Role of resistin in hepatic insulin resistance and the underlying mechanism

AIM: To investigate the role of resistin in hepatic insulin resistance and its mechanism. METHODS: A mouse model of hyperresistinemia in C57BL/6 mice was established by intravenous administration of the recombinant adenovirus encoding mouse resistin. Using periodic acid-Schiff staining we observed the effects of resistin on hepatic glycogen storage. Western blotting was used to measure AMPK-α protein and phosphrylated AMPK-α (Thr172) protein. mRNA levels of phosphoenolpyruvate carboxykinase gene and glucose-6-phosphatase gene were measured by real-time PCR. RESULTS: On day 5 after Adv injection, the concentration of plasma resistin was much higher in Adv-resistin-EGFP-treated mice than that in saline- or Adv-EGFP-treated mice. Semiquantitation of hepatic glucogen storage by PAS showed that the mice with hyperresistinemia had decreased glycogen particles compared to normal control and Adv-EGFP groups. The ratio of phosphorylated AMPK (Thr172)-α to total AMPK-α was used to evaluate hepatic AMPK activation. Compared with normal control and Adv-EGFP groups, the Adv-resistin-EGFP-treated mice had significantly lower ratio of p-AMPK/AMPK, and higher expression levels of G6Pase and PEPCK mRNA in liver. CONCLUSION: Resistin may decrease AMPK activation with downregulating expression of gluconeogenic enzymes, resulting in increased glucose production and decreased hepatic glycogen storage. Resistin may play an important role in hepatic insulin resistance.     

Correlation among plasma leptin,resistin an type 2 diabetes mellitus

AIM：To assess the association of resistin,obesity,serum lipid levels and insulin resistance with plasma leptin.METHODS：The concentrations of fasting serum glucose,insulin,lipid profiles,plasma resistin and leptin were assayed in 80 cases (including 37 controls with normal glucose tolerance and 43 patients with type 2 diabetes mellitus).RESULTS：Fasting plasma leptin level was positively correlated with sex,body mass index (BMI),waist circumference (WC),waist-hip ratio and fasting serum insulin (F Ins) （P<0.01）.Fasting plasma leptin level was negatively correlated with insulin sensitivity index (r=-0.373,P<0.01).There was no correlation between the concentrations of plasma leptin and FPG,TG,TC and resistin （P>0.05）.CONCLUSION：Fasting plasma leptin level is positively correlated with obesity and insulin resistance,not resistin.Leptin may play a role in the pathogenesis of type 2 diabetes mellitus.     

“2006中国蔬菜产业可持续发展研讨会”在武汉召开

由中国园艺学会主办，湖北省园艺学会、国家蔬菜改良中心华中分中心、华中农业大学园艺林学学院共同承办的“2006中国蔬菜产业可持续发展研讨会”于2006年4月13-15日在湖北武汉华中农业大学举行，来自全国17个省（市）和自治区47个单位的近120名代表出席这次全国性会议。会议开幕式由中国园艺学会蔬菜专业委员会主任、中国农业科学院蔬菜花卉研究所副所长孙日飞研究员主持。华中农业大学副校长李名家教授、武汉市张学忙副市长、科技部农村司魏勤芳处长、农业部科技司刘艳处长、武汉市科协黄和平主席和中国园艺学会理事长方智远院士先后致辞。     

Water decoction of Platycladus orientalis reduces blood glucose and lipids in mice

AIM: To study the effects of water decoction of Platycladus orientalis on reducing high blood glucose and lipids in diabetic and hyperlipidemia mice, respectively. METHODS: The mouse model of hyperlipidemia was established. The increasing rate of weight, hepatic index, lipid content of liver and atherosclerosis index (AI) were measured. Blood levels of total cholesterol (TC), triglyceride (TG), high-density lipoprotein cholesterol (HDL-C), adiponectin (ADP) and interleukin-6 (IL-6) were detected. According to these indexes, the function of water decoction of Platycladus orientalis for reducing high blood lipids caused by hyperlipidemia was evaluated. At the same time, the model of diabetes mellitus was established. The levels of fasting plasma glucose, fasting plasma insulin and blood tumor necrosis factor-α (TNF-α) were measured. The glucose tolerance test was performed. The insulin resistance index was calculated. The function of water decoction of Platycladus orientalis to reduce high blood glucose caused by diabetes was observed by these above indexes. RESULTS: Water decoction of Platycladus orientalis lowered the increasing rate of weight, hepatic index and lipid content of liver, reduced the blood content of TC, TG and IL-6, and elevated the blood HDL-C and ADP content caused by high-fat diet apparently (P<0.05). The effect of water decoction of Platycladus orientalis at high dose was more effective than that at low dose. Water decoction of Platycladus orientalis obviously alleviated the abnormity of glucose tole-rance test, reduced blood TNF-α and insulin levels, and decreased insulin resistance index caused by diabetes apparently (P<0.05). The effect of the drug at high dose was stronger than that at low dose. CONCLUSION: The water decoction of Platycladus orientalis dose-dependently reduces blood glucose caused by diabetes and blood lipids caused by hyperlipidemia in the mice. The mechanism to reduce blood lipids may be related with elevating blood ADP content and reducing blood IL-6 content, and the mechanism to reduce blood glucose may be related with lowering blood TNF-α content.     

Effects of TNF-α-mediated insulin resistance on INSIG2 and SCAP expressions in vivo

AIM: To investigate the effects of TNF-α induced insulin resistance (IR) on INSIG1, INSIG2, SCAP and SREBP expressions in mice. METHODS: Male C57BL/6J mice were randomly divided into 4 groups. The mice were given an intraperitoneal injection of TNF-α (6 μg·kg-1·d-1; 3 μg·kg-1·d-1 and 1 μg·kg-1·d-1) and saline (NC group) twice daily for 7 d. The insulin sensitivity and glucose metabolism in awaken mice were evaluated by intravenous glucose tolerance test (IVGTT). The mRNA expression and protein levels of gene were measured by RT-PCR and Western blotting. RESULTS: After TNF-α treatment, fasting blood glucose (FBG), plasma insulin and free fatty acids (FFA) were significantly elevated in TNF-α (6 μg·kg-1·d-1) group compared to NC, TNF-α (1 μg·kg-1·d-1) and TNF-α (3 μg·kg-1·d-1) groups (P<0.01 and P<0.05, respectively). There was a lower glucose tolerance in TNF-α (6 μg·kg-1·d-1) group than that in other three groups during IVGTT. In TNF-α (6 μg·kg-1·d-1) group, the insulin release of glucose-stimulation was higher than that in NC and TNF-α (1 μg·kg-1·d-1) groups (P<0.01 and P<0.05). The INSIG2 mRNA expression of adipose tissues in TNF-α (6 μg·kg-1·d-1) group was significantly increased compared with NC group (P<0.01), and INSIG2 protein levels were also increased (P<0.05). In TNF-α treatment mice, SCAP mRNA level in adipose tissues was significantly up-regulated than that in the controls (P<0.05). The mRNA expressions of INSIG1 and SREBP1 in two groups were not significantly changed (P>0.05). CONCLUSION: In TNF-α induced insulin resistance, INSIG2 and SCAP may be involved in the pathways of lipid metabolism.     

Impaired function of pancreatic β cells in the R6/2 transgenic mouse model of Huntington's disease

AIM: To study the function of pancreatic β cells in the R6/2 transgenic mouse of Huntingtons disease(HD), and to elucidate the pathogenetic mechanisms underlying diabetes mellitus in transgenic mice of HD. METHODS: By using the R6/2 transgenic mouse model of HD, fasting blood glucose and fasting insulin concentration in plasma of normal and HD mice were detected. Further, HE staining and immunofluorescence technique were used for morphometric analysis of islets in normal and HD mice. RESULTS: In contrast to normal mouse, R6/2 HD mouse showed hyperglycemia and hypoinsulinemia in fasting state. Pancreatic islets morphology showed that islets atrophied and cell number decreased in HD mouse. Poor functional index was observed in these mice, but insulin resistance index was normal. CONCLUSION: Impaired function of pancreatic cells may be the key factor contributing to the pathogenesis of diabetes in the R6/2 transgenic mouse model of HD.     

Protective effect on hepatic insulin resistance of Astragalus polysaccharide in the high fat-fed mice model

AIM: To investigate the effect and mechanism of Astragalus polysaccharide (APS) on the amelioration of hepatic insulin resistance in high fat-fed mouse model.METHODS: C57BL/6J mice (n=26) were divided into three groups randomly: C group (an animal model for control,n=10);IR group ( an animal model of insulin resistance,n=8) and IA group (an animal model in high-fat diet with APS treatment for12 weeks,700mg·kg^-1·d^-1,ig).High-fat diet was used to induce the formation of insulin resistant.The parameters and insulin sensitivity of the animals were observed.The pathological features of the liver were presented through microscope and TEM.The expression changes of hepatic GSK3β were measured by Western blotting.RESULTS: In this study,the fat-fed mouse model of insulin resistance was established successfully.The mice in IA group responded to the 12-week APS therapy with a significant decrease in the level of blood glucose,plasma insulin,body weight,hepatic TG/FFA and improved glucose tolerance compared with those in IR group.In addition,the expression and the activity of GSK3β were lower in IA group (vs IR group，P<0.05).We also found the hepatic steatosis could be significantly alleviated with APS therapy.CONCLUSION: These results indicate that APS prevents the occurrence of insulin resistance and the hepatic steatosis induced by high-fat diet,at least in part by inhibiting the expression and activity of the hepatic GSK3β.     

Protective effect of adiponectin on glucose and lipid metabolism by suppressing MHCⅡ expression

AIM: To investigate whether the protective effect of adiponectin on glucose and lipid metabolism is achieved through down-regulating major histocompatibility complex class Ⅱ (MHCⅡ) in the adipose tissue. METHODS: Adiponectin knockout (KO) mice and C57BL/6(WT) mice were fed with high-fat diet and standard diet for 24 weeks, respectively. The body weight, fasting blood glucose (FBG), fasting insulin (FINS), homeostasis model assessment of insulin resistance (HOMA-IR), triglyceride (TG), total cholesterol (TC), low-density lipoprotein cholesterol (LDL-C), high-density lipoprotein cholesterol (HDL-C), hepatic histology, and class Ⅱ trans-activator (CⅡTA), histocompatibility 2 class Ⅱ antigen E beta (H2-Eb1) and cluster of differentiation 74(CD74) mRNA and MHC Ⅱ protein levels in adipose tissue were measured at sacrifice. siRNA targeting MHC Ⅱ and overexpression vector was used in 3T3-L1 cells to explore the effect of adiponectin on the protein level of MHCⅡ. RESULTS: The levels of body weight, FBG, FINS, HOMA-IR, TC, TG, LDL-C, hepatic steatosis, CⅡTA, H2-Eb1 and CD74 mRNA expression, and MHCⅡ protein expression in the KO mice were higher than those in the WT mice that fed with high-fat diet or standard diet. In 3T3-L1 cells, inhibition of adiponectin reversed MHC Ⅱ protein level induced by specific siRNA. The expression of MHC Ⅱ in adipocytes decreased after adiponectin was overexpressed. CONCLUSION: Adiponectin improves glucose and lipid metabolism through suppressing the expression of MHCⅡ in the adipose tissue.     

Radix Pseudostellariae polysaccharide attenuates high fat diet induced hepatic insulin resistance in mice

AIM: To study the role of Radix Pseudostellariae polysaccharide (RPP) in hepatic insulin resistance.METHODS: Six-week-old C57BL/6J mice were randomly divided into low-fat diet (LFD) control group and high-fat diet (HFD) model group. After 16 weeks, intraperitoneal pyruvate tolerance test (IPPTT) was performed to determine the establishment of the HFD-induced hepatic insulin resistance model. HFD containing RPP (500 mg/kg) was given for 4 consecutive weeks. IPPTT, liver malondialdehyde (MDA) level and liver mitochondrial MDA level were measured. The protein levels of p-AKT (Ser473/Thr308), p-AMPK, nuclear factor E2-related factor 2 (Nrf2), NQO1 and IκBα in the liver tissues were measured by Western blot.RESULTS: After administration of RPP, a significant reduction in the levels of blood glucose and hepatic mitochondrial MDA was observed. The levels of p-AKT (Ser473/Thr308) and p-AMPK were significantly elevated in the liver tissues. The hepatic IκBα levels were up-regulated. RPP also enhanced the expression of Nrf2 system-regulated proteins NQO1 and HO-1 in the liver tissues.CONCLUSION: Radix Pseudostellariae polysaccharides effectively reduce HFD-induced hepatic insulin resistance in C57BL/6J mice and improves liver glucose metabolism by ameliorating HFD-impaired hepatic transduction of insulin signaling, activating Nrf2-associated signaling and inhibiting the expression of inflammatory signaling proteins.     

Influence of obesity on first-phase insulin secretion in individuals with different glucose tolerance

AIM: To explore the influence of obesity on the first-phase insulin secretion in the individuals with different glucose tolerance. METHODS: Thirty-eight subjects with normal glucose tolerance without family history of diabetes (normal control,NC), 32 subjects with normal glucose tolerance (NGT) who were the first-degree relatives of type 2 diabetic patients, 67 patients with impaired glucose regulation (IGR) and 35 newly-diagnosed type 2 diabetic patients (T2DM) were enrolled in the study. The patients in the 4 groups were further divided into 2 subgroups: overweight/obesity and normal weight subgroups. All subjects received oral glucose-insulin release test (OG-IRT) and intravenous glucose tolerance test (IVGTT). Acute insulin response at 3~5 min (AIR3-5) was used to reflect first-phase insulin secretion,and insulin sensitivity index (ISI) was used to reflect insulin sensitivity. The influence of obesity on the islet β-cell function and insulin sensitivity in the individuals with different glucose tolerance was observed. RESULTS: The level of AIR3-5 in NC overweight/obesity subgroup was significantly higher than that in normal weight subgroup (P<0.05), while there was no significant difference between other subgroups (P>0.05). No significant difference in the level of ISI between the patients of IGR in overweight/obesity subgroup and normal weight subgroup was observed, while in the other 3 overweight/obesity subgroups, ISI was lower than that in normal weight subgroups (P<0.05). ISI was negatively correlated with body mass index and waist circumference in all groups (P<0.05). ISI was also positively correlated with AIR3-5 in NC group and negatively correlated in the other 3 groups (P<0.05). CONCLUSION: The impact of obesity on insulin secretion is not the same in the subjects with different glucose tolerance. With the aggravation of insulin resistance, the first-phase insulin secretion in the subjects with normal glucose tolerance without family history of diabetes increases for compensation, while that in the normal glucose tolerance subjects who are the first-degree relatives of type 2 diabetic patients, the patients with impaired glucose regulation and the type 2 diabetic patients could not increase for compensation.     

Effect of L-glutamine on obesity and insulin resistance in high-fat diet-induced C57BL/6J mice

AIM:To explore the effect of L-glutamine (Gln) on obesity and insulin resistance in high-fat diet(HFD)-induced C57BL/6J mice. METHODS:Male C57BL/6J mice (n=60) were randomly divided into normal control (NC) group, HFD group, HFD+L-alanine (Ala) group and HFD+Gln group. Each group had 15 mice. The body weight of the mice was recorded weekly. Fasting blood glucose (FBG) of the mice was tested after 12-h fasting only with water-drinking at the end of the 16th week. The mice were sacrificed and epididymal fat pad was measured. The levels of insulin (INS), leptin (LEP), adiponectin (APN) and glucagon-like peptide-1 (GLP-1) were measured by ELISA. The insulin resistance index (IRI) and insulin sensitivity index (ISI) were calculated. RESULTS:Compared with NC group, the body weight, epididymal fat pad weight, and the levels of FBG, INS, IRI and LEP increased significantly in HFD group (P<0.05), while the levels of ISI and APN decreased significantly (P<0.05). Compared with HFD group, the body weight, and the levels of FBG, IRI and LEP decreased significantly in HFD+Gln group (P<0.05), while the levels of ISI and APN increased significantly (P<0.05). No significant difference of serum GLP-1 levels the four groups was observed. CONCLUSION:L-glutamine reduces the body weight and attenuates the insulin resistance of HFD-induced mice.     

Oxymatrine ameliorates high fat-induced insulin resistance in ApoE-/- mice

ATM: To investigate the effect of oxymatrine (OXY) on high fat-induced insulin resistance in mice, and to investigate the mechanism. METHODS: ApoE^-/-mice with high-fat diet for 16 weeks were divided into insulin resistance group, and OXY groups at concentrations of 25, 50 and 100 mg/kg. C57BL/6J mice served as normal control group. The mice in OXY groups were gavaged with OXY for 8 weeks. Glucose tolerance test in the mice was performed. Fasting blood glucose (FBG), total cholesterol (TC), triglyceride (TG), fatty acid (FFA) and fasting insulin (FINS) in the plasma were measured. The mRNA expression of insulin receptor (INSR), insulin receptor substrate-2 (IRS-2), glucose transporter 2 (GLUT2) in the liver tissues was examined by RT-qPCR. The protein levels of GLUT2, INSR, IRS-2, p-INSR, p-IRS-2, PI3K, p-PI3K, serine/threonine protein kinase (AKT) and p-AKT were examined by Western blot.RESULTS: OXY reduced the levels of FBG, TC, TG, FFA and FINS, and attenuated insulin resistance. Compared with insulin resistance group, the mRNA expression of INSR, IRS-2 and GLUT2 significantly increased in OXY groups (P<0.05). The protein levels of p-INSR/INSR, p-IRS-2/IRS-2, p-PI3K/PI3K, p-AKT/AKT and GLUT2 also increased in OXY groups (P<0.05).CONCLUSION: OXY ameliorates high fat-induced insulin resistance in mice via PI3K/AKT pathway.     

Protective effect of chrysin on mice with insulin resistance

AIM: To explore the effects of chrysin on insulin resistance (IRe) in a mouse model. METHODS: Male C57 mice were randomly divided into control group, IRe group, low-dose chrysin group (IRe+chrysin-low) and high-dose chrysin group (IRe+chrysin-high). After 24 weeks, the body weight, liver index and fat mass in all mice were detected. The blood glucose, insulin level and HOMA-IR were measured to determine the changes of the insulin resistance in the animals. The oxidative stress (SOD, GSH-Px and MDA) was also measured. The mRNA expression of insulin signaling pathway molecules (IR, IRS1, IRS2, Glut2 and Glut4) and inflammatory factors (TNF-α, IL-1β, IL-6 and NF-κB) was analyzed by real-time PCR. The protein levels of IRS1 and p65, and their phosphorylation were detected by Western blot. RESULTS: After 24-week intervention, the indicators in IRe group were higher than those in control group, including body fat deposition, serum glucose, serum insulin, HOMA-IR and liver oxidative stress (P<0.01), indicating that the model of insulin resistance was successfully established. Low dose and high dose of chrysin decreased the body weight, serum glucose, serum insulin and HOMA-IR in the IRe mice (P<0.05). The liver oxidative stress was also reduced in both groups (P<0.05). However, no statistical difference of the indexes between IRe+chrysin-low group and IRe+chrysin-high group was observed. Chrysin upregulated the mRNA expression of IR, IRS1, IRS2, Glut2 and Glut4 (P<0.05), and down-regulated the mRNA expression of various inflammatory factors. The inhibitory effect of chrysin on the mRNA expression of NF-κB was observed (P<0.05), especially in high dose group (P<0.05). It was confirmed that the effect of chrysin on liver IRe was related with the increase in the p-IRS1 levels and decrease in the p-p65 levels by Western blot. CONCLUSION: Chrysin inhibits obesity, hyperglycemia and hyperinsulinemia, and relieves insulin resistance and oxidative stress, which might be closely related to the regulation of insulin signaling pathway and the inhibition of inflammatory factor expression.     

Severe hypertriglyceridemia appears in apolipoprotein CⅢ transgenic mouse

AIM: To investigate the biological functions of human apolipoprotein CⅢ (hApoCⅢ) on metabo-lic regulation, hApoCⅢ transgenic mouse model was established.METHODS: hApoCⅢ transgenic mice were generated by injecting hApoCⅢ genomic fragment into fertilized mouse oocytes, and transgenic line was screened using PCR and Southern blotting. Plasma lipid analysis, fast protein liquid chromatography(FPLC), oral fat load test, post-heparin plasma lipoprotein lipase(LPL) activity analysis and glucose tolerance test were performed to characterize the phenotypic changes of lipid and glucose metabolism in the transgenic mice.RESULTS: We successfully generated the hApoCⅢ transgenic mouse model, which showed evident hypertriglyceridemia(HTG) , delayed triglyceride(TG) clearance and reduced LPL activity in post-heparin plasma while no change was detected in glucose tolerance test.CONCLUSION: It is suggested that the manifested evident HTG in hApoCⅢ transgenic mice may be due to inhibition of LPL activity in post-heparin plasma, which would lead to a prolonged clearance of plasma TG.     

Hypolipemic and hypoglycemic effects of total triterpenoids from Psidium guajava leaves on type 2 diabetic rats

AIM: To investigate the effects of total triterpenoids from Psidium guajava leaves (TTPGL) on blood glucose and lipids in type 2 diabetic rats. METHODS: The diabetic rats were induced by intraperitoneal injection of streptozotocin at dose of 35 mg/kg and feeding with high-fat diet. The animals were divided into 5 groups: diabetic model control group (model), TTPGL treatment groups (with the doses of 60, 120 and 240 mg/kg, respectively) and rosiglitazone treatment group (3 mg/kg). Another 12 normal SD rats were used as the normal controls. The rats received daily treatment for 6 weeks, and then the levels of fasting blood glucose (FBG), fasting insulin (FINS), triglyceride (TG), total cholesterol (TCH), free fatty acid (FFA), glycosylated hemoglobin (GHb) and glycosylated serum proteins (GSP) were measured. The protein expression of peroxisome proliferator-activated receptor γ (PPARγ) in adipose tissues was detected by Western blotting. RESULTS: Compared with normal control group, the levels of FBG, GHb and blood lipids were increased in type 2 diabetic rats. The FINS, insulin sensitivity index, and the protein expression of PPARγ in adipose tissues were decreased. Compared with model group, the levels of FBG and GSP were decreased,and the FINS, insulin sensitivity index, and the protein expression of PPARγ in adipose tissues significantly increased in TTPGL treatment groups (with the doses of 120 and 240 mg/kg). The levels of serum TG,TCH and FFA were significantly lower in TTPGL treatment groups (P<0.01 or P<0.05) as compared with the model controls. CONCLUSION: TTPGL decreases the levels of blood glucose and lipids in diabetic rats. TTPGL also increases serum insulin level and improves insulin sensitivity. The action mechanism of TTPGL may be related to the increase in the protein expression of PPARγ.     

Establishment of insulinoma animal models and observation of tumor characteristics

AIM：To establish the animal model of insulinoma and to analyze the properties of insulinoma for further study. METHODS：The hormone-releasing ability of rat insulinoma cell line INS-1 was detected in vitro. INS-1 cells were transplanted into the left kidney capsule of nude mice. The islets of the animals were destroyed by intraperitoneal injection of streptozocin (STZ) 3 d before transplantation or 2 weeks after transplantation. The venous blood was sampled, and the level of blood glucose less than 2.8 mmol/L was defined as successful establishment of insulinoma model. Different irritants were given to the model animals, and the changes of blood glucose and insulin content in serum were observed. The pancreatic tissues and the renal tissues in the injecting sites were taken from all mice for detecting insulin and glucagon by immunohistochemical staining. RESULTS：Insulinoma cells expressed insulin and glucagon at the same time. The blood glucose was less than 2.8 mmol/L 3 to 4 weeks after inoculation of INS-1 cells. Apparent tumor formed in the left kidneys where INS-1 cells were transplanted and the tumor diameters were more than 1 cm. The level of blood glucose transiently increased to higher than the normal level in the mice with tumor cell transplantation after intraperitoneal injection of STZ, and then decreased gradually and returned to less than 2.8 mmol/L after 2 weeks. The level of blood glucose in the normal nude mice after administration of STZ increased significantly. After transplantation of INS-1 cells, the level of blood glucose decreased gradually, and returned to less than 2.8 mmol/L after 4 weeks. After stimulated with high glucose, the blood glucose levels in the mice with 3 methods to establish the insulinoma models showed lower glucose peaks than that in the normal control mice. After stimulated with high glucose plus arginine or acetylcholine in the normal animals, the blood glucose peak was lower than that in the normal animals only stimulated with high glucose, and rapidly recovered to the normal level. However, the levels of blood glucose in the mice with 3 methods to establish the insulinoma models under the same stimulations were significantly higher than that in the mice only stimulated with high glucose. After stimulated with high glucose plus norepinephrine, the blood glucose peak time in the normal animals delayed, and the blood glucose level declined slowly. After stimulated with high glucose plus norepinephrine, the levels of blood glucose in the mice with 3 methods to establish the insulinoma models increased as compared with that in the mice only stimulated with high glucose. Compared with normal control group, serum insulin in insulinoma mice increased significantly. CONCLUSION：The insulinoma animal model is successfully established by transplantation of INS-1 cells into the renal capsule of nude mice. The insulinoma cells express both insulin and glucagon, and are not easily damaged by STZ.     

PPARs signaling pathway is involved in diabetic hepatopathy in mice

AIM: To investigate the role of peroxisome proliferator-activated receptors (PPARs)-inflammation signaling pathways in diabetic hepatopathy. METHODS: Diabetic mouse model was established by feeding the mice with a high-energy diet for 4 weeks combined with intraperitoneal injection of streptozotocin (STZ; 40 mg·kg^-1·d^-1 for 5 d). The hepatopathy model was confirmed by histopathological observation and the indexes of liver function, such as alanine aminotransferase (ALT), aspartate aminotransferase (AST) and alkaline phosphatase (ALP), after another 4 weeks. Moreover, fasting blood glucose (FBG), and serum levels of total cholesterol (TC), triglyceride (TG) and insulin were measured, and the HOMA insulin resistance index (HOMA-IR) was calculated. The mRNA and protein expression levels of PPARs and inflammation-related factors were measured by qPCR and Western blot, respectively. RESULTS: After treatment with STZ for 7 d, the FBG of mice exceeded 11.1 mmol/L, suggesting that the diabetic model was established. After 4 weeks, the structural deformation of the hepatocytes (including hepatocytes containing abundant fat vacuoles, and inflammatory cell infiltration), and the increases in the serum levels of insulin, HOMA-IR, TC, TG, ALT, AST and ALP were observed (P<0.01), indicating the occurrence and progression of hepatopathy in diabetic mice. Meanwhile, compared with the control group, the mRNA and protein expression of PPARα, PPARβ and PPARγ decreased, but the expression of nuclear factor-κB (NF-κB), cyclooxygenase 2 (COX-2) and inducible nitric oxide synthase (iNOS) significantly increased in the diabetic hepatopathy mice (P<0.01). CONCLUSION: Down-regulation of PPARα, PPARβ and PPARγ and activation of NF-κB-COX-2/iNOS signaling pathways may be involved in the diabetic hepatopathy in mice induced by long-term high-energy diet feeding combined with intraperitoneal injection of STZ.     

Expression of scavenger receptor B1 (SR-B1) in the liver of high fat and sugar diet-induced diabetic mice

AIM: To investigate serum lipid and the expression of SR-B1 in the livers of diabetic mice. METHODS: Ten normal diet, female C57BL/6J mice, fifteen high fat and sugar diet female C57BL/6J mice, five fed 8 weeks and ten fed 16 weeks were used in the experiment. Serum total cholesterol (TC), triglycerides (TG), high density lipoprotein cholesterol (HDL-C), fasting blood glucose (FBG), insulin (INS) and the expression of SR-B1 in the livers were measured. RESULTS: 1. In the high fat and sugar diet mice, serum TC and FBG at 16 weeks were significantly higher than that in normal diet mice (P<0.05). 2. The expression of SR-B1 protein in the liver of high fat and sugar diet mice was the higher than that in normal mice, and the SR-B1 expression in the liver of the mouse fed 16 weeks was also higher than that fed 8 weeks. CONCLUSION: The expression of SR-B1 protein in the liver of type 2 diabetes mice is higher than that in normal mice, perhaps it is related to the decrease in serum HDL-C.     

Early high-protein diet improves glucose metabolism in SGA rats via adiponectin-AMPK signaling

AIM: To investigate the effect of early high-protein diet on glucose metabolism in small-for-gestational-age (SGA) rats and the role of adiponectin-AMP-activated protein (AMPK) signaling in this process. METHODS: Forty-eight neonatal male SGA rats were established by maternal food restriction throughout the period of pregnancy. The animals were randomly divided into SGA control group (CS group, 24 rats) and high-protein intervention SGA group (HPS group, 24 rats) when born. Twenty-four normal neonatal male rats were used as normal control group (CN group). The rats in CN group and CS group were breastfed for 3 weeks and their mothers were provided free access to basic diet. After weaning, they were provided free access to basic diet until 12 weeks of age. The rats in HPS group were breastfed for 3 weeks and their mothers were provided free access to high-protein diet. After weaning, they were provided free access to high-protein diet until 4 weeks of age. At the 4th week of age, they were provided free access to basic diet until 12 weeks of age. From 4 to 12 weeks of age, fasting blood glucose and insulin were measured and homeostasis model assessment of insulin resistance index (HOMA-IR) was calculated. The serum adiponectin level and the visceral fat mass (VFM) were detected. The percentage of VFM to body weight (VFM%) was calculated. The visceral fat was paraffin-embedded and the adipocyte area was determined. The expression of AMPKα,phosphorylated AMPKα (p-AMPKα) and glucose transporter 4 (GLUT4) in skeletal muscle was analyzed. At the 12th week of age, the oral glucose tolerance test (OGTT) was performed. RESULTS: At the 4th week of age, no significant difference of HOMA-IR among groups was observed. At the 12th week of age, HOMA-IR in CS group was significantly higher than that in CN group and HPS group. No significant difference of HOMA-IR between CN group and HPS group was found. From 4 to 12 weeks of age, VFM% and adipocyte area of visceral fat in CS group were significantly higher than those in CN group and HPS group. No significant difference of VFM% and adipocyte area of visceral fat between CN group and HPS group was observed. The level of serum adiponectin, and the expression of p-AMPKα and GLUT4 in skeletal muscle were significantly lower in CS group than those in CN group and HPS group. The protein levels of p-AMPKα and GLUT4 were not significantly different between CN group and HPS group. No significant difference in the expression of AMPKα in skeletal muscle among groups was observed. At the 12th week of age, the area under the curve (AUC) of glucose level in OGTT was higher in CS group than that in CN group and HPS group. No significant difference of AUC of glucose level between CN group and HPS group was found. CONCLUSION: Early high protein diet may improve glucose metabolism in SGA rats, partly by avoiding excessive accumulation of visceral fat and possibly by activating adiponectin-AMPK signal pathway to increase GLUT4 expression.     

Differentiation of mouse iPSCs into insulin-producing cells and experimental study on treating diabetes

AIM: To induce mouse induced pluripotent stem cells (iPSCs) to differentiate into insulin-producing cells (IPCs) by a new 3-step method, and to detect the efficiency and maturity for the treatment of diabetic mice. METHODS: We constructed iPSCs from mouse embryonic fibroblasts of male C57/C mouse by piggyBac transposon, then induced the iPSCs into IPCs by a 3-step method. The cell morphological change was traced by microscopy during the process of differentiation. The expression of mRNA and protein associated with islet β cell development was determined by real-time PCR and immunofluorescence staining. Flow cytometry was used to analysis the efficiency of differentiation. Insulin and C-peptide secretions of IPCs in response to glucose at high (25 mmol/L) or low (5.5 mmol/L) level were measured by ELISA. The IPCs were transplanted into the capsul of left kidney in the male C57/C diabetic mouse model. Blood glucose was continuously monitored for 28 day, serum insulin was tested by ELISA in different stages. The glucose tolerance test was performed on the 28th day, and the left kidney was excised. RESULTS: IPCs were obtained from mouse iPSCs by the 3-step method. The cells expressed the marker genes (Pdx1, Ngn3, Pax6 and Ins2) and proteins (Pdx1, Nkx6.1 and insulin) of β cells. The glucose stimulation induced the secretion of insulin and C-peptide. The efficiency of differentiation was 28% detected by flow cytometry. After transplantation of IPCs to the diabetic mice, the blood glucose was decreased to normal level on the 3rd day,and serum insulin level and the ability of regulating glucose were improved. IPCs were still alive after 28 d of transplantation by pathological observation. CONCLUSION: iPSCs is efficiently induced into IPCs by a 3-step method , and the induction time is shortened significantly. The hyperglycemia of diabetes mice is reversed after transplanting IPCs to same sex inbred strain mice.