Ping Han

Amelioration of Lipid Abnormalities by α-Lipoic acid Through Antioxidative and Anti-Inflammatory Effects

Recent data have revealed that oxidative products and inflammatory mediators are increased in the insulin‐resistant states of obesity and type 2 diabetes mellitus (T2DM). Obese patients with impaired glucose tolerance (IGT) are at high risk for developing T2DM and have high incidence of dyslipidemia. α‐Lipoic acid (ALA) is a potent antioxidant with insulin sensitizing activity. However, it is not clear whether ALA is effective on lipid parameters in humans. This study has investigated 22 obese subjects with IGT (obese‐IGT), 13 of whom underwent 2‐week ALA treatment, 600 mg intravenously once daily. Before and after the treatment, euglycemic‐hyperinsulinemic clamps were used to measure insulin sensitivity. Meanwhile, plasma lipids, oxidative products, and chronic inflammatory markers were measured. After treatment of ALA in obese‐IGT patients, insulin sensitivity was improved, insulin sensitivity index (ISI) impressively enhanced by 41%. Plasma levels of free fatty acids (FFAs), triglyceride (TG), total cholesterol (T‐Chol), low density lipoprotein‐cholesterol (LDL‐Chol), small dense LDL‐Chol (sd‐LDL), oxidized LDL‐Chol (ox‐LDL‐Chol), very low density lipoprotein‐cholesterol (VLDL‐Chol) were all significantly decreased (P < 0.01). At the same time, both plasma oxidative products (malondialdehyde (MDA), 8‐iso‐prostaglandin) and inflammatory markers (tumor necrosis factor‐α (TNF‐α), interleukin‐6 (IL‐6)) were remarkably decreased (P < 0.01), while adiponectin was increased (P < 0.01). There are significant negative correlations between ISI and plasma FFAs, sd‐LDL‐Chol, ox‐LDL‐Chol, MDA, 8‐iso‐prostaglandin, TNF‐α, and IL‐6, and positive correlations with HDL‐Chol and adiponectin in obese‐IGT patients. The results indicate that short‐term treatment with ALA can improve insulin sensitivity and plasma lipid profile possibly through amelioration of oxidative stress and chronic inflammatory reaction in obese patients with IGT.

Epigallocatechin-3-O-gallate (EGCG) attenuates FFAs-induced peripheral insulin resistance through AMPK pathway and insulin signaling pathway in vivo

We aimed to investigate the effects and possible mechanisms of Epigallocatechin-3-O-gallate (EGCG) on free fatty acids (FFAs)-induced peripheral insulin resistance in vivo. Overnight-fasted Wistar rats were subjected to 48-h intravenous infusion of either saline or Intralipid plus heparin (IH) with or without different doses of EGCG co-injection. Hyperinsulinemic-euglycemic clamp was performed in awake rats to assess peripheral insulin sensitivity. Co-injection with EGCG significantly prevented FFAs-induced peripheral insulin resistance, decreased plasma markers of oxidative stress: malondialdehyde (MDA) and 8-isoprostaglandin, and increased antioxidant enzymes: superoxide dismutases (SOD) and Glutathione peroxidase (GPx). Furthermore, EGCG treatment reversed IH-induced: (1) decrease in Thr172 phosphorylation of AMP activated protein kinase (AMPK); (2) increase in protein kinase Cθ(PKCθ) membrane translocation and Ser307 phosphorylation of insulin receptor substrate-1 (IRS-1); (3) decrease in Ser473 phosphorylation of Akt and Glucose transporter 4 (GLUT4) translocation in skeletal muscle and adipose tissue. Our data suggest that EGCG treatment ameliorated FFAs-induced peripheral insulin resistance in vivo, and this might be through decreasing oxidative stress and PKCθ membrane translocation, activating the AMPK pathway and improving insulin signaling pathway in vivo. This study suggests the therapeutic value of EGCG in protecting from insulin resistance caused by elevated FFAs.

Taurine prevents free fatty acid-induced hepatic insulin resistance in association with inhibiting JNK1 activation and improving insulin signaling in vivo.

We infused the 48 h intralipid plus heparin (IH) to normal rats to elevate plasma free fatty acids (FFAs). Co-infusion of taurine was designed for the purpose of studying the effects of taurine on insulin sensitivity, oxidative stress, c-Jun NH-terminal kinase (JNK)1 activity and insulin signaling in livers of prolonged IH-infused rats. Cannulated rats were infused for 48 h intravenously with either saline or IH, with or without taurine. Hyperinsulinemic-euglycemic clamps with [6-3H] glucose infusion were performed to assess hepatic insulin sensitivity. IH infusion increased plasma 8-isoprostaglandin and hepatic malondialdehyde (MDA). IH also increased JNK1 activity and insulin receptor substrate 1/2 (IRS-1/2) serine phosphorylation, reduced insulin-stimulated IRS-1/2 tyrosine phosphorylation and Akt serine 473 phosphorylation, and induced hepatic insulin resistance. Taurine co-infusion with IH prevented the rise in 8-isoprostaglandin and MDA, inhibited the activation of JNK1, and improved insulin signaling and insulin resistance in liver. The present study has demonstrated that taurine, as an antioxidant, prevented hepatic oxidative stress and ameliorated hepatic insulin resistance. And this effect may be associated with the inhibition of JNK1 activation and the improvement of insulin signaling. This study suggests the therapeutic value of taurine in protecting from hepatic insulin resistance caused by elevated FFAs.

Amelioration in Hepatic Insulin Sensitivity by Reduced Hepatic Lipid Accumulation at Short-Term After Roux-en-Y Gastric Bypass Surgery in Type 2 Diabetic Rats

BackgroundPrevious studies showed that early after Roux-en-Y gastric bypass (RYGB), there is a remarkable improvement in type 2 diabetes, which is characterized by insulin resistance. This study aims to gain insight into the underlying mechanisms of this effect. We determined the acute effects of RYGB on hepatic and peripheral insulin sensitivity.MethodsA rat model of type 2 diabetes was established using high-fat diet combined with streptozotocin (30xa0mg/kg, ip). Animals were divided into four groups: diabetic, diabetic RYGB, diabetic RYGB sham, and control rats. Hyperinsulinemic-euglycemic clamps with tracer infusion were completed at 2xa0weeks postoperatively to assess insulin sensitivity. Triglyceride concentration in liver and muscle tissues was determined. Protein kinase C (PKC) membrane translocation, protein expression of phospho-c-Jun NH2-terminal kinase (JNK), and phospho-IκB kinase β (IKKβ) were assessed with western blot. Malondialdehyde (MDA) and superoxide dismutase (SOD) activities in the liver were also measured.ResultsRYGB surgery significantly improved hepatic insulin sensitivity index and decreased hepatic triglyceride concentration (Pu2009<u20090.05), without an improvement in peripheral insulin sensitivity. Membrane translocation of PKC-ε, PKC-δ, and PKC-θ; the ratio of MDA to SOD; and the expression of p-JNK and p-IKKβ in the liver were lower in the diabetic RYGB group than in the diabetic group.ConclusionsDiabetes remission was induced at short term after RYGB. The improvement of hepatic tissue lipotoxicity decreased the activation of certain PKC isoforms, the activity of JNK and IKK inflammatory signaling pathways, and the degree of oxidative stress. Furthermore, the hepatic insulin sensitivity was ameliorated, which is possibly a mechanism for early diabetes remission.

Salicylate prevents hepatic oxidative stress activation caused by short-term elevation of free fatty acids in vivo

It has been reported that high-dose salicylates, an IKKss inhibitor, may prevent FFAs-induced insulin resistance. In previous study, we found that in FFAs-induced insulin resistant rats, administration of salicylate was associated with a reduction of plasma malondialdehyde (MDA). In the present study, we investigated the effects of sodium salicylate on FFAs-induced insulin resistance and on oxidative stress in liver. Overnight-fasted Wistar rats were subject to 7h i.v. infusion of either saline or Intralipid plus 20U/ml heparin (IH) with or without salicylate. Hyperinsulinemic-euglycemic clamp with tracer infusion was performed to assess insulin-induced suppression of endogenous glucose production (EGP). Oxidative antioxidant markers, immunohistochemical inducible nitric oxide synthase (iNOS) stain, nitric oxide (NO), MDA, Superoxide dismutase (SOD) activity in liver was measured. Infusion of IH markedly decreased insulin-induced suppression of EGP, which were completely prevented by salicylate co-infusion. Furthermore, salicylate reversed IH-induced (1) increase in iNOS and NO expression in the liver; (2) increase in MDA/SOD in the liver. This study provides preliminary assessments of efficacy of sodium salicylate as a new treatment for FFAs-induced insulin resistances. The effect of increasing insulin sensitivity by salicylate in part may be secondary to reduce the oxidative stress in liver.

Acute blood glucose fluctuation induces myocardial apoptosis through oxidative stress and nuclear factor-ĸB activation.

Objective: It was the aim of this study to investigate whether acute blood glucose fluctuation induces myocardial apoptosis and to examine the potential mechanisms. Methods: Wistar rats were infused intermittently or continually with 50% glucose solution for 48 h. Serum and myocardium were taken to measure the levels of malondialdehyde and glutathione peroxidase. The expression of nuclear factor (NF)-ĸB and apoptosis in myocardial cells was determined with immunohistochemisty and terminal deoxynucleotidyl transferase dUTP nick end labeling, respectively. Expressions of B-cell lymphoma/leukemia-2-associated X protein and B-cell lymphoma/leukemia 2 in myocardium were tested with Western blot analysis. Results: The levels of malondialdehyde and B-cell lymphoma/leukemia-2-associated X protein in the acute blood glucose fluctuation group (AFG) were enhanced, but glutathione peroxidase and B-cell lymphoma/leukemia-2 were reduced compared with levels in the continually high blood glucose group (p < 0.05). The expression of NF-ĸB in the nuclei of myocardial cells in the AFG was significantly higher than that in the continually high blood glucose group (p < 0.05). Apoptotic myocytes were observed in myocardium of the AFG. Conclusion: Acute blood glucose fluctuation induces myocardial apoptosis, apparently associated with enhanced oxidative stress and activation of NF-ĸB.

The effect of LXRα, ChREBP and Elovl6 in liver and white adipose tissue on medium- and long-chain fatty acid diet-induced insulin resistance

AIMSnWe aimed to investigate the effects of LXRα, ChREBP and Elovl6 in the development of insulin resistance-induced by medium- and long-chain fatty acids.nnnMETHODSnSprague Dawley rats were fed a standard chow diet (Control group) or a high-fat, high sucrose diet with different fat sources (coconut oil, lard, sunflower and fish oil) for 8 weeks. These oils were rich in medium-chain saturated fatty acids (MCFA group), long-chain saturated fatty acids (LCFA group), n-6 and n-3 long-chain polyunsaturated fatty acids (n-6 PUFA and n-3 PUFA groups), respectively, which had different chain lengths and degrees of unsaturation. Hyperinsulinemic-euglycemic clamp with [6-(3)H] glucose infusion was performed in conscious rats to assess hepatic insulin sensitivity.nnnRESULTSnLCFA and n-6 PUFA groups induced hepatic insulin resistance and increased liver X receptor α (LXRα), carbohydrate response element binding protein (ChREBP) and long-chain fatty acid elongase 6 (Elovl6) expression in liver and white adipose tissue (WAT). Furthermore, LCFA and n-6 PUFA groups suppressed Akt serine 473 phosphorylation in liver and WAT. By contrast, in liver and WAT, MCFA and n-3 PUFA groups decreased LXRα, ChREBP and Elovl6 expression and improved insulin signaling and insulin resistance, but Akt serine 473 phosphorylation was not restored by MCFA group in WAT.nnnCONCLUSIONSnThis study demonstrated that the mechanism of the different effects of medium- and long-chain fatty acids on hepatic insulin resistance involves LXRα, ChREBP and Elovl6 alternations in liver and WAT. It points to a new strategy for ameliorating insulin resistance and diabetes through intervention on Elovl6 or its control genes.

Roux-en-Y Esophagojejunostomy Ameliorates Renal Function Through Reduction of Renal Inflammatory and Fibrotic Markers in Diabetic Nephropathy.

BackgroundRoux-en-Y bariatric surgery has been shown to have a remarkable and sustainable improvement in type 2 diabetes. Recent clinical studies have shown that bariatric surgery can improve or halt the development of diabetic microvascular complications such as nephropathy. However, the exact underlying mechanisms of surgical procedures are unknown. Here, we have investigated the effects of Roux-en-Y esophagojejunostomy (RYEJ) on renal function and inflammation and fibrosis biomarkers for renal injury in type 2 diabetic rats.MethodsSprague–Dawley rats with high fat diet and streptozotocin (STZ)-induced diabetes were randomly assigned into four groups: diabetic nephropathy (DN), DN treated with food restriction (DN-FR), DN treated with RYEJ surgery (DN-RYEJ), and DN-RYEJ sham (nu2009=u20096/group). Age-matched normal rats were assigned as control group. RYEJ and sham surgeries were performed. Hyperinsulinemic–euglycemic clamps with tracer infusion were completed to assess insulin sensitivity. Twenty-four hour urine albumin excretion rate (UAER) and glomerular filtration rate (GFR) were measured. The renal pathological injury was assessed by hematoxylin and eosin (HE) staining. Kidney messenger RNA (mRNA) and/or protein content/distribution of phospho-c-Jun NH2-terminal kinase (JNK), monocyte chemoattractant protein (MCP)-1, transforming growth factor (TGF)-β1, and mitogen-activated protein kinase phosphatase 5 (MKP5) were evaluated by real-time PCR and/or Western blotting/immunohistochemistry.ResultsRoux-en-Y esophagojejunostomy improved insulin sensitivity. RYEJ ameliorated renal function by improving UAER and GFR and attenuated glomerular hypertrophy after surgery. RYEJ also significantly downregulated the levels of JNK-mediated inflammatory response and upregulated the level of the anti-inflammatory mediator MKP5.ConclusionRoux-en-Y esophagojejunostomy alleviates insulin resistance. RYEJ surgery ameliorated renal function and attenuated glomerular hypertrophy in a DN rat model. The considerable nephroprotective function may be mainly attributed to the reduced inflammatory and fibrotic biomarkers after RYEJ. The improvements in renal function and inflammation are not wholly dependent on the magnitude of weight loss.

Roux-en-Y Gastric Bypass Reduces Lipid Overaccumulation in Liver by Upregulating Hepatic Autophagy in Obese Diabetic Rats

BackgroundThe decrease in lipotoxicity is one of the crucial mechanisms by which Roux-en-Y gastric bypass (RYGB) improves insulin sensitivity. Little work, however, has been performed to elucidate the exact mechanism of RYGB reducing hepatic lipid overaccumulation in response to heavy lipid and glucose challenge. Here, we explored the effects of RYGB on hepatic autophagy in obese diabetic rats.MethodsSprague–Dawley rats were divided into five groups: diabetic RYGB, diabetic RYGB sham, diabetic food restriction (FR), diabetic rats, and non-diabetic controls (nu2009=u200912/group). At 4-week post-operation, genetic and protein expressions of autophagy markers including Atg7 and Beclin 1 and the conversion of LC3 were examined with quantitative RT-PCR and Western blotting. Plasma glucagon-like peptide-1 (GLP-1) and triglyceride and total cholesterol levels in liver tissue were tested.ResultsIn both genetic and protein levels, we observed a significant upregulated autophagy in liver at 4xa0weeks after RYGB. Restored autophagy in liver played a key role in reducing the hepatic lipid burden in obese diabetic rats. The marked increase of autophagy in liver after RYGB correlated well with the plasma GLP-1 level.ConclusionsOur data demonstrate that RYGB significantly upregulated hepatic autophagy. We suggest that the effects of RYGB on autophagy in liver may be due to the increased GLP-1 level after surgery. Moreover, the activated autophagy in liver might play a key role in reducing the hepatic lipid overaccumulation after RYGB.

Role of the PKCβII/JNK signaling pathway in acute glucose fluctuation-induced apoptosis of rat vascular endothelial cells

AimsThe purpose of this study was to investigate the mechanism of vascular endothelial cell apoptosis induced by acute blood glucose fluctuation.MethodsThirty rats were assigned to three groups: normal saline (SAL group), constant high glucose (CHG group) and acute blood glucose fluctuation (AFG) group. Other forty rats were assigned to SAL group, AFG group, LY group (PKCβ inhibitor LY333531 was injected intragastrically to the rats who were under acute blood glucose fluctuation) and SP group (JNK inhibitor SP600125 was injected intraperitoneally to the rats who were under acute blood glucose fluctuation). Oxidative stress and inflammatory cytokines were detected. TUNEL was performed to detect apoptosis. Pro-caspase-3, caspase-3 p17, JNK, PKC-βII and insulin signaling-related protein expression were tested by Western blotting.ResultsAfter administration of LY333531, AFG-induced membrane translocation of PKCβII protein was inhibited, but SP600125 failed to affect AFG-induced PKCβII membrane translocation. After administration of LY333531, the AFG-induced increase in JNK activity was significantly compromised. LY333531 inhibited AFG-induced oxidative stress. However, SP600125 only slightly inhibited AFG-induced oxidative stress reaction (Pxa0>xa00.05). Both LY333531 and SP600125 can reverse AFG-induced endothelial cell apoptosis increase, inflammatory cytokines levels rise and insulin signaling impairment.ConclusionsIt is necessary to actively control blood glucose and avoid significant glucose fluctuation. PKCβII/JNK may serve as a target, and inhibitors of PKCβII/JNK may be used to help prevent cardiovascular diseases in patients with poor glucose control or significant glucose fluctuation.