Improving the Effect of Ferulic Acid on Inflammation and Insulin Resistance by Regulating the JNK/ERK and NF-κB Pathways in TNF-α-Treated 3T3-L1 Adipocytes.

In this study, ferulic acid was investigated for its potential in suppressing TNF-α-treated inflammation and insulin resistance in adipocytes. Ferulic acid suppressed TNF-α, IL-6, IL-1ß, and MCP-1. TNF-α increased p-JNK and ERK1/2, but treatment with ferulic acid (1, 10, and 50 µM) decreased p-JNK and ERK1/2. TNF-α induced the activation of IKK, IκBα, and NF-κB p65 compared to the control, but ferulic acid inhibited the activation of IKK, IκBα, and NF-κB p65. Following treatment with TNF-α, pIRS-1ser307 increased and pIRS-1tyr612 decreased compared to the control. Conversely, as a result of treatment with 1, 10, and 50 µM ferulic acid, pIRS-1ser307 was suppressed, and pIRS-1tyr612 was increased. Therefore, ferulic acid reduced inflammatory cytokine secretion by regulating JNK, ERK, and NF-κB and improved insulin resistance by suppressing pIRS-1ser. These findings indicate that ferulic acid can improve inflammation and insulin resistance in adipocytes.

HM-chromanone isolated from Portulaca oleracea L. alleviates insulin resistance and inhibits gluconeogenesis by regulating palmitate-induced activation of ROS/JNK in HepG2 cells.

Oxidative stress is a major cause of hepatic insulin resistance. This study investigated whether (E)-5-hydroxy-7-methoxy-3-(2-hydroxybenzyl)-4-chromanone (HM-chromanone), a homoisoflavonoid compound isolated from Portulaca oleracea L., alleviates insulin resistance and inhibits gluconeogenesis by reducing palmitate (PA)-induced reactive oxygen species (ROS)/c-Jun NH2-terminal kinase (JNK) activation in HepG2 cells. PA treatment (0.5 mM) for 16 h resulted in the highest production of ROS and induced insulin resistance in HepG2 cells. HM-chromanone, like N-acetyl-1-cysteine, significantly decreased PA-induced ROS production in the cells. HM-chromanone also significantly inhibited PA-induced JNK activation, showing a significant reduction in tumor necrosis factor and interleukin expression levels. Thus, HM-chromanone decreased the phosphorylation of Ser307 in insulin receptor substrate 1, while increasing phosphorylation of serine-threonine kinase (AKT), thereby restoring the insulin signaling pathway impaired by PA. HM-chromanone also significantly increased the phosphorylation of forkhead box protein O, thereby inhibiting the expression of gluconeogenic enzymes and reducing glucose production in PA-treated HepG2 cells. HM-chromanone also increased glycogen synthesis by phosphorylating glycogen synthase kinase-3ß. Therefore, HM-chromanone may alleviate insulin resistance and inhibit gluconeogenesis by regulating PA-induced ROS/JNK activation in HepG2 cells.

Scopoletin protects INS-1 pancreatic ß cells from glucotoxicity by reducing oxidative stress and apoptosis.

This study investigated whether scopoletin could protect INS-1 pancreatic ß cells from apoptosis and oxidative stress caused by high glucose. Cells were pretreated with glucose (5.5 or 30 mM) and then treated with 0, 5, 10, 25, or 50 µM Scopoletin. Cell viability and insulin secretion were measured in addition to ROS, TBARS, NO and antioxidant enzymes. Western blot analysis and flow cytometric assessment of apoptosis were also carried out. High glucose of 30 mM caused glucotoxicity and cell death in INS-1 pancreatic ß cells. However, 5, 10, 25 or 50 µM scopoletin increased the level of cell viability as concentrations increased. The levels of ROS, TBARS, and NO increased by high glucose were significantly decreased after scopoletin treatment. Scopoletin also raised antioxidant enzyme activities up against oxidative stress produced by high glucose. These effects influenced the apoptosis pathway, raising levels of anti-apoptotic protein, Bcl-2, and reducing levels of pro-apoptotic proteins, including JNK, Bax, cytochrome C, and caspase 9. Annexin V/propidium staining indicated that scopoletin significantly lowered high glucose-produced apoptosis. These results indicate that scopoletin can protect INS-1 pancreatic ß cells from glucotoxicity caused by high glucose and have potential as a pharmaceutical material to protect the pancreatic ß cells.

HMC Ameliorates Hyperglycemia via Acting PI3K/AKT Pathway and Improving FOXO1 Pathway in ob/ob Mice.

Type 2 diabetes is a disease characterized by hyperglycemia and is a growing health problem worldwide. Since many known diabetes drugs are side effects, it is necessary to develop natural substances with guaranteed safety. HM-chromanone isolated from Portulaca oleracea L. is a homoisoflavonoid compound. We investigated the effects of HM-chromanone on hyperglycemia and its mechanism in C57BL/6J ob/ob mice. C57BL/6J-Jms Slc mice were used as the control group, and C57BL/6J-ob/ob mice were divided into three groups: ob/ob (control), metformin (Met; positive control), and HM-chromanone (HMC). Fasting blood glucose was lower in the HMC group than those in the ob/ob group. Insulin resistance was improved by reducing HbA1c, plasma insulin, and HOMA-IR levels in the HMC group. HMC administration decreased the phosphorylation of IRS-1ser307 and increased the phosphorylation of IRS-1tyr612, PI3K, phosphorylation of AKTser473, and PM-GLUT4 in the skeletal muscles of ob/ob mice, indicating improved insulin signaling. HMC administration also increased the phosphorylation of FOXO1 in the liver of ob/ob mice. This inhibited PEPCK and G6pase involved in gluconeogenesis and regulated phosphorylation of glycogen synthase kinase 3ß and glycogen synthase involved in glycogen synthesis. In conclusion, HM-chromanone ameliorates hyperglycemia by PI3K/AKT and improves the FOXO1 in ob/ob mice.

HM-chromanone from Portulaca oleracea L. inhibits protein tyrosine phosphatase 1B and mitigates glucose production in insulin-resistant HepG2 cells.

This study aimed to identify the effect of (E)-5-hydroxy-7-methoxy-3-(2'-hydroxybenzyl)-4-chromanone (HM-chromanone), isolated from Portulaca oleracea L., on tyrosine phosphatase 1B (PTP1B) and glucose production in insulin-resistant HepG2 cells. The results revealed that HM-chromanone significantly decreases PTP1B expression and glucose production in insulin-resistant HepG2 cells. Furthermore, a molecular docking stimulation showed HM-chromanone inhibits PTP1B by binding to its active site. Additionally, HM-chromanone was found to significantly modulate insulin receptor substrate-1 (IRS1) by decreasing phosphorylated serine 307 and increasing phosphorylated tyrosine 612 and activating phosphatidylinositol 3-kinase (PI3K) in insulin-resistant HepG2 cells. Furthermore, HM-chromanone augmented the phosphorylation of Akt and forkhead box protein O1 in insulin-resistant HepG2 cells in a dose-dependent manner at the concentrations of 15-60 µM. Additionally, it significantly reduced the expression of glucose 6-phosphatase and phosphoenolpyruvate carboxykinase, which are main enzymes included in hepatic gluconeogenesis. Consequently, HM-chromanone was confirmed to significantly decrease glucose production and increase glucose uptake in insulin-resistant HepG2 cells.

HM-Chromanone Alleviates Hyperglycemia by Protecting Pancreatic Islet Cells in Streptozotocin-Induced Diabetic Mice.

We examined the effects of HM-chromanone (HMC) on alleviating hyperglycemia and protecting pancreatic ß-cells from streptozotocin (STZ)-induced damage in C57BL/6J mice. HMC was administered to STZ-induced diabetic mice at 10 or 30 mg/kg, for 14 days. Thereafter, changes in fasting blood glucose levels, insulin-secretion, histopathological examination of pancreas islet cell and apoptotic protein levels, and terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay were determined. The results revealed that HMC dose-dependently improved blood glucose concentrations and alleviated pancreatic islet cells damage. In diabetic mice, degeneration of the islet cells was observed wherein they appeared shrunken, with hyaline deterioration, nuclear dissolution, and condensation. However, morphology of the islet cell was restored, and nuclei were visibly rounded in the HMC (30 mg/kg)-administered diabetic mice. In addition, ß-cell numbers were markedly increased in HMC mice compared to STZ-induced diabetic mice, and the number of cells stained with glucagon was decreased. HMC markedly decreased the expression of proapoptotic proteins and increased antiapoptotic proteins, and the number of apoptotic cells detected by TUNEL was elevated. HMC decreased expression of interleukin (IL)-1ß, IL-6, and tumor necrosis factor-α in diabetic mice. Moreover, HMC increased antioxidant-enzymes activity, and decreased reactive oxygen species generation. In conclusion, the results demonstrate the potential of HMC to alleviate hyperglycemia by protecting the pancreatic ß-cells in diabetic mice.

A bioactive component of Portulaca Oleracea L., HM-chromanone, improves palmitate-induced insulin resistance by inhibiting mTOR/S6K1 through activation of the AMPK pathway in L6 skeletal muscle cells.

Increased free fatty acid levels in the blood are common in obesity and cause insulin resistance associated with type 2 diabetes in the muscles. Previous studies have confirmed the antidiabetic and anti-obesity potential of (E)-5-hydroxy-7-methoxy-3-(2-hydroxybenzyl)-4-chromanone (HM-chromanone). However, it is unknown how HM-chromanone alleviates obesity-related insulin resistance in L6 skeletal muscle cells. Palmitate induced insulin resistance and reduced glucose uptake, whereas HM-chromanone significantly increased glucose uptake. In palmitate-treated L6 skeletal muscle cells, HM-chromanone stimulated liver kinase B1 (LKB1) and 5'-adenosine monophosphate-activated protein kinase (AMPK) phosphorylation. The AMPK inhibitor compound C, and the LKB1 inhibitor radicicol blocked the effects of HM-chromanone. Furthermore, HM-chromanone significantly inhibited mammalian target of rapamycin (mTOR) and ribosomal protein S6 kinase 1 (S6K1) activation, but there was no change in protein kinase C θ (PKC θ) expression. When pAMPK was inhibited with compound C, the effect of HM-chromanone on the inhibition of mTOR and S6K1 was significantly diminished. This indicates that HM-chromanone inhibits mTOR and S6K1 activation through pAMPK activation. Inhibition of mTOR and S6K1 by HM-chromanone significantly reduced IRS-1Ser307 and IRS-1Ser632 phosphorylation, leading to insulin resistance. This resulted in an increase in PM-GLUT4 (glucose transporter 4) expression, thereby stimulating glucose uptake in insulin-resistant muscle cells. HM-chromanone can improve palmitate-induced insulin resistance by inhibiting mTOR and S6K1 through activation of the AMPK pathway in L6 skeletal muscle cells. These results show the therapeutic potential of HM-chromanone for improving insulin resistance in type 2 diabetes.

HM-chromanone reverses the blockade of insulin signaling induced by high glucose levels in human HepG2 cells.

This study investigated whether (E)-5-hydroxy-7-methoxy-3-(2'-hydroxybenzyl)-4-chromanone (HM-chromanone) could counteract the high glucose level-induced blockade of insulin signaling in human HepG2 cells. Cells were pre-incubated with glucose (5.5 or 33 mM) and then incubated with a medium containing various concentrations of HM-chromanone. Assays for glucose uptake, glycogen synthesis, and glucose production were performed. Western blotting helped elucidate the underlying molecular mechanisms. High glucose concentration (33 mM) significantly increased p-IRS-1ser307 levels and decreased p-Akt levels. However, HM-chromanone significantly decreased p-IRS-1ser307 levels while increasing p-IRS-1tyr612 and Akt levels, which restored insulin signaling disturbed by high glucose concentration. HM-chromanone significantly increased p-AMPK levels, which were reduced by high glucose in HepG2 cells. Knockdown of AMPK using siRNA increased p-IRS-1ser307 and decreased p-Akt levels, even after treatment with HM-chromanone in high glucose concentration-treated cells. HM-chromanone stimulated glycogen synthesis by increasing p-GSK3ßser9 and decreasing p-GSser641 levels in HepG2 cells under high glucose concentration; this effect was blocked by AMPK siRNA. HM-chromanone significantly decreased PEPCK, G6Pase, and hepatic glucose production, which were also blocked by AMPK siRNA. These results suggest that HM-chromanone could reverse insulin signaling blockade (induced by high glucose levels) through the activation of AMPK and stimulation of glucose uptake and glycogen synthesis in HepG2 cells.

Pheophorbide A isolated from Gelidium amansii inhibits adipogenesis by regulating adipogenic transcription factors and AMPK in 3T3-L1 adipocytes.

Adipocyte lipid accumulation causes adipocyte hypertrophy and adipose tissue increment, leading to obesity. As part of our efforts to isolate antiobesity agents from natural products, we first isolated the active compound from the extract of Gelidium amansii through bioassay-guided fractionation. We then hypothesized that pheophorbide A isolated from G amansii inhibits adipogenesis by downregulating adipogenic transcription factors; therefore, the antiadipogenic effects of pheophorbide A were investigated in 3T3-L1 adipocytes. On differentiation of 3T3-L1 preadipocytes into adipocytes, they were treated with pheophorbide A (0-83 µM). Pheophorbide A inhibited triglyceride accumulation (half maximal inhibitory concentration = 114.2 µM) and stimulated glycerol release in a dose-dependent manner in 3T3-L1 adipocytes. In addition, pheophorbide A significantly decreased leptin concentrations in 3T3-L1 adipocytes. Pheophorbide A inhibited adipogenesis by suppressing the expression of adipogenic transcriptional factors including peroxisome proliferator-activated receptor γ, CCATT/enhancer binding protein α, sterol regulatory element binding protein 1c, and fatty acid synthase. It also induced the expression of phosphorylation of AMP-activated protein kinase. Therefore, these results suggest that pheophorbide A may be useful for preventing or treating obesity because of its inhibitory effect on adipogenesis.

Betulinic acid improves TNF-α-induced insulin resistance by inhibiting negative regulator of insulin signalling and inflammation-activated protein kinase in 3T3-L1 adipocytes.

CONTEXT: Obesity is related to insulin resistance, and adipose tissue-secreted TNF-α may play a role in inducing obesity. TNF-α activates inflammatory protein kinase and impairs insulin signalling. OBJECTIVES: We investigated the effect of betulinic acid on insulin resistance caused by TNF-α treatment in 3T3-L1 adipocytes. MATERIAL AND METHODS: 3T3-L1 was exposed to TNF-α in the presence and absence of betulinic acid. Various parameters such as glucose uptake assay, cell viability, expression of proteins involved in insulin resistance were studied. RESULTS: Betulinic acid increased glucose uptake in TNF-α pre-treated cells and inhibited the activation of PTP1B and JNK and reduced IκBα degradation. Tyrosine phosphorylation was increased, and serine phosphorylation was decreased in IRS-1. DISCUSSION: Betulinic acid restored TNF-α impaired insulin signalling and increased PI3K activation and phosphorylation of Akt and increased plasma membrane expression of GLUT 4, which stimulated glucose uptake concentration-dependently. CONCLUSION: These results suggest that betulinic acid is effective at improving TNF-α-induced insulin resistance in adipocytes via inhibiting the activation of negative regulator of insulin signalling and inflammation-activated protein kinase and may potentially improve insulin resistance.

HM-Chromanone, a Major Homoisoflavonoid in Portulaca oleracea L., Improves Palmitate-Induced Insulin Resistance by Regulating Phosphorylation of IRS-1 Residues in L6 Skeletal Muscle Cells.

This study investigated the effect of (E)-5-hydroxy-7-methoxy-3-(2-hydroxybenzyl)-4-chromanone (HM-chromanone) on palmitate-induced insulin resistance and elucidated the underlying mechanism in L6 skeletal muscle cells. Glucose uptake was markedly decreased due to palmitate-induced insulin resistance in these cells; however, 10, 25, and 50 µM HM-chromanone remarkably improved glucose uptake in a concentration-dependent manner. HM-chromanone treatment downregulated protein tyrosine phosphatase 1B (PTP1B) and phosphorylation of c-Jun N-terminal kinase (JNK) and inhibitor of nuclear factor kappa-B kinase subunit beta (IKKß), which increased because of palmitate mediating the insulin-resistance status in cells. HM-chromanone promoted insulin receptor substrate-1 (IRS-1) tyrosine phosphorylation and suppressed palmitate-induced phosphorylation of IRS-1 serine. This activated phosphoinositide 3-kinase (PI3K) and stimulated protein kinase B (AKT) phosphorylation. Phosphorylated AKT promoted the translocation of Glucose transporter type 4 to the plasma membrane and significantly enhanced glucose uptake into muscle cells. Additionally, HM-chromanone increased glycogen synthesis through phosphorylating glycogen synthase kinase 3 alpha/beta (GSK3 α/ß) via AKT. Consequently, HM-chromanone may improve insulin resistance by downregulating the phosphorylation of IRS-1 serine through inhibition of negative regulators of insulin signaling and inflammation-activated protein kinases in L6 skeletal muscle cells.

HM-chromanone suppresses hepatic glucose production via activation of AMP-activated protein kinase in HepG2 cell.

We investigated whether (E)-5-hydroxy-7-methoxy-3-(2-hydroxybenzyl)-4-chromanone (HM-chromanone) could suppress the transcription factors expression and enzymes involved in glucose production by activating AMPK in hepatocytes. HepG2 cells were treated with a medium containing HM-chromanone (5-100 µM), compound C (10 µM) and insulin (100 nM). Glucose production and glycogen synthesis assay were determined using a glucose assay kit and glycogen assay kit, respectively. Activities of AMP-activated protein kinase (AMPK), acetyl CoA carboxylase (ACC), cAMP response element-binding protein (CREB), PPAR coactivator-1α (PGC1α), CREB-regulated transcription coactivator 2 (CRTC2), Glycogen synthase kinase (GSK3ß), Phosphoenolpyruvate carboxykinase (PEPCK), glycogen synthase (GS), Glucose 6-phosphatase (G6pase) and ß-actin were determined by Western blot analysis. HM-chromanone significantly inhibited hepatic glucose production and increased glycogen synthesis by activating glycogen synthase. HM-chromanone induced the phosphorylation of CRTC2 and GSK-3ß by phosphorylating AMPK in HepG2 cells, which was confirmed by compound C. Furthermore, it significantly decreased the phosphorylation of CREB in a time- and concentration-dependent manner, and the effect was reversed in the presence of compound C. Therefore, the complex formation of CRTC2 and CREB was inhibited. HM-chromanone inhibited the expression of PGC-1α, PEPCK, and G6Pase genes involved in production of hepatic glucose. The results showed that HM-chromanone activates AMPK in a time and concentration dependent manner, thus suppressing hepatic glucose production and increasing glycogen synthesis in HepG2 cells.

HM-Chromanone Ameliorates Hyperglycemia and Dyslipidemia in Type 2 Diabetic Mice.

The effects of (E)-5-hydroxy-7-methoxy-3-(2-hydroxybenzyl)-4-chromanone (HMC) on hyperglycemia and dyslipidemia were investigated in diabetic mice. Mice were separated into three groups: db/db, rosiglitazone and HMC. Blood glucose or glycosylated hemoglobin values in HMC-treated mice were significantly lower compared to db/db mice. Total cholesterol, LDL-cholesterol, and triglyceride values were lower, and HDL-C levels were higher, in the HMC group compared to the diabetic and rosiglitazone groups. HMC markedly increased IRS-1Tyr612, AktSer473 and PI3K levels and plasma membrane GLUT4 levels in skeletal muscle, suggesting improved insulin resistance. HMC also significantly stimulated AMPKThr172 and PPARα in the liver, and ameliorated dyslipidemia by inhibiting SREBP-1c and FAS. Consequently, HMC reduced hyperglycemia by improving the expression of insulin-resistance-related genes and improved dyslipidemia by regulating fatty acid synthase and oxidation-related genes in db/db mice. Therefore, HMC could ameliorate hyperglycemia and dyslipidemia in type 2 diabetic mice.

HM-chromanone attenuates TNF-α-mediated inflammation and insulin resistance by controlling JNK activation and NF-κB pathway in 3T3-L1 adipocytes.

Obesity is a major public health problem worldwide and causes inflammation and insulin resistance in adipose tissue. We investigated the ability of (E)-5-hydroxy-7-methoxy-3-(2'-hydroxybenzyl)-4-chromanone (HM-chromanone) isolated from Portulaca oleracea to attenuate the activation of inflammatory cytokines and signaling pathways associated with tumor necrosis factor (TNF)-α-mediated inflammation and insulin resistance in 3T3-L1 adipocytes. TNF-α triggers the release of inflammatory cytokines and activation of the mitogen-activated protein kinase and nuclear factor (NF)-κB signaling pathways. In this study, HM-chromanone inhibited the production of inflammatory cytokines and chemokines [TNF-α, interleukin (IL)-6, IL-1ß, and monocyte chemoattractant protein 1] involved in inflammation and insulin resistance. Furthermore, TNF-α treatment increased c-Jun-NH2 terminal kinase (JNK) phosphorylation, whereas HM-chromanone significantly decreased JNK phosphorylation in a dose-dependent manner. TNF-α treatment increased the activation of inhibitor kappa B (IκB) kinase (IKK), IκBα, and NF-κBp65 compared with that of the control. However, HM-chromanone significantly blocked IKK, IκBα, and NF-κBp65 activation. Upon adipocyte stimulation with TNF-α, phosphorylated insulin receptor substrate (pIRS)-1 serine 307 levels increased and pIRS-1 tyrosine 612 levels decreased compared with those of the control. Upon treatment with HM-chromanone, serine 307 phosphorylation of IRS-1 was inhibited and tyrosine 612 phosphorylation of IRS-1 was increased. Thus, HM-chromanone improved TNF-α-mediated inflammation and insulin resistance by regulating JNK activation and the NF-κB pathway, thereby reducing inflammatory cytokine secretion and inhibiting serine phosphorylation of IRS-1 in the insulin signaling pathway. These results suggest the potential of HM-chromanone to improve inflammatory conditions and insulin resistance in adipocytes.

Scopoletin stimulates the secretion of insulin via a KATP channel-dependent pathway in INS-1 pancreatic ß cells.

OBJECTIVES: In this study, we investigated whether scopoletin stimulated the secretion of insulin in pancreatic ß cells as well as the underlying mechanism involved in this process. METHODS: We incubated the INS-1 pancreatic ß cells with various concentrations of glucose (1.1, 5.6 or 16.7 mM) in the presence or absence of scopoletin. We then analysed the secretion of insulin in the cells treated with insulin secretion inhibitors or secretagogues. The intracellular influx of calcium induced by scopoletin was also analysed using the Fluo-2 AM dye. KEY FINDINGS: We found that scopoletin (1-20 µM) markedly induced the secretion of insulin in a glucose concentration-dependent manner compared with the control. At depolarizing concentrations of potassium chloride (KCl), scopoletin markedly enhanced the insulin secretion compared with the cells which were treated only with KCl. Moreover, the treatment with diazoxide-opening K+ATP channel and verapamil blocking Ca2+ channel significantly decreased the scopoletin-induced increase in insulin secretion. After the pre-treatment of cells with a Ca2+ fluorescent dye, treatment with 20 µM scopoletin resulted in a significant increase in the influx of intracellular Ca2+, exhibiting fluorescence changes in various spectra. CONCLUSIONS: Scopoletin stimulates the secretion of insulin via a K+ATP channel-dependent pathway in the INS-1 pancreatic ß cells.

(E)-5-hydroxy-7-methoxy-3-(2'-hydroxybenzyl)-4-chromanone isolated from Portulaca oleracea L. suppresses LPS-induced inflammation in RAW 264.7 macrophages by downregulating inflammatory factors.

CONTEXT: Portulaca oleracea L. is herbaceous succulent annual plant, which belongs to the Portulacaceae family. Many studies have shown its wide spectrum of pharmacological activities such as anti-cancer and anti-diabetic effects. OBJECTIVES: The objective of this study was to identify the anti-inflammatory effects of HM-chromanone isolated from Portulaca oleracea L. in LPS-stimulated RAW 264.7 macrophages. MATERIALS AND METHODS: LPS (1 µg/ml)-stimulated mouse RAW 264.7 macrophages were used to assess the anti-inflammatory effect of HM-chromanone (10-50 µM). Cell viability was evaluated by MTT assay. In addition, the production of intracellular ROS, superoxide anion, lipid peroxide, NO, and PGE2, and activity of antioxidant enzymes were analyzed. The expressions of iNOS, COX-2, IκB, NF-κB, TNF-α, IL-1ß and IL-6 were evaluated by western blot analysis. RESULTS: HM-chromanone has demonstrated that there is no significant cytotoxic effect on the viability of RAW 264.7 macrophages. In LPS-stimulated RAW 264.7 cells, HM-chromanone treatment was found to significantly inhibit the production of intracellular ROS, superoxide anion and lipid peroxide, while enhancing the activity of antioxidant enzymes such as SOD, catalase, and GSH-px. Additionally, HM-chromanone treatment was observed to inhibit NO and PGE2 production by inhibiting the expression of iNOS and COX-2. Subsequently, HM-chromanone was observed to significantly suppress LPS-induced expression of IκB, NF-κB, TNF-α, IL-1ß and IL-6. DISCUSSION AND CONCLUSION: Overall, our results suggested that HM-chromanone suppresses LPS-induced inflammation in RAW 264.7 macrophages by downregulating the expression of inflammatory factors.

Alleviating effects of lupeol on postprandial hyperglycemia in diabetic mice.

This study aimed to investigate the inhibition activities of lupeol on carbohydrate digesting enzymes and its ability to improve postprandial hyperglycemia in streptozotocin (STZ)-induced diabetic mice. α-Glucosidase and α-amylase inhibitory assays were executed using a chromogenic method. The effect of lupeol on hyperglycemia after a meal was measured by postprandial blood glucose in STZ-induced diabetic and normal mice. The mice were treated orally with soluble starch (2 g/kg BW) alone (control) or with lupeol (10 mg/kg BW) or acarbose (10 mg/kg BW) dissolved in water. Blood samples were taken from tail veins at 0, 30, 60, and 120 min and blood glucose was measured by a glucometer. Lupeol showed noticeable inhibitory activities on α-glucosidase and α-amylase. The half-maximal inhibitory concentrations (IC50) of lupeol on α-glucosidase and α-amylase were 46.23 ± 9.03 and 84.13 ± 6.82 µM, respectively, which were more significantly effective than those of acarbose, which is a positive control. Increase in postprandial blood glucose level was more significantly lowered in the lupeol-administered group than in the control group of both STZ-induced diabetic and normal mice. In addition, the area under the curve was significantly declined with lupeol administration in the STZ-induced diabetic mice. These findings suggest that lupeol can help lower the postprandial hyperglycemia by inhibiting carbohydrate-digesting enzymes.

HM-chromanone, a component of Portulaca oleracea L., stimulates glucose uptake and glycogen synthesis in skeletal muscle cell.

BACKGROUND: Diabetes mellitus is a chronic metabolic disease characterized by increased blood glucose levels. In order to lower blood glucose, it is important to stimulate glucose uptake and glycogen synthesis in the muscle. (E)-5-hydroxy-7-methoxy-3-(2'-hydroxybenzyl)-4-chromanone (HM-chromanone), a constituent isolated from Portulaca oleracea L., exhibits anti-diabetic effects; however, its mechanisms are not yet clearly understood on glucose uptake and glycogen synthesis in muscle cells. PURPOSE: In the present study, we examined the effects of HM-chromanone on glucose uptake into L6 skeletal muscle cells and elucidated the underlying mechanisms. METHODS: The effects of HM-chromanone on glucose uptake into L6 skeletal muscle cells were assessed by 2-Deoxyglucose uptake assay. Western blot analysis was carried out to elucidate the underlying molecular mechanisms. RESULTS: We found that HM-chromanone promoted glucose uptake into L6 skeletal muscle cells in a dose-dependent manner. Moreover, HM-chromanone induced the phosphorylation of IRS-1Tyr612 and AKTSer473, and the activation of PI3K. HM-chromanone also stimulated the phosphorylation of AMPKThr172, AS160Thr642, TBC1D1Ser237, and ACC via the CaMKKß pathway. Furthermore, HM-chromanone increased glycogen synthesis through the inactivation of glycogen synthase kinase 3 α/ß. CONCLUSION: The results of this study indicate that HM-chromanone stimulates glucose uptake through the activation of the PI3K/AKT and CaMKKß-AMPK pathways and glycogen synthesis via the GSK3 α/ß pathway in L6 skeletal muscle cells.

HM-chromanone inhibits adipogenesis by regulating adipogenic transcription factors and AMPK in 3T3-L1 adipocytes.

Portulaca oleracea L. is used as a folk medicine in many countries because of its wide range of pharmacological effects. HM-chromanone, isolated from P. oleracea using bioassay-guided fractionation and HPLC, belongs to the homoisoflavonoid group and has been shown to exert several biological effects. In this study, we evaluated whether HM-chromanone inhibits adipogenesis by regulating adipogenic transcription factors in 3T3-L1 adipocytes. The results showed that HM-chromanone suppresses adipocyte differentiation and adipogenesis in a dose-dependent manner in 3T3-L1 adipocytes. The HM-chromanone-treated adipocytes exhibited lower triglyceride accumulation and leptin secretion, and higher glycerol and adiponectin secretion than the control adipocytes. Microscopic observation using oil red O staining revealed a dose-dependent reduction in the number of lipid droplets in the HM-chromanone-treated adipocytes compared to the control group. HM-chromanone significantly down-regulated the protein expression of major adipogenic transcription factors sterol regulatory element binding protein-1c (SREBP-1c), peroxisome proliferator-activated receptor γ (PPARγ), and CCAAT/enhancer binding protein α (C/EBPα) and markedly inhibited several key adipogenic enzymes including fatty acid synthase (FAS) and acetyl-CoA carboxylase (ACC). In addition, adipose triglyceride lipase (ATGL) and hormone-sensitive lipase (HSL) were both more activated in the HM-chromanone-treated adipocytes than in the control adipocytes. HM-chromanone also promoted the phosphorylation of 5' Adenosine monophosphate-activated protein kinase (AMPK), which inhibits adipogenesis through the regulation of adipogenic transcription factors. These results suggest that HM-chromanone may be an effective anti-adipogenesis agent that functions via the suppression of adipogenic transcription factors and the activation of AMPK.

Portulaca oleracea L. extract reduces hyperglycemia via PI3k/Akt and AMPK pathways in the skeletal muscles of C57BL/Ksj-db/db mice.

HEADINGS ETHNOPHARMACOLOGICAL RELEVANCE: Portulaca oleracea L. is a succulent annual herb, which has various pharmacological effects including antidiabetic property. However, in vivo the reducing effect of P. oleracea on hyperglycemia and its mechanism of action have not been clarified in a mouse model of type 2 diabetes. AIM OF THE STUDY: The effects of Portulaca oleracea L. extract (POE) on hyperglycemia were investigated in an animal model of type 2 diabetes. MATERIALS AND METHODS: C57BL/Ksj-db/db mice were randomly divided into three groups: db/db-control group was fed a standard semi-synthetic diet (AIN-93 G), db/db-RG group was fed AIN-93 G supplemented with rosiglitazone (RG) (0.005%, w/w), and db/db-POE group was fed AIN-93 G supplemented with POE (0.4%, w/w) for 6 weeks. Diabetes-related physical and biochemical indicators and the phosphorylation of components of PI3k/Akt and AMPK pathways were measured. RESULTS: The blood glucose and the glycosylated hemoglobin levels (HbA1c) in db/db-POE group were significantly lower than those in db/db-control group. In db/db-POE group, The homeostatic index of insulin resistance (HOMA-IR) decreased significantly, whereas the quantitative insulin sensitivity check index (QUICKI) was higher than those in db/db-control group. POE significantly elicited the phosphorylation of IRS-1Tyr612, AktSer473, and AS160Thr642, and the activation of PI3K in the skeletal muscle of mice. Additionally, POE significantly stimulated the phosphorylation of AMPKThr172, TBC1D1Ser231, and ACCSer79 and elevated the expression of plasma membrane-glucose transporter type 4 (GLUT4). CONCLUSIONS: These results indicate that POE reduces hyperglycemia by improving insulin resistance through the PI3k/Akt and AMPK pathways in the skeletal muscle of C57BL/Ksj-db/db mice.