PMID: 

J BUON. 2019 Sep-Oct;24(5):1964-1969. PMID: 31786862

Abstract Title: 

Inhibition of cancer cell growth by oleanolic acid in multidrug resistant liver carcinoma is mediated via suppression of cancer cell migration and invasion, mitochondrial apoptosis, G2/M cell cycle arrest and deactivation of JNK/p38 signalling pathway.

Abstract: 

PURPOSE: Liver cancer accounts for considerable mortality across the globe. The sharp upsurge in the incidence of liver cancer, unavailability of standard treatments and the adverse side effects associated with the existing drugs has made it compulsory to explore novel and more effective anticancer molecules. In this study the anticancer effects of a natural compound oleanolic acid were investigated in vitro.METHODS: The human HepG2 liver cancer cells were treated with various concentrations of oleanolic acid for 24 h. The antiproliferative effects of oleanolic acid were measured by CCK8 cell viability assay. DAPI and annexin V/propidium iodide (PI) assays were employed to examine the induction of apoptosis. Transwell assay was performed to examine the cell migration and invasion. Expression analysis was performed by western blot analysis.RESULTS: The results showed that oleanolic acid decreased the viability of the liver cancer HepG2 cells and exhibited an IC50 of 30µM. The cytotoxicity of oleanolic acid was also investigated on the normal liver cells AML12 and it was found that oleanolic acid and exerted very low toxic effects on these cells and exhibited an IC50 of 120 µM. Oleanolic acid also caused remarkable changes in the morphology of the HepG2 cells and inhibited their colony formation potential. Flow cytometry indicated oleanolic acid triggered G2/M arrest of the liver HepG2 cancer cells. PI and DAPI staining revealed that oleanolic acid prompted apoptosis of the HepG2 cells. The apoptotic cells increased from 2.2% in control to around 35% at 30µM concentration. Oleanolic acid also suppressed the migration and invasion of the liver cancer cells via blocking of the JNK/p38 signalling pathway.CONCLUSIONS: The results of the current research revealed that oleanolic acid can be a molecule that may be utilised in the treatment of liver cancer in the future.

read more

PMID: 

Biomolecules. 2019 Nov 1 ;9(11). Epub 2019 Nov 1. PMID: 31683841

Abstract Title: 

Oleanolic Acid Exerts a Neuroprotective Effect Against Microglial Cell Activation by Modulating Cytokine Release and Antioxidant Defense Systems.

Abstract: 

Microglia respond to adverse stimuli in order to restore brain homeostasis and, upon activation, they release a number of inflammatory mediators. Chronic microglial overactivation is related to neuroinflammation in Alzheimer's disease. In this work, we show that oleanolic acid (OA), a natural triterpene present in food and medicinal plants, attenuates the activation of BV2 microglial cells induced by lipopolysaccharide (LPS). Cell pretreatment with OA inhibited the release of IL-1β, IL-6, TNF-α, and NO, which was associated with the downregulation of the expression of genes encoding for these cytokines and inducible nitric oxide synthase (iNOS), and the reinforcement of the endogenous antioxidant cell defense. These findings advocate considering OA as a novel neuroprotective agent to inhibit oxidative stress and inflammatory response in activated microglia associated with Alzheimer's disease.

read more

PMID: 

Biomolecules. 2019 Nov 3 ;9(11). Epub 2019 Nov 3. PMID: 31684190

Abstract Title: 

Extract Alleviates Methotrexate-Induced Renal Injury: New Impact on PI3K/Akt/mTOR Signaling and MALAT1 Expression.

Abstract: 

Renal injury induced by the chemotherapeutic agent methotrexate (MTX) is a serious adverse effect that has limited its use in the treatment of various clinical conditions. The antioxidant activity ofextract (GB) was reported to mitigate renal injury induced by MTX. Our research was conducted to examine the nephroprotective role of GB versus MTX-induced renal injury for the first time through its impact on the regulation of phosphatidylinositol 3-kinase/protein kinase B/ mammalian target of rapamycin (PI3K/Akt/mTOR) signaling together with the renal level of TGF-β mRNA and long non-coding RNA-metastasis-associated lung adenocarcinoma transcript-1 (MALAT1) expression. A group of adult rats was intraperitoneally (ip) injected with MTX 20 mg/kg as a single dose to induce kidney injury (MTX group). The other group of rats was orally administered with GB 60 mg/kg every day for 10 days (GB+ MTX group). The MTX increased the serum creatinine and urea levels, renal TGF-β mRNA and MALAT1 expression, in addition to dysregulation of the PI3K/Akt/mTOR signaling when compared with normal control rats that received saline only (NC group). Moreover, renal damage was reported histopathologically in the MTX group. The GB ameliorated the renal injury induced by MTX and reversed the changes of these biochemical analyses. The involvement of PI3K/Akt/mTOR signaling and downregulation of TGF-β mRNA and MALAT1 renal expressions were firstly reported in the nephroprotective molecular mechanism of GB versus MTX-induced renal injury.

read more

PMID: 

Prog Neuropsychopharmacol Biol Psychiatry. 2019 Nov 9 ;98:109776. Epub 2019 Nov 9. PMID: 31707092

Abstract Title: 

Caffeine prevents neurodegeneration and behavioral alterations in a mice model of agitated depression.

Abstract: 

Longitudinal and some experimental studies have showed the potential of caffeine to counteract some depressive behaviors and synaptic dysfunctions. In this study, we investigated the potential of caffeine in preventing behavioral outcomes, neurodegeneration and synaptic proteins alterations in a mice model of agitated depression by bilateral olfactory bulbectomy (OB). For this purpose, bulbectomized mice received caffeine (0.3 g/L and 1.0 g/L, drinking water), during the active cycle, for seven weeks (two before the surgery and throughout five weeks after OB). Caffeine prevented OB-induced hyperactivity and recognition memory impairment and rescue self care and motivational behavior. In the frontal cortex, bulbectomized mice presented increase in the adenosine Areceptors (AR) and GFAP, while adenosine Areceptors (AR) increased in the hippocampus and striatum and SNAP-25 was decreased in frontal cortex and striatum. Caffeine increased AR in the striatum of bulbectomized mice and in SHAM-water group caffeine increased AR in the striatum and decreased SNAP-25 in the frontal cortex. Astrogliosis observed in the polymorphic layer of the dentate gyrus of OB mice was prevented by caffeine as well as the neurodegeneration in the striatum and piriform cortex. Based on these behavioral and neurochemical evidences, caffeine confirms its efficacy in preventing neurodegeneration associated with memory impairment and may be considered as a promising therapeutic tool in the prophylaxis and/or treatment of depression.

read more

PMID: 

Biomolecules. 2019 Nov 8 ;9(11). Epub 2019 Nov 8. PMID: 31717470

Abstract Title: 

Caffeine May Abrogate LPS-Induced Oxidative Stress and Neuroinflammation by Regulating Nrf2/TLR4 in Adult Mouse Brains.

Abstract: 

Herein, we assayed the antioxidant and anti-inflammatory potential of caffeine in a lipopolysaccharide (LPS)-injected mouse model of neurodegeneration and synaptic impairment. For this purpose, LPS was injected for two weeks on an alternate-day basis (250µg/kg/i.p. for a total of seven doses), while caffeine was injected daily for four weeks (30 mg/kg/i.p/four weeks). According to our findings, there was a significant increase in the level of reactive oxygen species (ROS), as evaluated from the levels of lipid peroxidation (LPO) and ROS assays. Also, we evaluated the expression of nuclear factor erythroid-2-related factor 2 (Nrf2) and the enzyme hemeoxygenase 1 (HO-1) in the mouse groups and found reduced expression of Nrf2 and HO-1 in the LPS-treated mice brains, but they were markedly upregulated in the LPS + caffeine co-treated group. We also noted enhanced expression of toll-Like Receptor 4 (TLR4), phospho-nuclear factor kappa B (p-NF-kB), and phospho-c-Jun n-terminal kinase (p-JNK) in the LPS-treated mice brains, which was significantly reduced in the LPS + caffeine co-treated group. Moreover, we found enhanced expression of Bcl2-associated X, apoptosis regulator (Bax), and cleaved caspase-3, and reduced expression of B-cell lymphoma 2 (Bcl-2) in the LPS-treated group, which were markedly reversed in the LPS + caffeine co-treated group. Furthermore, we analyzed the expression of synaptic proteins in the treated groups and found a marked reduction in the expression of synaptic markers in the LPS-treated group; these were significantly upregulated in the LPS + caffeine co-treated group. In summary, we conclude that caffeine may inhibit LPS-induced oxidative stress, neuroinflammation, and synaptic dysfunction.

read more

PMID: 

Clin Cancer Res. 2003 Jul ;9(7):2866-75. PMID: 12855667

Abstract Title: 

Betulinic acid-induced programmed cell death in human melanoma cells involves mitogen-activated protein kinase activation.

Abstract: 

Betulinic acid, a naturally occurring triterpene found in the bark of the white birch tree, has been demonstrated to induce programmed cell death with melanoma and certain neuroectodermal tumor cells. We demonstrate currently that treatment of cultured UISO-Mel-1 (human melanoma cells) with betulinic acid leads to the activation of p38 and stress activated protein kinase/c-Jun NH(2)-terminal kinase [widely accepted proapoptotic mitogen-activated protein kinases (MAPKs)] with no change in the phosphorylation of extracellular signal-regulated kinases (antiapoptotic MAPK). Moreover, these results support a link between the MAPKs and reactive oxygen species (ROS). As demonstrated previously, cells treated with betulinic acid generate ROS. Preincubation of cells with antioxidants blocks the process of programmed cell death, and prevents the phosphorylation of p38 and stress activated protein kinase/c-Jun NH(2)-terminal kinase. These data suggest that ROS act upstream of the MAPKs in the signaling pathway of betulinic acid. In addition to mediating these responses, treatment of cells with betulinic acid resulted in a gradual depolarization of mitochondrial membrane potential, a phenomenon established to contribute to the induction of programmed cell death. Interestingly, p38 was capable of partially modulating this perturbation, and investigations of mitochondria-associated apoptotic events indicate no involvement of known caspases. These data provide additional insight in regard to the mechanism by which betulinic acid induces programmed cell death in cultured human melanoma cells, and it likely that similar responses contribute to the antitumor effect mediated with human melanoma carried in athymic mice.

read more

PMID: 

Cancer Res. 2007 Mar 15 ;67(6):2816-23. PMID: 17363604

Abstract Title: 

Betulinic acid inhibits prostate cancer growth through inhibition of specificity protein transcription factors.

Abstract: 

Betulinic acid is a pentacyclic triterpene natural product initially identified as a melanoma-specific cytotoxic agent that exhibits low toxicity in animal models. Subsequent studies show that betulinic acid induces apoptosis and antiangiogenic responses in tumors derived from multiple tissues; however, the underlying mechanism of action is unknown. Using LNCaP prostate cancer cells as a model, we now show that betulinic acid decreases expression of vascular endothelial growth (VEGF) and the antiapoptotic protein survivin. The mechanism of these betulinic acid-induced antiangiogenic and proapoptotic responses in both LNCaP cells and in tumors is due to activation of selective proteasome-dependent degradation of the transcription factors specificity protein 1 (Sp1), Sp3, and Sp4, which regulate VEGF and survivin expression. Thus, betulinic acid acts as a novel anticancer agent through targeted degradation of Sp proteins that are highly overexpressed in tumors.

read more

PMID: 

J Vet Sci. 2014 ;15(1):141-8. Epub 2013 Dec 27. PMID: 24378582

Abstract Title: 

Betulinic acid prevents alcohol-induced liver damage by improving the antioxidant system in mice.

Abstract: 

Betulinic acid (BA), a pentacyclic lupane-type triterpene, has a wide range of bioactivities. The main objective of this work was to evaluate the hepatoprotective activity of BA and the potential mechanism underlying the ability of this compound to prevent liver damage induced by alcohol in vivo. Mice were given oral doses of BA (0.25, 0.5, and 1.0 mg/kg) daily for 14 days, and induced liver injury by feeding 50% alcohol orally at the dosage of 10 ml/kg after 1 h last administration of BA. BA pretreatment significantly reduced the serum levels of alanine transaminase, aspartate transaminase, total cholesterol, and triacylglycerides in a dose-dependent manner in the mice administered alcohol. Hepatic levels of glutathione, superoxide dismutase, glutathione peroxidase, and catalase were remarkably increased, while malondialdehyde contents and microvesicular steatosis in the liver were decreased by BA in a dose-dependent manner after alcohol-induced liver injury. These findings suggest that the mechanism underlying the hepatoprotective effects of BA might be due to increased antioxidant capacity, mainly through improvement of the tissue redox system, maintenance of the antioxidant system, and decreased lipid peroxidation in the liver.

read more

PMID: 

Nat Prod Commun. 2016 Dec ;11(12):1855-1858. PMID: 30508350

Abstract Title: 

Caffeic Acid Derivatives from Bacopa monniera Plants as Inhibitors of Pancreatic Lipase Activity and their Structural Requirements.

Abstract: 

The methanol extract of whole Bacopa monniera plants inhibited pancreatic lipase activity in vitro. From this extract we have reported the isolation of 11 triterpene glycosides and 5 phenylethanoid- and/or phenylpropanoid- glycosides. In this paper, we describe the effects of the methanol extract and/or its constituents on pancreatic lipase activity and the isolation of an active constituent, desrhamnosyl isoacteoside. In addition, the structural requirements for its inhibitory effects were examined. We also examined the effects on the elevation of plasma triglyceride (TG) levels in olive oil loaded mice. The major active constituents, desrhamnosyl isoacteoside and plantainoside B, reduced plasma TG levels in the mice. The inhibitory effects of B. monniera and its constituents on pancreatic lipase activity and plasma TG level are reported for the first time.

read more

PMID: 

Trials. 2019 Jun 10 ;20(1):345. Epub 2019 Jun 10. PMID: 31182153

Abstract Title: 

Efficacy of Cognition Support Formula® on cognitive function in older adults with subjective cognitive impairment: a protocol for a 26-week, randomised, double-blind, placebo-controlled trial.

Abstract: 

BACKGROUND: Due to an ageing population in Australia there has been an increase in the number of older adults with subjective cognitive impairment (SCI), a self-reported decline in cognitive function associated with an increased risk of mild cognitive impairment and dementia. There is no current, recommended treatment for SCI; therefore, the effectiveness of a supplement approved by the Therapeutic Goods Association that has the potential to enhance cognitive function in an at-risk cohort should be tested. The primary aim of this proposed research is to determine the efficacy of 6 months of treatment with BioCeuticals Cognition Support Formula® (containing Bacopa monniera (brahmi), Ginkgo biloba, Panax ginseng and alpha-lipoic acid) on cognition in older adults with SCI (utilising the CogState® one card learning and identification tests as co-primary outcome measures ofvisual short-term memory and attention; mean speed (ms), accuracy (%), and total number of hits, misses, and anticipations) compared with placebo. The secondary aims are to assess an improvement in other cognitive domains (executive functioning, processing speed, and working memory), evaluate safety, adverse effects, and determine efficacy on mood, fatigue, and neurocognition. It is expected that improvements across the study timepoints in the co-primary outcomes in the active treatment group (compared with placebo) will be evident.METHOD: One-hundred and twenty participants will be recruited for the randomised, double-blind, placebo-controlled study. Participants will be randomly assigned to one of the treatment groups (active or placebo) at a 1:1 ratio, and will be required to complete a series of cognitive (using CogState®), mood (using the Depression, Anxiety, Stress Scale (DASS-42) and Short Health Anxiety Inventory (SHAI)), and fatigue (using the Functional Assessment of Chronic Illness Therapy Fatigue Scale (FACIT-F)) tasks at baseline (0 months), the midpoint (3 months), and the endpoint (6 months). These tasks will be evaluated between timepoints (baseline vs. midpoint, midpoint vs. endpoint, and baseline vs. endpoint). Neurocognition will be measured by electroencephalography at baseline and at the endpoint in half of the participants. Adverse effects will be documented over the 6-month trial period.DISCUSSION: This is the first study to test the efficacy of Cognition Support Formula® on cognition in older adults with SCI. As people with SCI have an increased risk of dementia, and there are limited treatments options for this population, it is important to assess a supplement that has the potential to enhance cognitive function.TRIAL REGISTRATION: Universal Trial Number (UTN), U1111-1196-9548. Australian New Zealand Clinical Trials Registry, ACTRN12617000945325 . Registered on 30 June 2017.

read more