Calcium/calmodulin dependent protein kinase IV in trophoblast cells under insulin resistance: functional and metabolomic analyses.

BACKGROUND: Insulin resistance (IR) is an important determinant of glucose metabolic disturbance and placental dysplasia in gestational diabetes mellitus (GDM). Calcium/calmodulin dependent protein kinase IV (CAMK4) improves insulin IR induced by a high-fat diet (HFD). The current study sought to elucidate the role and potential mechanism of CAMK4 in GDM. METHODS: A GDM model was established in female C57BL/6J mice via HFD feeding for one week before mating and throughout gestation. The IR was elicited by 10-6 M insulin treatment for 48 h in HTR-8/SVneo cells and mouse primary trophoblast cells. The function of CAMK4 was investigated by transfection of overexpression plasmid in HTR-8/SVneo cells and infection of lentivirus loaded with CAMK4 encoding sequence in primary trophoblast cells. Real-time PCR, western blot, cell counting kit-8, transwell, wound healing, dual-luciferase reporter assay, and liquid chromatography/mass spectrometry-based untargeted metabolomics were performed to confirm the effects of CAMK4 on trophoblast cells. RESULTS: Decreased CAMK4 expression was found in the placenta of GDM mice. CAMK4 overexpression ameliorated IR-induced viability impairment, migratory and invasive capacity inhibition, autophagy blocking, insulin signaling inactivation and glucose uptake disorder in trophoblast cells. CAMK4 also transcriptionally activated orphan nuclear receptor NUR77, and the effects of CAMK4 were abrogated by silencing of NUR77. Metabolomics analysis revealed that CAMK4 overexpression caused alterations of amino acid, lipid and carbohydrate metabolism, which were important in GDM. CONCLUSION: Our results indicated that CAMK4/NUR77 axis may provide novel potential targets in GDM treatment.

Orphan nuclear receptor NUR77 relieves insulin resistance in HTR-8/SVneo trophoblast cells through activation of autophagy and insulin signaling

Gestational diabetes mellitus (GDM) is a common pregnancy complication with a high incidence in women. Orphan nuclear receptor NUR77 is involved in regulating glucose metabolism. However, its role in GDM has not been fully elucidated yet. In this study, an animal model of GDM was established by feeding mice with a high-fat diet (HFD) before and during pregnancy. NUR77 expression was abnormally upregulated in placenta tissues of GDM mice. We performed gain- and loss-of-function studies of NUR77 in HTR-8/SVneo cells. Cells were incubated with 1 × 10−6 M insulin for 48 h to induce insulin resistance (IR). The expression of NUR77 was downregulated in HTR-8/SVneo cells following IR induction. Overexpression of NUR77 promoted cell proliferation, migration, and invasion. Notably, NUR77 promoted glucose uptake and enhanced insulin sensitivity in vitro. NUR77 increased the ratio of p-insulin receptor β (IRβ)Tyr1361/IRβ, p-insulin receptor substrate (IRS)-1Tyr612/IRS-1, p-Akt/Akt and decreased p-IRS-1Ser307/IRS-1, as well as lowered the expression of glucose transport protein type 1 (GLUT1) and elevated GLUT4. These results suggest the involvement of IRβ/IRS/Akt/GLUT4 signaling activation in the regulatory effects of NUR77 on IR in HTR-8/SVneo cells. Silencing of NUR77 displayed opposite effects. Besides, NUR77 enhanced the expression of autophagy-related protein Beclin 1 and the ratio of LC3II/LC3I. Further study demonstrated that the inhibitory effect of NUR77 on IR was partially attributed to the activation of autophagy. Therefore, we demonstrate that NUR77 enhances insulin sensitivity in HTR-8/SVneo cells likely through activating IRβ/IRS/Akt/GLUT4 pathway and regulating autophagy. (AU)

Tortuosity of branch retinal artery is more associated with the genesis and progress of diabetic retinopathy.

Objective: The purpose of this study is to investigate the potential of using the tortuosity of branch retinal artery as a more promising indicator for early detection and accurate assessment of diabetic retinopathy (DR). Design and method: The diagnoses, consisting of whether DR or not as well as DR severity, were given by ophthalmologists upon the assessment of those fundus images from 495 diabetic patients. Meanwhile, benefiting from those good contrast and high optical resolution fundus images taken by confocal scanning laser ophthalmoscope, the branch arteries, branch veins, main arteries and main veins in retina can be segmented independently, and the tortuosity values of them were further extracted to investigate their potential correlations with DR genesis and progress based on one-way ANOVA test. Results: For both two comparisons, i.e., between non-DR group and DR group as well as among groups with different DR severity levels, larger tortuosity increments were always observed in retinal arteries and the increments in branch retinal vessels were even larger. Furthermore, it was newly found that branch arterial tortuosity was significantly associated with both DR genesis (p=0.030) and DR progress (p<0.001). Conclusion: Based on this cohort study of 495 diabetic patients without DR and with different DR severity, the branch arterial tortuosity has been found to be more closely associated with DR genesis as well as DR progress. Therefore, the branch arterial tortuosity is expected to be a more direct and specific indicator for early detection of DR as well as accurate assessment of DR severity, which can further guide timely and rational management of DR to prevent from visual impairment or even blindness resulting from DR.

Orphan nuclear receptor NUR77 relieves insulin resistance in HTR-8/SVneo trophoblast cells through activation of autophagy and insulin signaling.

Gestational diabetes mellitus (GDM) is a common pregnancy complication with a high incidence in women. Orphan nuclear receptor NUR77 is involved in regulating glucose metabolism. However, its role in GDM has not been fully elucidated yet. In this study, an animal model of GDM was established by feeding mice with a high-fat diet (HFD) before and during pregnancy. NUR77 expression was abnormally upregulated in placenta tissues of GDM mice. We performed gain- and loss-of-function studies of NUR77 in HTR-8/SVneo cells. Cells were incubated with 1 × 10-6 M insulin for 48 h to induce insulin resistance (IR). The expression of NUR77 was downregulated in HTR-8/SVneo cells following IR induction. Overexpression of NUR77 promoted cell proliferation, migration, and invasion. Notably, NUR77 promoted glucose uptake and enhanced insulin sensitivity in vitro. NUR77 increased the ratio of p-insulin receptor ß (IRß)Tyr1361/IRß, p-insulin receptor substrate (IRS)-1Tyr612/IRS-1, p-Akt/Akt and decreased p-IRS-1Ser307/IRS-1, as well as lowered the expression of glucose transport protein type 1 (GLUT1) and elevated GLUT4. These results suggest the involvement of IRß/IRS/Akt/GLUT4 signaling activation in the regulatory effects of NUR77 on IR in HTR-8/SVneo cells. Silencing of NUR77 displayed opposite effects. Besides, NUR77 enhanced the expression of autophagy-related protein Beclin 1 and the ratio of LC3II/LC3I. Further study demonstrated that the inhibitory effect of NUR77 on IR was partially attributed to the activation of autophagy. Therefore, we demonstrate that NUR77 enhances insulin sensitivity in HTR-8/SVneo cells likely through activating IRß/IRS/Akt/GLUT4 pathway and regulating autophagy.

Biological networks in gestational diabetes mellitus: insights into the mechanism of crosstalk between long non-coding RNA and N6-methyladenine modification.

BACKGROUND: Gestational diabetes mellitus (GDM) is one of the most common complications of pregnancy. The mechanism underlying the crosstalk between long non-coding RNAs (lncRNAs) and N6-methyladenine (m6A) modification in GDM remain unclear. METHODS: We generated a lncRNA-mediated competitive endogenous RNA (ceRNA) network using comprehensive data from the Gene Expression Omnibus database, published data, and our preliminary findings. m6A-related lncRNAs were identified based on Pearson correlation coefficient (PCC) analysis using our previous profiles. An integrated pipeline was established to constructed a m6A-related subnetwork thereby predicting the potential effects of the m6A-related lncRNAs. RESULTS: The ceRNA network was composed of 16 lncRNAs, 17 microRNAs, 184 mRNAs, and 338 edges. Analysis with the Kyoto Encyclopedia of Genes and Genomes database demonstrated that genes in the ceRNA network were primarily involved in the development and adverse outcomes of GDM, such as those in the fatty acid-metabolism pathway, the peroxisome proliferator-activated receptor signaling pathway, and thyroid hormone signaling pathway. Four m6A-related lncRNAs were involved in the ceRNA network, including LINC00667, LINC01087, AP000350.6, and CARMN. The m6A-related subnetwork was generated based on these four lncRNAs, their ceRNAs, and their related m6A regulators. Genes in the subnetwork were enriched in certain GDM-associated hormone (thyroid hormone and oxytocin) signaling pathways. LINC00667 was positively correlated with an m6A "reader" (YTHDF3; PCC = 0.95) and exhibited the highest node degree in the ceRNA network. RIP assays showed that YTHDF3 directly bind LINC00667. We further found that MYC possessed the highest node degree in a protein-protein interaction network and competed with LINC00667 for miR-33a-5p. qPCR analysis indicated that LINC00667, YTHDF3 and MYC levels were upregulated in the GDM placentas, while miR-33a-5p was downregulated. In a support-vector machine classifier, an m6A-related module composed of LINC00667, YTHDF3, MYC, and miR-33a-5p showed excellent classifying power for GDM in both the training and the testing dataset, with an accuracy of 76.19 and 71.43%, respectively. CONCLUSIONS: Our results shed insights into the potential role of m6A-related lncRNAs in GDM and have implications in terms of novel therapeutic targets for GDM.

Impact of Insulin Resistance on Cardiometabolic Risk Factors and an Anthropometry-Based Predictive Nomogram for Insulin Resistance Among Adolescents in China.

Objective: We aimed to investigate the impact of insulin resistance (IR), as determined by the homeostasis model assessment of insulin resistance (HOMA-IR), on cardiometabolic risk factors (CMRFs), and develop an anthropometry-based predictive nomogram for IR among adolescents in China. Design: Data were acquired from a cross-sectional study with a stratified cluster sampling method, conducted among adolescents in Northeast China. Participants: A total of 882 adolescents (aged 12-16 years, 468 boys) were included. Measurements: All participants underwent anthropometric and biochemical examinations. The thresholds of IR included the 90th percentile of the HOMA-IR for adolescents with a normal body mass index (BMI) and fasting plasma glucose (FPG) level within each sex group (Cutoff A), and the 75th percentile for all participants of the same sex (Cutoff B). Results: The HOMA-IR was associated with CMRFs. IR, as defined by both cutoffs A and B, was significantly associated with most CMRFs, except decreased HDL-C levels. Excellent concordance (κ = 0.825) was found between these two criteria in diagnosing IR. However, IR using cutoff A, was more closely associated with cardiometabolic risk. The incidence of IR, as defined by cutoff A, was 18.93% and increased from 10.99% to 43.87% based on the different BMI categories. Further, an anthropometry-based predictive model for IR, incorporating sex, age, waist-to-hip ratio, weight and BMI, was developed and presented as a nomogram. Conclusions: IR among adolescents is strongly related to cardiometabolic risk. We developed an anthropometry-based predictive nomogram for IR among adolescents, which may facilitate health counselling and self-risk assessments.

circMAP3K4 regulates insulin resistance in trophoblast cells during gestational diabetes mellitus by modulating the miR-6795-5p/PTPN1 axis.

BACKGROUND: Insulin resistance (IR) during gestational diabetes mellitus (GDM) has been linked to dysregulated insulin-PI3K/Akt pathway. A defective insulin-PI3K/Akt pathway and dysregulated circular RNA (circRNA) levels have been observed in the placentas of patients with GDM; however, the mechanisms underlying this association remain unclear. METHODS: circRNAs potentially associated with GDM were selected through bioinformatics analysis and initially identified by quantitative real-time PCR (qPCR) in 9 GDM patients and 9 healthy controls, of which circMAP3K4 was further validated in additional 84 samples by qPCR. circMAP3K4 identity and localization were verified. Pearson correlation analysis was applied to evaluate the correlation between circMAP3K4 expression in the placental tissues of GDM patients and IR-related indicators. An IR model of trophoblasts was constructed using glucosamine. Interactions between miR-6795-5p and circMAP3K4 or PTPN1 were confirmed using a dual-luciferase reporter assay. The circMAP3K4/miR-6795-5p/PTPN1 axis and key markers in the insulin-PI3K/Akt pathway in placentas and trophoblasts were evaluated through qRT-PCR, immunofluorescence, and western blotting. The role of circMAP3K4 in glucose metabolism and cell growth in trophoblasts was determined using the glucose uptake and CCK8 assay, respectively. RESULTS: circMAP3K4 was highly expressed in the placentas of patients with GDM and the IR trophoblast model; this was associated with a dysregulated insulin-PI3K/Akt pathway. circMAP3K4 in the placentas of GDM patients was positively correlated with weight gain during pregnancy and time-glucose area under the curve of OGTT. circMAP3K4 and PTPN1 could both bind to miR-6795-5p. miR-6795-5p and PTPN1 were downregulated and upregulated, respectively, in the placentas of GDM patients and the IR trophoblast model. circMAP3K4 silencing or miR-6795-5p overexpression partially reversed the decrease in glucose uptake, inhibition in cell growth, and downregulated IRS1 and Akt phosphorylation in IR-trophoblasts; this restoration was reversed upon co-transfection with an miR-6795-5p inhibitor or PTPN1. CONCLUSION: circMAP3K4 could suppress the insulin-PI3K/Akt signaling pathway via miR-6795-5p/PTPN1 axis, probably contributing to GDM-related IR.

N6-Methyladenosine-Related Gene Signature Associated With Monocyte Infiltration Is Clinically Significant in Gestational Diabetes Mellitus.

Objective: The objective of this study was to reveal the potential crosstalk between immune infiltration and N6- methyladenosine (m6A) modification in the placentas of patients with gestational diabetes mellitus (GDM), and to construct a model for the diagnosis of GDM. Methods: We analyzed imbalanced immune infiltration and differentially expressed m6A-related genes (DMRGs) in the placentas of patients with GDM, based on the GSE70493 dataset. An immune-related DMRG signature, with significant classifying power and diagnostic value, was identified using a least absolute shrinkage and selection operator (LASSO) regression. Based on the selected DMRGs, we developed and validated a nomogram model using GSE70493 and GSE92772 as the training and validation sets, respectively. Results: Infiltration of monocytes was higher in GDM placentas than in control samples, while the infiltration of macrophages (M1 and M2) in GDM placentas was lower than in controls. A total of 14 DMRGs were strongly associated with monocyte infiltration, seven of which were significant in distinguishing patients with GDM from normal controls. These genes were CD81, CFH, FABP5, GBP1, GNG11, IL1RL1, and SLAMF6. The calibration curve, decision curve, clinical impact curve, and receiver operating characteristic curve showed that the nomogram recognized GDM with high accuracy in both the training and validation sets. Conclusions: Our results provide clues that crosstalk between m6A modification and immune infiltration may have implications in terms of novel biomarkers and therapeutic targets for GDM.

Aberrantly Expressed Non-Coding RNAs in the Placenta and Their Role in the Pathophysiology of Gestational Diabetes Mellitus.

Gestational diabetes mellitus (GDM), one of the most common complications during pregnancy, is associated with a high risk of short- and long-term adverse effects on the mother and offspring. Placenta-derived hormones and cytokines aggravate maternal insulin resistance (IR) during pregnancy, which in turn contribute to GDM. The hyperglycemia and IR in GDM result in aberrant placental structure and function adversely affecting fetal growth and well-being. Therefore, it is reasonable to assume that structural and functional alterations in the placenta contribute to the pathogenesis of GDM and GDM-related complications. Increasing evidence suggests that multiple non-coding RNAs (ncRNAs), including microRNAs, long non-coding RNAs, and circular RNAs, are dysregulated in placentas of patients with GDM and linked to abnormal placental structure, metabolism, and function. Manipulation of ncRNA expression led to some key pathophysiological features of GDM, such as trophoblast dysfunction, changes in intracellular glucose metabolism, and inflammation. Moreover, placenta-specific ncRNAs may be potential diagnostic biomarkers and even therapeutic targets for GDM. This review summarizes data published on the involvement of aberrantly expressed placental ncRNAs in GDM and provides information on their role in the pathogenesis of GDM and GDM-associated complications.

Secreted Frizzled-Related Protein 5 is Associated with Glucose and Lipid Metabolism Related Metabolic Syndrome Components Among Adolescents in Northeastern China.

PURPOSE: Secreted frizzled-related protein 5 (SFRP5) is a novel anti-inflammatory adipokine that has been associated with various metabolic diseases. However, such relationship among adolescents remains unclear. The purpose of this study was to clarify the relationship between SFRP5 and the components of metabolic syndrome in Chinese adolescents. PATIENTS AND METHODS: In this cross-sectional study, we included a total of 684 adolescents aged 11-16 years old from Liaoyang city, Liaoning province, China. The ELISA kits were implemented to measure the plasma SFRP5 and high-sensitivity C-reactive protein. Serum total cholesterol (TC), low-density lipoprotein cholesterol (LDL-C), triglycerides (TG), and high-density lipoprotein cholesterol (HDL-C), serum uric acid (UA), alanine aminotransferase (ALT), aspartate aminotransferase (AST), fasting plasma glucose (FPG), and fasting serum insulin (FINS) were also measured. RESULTS: The multivariate logistic regression analysis showed that low SFRP5 level were an independent risk factor of high FPG [odds ratio (OR)=5.31, 95% confidence interval (CI): 1.85-15.22, P<0.01] and high TC (OR=1.73, 95% CI: 1.01-2.96, P<0.05) when adjusting for age, sex, family history of diabetes, body mass index, and high-sensitivity C-reactive protein. CONCLUSION: The lower level of SFRP5 is strongly related to lipid and glucose metabolism among adolescents in Northeast China. The risk of high fasting plasma glucose and high total cholesterol increases significantly as the plasma SFRP5 level decreases.

X-linked congenital adrenal hypoplasia: a case presentation.

BACKGROUND: Most patients with congenital adrenal hypoplasia (AHC) develop symptoms during infantile and juvenile periods, with varying clinical manifestations. AHC is a disease that is easily misdiagnosed as Addison's disease or congenital adrenal hyperplasia (CAH). There was also a significant time difference between the age at which patients developed symptoms and the age at which they were diagnosed with AHC. Most patients showed early symptoms during infantile and juvenile periods, but were diagnosed with AHC many years later. CASE PRESENTATION: We are currently reporting a male patient who developed systemic pigmentation at age 2 and was initially diagnosed with Addison's disease. At 22 years of age, he experienced a slipped capital femoral epiphysis (SCFE), a disease mostly seen in adolescents aged 8-15 years, an important cause of which is endocrine disorder. Testes evaluated using color Doppler Ultrasonography suggested microcalcifications. Further genetic testing and auxiliary examinations revealed that the patient had hypogonadotropic hypogonadism (HH) and DAX-1 gene disorders, at which time he was diagnosed with AHC complicated by HH. He was given hormone replacement therapy, followed by regular outpatient review to adjust the medication. CONCLUSIONS: The typical early symptoms of AHC are hyperpigmentation and ion disturbance during infantile and juvenile periods, while few patients with AHC develop puberty disorders as early symptoms. AHC is prone to being misdiagnosed as Addison's disease, and then gradually develops the symptoms of HH in adolescence. The definitive diagnosis of AHC ultimately is based on the patient's clinical presentation, laboratory results and genetic testing results.

Estimating the Risk of Insulin Requirement in Women Complicated by Gestational Diabetes Mellitus: A Clinical Nomogram.

PURPOSE: This study sought to develop a nomogram for the prediction of insulin requirement in a Chinese population with gestational diabetes mellitus (GDM). MATERIALS AND METHODS: We performed a retrospective cohort study involving 626 Chinese women with GDM, of whom 188 were treated with insulin. "Least absolute shrinkage and selection operator" regression was used to optimize the independent predictors of insulin requirement during pregnancies complicated with GDM. Cox proportional hazards regression analysis was performed to establish a prediction model incorporating the selected predictors, and the nomogram was constructed to achieve individual prediction. The C-index, calibration plot and decision curve analysis were used to validate the model. RESULTS: Maternal age, family history of type 2 diabetes mellitus in a first-degree relative, a prior GDM history, fasting plasma glucose, hemoglobin A1c, gestational age, and body mass index values at the time of GDM diagnosis were the risk factors for insulin treatment. The model displayed medium predictive power with a C-index of 0.77 (95% confidence interval: 0.73-0.81) and relatively good calibration accuracies. The decision curve demonstrated a positive net benefit with a threshold between 0.09 and 0.70. CONCLUSION: The findings suggest that our nomogram, incorporating seven indicators, is useful in predicting individualized survival probabilities of insulin requirement.

Fasudil alleviated insulin resistance through promotion of proliferation, attenuation of cell apoptosis and inflammation and regulation of RhoA/Rho kinase/insulin/nuclear factor-κB signalling pathway in HTR-8/SVneo cells.

OBJECTIVES: The aim of this study was to evaluate the effects of fasudil on insulin resistance (IR) in HTR-8/SVneo cells. METHODS: HTR-8/SVneo cells were treated with insulin or/and fasudil. Cell proliferation, apoptosis, inflammation and related signalling pathways were assessed. KEY FINDINGS: Insulin treatment significantly enhanced the protein expressions of RhoA and Rho kinase (ROCK1 and ROCK2), but decreased glucose consumption. Administration of fasudil effectively promoted glucose uptake. Moreover, fasudil enhanced cell viability and the level of proliferating cell nuclear antigen (PCNA). Insulin-mediated cell apoptosis was inhibited by fasudil via the down-regulation of bax and cleaved-caspase-3, and the up-regulation of bcl-2. At the same time, fasudil led to the reduction of IL-1ß, TNF-α, IL-6 and IL-8 mRNA levels in insulin-treated cells. In addition, RhoA, ROCK2 and phosphorylated myosin phosphatase target subunit-1 (p-MYPT-1) expressions were down-regulated by fasudil. Importantly, fasudil activated insulin receptor substrate-1 (IRS-1) through increasing p-IRS-1 (Tyr612) and p-Akt expressions. The nuclear NF-κB p65 and p-IκB-α levels were reduced via the administration of fasudil in insulin-treated cells. CONCLUSIONS: Fasudil mitigated IR by the promotion of cell proliferation, inhibition of apoptosis and inflammation and regulation of RhoA/ROCK/insulin/NF-κB signalling pathway through in vitro studies.

The role of Smad4 in the regulation of insulin resistance, inflammation and cell proliferation in HTR8-Svneo cells.

Gestational diabetes mellitus (GDM) is a metabolic disorder whose major pathophysiological basis is demonstrated as placental insulin resistance (IR), while Smad4 always functions in the signal transduction of transforming growth factor beta (TGF-ß) pathway. Our study aims to figure out the role of Smad4 in an insulin resistance (IR) cellular model using placental trophoblast cell line. Importantly, HTR8-Svneo cells, in the status of IR, indicated a significant increase in the expression of Smad4. Subsequently, the HTR8-Svneo cell line with up-regulated or depleted Smad4 was respectively achieved by the effective over-expressed plasmid or siRNA of Smad4. We found out that the deficiency of Smad4 could promote the insulin sensitivity and restrict the inflammatory response in IR group of cells with significant augment in glucose uptake, up-regulation of insulin signalling-related molecules and attenuation in inflammatory biomarker expressions. On the contrary, the over-expression of Smad4 showed a reversal effect on these alterations in IR group of cells. Besides, the positive effect of Smad4 on cell viability was also observed in our study. SIGNIFICANCE OF THE STUDY: Gestational diabetes mellitus (GDM) is a metabolic disorder whose major pathophysiological basis is demonstrated as insulin resistance (IR). Importantly, our findings indicate that the deficiency of Smad4 significantly improves the insulin sensitivity and relieves the inflammation in the cellular model of IR. Besides, the positive effect of Smad4 on cell viability was also observed in our study. Our present findings provide novel insights for the investigation on molecular details about the GDM pathogenesis.

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

AIMS: The purpose of this study was to investigate the mechanism of vascular endothelial cell apoptosis induced by acute blood glucose fluctuation. METHODS: Thirty 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 (PKCß 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-ßII and insulin signaling-related protein expression were tested by Western blotting. RESULTS: After administration of LY333531, AFG-induced membrane translocation of PKCßII protein was inhibited, but SP600125 failed to affect AFG-induced PKCßII 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 (P > 0.05). Both LY333531 and SP600125 can reverse AFG-induced endothelial cell apoptosis increase, inflammatory cytokines levels rise and insulin signaling impairment. CONCLUSIONS: It is necessary to actively control blood glucose and avoid significant glucose fluctuation. PKCßII/JNK may serve as a target, and inhibitors of PKCßII/JNK may be used to help prevent cardiovascular diseases in patients with poor glucose control or significant glucose fluctuation.

Roux-en-Y esophagojejunostomy reduces serum and aortic inflammatory biomarkers in type 2 diabetic rats.

BACKGROUND: Recent studies have shown the reduction in serum inflammatory biomarkers by bariatric surgery. However, few studies have reported its effects on local vascular inflammation. We have investigated the effects of Roux-en-Y esophagojejunostomy on both serum and aortic tissue inflammation biomarkers in type 2 diabetic rats. METHODS: Sprague-Dawley rats were divided into five groups: diabetic RYEJ, diabetic RYEJ sham, diabetic food restriction, diabetic rats, and non-diabetic control (n = 6/group). At 4 weeks after surgery, serum leptin, interleukin-6, C-reactive protein, interleukin-1ß (IL-1ß), tumor necrosis factor-α (TNF-α), chemerin, and adiponectin were measured. Aortic expression of c-Jun NH2-terminal kinase and p38 mitogen-activated protein kinases was examined with immunohistochemistry; aortic expression of IL-1ß, TNF-α, intercellular adhesion molecule-1, and vascular adhesion molecule-1 was assessed with western blotting. Hyperinsulinemic-euglycemic clamps with tracer infusion were completed to assess insulin sensitivity. RESULTS: Roux-en-Y esophagojejunostomy significantly decreased serum inflammatory parameters and increased the concentration of the anti-inflammatory mediator adiponectin. The aortic inflammatory protein expression was markedly decreased in the diabetic RYEJ group as compared with that of the diabetic group (P < 0.05). Additionally, Roux-en-Y esophagojejunostomy improved insulin sensitivity and dyslipidemia and decreased body weight and total body fat. CONCLUSIONS: Roux-en-Y esophagojejunostomy reduces the systemic and local vascular inflammation. The improvements in systemic and vascular inflammation are not wholly dependent on the magnitude of weight loss. Moreover, Roux-en-Y esophagojejunostomy alleviates insulin resistance and improves the features of metabolic syndrome, leading to a reduction in multiple cardiovascular risks.