Effects Of Oleanolic Acid Research Articles

인간 기도 상피세포에서 IKK/IκBα/NF-κB p65 신호전달 경로를 경유한 호흡기 MUC5AC 뮤신 유전자의 발현에 미치는 올레아놀산의 영향

In the present study, we investigated whether oleanolic acid, an anti-inflammatory natural product, significantly affects the phorbol 12-myristate 13-acetate (PMA)-induced gene expression of airway MUC5AC mucin. Confluent NCIH292 cells were pretreated with oleanolic acid for 30 min and then stimulated with PMA for 24 h or the indicated periods. The MUC5AC mucin mRNA expression was measured by RT-PCR. Production of MUC5AC mucin protein was measured by ELISA. To elucidate the action mechanism of oleanolic acid, effect of oleanolic acid on PMA-induced NF-B signaling pathway was investigated by western blot analysis. The results were as follows: (1) Oleanolic acid significantly inhibited the production of MUC5AC mucin protein and down-regulated the expression of MUC5AC mucin mRNA; (2) Oleanolic acid inhibited PMA-induced activation (phosphorylation) of inhibitory kappa B kinase (IKK), and thus phosphorylation and degradation of inhibitory kappa Ba (IB); (3) Oleanolic acid inhibited PMA-induced nuclear translocation of nuclear factor kappa B (NF-B) p65, led to the down-regulation of MUC5AC mucin gene expression in NCI-H292 cells. These results suggest that oleanolic acid can regulate the gene expression of mucin by acting on airway epithelial cells through regulation of NF-B signaling pathway.

Effects of oleanolic acid on acute liver injury triggered by lipopolysaccharide in broiler chickens

ABSTRACT 1. Infectious injury caused by lipopolysaccharide (LPS), a metabolite of gram-negative bacteria, can induce stress responses in animals and is a significant cause of morbidity and mortality in young birds. The purpose of this study was to investigate the effects of dietary supplementation with oleanolic acid (OA) on acute liver injury in broiler chickens challenged with LPS. 2. In total, 120 broiler chickens were randomly divided into six groups and fed a basal diet containing 0, 50, 100, or 200 mg/kg OA or 100 mg/kg aureomycin. On d 15, broiler chickens were injected with either LPS or an equivalent volume of normal saline. Six hours after LPS injection, two broiler chicks were randomly selected for sampling in each replicate. 3. The results indicated that dietary aureomycin was ineffective in alleviating LSP-associated liver injury, but protected broiler chickens from LPS-induced liver damage. This promoted a significant reduction in the levels of malondialdehyde and an increase in the levels of superoxide dismutase in liver. In addition, OA was found to cause significant reductions in the relative expression of IL-1β, IL-6, and TNF-α in broiler liver tissues, whereas the relative expression of IL-10 was significantly increased. 4. In conclusion, oleanolic acid can alleviate oxidative stress and injury in the livers of broiler chickens induced by lipopolysaccharide. Consequently, oleanolic acid has potential utility as a novel anti-inflammatory and antioxidant feed additive.

Effects of oleanolic acid and ursolic acid on depression-like behaviors induced by maternal separation in mice

Oleanolic acid (OA) and ursolic acid (UA) are structural isomeric triterpenoids. Both triterpenoids have been reported to be able to improve depression. However, no studies have compared their effects in the same system. Whether OA or UA could ameliorate depression-like behaviors in maternal separation (MS)-induced depression-like model was investigated. MS model is a well-accepted mouse model that can reflect the phenotype and pathogenesis of depression. Depression is a mental illness caused by neuroinflammation or changes in neuroplasticity in certain brain regions, such as the prefrontal cortex and hippocampus. Depression-like behaviors were measured using splash test or forced swimming test. In addition, anxiety-like behaviors were also measured using the open field test or elevated plus-maze test. MS-treated female mice showed greater depression-like behaviors than male mice, and that OA improved several depression-like behaviors, whereas UA only relieved anxiety-like behavior of MS-treated mice. Microglial activation, expression levels of TNF-α, and mRNA levels of IDO1 were increased in the hippocampi of MS-treated female mice. However, OA and UA treatments attenuated such increases. In addition, expression levels of synaptophysin and PSD-95 were decreased in the hippocampi of MS-treated female mice. These decreased expression levels of synaptophysin were reversed by both OA and UA treatments, although decreased PSD-95 expression levels were only reversed by OA treatment. Our findings suggest that MS cause depression-like behaviors through female-specific neuroinflammation, changes of tryptophan metabolism, and alterations of synaptic plasticity. Our findings also suggest that OA could reverse MS-induced depression-like behaviors more effectively than UA.

Effects of oleanolic acid on hair growth in mouse dorsal skin mediated via regulation of inflammatory cytokines.

Oleanolic acid (OA) is a pentacyclic triterpenoid with favourable physiological activity. It is widely distributed in more than 200 species of plants. OA has garnered significant interest because of its potential biological activities, such as antioxidant, bacteriostatic, and hair growth-promoting effects. To study the effect of OA on hair growth and related mechanisms, we investigated hair growth in mice with testosterone-induced androgenetic alopecia (AGA) that were treated with three different concentrations of OA. The antioxidant, bacteriostatic, and cytotoxic effects of OA were evaluated. We found that mice with testosterone-induced AGA treated with 1% or 0.5% OA showed significantly enhanced hair growth and increased vascular endothelial growth factor/glyceraldehyde-3-phosphate dehydrogenase ratio and levels of fibroblast growth factor receptor and insulin-like growth factor 1. Using an immunofluorescence staining assay, we demonstrated that β-catenin, a key Wnt signalling transducer, was highly expressed in the OA-treated groups. These results suggest that OA may promote hair growth by stimulating hair matrix cell proliferation via the Wnt/β-catenin pathway and lowering the levels of tumour necrosis factor-alpha, and transforming growth factor-beta 1, dihydrotestosterone, and 5α-reductase.

Effects of Oleanolic Acid on Myocardial Injury in Diabetic Mice by Regulating Phosphatidylinositol 3 Kinase/Protein Kinase B/Glucose Transporter Type 4 Signalling Pathway

One of the leading causes of diabetes-related deaths is myocardial damage, which may be the cause of heart failure in people with type 2 diabetes mellitus. The objective of the research was to look at the impact of coronary failure with kind a pair of type 2 diabetes mellitus. This project aimed to explore the protective impact of oleanolic acid on myocardial damage in type 2 diabetes mellitus and to investigate the connected mechanism. Specific pathogen free grade db/db male mice were elected as model, while the Db/m mice were opted for control. Different doses of drug intervention were performed and the general condition, cardiac function, blood glucose, blood lipids, degree of myocardial injury, and degree of oxidative stress were examined by morphological examination of myocardial tissues, biochemical examination, and gene and protein amount detection. The results showed that lactate dehydrogenase, creatine kinase isoenzyme, total cholesterol, triglycerides and malondialdehyde levels in serum of Db/db mice were increased, while phosphatidylinositol 3 kinase, protein kinase B, glucose transporter 4 expression and superoxide dismutase level in myocardial tissue were decreased. After using oleanolic acid, the serum concentrations of lactate dehydrogenase, creatine kinase isoenzyme, total cholesterol, triglycerides and malondialdehyde were declined in Db/db mice, while phosphatidylinositol 3 kinase, protein kinase B, glucose transporter 4 expression and superoxide dismutase level were rised. The results recommend that oleanolic acid has protective impact on cardiac muscle injury in Db/db mice and conjointly the mechanism could also be correlated with promoting the activation of phosphatidylinositol 3 kinase/protein kinase B/glucose transporter 4 signal transduction.

Pentacyclic triterpenoid ursolic acid induces apoptosis with mitochondrial dysfunction in adult T-cell leukemia MT-4 cells to promote surrounding cell growth.

We investigated the antitumor effects of oleanolic acid (OA) and ursolic acid (UA) on adult T-cell leukemia cells. OA and UA dose-dependently inhibited the proliferation of adult T-cell leukemia cells. UA-treated cells showed caspase 3/7 and caspase 9 activation. PARP cleavage was detected in UA-treated MT-4 cells. Activation of mTOR and PDK-1 was inhibited by UA. Autophagosomes were detected in MT-4 cells after UA treatment using electron microscopy. Consistently, mitophagy was observed in OA- and UA-treated MT-4 cells by confocal microscopy. The mitochondrial membrane potential in MT-4 cells considerably decreased, and mitochondrial respiration and aerobic glycolysis were significantly reduced following UA treatment. Furthermore, MT-1 and MT-4 cells were sorted into two regions based on their mitochondrial membrane potential. UA-treated MT-4 cells from both regions showed high activation of caspase 3/7, which were inhibited by Z-vad. Interestingly, MT-4 cells cocultured with sorted UA-treated cells showed enhanced proliferation. Finally, UA induced cell death and ex vivo PARP cleavage in peripheral blood mononuclear cells from patients with adult T-cell leukemia. Therefore, UA-treated MT-4 cells show caspase activation following mitochondrial dysfunction and may produce survival signals to the surrounding cells.

OLEANOLIC ACID, A PROSPECTIVE PROTECTIVE AGENT AGAINST BRAIN ENERGY METABOLISM AND OXIDATIVE DYSFUNCTIONS FOLLOWING HEXAVALENT CHROMIUM EXPOSURE IN MICE

Objective: Effect of oleanolic acid against hexavalent chromium-induced altered brain energy metabolism associated with oxidative stress was evaluated in the present study. Methods: Swiss albino mice were divided into three groups, Control (n=6), chromium-treated (n=6), and oleanolic acid (OA) supplemented (n=6). The chromium treated group was orally administered with K2Cr2O7 for 30 days at a dose of 10 mg/kg b.w/day. OA supplementation was given at a dose of 5 mg/kg bw/day for the past 14 days of chromium treatment. Control group received the vehicle only. After the treatment, whole brain was removed for examining the parameters such as pyruvic acid, free amino nitrogen, tissue protein, TCA cycle enzyme activities, NADH dehydrogenase function, and oxidative stress markers. Results: Significant decrease in cerebral pyruvic acid content associated with suppressed malate dehydrogenase and succinate dehydrogenase activities were observed. The NADH dehydrogenase activity was inhibited owing to enhanced accumulation of chromium in cerebral tissue. Depletion of proteins and increased free amino acid nitrogen were accompanied with inhibited cathepsin, pronase and trypsin activities, and increased transaminase function. In addition, GSH content was decreased along with increased lipid peroxidation, oxidized GSSG content, TG/GSSG ratio, carbonylated protein content, and tissue free hydroxyl radical formation. Superoxide dismutase, catalase, glutathione reductase, and glutathione peroxidase were also inhibited by hexavalent chromium. Oleanolic acid supplementation was found to have significant protective effect against brain metabolic and oxidative dysfunctions. Conclusion: The present study elucidated therapeutic efficacy of oleanolic acid against hexavalent chromium toxicity in brain tissue of mice.

Oleanolik asidin çekal ligasyon ve punksiyon ile sepsis oluşturulan ratlarda kalp hasarı üzerine etkileri

Bu çalışmada zeytin ağacı (Olea europeae) ağacının yaprağından saflaştırılan oleanolik asidin (OEA) ratlarda çekal ligasyon ve punksiyonla (CLP) indüklenen sepisisin oluşturduğu kalp hasarı üzerine etkileri incelendi. Her gruba eşit olacak şekilde 30 adet Wistar erkek rat sham, sepsis, OEA-150, OEA-300 ve CEFT olarak 5 gruba ayrıldı. Kromatografik metotlarla zeytin yaprağından saflaştırılan oleanolik asit OEA gruplarına 150 ve 300 mg/kg dozlarda oral yolla uygulandı. CEFT grubuna seftriakson 150 mg/kg dozda intraperitoneal yolla uygulandı. Uygulamadan 24 saat sonra ratlardan kan serumu ve kalp dokuları, histopatolojik, immünohistokimyasal ve biyokimyasal analizler için alındı. Yapılan histopatolojik ve immünohistokimyasal incelemede, sepsis grubunun doku tümör nekrozis faktör (TNF)- ve interlökin (IL)-1β ekspresyonlarında artış, myokartta kas liflerinde şiddetli düzeyde hyalin dejenerasyon, Zenker nekrozu ve damarlarda şiddetli düzeyde hiperemi gözlendi. OEA ve seftriakson (CEFT) sitokin ekspresyonlarında ve histopatolojik bulgularda azalma sağladı. Sepsis, doku superoksit dismutaz aktivitesini (SOD) azalttı. Ayrıca lipid peroksidasyonu (LPO) ve glutatyon (GSH) seviyeleri ile doku katalaz (KAT), serum kreatin kinaz (CK-MB) ve serum laktat dehidrojenaz (LDH) aktivitelerini önemli şekilde arttırdı. OEA ve CEFT, SOD aktivitesini önemli derecede arttırırken diğer parametrelerde belirgin düşüşe neden oldu. Bu çalışmadan elde edilen bulgular ışığında oleanolik asidin sepsis sırasında oluşan kalp dokusu hasarının azaltılmasında faydalı olabileceği düşünülmektedir.

Oleanolic acid causes reversible contraception in male mice by increasing the permeability of the germinal epithelium.

The effects of oleanolic acid (OA) on the fertility of male mice were investigated using both invivo and invitro experimental models. The experimental group (n=12) was treated with a daily dose of 30mgOAkg-1 bodyweight (i.p.), while the control group (n=6) received a daily dose of 10% ethanol solution (1mLkg-1 bodyweight). The effect of OA on the permeability status of TM4 Sertoli monolayers was investigated by measuring the transepithelial electrical resistance (TER), intracellular electrical resistance and semiquantitative RT-PCR. After 45 days, OA-treated males produced no pregnancies but in the control group, all 12 females were impregnated (69 offspring). Male mice, which demonstrated sterility when exposed to OA, recovered their fertility after 30 days (78 offspring). Testicular histological observations of OA-treated mice showed detachment of adjacent Sertoli-Sertoli cells. A control monolayer developed TER of 300-400 Ω.cm2, but OA (50, 100, 200µgL-1) treated monolayers developed TER of approximately 100Ω.cm2. Intracellular electrophysiological and RT-PCR data supported the premise that OA compromised tight junctional permeability. The study demonstrated reversible contraception in male mice by increasing the permeability of the germinal epithelium and further postulates that contraceptive reversibility is brought about by the reconstitution of the paracellular junctions between adjacent Sertoli cells.

PSV-5 Effects of oleanolic acid on in vitro fermentation of bahiagrass hay and a high-grain substrate.

PSV-5 Effects of oleanolic acid on in vitro fermentation of bahiagrass hay and a high-grain substrate.

The Protective effect of oleanolic acid on acute cholestatic liver injury in rats

Objective The protective effect of oleanolic acid on acute cholestatic liver injury in rats. Methods Thirty male SD rats were randomly divided into 3 groups ( n= 10 per group): sham group, bile duct ligated (BDL) group, and bile duct ligated with oleanolic acid (BDL+OA ) group. After 7 days, liver samples in all rats were collected. Expressions of bile acids pump and nuclear receptors at mRNA and protein levels were detected by RT- qPCR and Western blotting. Results At mRNA level, the expression of Mrp4 and Oatp1 expression in the BDL and BDL+OA groups were increased as compared with that in the sham group. The expression of Mrp4 increased 1.8 times in the BDL group and increased 2.3 times in the BDL+OA group (P<0.05), but the expression of Oatp1 was not statistically significant; AhR was increased 1.7-fold in the BDL group and 2.8 times in the BDL+OA group, Nrf2 was increased 1.5-fold in the BDL group and 2.1 times in the BDL+OA group with statistically significant difference. At the protein level, in the BDL group, Mrp4 expression increased 1.3 times, Oatp1 expression increased 1.5 times, AhR expression increased 1.3 times, Nrf2 expression increased 1.4 fold; in the BDL+OA group, Mrp4 expression increased 1.8 fold, AhR expression increased 1.9 fold, with statistical significance between the two groups. Oatp1 expression increased 1.4-fold in the BDL+OA group as compared with the BDL group showing no statistical significance. Conclusions Oleanolic acid stimulates the hepatic expression of bile acids pump Mrp4 associated with the activation of nuclear receptors AhR and Nrf2 in acute bile duct-ligated rats. Key words: Oleanolic acid; Cholestasis; MRP4; Nuclear receptors

PL - 023 Effects of Oleanolic Acid on Omega-3 and 6 Fatty Acids Metabolism in Rats fed with High Fructose Diet

Objective The present study investigated the effect of OA on metabolism of omega 3 and 6 together with genes associated with lipid metabolism in high fructose diet fed Sprague Dawley rats. &#x0D; Methods Real Time PCR qPCR was conducted to evaluate gene expressions levels.&#x0D; Gas Chromatography Mass Spectrometry (GS-MS): Lipid metabolism metabolites were assesed.&#x0D; Measurement of gene expression using qPCR analysis: Expressions levels of glucose transport and lipid metabolism genes were assessed. &#x0D; Measurements of inflammatory markers concentration in Blood plasma:TNF-α and adiponectin concentrations were determined.&#x0D; Results It was found that high fructose diet decreased gene ex pression levels of NRF-1 and GLUT-4 genes, however administration of OA increased expression levels of these genes. Furthermore, adiponectin concentration was lowered in HFD group however this was reversed when HFD was accompanied by OA administration. This increase in concentration of the anti-inflammatory molecule followed the increase in adiponectin receptor, AdipoR1 gene expression in skeletal muscle tissue. The results on lipid metabolism metabolites were consistent the observed molecular events, the anti-inflammatory signaling molecules (EPA and DHA) were five times higher in HF+OA than they were in HFD. High FAS,ACC-2 gene expression levels in HFD group, low 2 expression levels these genes together with increased CPT-1 in OA treated groups coincided with increased levels of arachidonic acid and linoleic acid levels in HFD, threefold increase compared to the control, and decreased levels in OA treated rats. &#x0D; Conclusions Supplementation of diet with oleanolic acid produces DHA and EPA, thus modulating anti-inflammatory response and modulating effects of exercise in reversing diabetes. We provided evidence to effect that diet has a therapeutic potential in managing metabolic syndrome related diseases.

Oleanolic Acid Alters Multiple Cell Signaling Pathways: Implication in Cancer Prevention and Therapy.

Nowadays, much attention has been paid to diet and dietary supplements as a cost-effective therapeutic strategy for prevention and treatment of a myriad of chronic and degenerative diseases. Rapidly accumulating scientific evidence achieved through high-throughput technologies has greatly expanded the understanding about the multifaceted nature of cancer. Increasingly, it is being realized that deregulation of spatio-temporally controlled intracellular signaling cascades plays a contributory role in the onset and progression of cancer. Therefore, targeting regulators of oncogenic signaling cascades is essential to prevent and treat cancer. A plethora of preclinical and epidemiological evidences showed promising role of phytochemicals against several types of cancer. Oleanolic acid, a common pentacyclic triterpenoid, is mainly found in olive oil, as well as several plant species. It is a potent inhibitor of cellular inflammatory process and a well-known inducer of phase 2 xenobiotic biotransformation enzymes. Main molecular mechanisms underlying anticancer effects of oleanolic acid are mediated by caspases, 5′ adenosine monophosphate-activated protein kinase, extracellular signal–regulated kinase 1/2, matrix metalloproteinases, pro-apoptotic Bax and bid, phosphatidylinositide 3-kinase/Akt1/mechanistic target of rapamycin, reactive oxygen species/apoptosis signal-regulating kinase 1/p38 mitogen-activated protein kinase, nuclear factor-κB, cluster of differentiation 1, CKD4, s6k, signal transducer and activator of transcription 3, as well as aforementioned signaling pathways . In this work, we critically review the scientific literature on the molecular targets of oleanolic acid implicated in the prevention and treatment of several types of cancer. We also discuss chemical aspects, natural sources, bioavailability, and safety of this bioactive phytochemical.

Effect of oleanolic acid on the activity, secretion and gene expression of matrix metalloproteinase-3 in articular chondrocytes in vitro and the production of matrix metalloproteinase-3 in vivo.

In the present study, we tried to examine whether oleanolic acid regulates the activity, secretion and gene expression of matrix metalloproteinase-3 (MMP-3) in primary cultured rabbit articular chondrocytes, as well as the production of MMP-3 in the knee joint of rat to evaluate the potential chondroprotective effect of oleanolic acid. Rabbit articular chondrocytes were cultured in a monolayer, and reverse transcription-polymerase chain reaction (RT-PCR) was used to measure interleukin-1β (IL-1β)-induced gene expression of MMP-3, MMP-1, MMP-13, a disintegrin and metalloproteinase with thrombospondin motifs-4 (ADAMTS-4), ADAMTS-5 and type II collagen. In rabbit articular chondrocytes, the effects of oleanolic acid on IL-1β-induced secretion and proteolytic activity of MMP-3 were investigated using western blot analysis and casein zymography, respectively. The effect of oleanolic acid on in vivo MMP-3 protein production was also examined, after intra-articular injection to the knee joint of rat. The results were as follows: (1) oleanolic acid inhibited the gene expression of MMP-3, MMP-1, MMP-13, ADAMTS-4, and ADAMTS-5, but increased the gene expression of type II collagen; (2) oleanolic acid reduced the secretion and proteolytic activity of MMP-3; (3) oleanolic acid suppressed the production of MMP-3 protein in vivo. These results suggest that oleanolic acid can regulate the activity, secretion and gene expression of MMP-3, by directly acting on articular chondrocytes.

Oleanolic acid attenuates liver ischemia reperfusion injury by HO-1/Sesn2 signaling pathway

Ischemia reperfusion injury (IRI) is unavoidable in liver transplantation and hepatectomy. The present study aimed to explore the possible mechanism and the effect of oleanolic acid (OA) in hepatic IRI. Mice were randomly divided into 6 groups based on different treatment. IRI model: The hepatic artery, portal vein, and bile duct to the left and median liver lobes (70% of the liver) were occluded with an atraumatic bulldog clamp for 90 minutes and then the clamp was removed for reperfusion. The mice were sacrificed 6 hours after reperfusion, and blood and liver tissues were collected. Liver injury was evaluated by biochemical and histopathologic examinations. The expressions of Sesn2, PI3K, Akt and heme oxygenase-1 (HO-1) were measured with quantitative real-time RT-PCR and Western blotting. The serum aminotransferases level and scores of hepatic histology were increased after reperfusion. The increase was attenuated by pretreatment with OA (P<0.01). Compared with the IR group, OA pretreatment significantly up-regulated the expression of Sesn2, PI3K, Akt and HO-1 in IR livers (P<0.05). Administration of zinc protoporphyrin (ZnPP), an inhibitor of HO-1, diminished the OA effect on HO-1 and Sesn2 expressions (P<0.05) and the protective effect of OA on IRI. Our results demonstrate that OA can attenuate hepatic IRI. The protective mechanism may be related to the OA-induced HO-1/Sesn2 signaling pathway.

Cytoprotective Effects of Oleanolic Acid in Human Umbilical Vascular Endothelial Cells is Mediated Via UCP2/ROS/Cytochrome C/AIF Pathway.

The aim of this study is to assess the potential protective effect of oleanolic acid (OA) against ox-LDL induced damage in human umbilical vascular endothelial cells (HUVECs) and investigate potential mechanism of action including antioxidative effects and inhibition of mitochondria apoptosis pathway. Cell counting kit 8 was used to evaluate the viability of HUVECs. 2', 7'-DCFH-DA staining and flow cytometry was used to assess the levels of intracellular reactive oxygen species in HUVECs. The protein expression levels of uncoupling protein 2, cytochrome C, and apoptosis induction factors were measured by western blotting. The results indicated that OA treatment alleviated ox-LDL induced cytotoxicity in HUVECs and ameliorated the reactive oxygen species levels. Western blotting results demonstrated that OA treatment increased the expression level of uncoupling protein 2 and decreased the release of cytochrome C and apoptosis induction factors from mitochondria to cytoplasm, suggesting inhibition of mitochondria apoptosis pathway. In conclusion, OA could protect HUVECs from ox-LDL-induced cytotoxicity; its antioxidant property and inhibition of mitochondria apoptosis are likely crucial contributors.

마우스에서 Oleanolic Acid와 Hederagenin이 급성 알코올성 간독성에 미치는 효과

본 연구는 선행연구를 통한 으름 추출물의 간 보호 효과에 대하여 으름 추출물 내 유효물질로 알려진 oleanolic acid와 hederagenin을 생쥐에 투여하였을 때 알코올로 인한 급성 간독성 상태에서 간 보호 효과를 조사하였다. 그 결과 oleanolic acid와 hederagenin의 투여는 알코올로 유발된 급성 간독성 상태에서 혈중 알코올 농도 감소 및 alcohol dehydrogenase mRNA 발현을 증가시키는 것으로 나타났으며, 간 조직 내 항산화 물질인 glutathione(GSH)의 농도를 증가시키고, GSH synthetic enzyme의 mRNA 발현 역시 증가시키는 것으로 확인되었다. 또한 염증반응 지표인 tumor necrosis factor-alpha의 mRNA 발현도 억제하는 것으로 분석되었다. 위의 결과들과 기존의 실험에서 확인한으름 추출물의 간 보호 효과를 종합하였을 때 으름 추출물의 알코올로 유발된 산화 스트레스, 염증반응에 대한 간 보호 효과는 추출물 내 유효물질로 알려진 oleanolic acid 및 hederagenin의 작용에 의한 것이라 생각한다.

Anticancer and apoptotic activities of oleanolic acid are mediated through cell cycle arrest and disruption of mitochondrial membrane potential in HepG2 human hepatocellular carcinoma cells.

Hepatocellular carcinoma (HCC) is an aggressive form of cancer, with high rates of morbidity and mortality, a poor prognosis and limited therapeutic options. The objective of the present study was to demonstrate the anticancer activity of oleanolic acid in HepG2 human HCC cells. Cell viability was evaluated using an MTT assay, following administration of various doses of oleanolic acid. The effect of oleanolic acid on cell cycle phase distribution and mitochondrial membrane potential was evaluated using flow cytometry with propidium iodide and rhodamine-123 DNA-binding cationic fluorescent dyes. Fluorescence microscopy was employed to detect morphological changes in HepG2 cells following oleanolic acid treatment. The results revealed that oleanolic acid induced a dose-dependent, as well as time-dependent inhibition in the growth of HepG2 cancer cells. Following acridine orange and ethidium bromide staining, treatment with various doses (0, 5, 25 and 50 µM) of oleanolic acid induced typical morphological changes associated with apoptosis, including cell shrinkage, membrane blebbing, nuclear condensation and apoptotic body formation. Cell cycle analysis revealed that oleanolic acid induced cell cycle arrest in HepG2 cells at the sub-G1 (apoptotic) phase of the cell cycle, in a dose-dependent manner. Staining with Annexin V-fluorescein isothiocyanate and propidium iodide revealed that apoptosis occurred early in these cells. Oleanolic acid treatment also resulted in fragmentation of nuclear DNA in a dose-dependent manner, producing the typical features of DNA laddering on an agarose gel. The results also demonstrated that oleanolic acid treatment resulted in a potent loss of mitochondrial membrane potential, which also occurred in a dose-dependent manner. Therefore, oleanolic acid may be used as a therapeutic agent in the treatment of human HCC.

18F-FDG PET/CT in evaluating the radiosensitizing effect of oleanolic acid

Objective To evaluate the feasibility of 18F-FDG PET/CT in assessing the radiosensitizing effect of oleanolic acid (OA) in C6 rat glioma model, and to explore the possible mechanism of radiosensitizing effect of OA. Methods Thirty-two C6 glioma-bearing SD rats were divided into 4 groups by random number table: group A without OA and radiotherapy, group B with OA alone, group C with radiotherapy alone, group D with OA and radiotherapy. 18F-FDG PET/CT was performed before radiotherapy, at 24 h and 7 d after radiotherapy. The tumor SUVmax was measured. All rats were sacrificed and tumor tissues were excised for HE and immunohistochemistry staining. Paired t test, one-way analysis of variance, LSD-t test and Spearman correlation analysis were performed to analyze the data using SPSS 13.0. Results There was no statistically significant difference in SUVmax among the 4 groups (SUVmax of groups A, B, C, D: 5.252±0.536, 5.261±0.544, 5.273±0.520, 5.232±0.507) before treatment (F=0.008, P>0.05). At 24 h post-radiotherapy, SUVmax of groups C and D (4.766±0.511 and 4.403±0.486) decreased significantly (t=14.788, 13.366, both P 0.05). At 7 d post-radiotherapy, SUVmax of groups C and D decreased significantly (t=9.750, 10.530, both P 0.05). SUVmax of Group D decreased significantly compared with that of group C (t=5.329, P>0.05). Less tumor cells and more areas of necrosis were observed in group D compared with those in other groups by HE staining. The expression of HIF-1α was lower in group D than that in other groups by immunohistochemistry staining. HIF-1α expression was positively correlated with SUVmax (r=0.853, P<0.05). Conclusion 18F-FDG PET/CT has potential to evaluate the radiosensitizing effect of OA in vivo, and the radiosensitization of OA might be related to the down regulation of HIF-1α. Key words: Glioma; Radiotherapy; Radiation-sensitizing agents; Oleanolic acid; Tomography, emission-computed; Tomography, X-ray computed; Deoxyglucose; Rats

Effects of oleanolic acid on learning and memory, synaptic morphology and the expression of CREB in the hippocampus and cereberal cortex of SAMP8 mice

Objective To observe the influence of oleanolic acid on the ethology of 9-month-old mice, the completeness of synapsis structure and the expression of cAMP-response element binding protein(CREB) in cortex and hippocampus. Methods Thirty 9-month-old healthy male SMAP8 mice were randomly divided into model group, oleanolic acid group and aricept group, and with 10 rats in each group, while 10 healthy male mice of the same age and species as normal group.Oleanolic acid group and aricept group were given intragastric administration with corresponding drugs, while the normal group and model group were given the same amount of physiological saline. 4 weeks later, the ethology changes were observed by Morris water maze and morphology changes of hippocampus neurons were viewed by electron microscope and the expression of CREB was detected by Western Blot. Results (1)Morris water maze results suggested that compared with the normal group, the latency time in the model group mice was longer, which were ((83.33±4.96) s, (75.13±6.01) s, ( 71.75±7.77) s, (63.40±8.93) s, (60.97±8.38) s), while compared with the model group, the latency time in the oleanolic acid group and the aricept group was remarkably shorter(P<0.05), which were (75.97±4.49)s, (64.98±4.93)s, (64.16±6.23)s, (53.47±5.99)s, (47.91±7.64)s and (71.30±7.65)s, (63.32±7.57)s, (59.82±4.69)s, (52.28±5.90)s, (46.22±7.27)s respectively. In the spatial probe trial, compared with the normal group, the crossing times of the model was less, while compared with the model one, the crossing times of the oleanolic acid group and the aricept group was more(P<0.05). (2)Compared with the normal group, the number of synapses in the model group was smaller, in which the synaptic cleft was mixed with the front of synapses severely swollen, uninform synaptic vesicles and few clear outlines of mitochondrias. While the oleanolic acid group and the aricept group had clear synapses outlines with the front of synapses slightly swollen, intensive and uniform synaptic vesicles and clear mitochondrias with their cristae not easy to be seen. (3)The Western Blot showed that compared with the normal group, there was a decline in the CREB expression both in the cortex and hippocampus in the model group, while compared with the model group, there was a rise in the oleanolic acid group as well as the aricept group(P<0.05). Conclusion Oleanolic acid can improve the learning and memorizing of model rats, which is possibly related to the increased expression of CREB protein to protect the synapses structure of model mice. Key words: Oleanolic acid; Alzheimer's Disease; SAMP8 mice; Synapsis electron microscope; cAMP-response element binding protein