicated by the results of elevated plus maze test, open field test, and light-dark box test. It also demonstrated the anti-depressant like effect by significantly reducing the immobility time in tail suspension test and forced swim test. The oxidative stress was reduced by pretreatment with 25-MHA, improving the antioxidant enzymes level such as glutathione, glutathione sulfo-transferase, and catalase. Similarly, 25-MHA attenuated the bacterial infection induced neuroinflammation by inhibiting the interleukin- 6 (IL-6), interleukin-1 beta (IL-1ß), and tumor necrosis factor-α (TNF-α) levels in prefrontal cortex and hippocampus regions. Pretreatment of 25-MHA also decreased the cortisol level and prevented changes in the thickness of the granular layer in the dentate gyrus. It also inhibited the DNA damage in hippocampus region as analyzed by comet assay. Hence, present results demonstrated that 25-MHA possesses anti-anxiety and anti-depressant activities due to the ability to reduce neuroinflammatory, oxidative stress and apoptosis induced by the administration of LPS, E. coli, and S. aureus.

Diadzein ameliorates 5-fluorouracil-induced intestinal mucositis by suppressing oxidative stress and inflammatory mediators in rodents.

5-Fluorouracil (5-FU) is one of the most commonly prescribed anti-cancer agent. However, its use is associated with several debilitating adverse effects such as intestinal mucositis (IM) and myelosuppression. Oxidative stress and inflammation are major contributors in the development of mucositis. Diadzein is known for its potent anti-inflammatory and anti-oxidative activities from decades. The present study focused on investigating the effects of diadzein on intestinal mucositis induced by 5-FU by mainly focusing on oxidative stress and inflammatory markers in mice. Mucositis was induced in mice by administration of 5-FU (50 mg/kg, i.p.), once daily for three days and diadzein (1, 5, 10 mg/kg) was administered once daily for seven days. Diadzein pretreatment was found to reduce the severity of mucosal injury in a dose-dependent manner. Diadzein significantly reversed weight loss, relieved diarrhea, and improved histopathological deformities associated with inflammation. Moreover, diadzein remarkably improved the intestinal wall histopathology by reducing inflammatory mediators infiltration and prevented suppression of antioxidants (glutathione, glutathione sulfo-transferase, and catalase) by 5-FU administration. Furthermore, nitrite production in intestinal tissue was reduced by diadzein consistent with the observed modulation of inflammatory markers. Additionally, diadzein also improved the amended microflora profile, by reducing the number of pathogenic bacteria and increasing the abundance of probiotics. Taken together, the behavioral, biochemical and histological outcomes of the present study demonstrates that diadzein has significant anti-mucositis properties in 5-FU induced mucositis model, and the attenuative potential of diadzein might be due to inhibition of oxidative stress and inflammatory mediators.

Ginsenosides from Korean Red Ginseng ameliorate lung inflammatory responses: inhibition of the MAPKs/NF-κB/c-Fos pathways.

BACKGROUND: Korean Red Ginseng (steamed and dried white ginseng, Panax ginseng Meyer) is well known for enhancing vital energy and immune capacity and for inhibiting cancer cell growth. Some clinical studies also demonstrated a therapeutic potential of ginseng extract for treating lung inflammatory disorders. This study was conducted to establish the therapeutic potential of ginseng saponins on the lung inflammatory response. METHODS: From Korean Red Ginseng, 11 ginsenosides (Rb1, Rb2, Rb3, Rc, Rd, Re, Rf, Rg1, Rg2, Rg3, and Rh2) were isolated. Their inhibitory potential and action mechanism were evaluated using a mouse model of lung inflammation, acute lung injury induced by intranasal lipopolysaccharide administration. Their anti-inflammatory activities were also examined in lung epithelial cell line (A549) and alveolar macrophage (MH-S). RESULTS: All ginsenosides orally administered at 20 mg/kg showed 11.5-51.6% reduction of total cell numbers in bronchoalveolar lavage fluid (BALF). Among the ginsenosides, Rc, Re, Rg1, and Rh2 exhibited significant inhibitory action by reducing total cell numbers in the BALF by 34.1-51.6% (n = 5). Particularly, Re showed strong and comparable inhibitory potency with that of dexamethasone, as judged by the number of infiltrated cells and histological observations. Re treatment clearly inhibited the activation of mitogen-activated protein kinases, nuclear factor-κB, and the c-Fos component in the lung tissue (n = 3). CONCLUSION: Certain ginsenosides inhibit lung inflammatory responses by interrupting these signaling molecules and they are potential therapeutics for inflammatory lung diseases.

Sesquiterpene lactones-enriched fraction of Inula helenium L. induces apoptosis through inhibition of signal transducers and activators of transcription 3 signaling pathway in MDA-MB-231 breast cancer cells.

Inula helenium L., commonly known as Elecampane, has been extensively used for many countries in the folk medicine. Its root is a rich source of sesquiterpene lactones, which possess various pharmacological activities. To develop the phytomedicine including sesquiterpene lactones, we prepared hexane fraction from I. helenium (HFIH) and examined the inhibitory effect of HFIH on signal transducers and activators of transcription 3 (STAT3) activation in human breast cancer MDA-MB-231 cells. Additionally, detailed chemical investigation was done to pinpoint the most active sesquiterpene lactones responsible for its anticancer activity. HFIH selectively suppressed STAT3 phosphorylation at tyrosine 705, not affecting its upstream kinases. HFIH downregulated the expression of STAT3 target genes including cyclin D1 , c-myc, and bcl-2 and induced caspase-mediated apoptosis. Moreover, sesquiterpene lactones of HFIH clearly suppressed STAT3 activation. The in vivo results further supported that HFIH inhibits the growth of human breast xenograft tumors. Our results suggest that HFIH possesses potential anticancer activity, which is mainly mediated through STAT3 signaling pathway. These findings provide the potential of HFIH as a promising phytomedicine for the treatment and prevention of triple-negative breast cancer.

Annona muricata Leaf Extract Triggered Intrinsic Apoptotic Pathway to Attenuate Cancerous Features of Triple Negative Breast Cancer MDA-MB-231 Cells.

Annona muricata L., known as graviola, is an evergreen plant of the tropical regions and is a rich source of natural products. Graviola has various biological activities, and it is best known for its anticancer activity. This study aimed to investigate the effects of crude graviola extract in vitro on breast cancer cells; in particular, we aimed to identify an agent against triple negative breast cancer (TNBC). We used the TNBC MDA-MB-231 cell line as the experimental model and the ER(+) non-TNBC MCF-7 breast cancer cell line as the control. We identified annonaceous acetogenins, including annonacin isomers, characteristic to this plant by using liquid chromatography tandem mass spectrometry (LC/MS/MS). We observed a significant decrease in the cell viability in both cell lines within 48 h, whereas impaired cell motility and invasiveness were observed only in the MDA-MB-231 cell line. While the MCF-7 cells showed an ER-dependent mechanism of apoptosis, the apoptosis of MDA-MB-231 cells was governed by an intrinsic apoptotic pathway triggered by graviola leaf extract (GLE).

Bioassay-guided isolation of cantharidin from blister beetles and its anticancer activity through inhibition of epidermal growth factor receptor-mediated STAT3 and Akt pathways.

Cantharidin is an active constituent of blister beetles (cantharides) which have traditionally been used for cancer treatment. Several studies have shown that cantharidin has a cytotoxic effect on various cancer cells. However, few studies have examined the effect of cantharidin on signal transducer and activator of transcription 3 (STAT3) signaling in cancer. In this study, we isolated cantharidin from cantharides by bioassay-guided fractionation and examined its inhibitory effect on STAT3 activation in human breast cancer MDA-MB-231 cells, expressing high level of phosphorylated STAT3. Cantharides were extracted with acetonitrile and separated into hexane, methylene chloride/acetonitrile, and water fractions. The methylene chloride/acetonitrile fraction was further separated into four fractions by preparative high-throughput high-performance liquid chromatography. Cantharidin was then isolated from the third fraction by countercurrent chromatography and structurally determined by comparing nuclear magnetic resonance and high-resolution mass spectrometry data. Cantharidin inhibited STAT3 tyrosine phosphorylation in MDA-MB-231 cells. Cantharidin suppressed epidermal growth factor (EGF)-induced STAT3 and PI3K/Akt signaling pathways through inhibition of EGF receptor phosphorylation. Moreover, cantharidin reduced cell proliferation and induced apoptosis with downregulation of STAT3 target genes, such as Bcl-2, COX-2, and cyclin D1. Taken together, this study provides evidence that cantharidin may be a potential therapeutic agent for triple-negative breast cancer by reducing EGFR-mediated STAT3 and Akt signaling pathways.

Highly stable gold nanoparticles green-synthesized by upcycling cartilage waste extract from yellow-nose skate (Dipturus chilensis) and evaluation of its cytotoxicity, haemocompatibility and antioxidant activity.

Skate (Dipturus chilensis) cartilage extract was utilized as a green reducing agent for the synthesis of spherical gold nanoparticles with an average size of 16.7 ± 0.2 nm. The gold nanoparticle solution showed a surface plasmon resonance at 543 nm with a wine-red colour. A strong X-ray diffraction pattern and clear lattice structure in high-resolution transmission electron microscopy indicated a face-centred cubic structure of the gold nanoparticles. The gold nanoparticles retained excellent colloidal stability. Gold nanoparticles showed strong antioxidant activity in terms of 2,2-diphenyl-1-picrylhydrazyl radical scavenging activity. In vitro cytotoxicity was observed for seven cancer cells assessed by the water-soluble tetrazolium assay. Among the seven cancer cells, the highest cytotoxicity was observed for MDA-MB-231 (human breast adenocarcinoma cell) followed by HeLa (human epithelial cervix adenocarcinoma cell) and lastly by HT-29 (human colorectal adenocarcinoma cell). Furthermore, gold nanoparticles showed excellent haemocompatibility, indicating the possibility of their use as a future nanomedicine. These results strongly suggest that gold nanoparticles green-synthesized by upcycling skate cartilage waste extract will be valuable carriers or vehicles for the delivery of drugs or bioactive molecules, such as anti-cancer agents, for the treatment of cancers.

Attenuation of inflammatory pain by puerarin in animal model of inflammation through inhibition of pro-inflammatory mediators.

In the current study, the puerarin was investigated for both acute Carrageenan and chronic CFA-induced inflammatory pain models. The Puerarin treatment significantly attenuated (P < 0.001) the mechanical hyperalgesia and mechanical allodynia in both Carrageenan and CFA-induced hyperalgesia. The Puerarin treatment also remarkably reduced (p < 0.001) the thermal hyperalgesic responses in both acute Carrageenan as well as chronic CFA-induced models. Furthermore, the Puerarin administration was also associated with significant inhibition of (p < 0.001) paw edema in both Carrageenan and CFA-induced models. The inflammatory mediators such as IL-1ß, IL-6, TNF-α and vascular endothelial growth factor (VEGF) are significantly enhanced during inflammatory conditions, however, the Puerarin administration significantly altered (P < 0.001) the mRNA expression levels of these mediators. Additionally, the Puerarin treatment also significantly enhanced (P < 0.001) the mRNA expressions levels of the anti-oxidant enzymes such as Nrf2, HO-1 and SOD2. The Puerarin treatment is associated with significant (P < 0.001) inhibition of the acetic acid-induced Evans blue vascular permeability. Moreover, the concentration of Puerarin in various tissues was analyzed using High-performance liquid chromatography (HPLC) and the results showed that the Puerarin was significantly distributed towards the peripheral tissues such as liver and kidney and less distributed towards the brain.