Inhibition of Free fatty acid-Induced Insulin Resistance by Rosemary Extract: Investigation of the Mechanisms Involved

Excess of plasma free fatty acids (FFA) are highly associated with insulin resistance and are a major risk factor for the development of type 2 diabetes mellitus. This thesis investigates the effect of rosemary extract and rosemary extract polyphenols carnosic acid (CA), rosmarinic acid (RA) and carnosol (COH) on recovering/blunting FFA-induced insulin resistance in skeletal muscle cells and the mechanism(s) involved. Exposure of L6 myotubes with the FFA palmitate significantly reduced the insulin-stimulated glucose uptake. Most importantly, the insulin-stimulated glucose uptake was restored in the presence of RE and its polyphenols CA, RA and COH. Furthermore, treatment with palmitate increased serine phosphorylation of IRS-1 and significantly decreased the insulin-stimulated phosphorylation of Akt. These effects were completely abolished in the presence of rosemary extract. Additionally, we investigated the effect of palmitate and rosemary extract on the phosphorylation and of JNK, mTOR, P70S6K and AMPK kinases. Our results indicate that palmitate treatment increased the phosphorylation of JNK, mTOR, p70S6K whereas rosemary extract completely abolished this effect. Additionally, rosemary extract increased the phosphorylation of AMPK even in the presence of palmitate. The expression levels of these proteins were not affected by any of the treatments. These results indicate that treatment with rosemary extract attenuated the palmitate-induced phosphorylation of the serine residues of IRS-1, mTOR, p70 S6K and JNK while increasing the phosphorylation of AMPK. Additionally, treatment with RE restored the insulin-stimulated glucose uptake in palmitate-induced insulin resistant cells

Rosemary Extract as a Potential Anti-Hyperglycemic Agent: Current Evidence and Future Perspectives

Type 2 diabetes mellitus (T2DM), a disease on the rise and with huge economic burden to health care systems around the globe, results from defects in insulin action (termed insulin resistance) combined with impaired insulin secretion. Current methods of prevention and treatments for insulin resistance and T2DM are lacking in number and efficacy and, therefore, there is a need for new preventative measures and targeted therapies. In recent years, chemicals found in plants/herbs have attracted attention for their use as functional foods or nutraceuticals for preventing and treating insulin resistance and T2DM. Rosemary is an evergreen shrub indigenous to the Mediterranean region and South America, which contains various polyphenols. Rosemary extract and its polyphenolic constituents have been reported to have antioxidant, anti-inflammatory, anticancer, and anti-hyperglycemic properties. The current review summarizes the existing in vitro and in vivo studies examining the anti-diabetic effects of rosemary extract and its polyphenolic components and highlights the known mechanism of action

Inhibition of Free fatty acid-Induced Insulin Resistance by Rosemary Extract: Investigation of the Mechanisms Involved

Excess of plasma free fatty acids (FFA) are highly associated with insulin resistance and are a major risk factor for the development of type 2 diabetes mellitus. This thesis investigates the effect of rosemary extract and rosemary extract polyphenols carnosic acid (CA), rosmarinic acid (RA) and carnosol (COH) on recovering/blunting FFA-induced insulin resistance in skeletal muscle cells and the mechanism(s) involved. Exposure of L6 myotubes with the FFA palmitate significantly reduced the insulin-stimulated glucose uptake. Most importantly, the insulin-stimulated glucose uptake was restored in the presence of RE and its polyphenols CA, RA and COH. Furthermore, treatment with palmitate increased serine phosphorylation of IRS-1 and significantly decreased the insulin-stimulated phosphorylation of Akt. These effects were completely abolished in the presence of rosemary extract. Additionally, we investigated the effect of palmitate and rosemary extract on the phosphorylation and of JNK, mTOR, P70S6K and AMPK kinases. Our results indicate that palmitate treatment increased the phosphorylation of JNK, mTOR, p70S6K whereas rosemary extract completely abolished this effect. Additionally, rosemary extract increased the phosphorylation of AMPK even in the presence of palmitate. The expression levels of these proteins were not affected by any of the treatments. These results indicate that treatment with rosemary extract attenuated the palmitate-induced phosphorylation of the serine residues of IRS-1, mTOR, p70 S6K and JNK while increasing the phosphorylation of AMPK. Additionally, treatment with RE restored the insulin-stimulated glucose uptake in palmitate-induced insulin resistant cells

Rosmarinic Acid, a Rosemary Extract Polyphenol, Increases Skeletal Muscle Cell Glucose Uptake and Activates AMPK

Skeletal muscle is a major insulin-target tissue and plays an important role in glucose homeostasis. Impaired insulin action in muscles leads to insulin resistance and type 2 diabetes mellitus. 5′ AMP-activated kinase (AMPK) is an energy sensor, its activation increases glucose uptake in skeletal muscle and AMPK activators have been viewed as a targeted approach in combating insulin resistance. We previously reported AMPK activation and increased muscle glucose uptake by rosemary extract (RE). In the present study, we examined the effects and the mechanism of action of rosmarinic acid (RA), a major RE constituent, in L6 rat muscle cells. RA (5.0 μM) increased glucose uptake (186 ± 4.17% of control, p < 0.001) to levels comparable to maximum insulin (204 ± 10.73% of control, p < 0.001) and metformin (202 ± 14.37% of control, p < 0.001). Akt phosphorylation was not affected by RA, while AMPK phosphorylation was increased. The RAstimulated glucose uptake was inhibited by the AMPK inhibitor compound C and was not affected by wortmannin, an inhibitor of phosphoinositide 3-kinase (PI3K). The current study shows an effect of RA to increase muscle glucose uptake and AMPK phosphorylation. RA deserves further study as it shows potential to be used as an agent to regulate glucose homeostasis.Brock University Library Open Access Publishing Fun

Attenuation of Free Fatty Acid-Induced Muscle Insulin Resistance by Rosemary Extract

Elevated blood free fatty acids (FFAs), as seen in obesity, impair muscle insulin action leading to insulin resistance and Type 2 diabetes mellitus. Serine phosphorylation of the insulin receptor substrate (IRS) is linked to insulin resistance and a number of serine/threonine kinases including JNK, mTOR and p70 S6K have been implicated in this process. Activation of the energy sensor AMP-activated protein kinase (AMPK) increases muscle glucose uptake, and in recent years AMPK has been viewed as an important target to counteract insulin resistance. We reported recently that rosemary extract (RE) increased muscle cell glucose uptake and activated AMPK. However, the effect of RE on FFA-induced muscle insulin resistance has never been examined. In the current study, we investigated the effect of RE in palmitate-induced insulin resistant L6 myotubes. Exposure of myotubes to palmitate reduced the insulin-stimulated glucose uptake, increased serine phosphorylation of IRS-1, and decreased the insulin-stimulated phosphorylation of Akt. Importantly, exposure to RE abolished these effects and the insulin-stimulated glucose uptake was restored. Treatment with palmitate increased the phosphorylation/activation of JNK, mTOR and p70 S6K whereas RE completely abolished these effects. RE increased the phosphorylation of AMPK even in the presence of palmitate. Our data indicate that rosemary extract has the potential to counteract the palmitate-induced muscle cell insulin resistance and further studies are required to explore its antidiabetic properties

Attenuation of Free Fatty Acid-Induced Muscle Insulin Resistance by Rosemary Extract

Elevated blood free fatty acids (FFAs), as seen in obesity, impair muscle insulin action leading to insulin resistance and Type 2 diabetes mellitus. Serine phosphorylation of the insulin receptor substrate (IRS) is linked to insulin resistance and a number of serine/threonine kinases including JNK, mTOR and p70 S6K have been implicated in this process. Activation of the energy sensor AMP-activated protein kinase (AMPK) increases muscle glucose uptake, and in recent years AMPK has been viewed as an important target to counteract insulin resistance. We reported recently that rosemary extract (RE) increased muscle cell glucose uptake and activated AMPK. However, the effect of RE on FFA-induced muscle insulin resistance has never been examined. In the current study, we investigated the effect of RE in palmitate-induced insulin resistant L6 myotubes. Exposure of myotubes to palmitate reduced the insulin-stimulated glucose uptake, increased serine phosphorylation of IRS-1, and decreased the insulin-stimulated phosphorylation of Akt. Importantly, exposure to RE abolished these effects and the insulin-stimulated glucose uptake was restored. Treatment with palmitate increased the phosphorylation/activation of JNK, mTOR and p70 S6K whereas RE completely abolished these effects. RE increased the phosphorylation of AMPK even in the presence of palmitate. Our data indicate that rosemary extract has the potential to counteract the palmitate-induced muscle cell insulin resistance and further studies are required to explore its antidiabetic properties

Antidiabetic Effects of Hydroxytyrosol: In Vitro and In Vivo Evidence

Insulin resistance, a pathological condition characterized by defects in insulin action leads to the development of Type 2 diabetes mellitus (T2DM), a disease which is currently on the rise that pose an enormous economic burden to healthcare systems worldwide. The current treatment and prevention strategies are considerably lacking in number and efficacy and therefore new targeted therapies and preventative strategies are urgently needed. Plant-derived chemicals such as metformin, derived from the French lilac, have been used to treat/manage insulin resistance and T2DM. Other plant-derived chemicals which are not yet discovered, may have superior properties to prevent and manage T2DM and thus research into this area is highly justifiable. Hydroxytyrosol is a phenolic phytochemical found in olive leaves and olive oil reported to have antioxidant, anti-inflammatory, anticancer and antidiabetic properties. The present review summarizes the current in vitro and in vivo studies examining the antidiabetic properties of hydroxytyrosol and investigating the mechanisms of its action

Amelioration of High-Insulin-Induced Skeletal Muscle Cell Insulin Resistance by Resveratrol Is Linked to Activation of AMPK and Restoration of GLUT4 Translocation

Insulin resistance, the hallmark of type 2 diabetes mellitus (T2DM), is linked to hyperinsulinemia, which develops to counterbalance initial peripheral hormone resistance. Studies indicate that chronically elevated levels of insulin lead to skeletal muscle insulin resistance by deregulating steps within the insulin signaling cascade. The polyphenol resveratrol (RSV) has been shown to have antidiabetic properties in vitro and in vivo. In the present study, we examined the effect of RSV on high insulin (HI)-induced insulin resistance in skeletal muscle cells in vitro and investigated the mechanisms involved. Parental and GLUT4myc-overexpressing L6 rat skeletal muscle cells were used. [3H]2-deoxyglucose (2DG) uptake was measured, and total and phosphorylated levels of specific proteins were examined by immunoblotting. Exposure of L6 cells to HI levels (100 nM) for 24 h decreased the acute-insulin-stimulated 2DG uptake, indicating insulin resistance. HI increased ser307 and ser636/639 phosphorylation of IRS-1 (to 184% &plusmn; 12% and 225% &plusmn; 28.9% of control, with p &lt; 0.001 and p &lt; 0.01, respectively) and increased the phosphorylation levels of mTOR (174% &plusmn; 6.7% of control, p &lt; 0.01) and p70 S6K (228% &plusmn; 33.5% of control, p &lt; 0.01). Treatment with RSV abolished these HI-induced responses. Furthermore, RSV increased the activation of AMPK and restored the insulin-mediated increase in plasma membrane GLUT4 glucose transporter levels. These data suggest that RSV has a potential to counteract the HI-induced muscle insulin resistance

Rosemary Extract Activates AMPK, Inhibits mTOR and Attenuates the High Glucose and High Insulin-Induced Muscle Cell Insulin Resistance

Impaired action of insulin in skeletal muscle, termed insulin resistance, leads to increased blood glucose levels resulting in compensatory increase in insulin levels. The elevated blood glucose and insulin levels exacerbate insulin resistance and contribute to the pathogenesis of type 2 diabetes mellitus (T2DM). In previous studies we found attenuation of free fatty acid-induced muscle cell insulin resistance by rosemary extract (RE). In the present study we investigated the effects of RE on high glucose (HG) and high insulin (HI)-induced muscle cell insulin resistance. Exposure of L6 myotubes to 25 mM glucose and 100 nM insulin for 24 h, to mimic hyperglycemia and hyperinsulinemia, abolished the acute insulin-stimulated glucose uptake, increased the serine phosphorylation of IRS-1 and the phosphorylation/ activation of mTOR and p70S6K. Treatment with RE significantly improved the insulin-stimulated glucose uptake and increased the acute insulin-stimulated tyrosine phosphorylation while reduced the HG+HI-induced serine phosphorylation of IRS-1 and phosphorylation of mTOR and p70S6K. Additionally, treatment with RE significantly increased the phosphorylation of AMPK, its downstream effector ACC and the plasma membrane GLUT4 levels. Our data indicate a potential of RE to counteract muscle cell insulin resistance and more studies are required to investigate its effectiveness in vivo. Novelty: • Rosemary extract (RE) phosphorylated muscle cell AMPK and ACC under both normal and high glucose (HG)/high insulin (HI) conditions. • The HG/HI-induced serine phosphorylation of IRS-1 and activation of mTOR and p70S6K were attenuated by RE. • RE increased the insulin-stimulated glucose uptake by enhancing GLUT4 glucose transporter translocation to plasma membrane.The accepted manuscript in pdf format is listed with the files at the bottom of this page. The presentation of the authors' names and (or) special characters in the title of the manuscript may differ slightly between what is listed on this page and what is listed in the pdf file of the accepted manuscript; that in the pdf file of the accepted manuscript is what was submitted by the author

Attenuation of Free Fatty Acid (FFA)-Induced Skeletal Muscle Cell Insulin Resistance by Resveratrol is Linked to Activation of AMPK and Inhibition of mTOR and p70 S6K

Insulin resistance, a main characteristic of type 2 diabetes mellitus (T2DM), is linked to obesity and excessive levels of plasma free fatty acids (FFA). Studies indicated that significantly elevated levels of FFAs lead to skeletal muscle insulin resistance, by dysregulating the steps in the insulin signaling cascade. The polyphenol resveratrol (RSV) was shown to have antidiabetic properties but the exact mechanism(s) involved are not clearly understood. In the present study, we examined the effect of RSV on FFA-induced insulin resistance in skeletal muscle cells in vitro and investigated the mechanisms involved. Parental and GLUT4myc-overexpressing L6 rat skeletal myotubes were used. [3H]2-deoxyglucose (2DG) uptake was measured, and total and phosphorylated levels of specific proteins were examined by immunoblotting. Exposure of L6 cells to FFA palmitate decreased the insulin-stimulated glucose uptake, indicating insulin resistance. Palmitate increased ser307 (131% &plusmn; 1.84% of control, p &lt; 0.001) and ser636/639 (148% &plusmn; 10.1% of control, p &lt; 0.01) phosphorylation of IRS-1, and increased the phosphorylation levels of mTOR (174% &plusmn; 15.4% of control, p &lt; 0.01) and p70 S6K (162% &plusmn; 20.2% of control, p &lt; 0.05). Treatment with RSV completely abolished these palmitate-induced responses. In addition, RSV increased the activation of AMPK and restored the insulin-mediated increase in (a) plasma membrane GLUT4 glucose transporter levels and (b) glucose uptake. These data suggest that RSV has the potential to counteract the FFA-induced muscle insulin resistance