[Effects of AdipoRon orally on the functions of spleen and pancreas in type 2 diabetic mice].

OBJECTIVE: To observe the effects of AdipoRon orally on the functions of spleen and pancreas in type 2 diabetic mice, in order to present data for clinical application. METHODS: Forty C57/BL6 male mice were randomly divided into 2 groups: normal control group (n=10) and model group (n=30), the former group was fed normally, while the later group was fed with high fat and sugar for 4 weeks.After that, type 2 diabetes model was established in DM group induced by intraperitoneal injection of streptozotocin (STZ, 40 mg/kg).As type 2 diabetes model established successfully, the model mice were randomly divided into three groups (n=10): diabetes mellitus (DM) group, high dose of AdipoRon group (DM + H) and low dose of adiponRon group (DM + L).All the four groups were treated with saline, saline, AdipoRon at the doses of 20 mg/kg and 50 mg/kg by gavages respectively, once a day for 10 days.And then put them to death for collecting blood, pancreas and spleen.Pathological changes of pancreas were observed with a light microscope after HE staining.Protein contents of insulin receptor (INSR), insulin receptor substrate 1( IRS-1) and tumor necrosis factor-α(TNF-α) in pancreatic and spleen tissues were detected by ELISA.The protein level of phosphorylation insulin receptor substrate 1(p-IRS-1) in pancreas was determined by Western blot, and the expression of insulin mRNA in pancreas was tested by RT-PCR. RESULTS: Under the light microscope, it was visible that the pancreatic tissue in NC group was full and closely packed, and the islet was big.Pancreatic tissue of DM mice was incompact and the islet of DM mice was smaller than that of normal mice.As for the mice treated with AdipoRon orally, the pancreatic tissue was full and closely arranged, and the islet was slightly smaller.Compared with NC group, the levels of TNF-α in pancreas and spleen of DM group were increased markedly, the levels of INSR and IRS-1 were decreased, the spleen coefficient, p-IR-1 protein level and insulin mRNA expression in pancreas were decreased, all were significant statistically (P＜0.05).Compared with DM group, the levels of TNF-α in pancreas and spleen of AdipoRon groups were decreased, the levels of INSR and IRS-1 in pancreas and spleen of AdipoRon groups were increased, while the spleen coefficient was increased (P＜0.05).The p-IRS-1 protein level and insulin mRNA expression in pancreas in DM+H group were increased (P＜0.05).Compared with DM + L group, the level of TNF-α was decreased, and the levels of INSR and IRS-1 were significantly increased (P＜0.05) in DM + H group (P＜0.05). CONCLUSION: Oral administration of AdipoRon can protect the spleen and pancreas of diabetic mice by decreasing the inflammatory response, up-regulating the expression of INSR, and increasing p-IRS-1 level in diabetic mice.

[The intervention effects of AdipoRon on renal injury in type 2 diabetic mice].

OBJECTIVE: To study the effects of adiponin receptor agonist (AdipoRon) on renal injury in type 2 diabetic mice. METHODS: The experiment was carried out on 40 SPF C57/BL6 male mice and they were randomly divided into normal control group (n=10) and experimental group (n=30). Mice in experimental group were given with high sugar and high fat feed in combination with only an intraperitoneal injection of small dose of streptozotocin to build the model of type 2 diabetes (T2DM), which were randomly divided into three groups, model control group (DM), low dose AdipoRon group (DM + L) and high dose AdipoRon group (DM+H)(n=10). Then the change of blood glucose was detected. The serum levels of insulin receptor (INSR), insulin receptor substrate-1 (IRS-1) and tumor necrosis factor-α (TNF-α) in mice were measured by ELASA. Pathological changes of renal tissues were observed with a light microscope after HE staining. The expressions of pancreatic duodenal homebox-1 (PDX-1) and insulin mRNA in renal tissues were detected by RT-PCR. The content of phosphated insulin receptor substrate-1 (p-IRS-1) protein in the kidney was determined by Western blot. RESULTS: Compared with DM mice, blood glucose and TNF-α levels in DM + H mice and DM + L mice were significantly reduced (P<0.05), while the expressions of INSR,IRS-1 and the content of p-IRS-1 were increased markedly(P<0.05), and the expressions of PDX-1 and insulin mRNA in renal tissue were increased significantly(P<0.05, P<0.01). CONCLUSIONS: Mice treated with AdipoRon have lower blood glucose and TNF-α levels, and higher protein expression levels of INSR, IRS-1, and higher mRNA expression levels of PDX-1 and insulin, and the content of p-IRS-1. All of these indicate that AdipoRon has a certain effects on renal injury in type 2 diabetic mice.

[Intervention effects of oral active AdipoRon on liver oxidative stress in type 2 diabetic mice].

OBJECTIVE: To explore the intervention effects of oral active AdipoRon on liver oxidative stress in type 2 diabetic mice, which provides basic data for clinical application. METHODS: Thirty-two healthy male C57BL/6 mice were divided into 4 groups:normal group (NC, n=8), diabetes mellitus group (DM, n=8), high dose AdipoRon treatment group (DM + H, n=8) and low dose AdipoRon treatment group (DM + L, n=8). Following six weeks high fat feed, mice of DM, DM + H and DM + L were intraperitoneally injected with 40 mg/kg streptozocin (STZ), leading to type 2 diabetes. Afterwards, DM + H group and DM + L group were continuously treated with high dose and low doses of oral AdipoRon respectively for 10 days, following which, related biochemical indicators were detected. Western blot method was used to detect the p-IRS-1 protein expression in liver tissue and RT-PCR method to detect PDX-1 mRNA expression in the pancreas. RESULTS: The blood glucose of DM group was obviously higher than that of NC group (P < 0.05). Compared to that of DM group, blood glucose of DM + H group as well as DM + L group was significantly lower. Activity of superoxide dismutase (SOD), catalase (CAT) in liver tissue of DM mice was significantly lower than that of NC group (P < 0.05); activity of malondialdehyde (MDA) and nitric oxide synthase (NOS) in DM group significantly higher than that of NC group (P < 0.05); activity of SOD and CAT in DM + L group and DM + H group obviously higher than DM group (P < 0.05); activity of MDA and NOS in DM + L group and DM + H group significantly lower than DM group (P < 0.05). And the p-IRS-1 protein expression in liver tissue and PDX-1 mRNA level in pancreas increased significantly (P < 0.05). CONCLUSIONS: Oral active Adi-poRon which reduced the blood glucose levels of mice had a certain intervention effect on liver tissue oxidative stress in type 2 diabetes mice.

[AdipoRon for the treatment of type 2 diabetes in mice and its possible mechanism of the liver].

OBJECTIVE: To observe the effect of AdipoRon for the treatment of type 2 diabetes (T2DM)in mice and its effect on the liver. METHODS: Forty male C57/BL6 mice (SPF) were randomly divided to normal control (NC) group and the experimental group. To establish the T2DM mice model, mice in the experimental group were fed with high fat and high glucose, combined with intraperitoneal injection of streptozotocin (STZ) in small doses, and mice were further subdivided into model control (DM) group, model control with low AdipoRon (DM+L) group and model control with high AdipoRon (DM+H) group (n=10). Serum indexes, such as levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP) were detected biochemically and the morphological changes of liver cells were observed with HE staining and expression of liver carbohydrate related gene (PEPCK) were determined by real-time fluorescence quantitative PCR (real time FQ-PCR). RESULTS: Compared with mice in the DM group, levels of ALT, AST, ALP, triglyceride (TG), glucose (GLU) reduced in DM+L and DM+H group (P<0.05). Concentrations of serum free fatty acids (FFA) in DM+L and DM+H group reduced significantly (P<0.05). Besides, concentrations of liver glucose-6-phosphatase (G-6-P) in the mice of DM+L group reduced significantly, while there was no significant difference in the content of G-6-P between the mice of DM+H group and the mice of DM group. Furthermore, the expression of the liver phosphoenolpyruvate carboxylase (PEPCK) in the DM+H group reduced significantly (P<0.05). Compared with the DM group no significant change was found in the PEPCK expression between DM+L and DM group. CONCLUSIONS: The serum indexes such as levels of ALT, AST, ALP, TG, Glu, G-6-P and PEPCK were all reduced in DM mice treated with AdipoRon, indicating the obvious protecting effect of AdipoRon on the liver in DM mice.

[Effects of pirfenidone on hepatic fibrosis in mice induced by carbon tetrachloride].

OBJECTIVE: To investigate the effects of pirfenidone on CCl4-induced liver fibrosis in mice. METHODS: After 8-week feeding, 40 healthy male SPF ICR mice were randomly divided into 4 groups:liver fibrosis group (CCL4 group), low doses of Pirfenidone group (PFD-L group), high doses of Pirfenidone group (PFD-H group) and control group. The mice in CCL4 group, low doses of Pirfenidone group (PFD-L group), high doses of Pirfenidone group (PFD-H group) were injected intraperitoneally with 0.4 ml 10% CCL4 solution dissolved in soybean oil. Then the PFD-L and PFD-H groups were treated with 120 mg and 240 mg PFD via gastric gavage, respectively. Control group was injected with same volume of saline. Alanine aminotransferase(ALT), aspartate aminotransferase(AST), alkaline phosphatase(ALP) in serum were tested with automatic biochemistry analyzer and the pathologic changes of liver tissue were examined by HE staining. Furthermore, we identi-fied hyaluronic acids(HA), laminin(LN), collagentype IV(IV-C) in serum using radioimmunoassay and the expression of smooth muscle acti-nalpha(α-SMA) related gene in liver was tested by real-time fluorescence quantitative PCR. RESULTS: Compared with control group, hepatic lobules in CCL4 mice were damaged significantly, collagenous fiber was deposited obviously, and counterfeit hepatic lobules formed. The serum levels of ALT, AST, ALP were increased obviously (P<0.05) with the enhancement of HA, LN, IV-C in serum (P<0.05) and the ex-pression of α-SMA related gene (P<0.05). Compared to CCL4-treated mice, the serum levels of ALT, AST, ALP in PFD-L and PFD-H groups were decreased, HA, LN, IV-C in PFD-L and PFD-H mice went down obviously,and the expression of α-SMA related gene was con-trolled (P<0.05). From pathological observation, we found the degree of liver fibrosis in PFD-L mice was reduced and collagenous fiber was decreased, only a little counterfeit hepatic lobule could be found. Cell arrangement in PFD-H mice recovered, the structural of hepatic lobules disordered and no obvious counterfeit hepatic lobules were found. Therefore, the recovery of PFD-H group was better than PFD-L group. CONCLUSIONS: Pirfenidone has a protective role in improving the outcome of the liver fibrosis and it may become a new direction of early intervention in liver fibrosis.

[Effect of ligustrazine and L-arginine on function of mitochondria in myocardium after ischemia/reperfusion injury of myocardium in rabbits].

AIM: To study the effect of ligustrazine (LGT) and L-arginine(L-Arg)on function of mitochondria in myocardium after myocardial ischemia/reperfusion injury (MI/RI). METHODS: 50 rabbits were randomly divided into five groups (n=10): Control group(A), MI/R group(B), MI/R + LGT group (C), MI/R+ L-Arg group (D), MI/R+ LGT + L-Arg group (E). The mitochondrial respiratory function, Ca2+ concentration ([Ca2+]m), malondialdehyde (MDA) content and superoxide dismutase (SOD) activity were deter mined. Meanwhile, the contents of ATP and EC in the myocardial tissue were measured, respectively. RESULTS: It was found that mitochondrial respiratory control rate (RCR), state 3 (ST3), SOD in C, D, E group were higher than those of B group, state 4 (ST4), [Ca2+]m, MDA were lower than those of B group, ATP and EC levels of myocardial tissue were higher than those in B group; and there were not significant differences between E and A group of above. CONCLUSION: LGT and IL-Arg can improve function of mitochondria in myocardium after ischemia/reperfusion injury of myocardium in rabbits by decreasing oxygen free radical level and Ca" overload in the mitochondria.

[Protective effect of propofol on liver during ischemia-reperfusion injury in patients undergoing liver surgery].

OBJECTIVE: To explore the protective effect of propofol on liver during hepatic ischemia/reperfusion injury (HIRI) and its mechanisms in patients undergoing liver surgery. METHODS: Eighteen patients who were scheduled for selective hepatic surgery were randomly divided into control group (n=9) and propofol treatment group (n=9). Changes of several parameters in plasma and effects of propofol on them were observed before liver ischemia, at end of ischemia and at reperfusion for 25 minutes, parameters of which included superoxide dismutase (SOD), xanthine oxidase (XO), lipid peroxide (LPO) and alanine aminotransferase (ALT) activity, and the ultrastructure changes in liver tissue were observed under electron microscope at 25 minutes after reperfusion. RESULTS: SOD activity decreased remarkably (P<0.01); XO activity, LPO concentration and ALT value increased significantly (P<0.01) during HIRI, and there were abnormal changes of the hepatic ultrastructure at 25 minutes after reperfusion. Afer treatment with propofol, the variation of all parameters were alleviated markedly (P<0.05 and P<0.01). CONCLUSION: Propofol has protective effects on HIRI by reducing oxygen free radical level and inhibiting lipid peroxidation after hepatic ischemia/reperfusion in patients undergoing liver cancer surgery.