Mechanism of Astragaloside Ⅳ on db/db Mice with Type 2 Diabetes Mellitus and Non-alcoholic Fatty Liver Disease Based on AMPK Signaling Pathway / 中国实验方剂学杂志

ObjectiveTo study the mechanism of astragaloside Ⅳ （AS Ⅳ） on db/db mice with type 2 diabetes mellitus （T2DM） and non-alcoholic fatty liver disease （NAFLD） based on network pharmacology and experimental validation. MethodA total of 24 db/db mice were randomly divided into four groups： model group， metformin group， and low-dose and high-dose AS Ⅳ groups. Six C57 mice were used as the blank group. The low-dose and high-dose AS Ⅳ groups were given AS Ⅳ of 0.015 and 0.030 g·kg-1 by gavage， and the metformin group was given 0.067 g·kg-1 by gavage. The blank and model groups were given equal volumes of distilled water by gavage. After intragastric administration， fasting blood glucose （FBG） was detected， and an oral glucose tolerance test was performed. Serum lipid level and liver histopathology were detected. The target and enrichment pathway of AS Ⅳ for treating T2DM and NAFLD were predicted by network pharmacology， and the main enrichment pathway was verified by molecular biology techniques. The protein expressions of AMPK， p-AMPK， sterol regulatory element-binding protein-1 （SREBP-1）， and fatty acid synthetase （FAS） in liver tissue were detected by Western blot. ResultCompared with the blank group， the levels of body mass， liver weight coefficient， fasting blood glucose， serum total cholesterol， triglyceride， and low-density lipoprotein cholesterol in mice treated with AS Ⅳ were decreased （P<0.05， P<0.01）. The pathology of liver tissue showed significant improvement in lipid accumulation， and imaging results showed that the degree of fatty liver was reduced after AS Ⅳ therapy. Network pharmacological prediction results showed that vascular endothelial growth factor α （VEGFA）， galactoagglutinin 3 （LGALS3）， serine/threonine kinase B2 （Akt2）， RHO-associated coiled-coil protein kinase 1 （ROCK1）， serine/threonine kinase B1 （Akt1）， signaling and transcriptional activator protein （STAT3）， and messtimal epidermal transformation factor （MET） were key targets in "drug-disease" network. The results from the Kyoto encyclopedia of genes and genomes （KEGG） enrichment showed that the AMP-dependent protein kinase （AMPK） signaling pathway was strongly associated with T2DM and NAFLD. Western blot results showed that compared with the blank group， the expression levels of p-AMPK/AMPK in the model group were significantly down-regulated， while those of SREBP-1 and FAS proteins were significantly up-regulated （P<0.01）. Compared with the model group， the expression levels of p-AMPK/AMPK in the metformin group and high-dose AS Ⅳ group were significantly up-regulated， while those of SREBP-1 and FAS proteins were significantly down-regulated （P<0.05， P<0.01）. ConclusionAS Ⅳ regulates the expression of lipid proteins by activating the AMPK signaling pathway， thereby improving lipid metabolism.

Effect of South African Herb Hoodia gordonii on Liver Glucolipid Metabolism and PI3K/Akt/FoxO1 Signalling Pathway in db/db Mice / 中国实验方剂学杂志

ObjectiveTo observe the effects of the South African herb Hoodia gordonii （HG） on glucolipid metabolism in diabetic db/db mice and explore the possible mechanisms of HG on the liver of db/db mice based on the phosphoinositide-3 kinase （PI3K）/protein kinase B （Akt）/factor forkhead protein O1 （FoxO1） signaling pathway. MethodA total of 30 db/db mice were randomly divided into five groups according to fasting blood glucose： model group， metformin group （0.195 g·kg-1）， and low dose （0.39 g·kg-1）， medium dose （0.78 g·kg-1）， and high dose （1.56 g·kg-1） HG groups， with six m/m mice in each group， and another six m/m mice were set as normal group. The mice in the normal and model groups were given saline of 9 mL·kg-1 by gavage. Body weight， water intake， and fasting blood glucose of the mice in each group were measured weekly. After six weeks of continuous administration， serum insulin （FINS）， low-density lipoprotein cholesterol （LDL）， total cholesterol （TC）， triglyceride （TG）， alanine aminotransferase （ALT）， aspartate aminotransferase （AST）， urea， and creatinine （CREA） were measured， and liver sections were embedded and stained with hematoxylin-eosin （HE）， periodic acid-Schiff （PAS）， and oil red O. Protein expression of PI3K p85， p-Akt， and p-FoxO1 in liver was detected by immunohistochemistry. The mRNA expression of PI3K， Akt， and FoxO1 in liver tissue was detected by real-time polymerase chain reaction （Real-time PCR）. ResultAfter six weeks of administration intervention， it was found that fasting blood glucose was significantly downregulated in mice in the three HG groups （P<0.05）. The level of islet resistance index was significantly reduced in both the low and medium dose HG groups （P<0.05）. The expression levels of TC， TG， and LDL were reduced in all HG groups （P<0.05， P<0.01）. Pathologically， HG could alleviate hepatocyte steatosis， reduce the volume and content of lipid droplets in liver， and increase the distribution of glycogen granules in liver to some extent in mice. Immunohistochemical assays revealed that PI3K p85 protein expression was significantly increased in the low， medium， and high dose HG groups compared with the model group （P<0.01）. p-Akt protein expression was significantly increased in the medium and high dose HG groups （P<0.05， P<0.01）. p-FoxO1 protein expression was significantly increased in the low， medium， and high dose HG groups （P<0.05， P<0.01）. Compared with the model group， PI3K mRNA was increased in low dose， medium dose， and high dose HG groups （P<0.05）， and Akt mRNA was increased in high dose HG group （P<0.05）. FoxO1 mRNA was decreased in low dose， medium dose， and high dose HG groups （P<0.05）. ConclusionHG can ameliorate the disorder of glucolipid metabolism in db/db mice， which may be related to its activation of the hepatic PI3K/Akt/FoxO1 signaling pathway.

Design, synthesis and biological activity of DB02 amino acid derivatives as HIV-1 non-nucleoside reverse transcriptase inhibitors / 药学学报

To improve the stability of amino acid ester derivatives of DB02, a series of 24 amide derivatives of DB02 amino acids as non-nucleoside HIV-1 reverse transcriptase inhibitor were designed and synthesized based on bioisosterism by replacing amino acid ester scaffold with more stable amide bond. The anti-HIV-1 activity of these compounds was evaluated by MTT assay and counting the number of syncytia. Most of the target compounds showed a potential anti-HIV-1 activity, among which compounds 2d, 2i, 2l, 2s, and 2w had better antiviral effect than lead compound DB02, with a therapeutic index > 1 000.00. Finally, the structure-activity relationship of these compounds was discussed, which provided new ideas for the further development of DB02 derivatives.

Effect of Loulianwan on Gut Microbiota in db/db Mice with Type 2 Diabetes Mellitus Based on 16S rRNA Sequencing Technology / 中国实验方剂学杂志

ObjectiveTo investigate the effect of Loulianwan on the gut microbiota of db/db mice with type 2 diabetes mellitus （T2DM）. MethodMale db/m+ mice aged 4-5 weeks were assigned to the normal group， and male db/db model mice of the same age were randomly divided into model group， metformin group （0.25 g·kg-1·d-1）， and Loulianwan group （13 g·kg-1·d-1）， with six mice in each group. Drug intervention lasted five weeks. The body weight， water intake， and fasting blood glucose （FBG） of the mice were recorded every week. After five weeks， the FBG， liver triglyceride （TG）， liver total cholesterol （TC）， glycated serum protein （GSP）， and fasting serum insulin （FINS） were detected， and the insulin resistance index （HOMA-IR） was calculated. The feces in the mouse intestines were collected， and the 16S rRNA sequencing technology was used to detect the structural changes in the fecal gut microbiota of mice in each group. ResultCompared with the normal group， the model group showed increased body weight， water intake， FBG， liver TG， liver TC， GSP， FINS， and HOMA-IR （P<0.01）. Compared with the model group， the Loulianwan group showed reduced water intake， FBG， liver TG， liver TC， GSP， FINS， and HOMA-IR （P<0.01）. The gut microbiota in the Loulian Lills group changed from phylum to genus level. The relative abundance of beneficial bacteria increased and the relative abundance of harmful bacteria decreased. Among them， the abundance of Akkermansia muciniphila， Blautia， Ruminococcus， and Parabacteroides increased （P<0.01）. ConclusionLoulianwan can significantly improve glucose and lipid metabolism in db/db mice with T2DM， and its mechanism may be related to the increase in the abundance of Akkermansia muciniphila， Blautia， Ruminococcus， and Parabacteroides in the intestine.

Adverse childhood experiences and delinquent behaviour: Predictors and mediating variables

ABSTRACT Background: Adverse childhood experiences (ACEs) have been identified as a risk factor for the development of mental health and behavioural outcomes throughout life, including delinquent behaviours. This article focuses on the relationship between ACEs and delinquent behaviour (DB), seeking to identify predictors and mediating variables. Methods: The quantitative study comprised 175 Portuguese adolescents, aged 12 and 17 years of age (M = 14.99, SD = 2.26). Results: ACEs and exposure to traumatic events (ETE) are predictive of DB. Antisocial traits (AT) was found to be mediating the relationship between ACEs and DB, as well as the relation between ETE and DB. Conclusion: The results indicate that it is necessary that professionals in health behaviour field prevent and intervene in ACEs and in ETE, both predictors of DB. The results of this study allow to understand the role of ACEs in DB and its mediating variables, which must be considered to mitigate the harmful impact of ACEs in DB.

Hypoglycemic Effect of Total Flavonoids from Potentillae Discoloris Herba in Type 2 Diabetic db/db Mice / 中国实验方剂学杂志

Objective:The hypoglycemic effects and mechanisms of total flavonoids from Potentillae Discoloris Herba（TFE） on insulin resistance through the phosphatidylinositol-3 kinase （PI3K）/protein kinase B （Akt） signaling pathway in db/db mice were investigated. Method:The 24 db/db mice were randomly divided into four groups， model group， metformin group and TFE 100，400 mg·kg^-1 group respectively. The 6 db/m mice as normal control group. After 4 weeks treatment， the mice were processed and the levels of fasting blood glucose（FBG）， glycated serum protein（GSP），fasting blood insulin（FINS），triglyceride（TG）， total cholesterol（TC）， low density lipoprotein cholesterol （LDL-C），high density lipoprotein cholesterol （HDL-C） in serum were detected. Homeostatic model assessments of insulin resistance（HOMA-IR）were quantified. Hematoxylin-eosin（HE）staining of liver and pancreatic tissues were examined. The expression of IR<italic>β</italic>， IRS-1，PI3K，phosphorylation-PI3K （p-PI3K）， Akt， phosphorylation-Akt（p-Akt） and glucose transporter 4（GLUT4） in livers were assessed by Western blot. Result:Compared with normal group， model group showed liver and pancreas injury. FBG， GSP， TC， TG， LDL-C， FINS and MDA levels in serum were significantly increased （<italic>P</italic><0.01）， HDL-C and SOD levels in serum were significantly decreased （<italic>P</italic><0.05）， liver glycogen content was significantly decreased （<italic>P</italic><0.01）， as well as expression of IR， IRS-1， p-PI3K/PI3K， p-Akt/Akt and GLUT4 protein in liver tissues were significantly decreased （<italic>P</italic><0.01）. Compared with model group， TFE was able to relieve liver and pancreas injury，while the levels of FBG， GSP， TC， TG， FINS and MDA in serum were significantly decreased （<italic>P</italic><0.05）， HDL-C and SOD levels liver were significantly increased （<italic>P</italic><0.05）， liver glycogen content was significantly increased （<italic>P</italic><0.01）， and the expressions of IRS-1， p-PI3K/PI3K， p-Akt/Akt and GLUT4 protein in liver tissues were significantly up-regulated （<italic>P</italic><0.05）. Conclusion:These findings indicate that TFE has the potential to reduce blood sugar and alleviates insulin resistance through the PI3K/Akt signaling pathway in the livers of db/db mice.

Rosiglitazone promotes phosphorylation of AMPK protein and improves insulin resistance in db/db mice / 上海预防医学

Objective To observe the effect of rosiglitazone on the protein expression of AMPK and GLUT4 in peripheral tissue (liver, skeletal muscle and fat) of type 2 diabetic db/db mice and to prove that rosiglitazone can regulate the glucose metabolism in db/db mice partly through the AMPK pathway. Methods db/db mice were randomly divided into model group and rosiglitazone group according to their blood glucose.The db/m mice were normal control group.After 4 weeks of administration, fasting blood glucose was detected in each group.Western blot was used to detect the contents of AMPK, p-AMPK and GLUT4 in liver, skeletal muscle and adipose tissue. Results (1) Rosiglitazone significantly reduced the fasting blood glucose of db/db mice; (2)Rosiglitazone increased the level of AMPK phosphorylation in the liver, skeletal muscle and adipose tissue of db/db mice, and increased the content of GLUT4 protein in skeletal muscle and adipose tissue. Conclusion Rosiglitazone can increase the phosphorylation of AMPK and the expression of GLUT4 protein in the liver, muscle and fat tissue of db/db mice, and promote the uptake and utilization of glucose in peripheral tissue, suggesting that it can regulate glucose metabolism in db/db mice partly through the AMPK pathway.

Study on mechanism of berberine on regulating glycogen metabolism by repairing glycogen structure / 中草药

Objective: To investigate the mechanism of berberine regulating glycogen metabolism and its effect on glycogen structure. Methods: The db/db mice were used as diabetic mice to investigate the effect of berberine on blood glucose levels in db/db mice. The liver glycogen was extracted for SEC and TEM analysis to investigate the effect of berberine on liver glycogen and the mechanism of its effect. Results: Berberine significantly decreased the fasting blood glucose level and the insulin level in serum of db/db mice, and berberine repaired the damaged glycogen structure. Meanwhile, berberine decreased the expression of GP, GDBE, cAMP in liver tissue and the glucagon level in serum of db/db mice. Conclusion: Berberine regulated the cAMP/GP signaling pathway, improved hepatic glycogen structure in db/db mice, and repaired damaged glycogen structure, which may be one of the mechanisms for regulating hepatic glucose metabolism and improving symptoms of diabetes.

Effects of artemether on glucose and lipid metabolism in C57BL/KsJ-db/db mice / 中草药

Objective To investigate the effects of different doses of artemether on glycolipid metabolism in C57BL/KsJ-db/db mice. Methods Eight-week-old male C57BL/KsJ-db/db mice were divided into model group (ig given 1% methylcellulose) and artemether 400, 200, 100, 50 groups (ig given 400, 200, 100, 50 mg/kg artemether respectively + 1% methylcellulose), with six mice in each group. Another six male C57BL/KsJ-db/+ mice were selected as the control group. All groups were administered for 4 weeks. The quality of the mice was measured every 2 d; The food intake of the mice was measured every 3 d and the average daily food intake and body mass changes were evaluated; The amount of water in mice was measured every 2 d; The urine volume of the mice was measured every 3 d; After 8 h of fasting, blood was collected from the tail vein, and the fasting blood glucose of the mice was measured by Roche blood glucose meter and matching test paper every 7 d. Mice were assessed for glucose tolerance and sensitivity to insulin by ip glucose tolerance test (IPGTT) and ip insulin tolerance test (IPITT). The serum total cholesterol (TC), triglyceride (TG), and free fatty acid (FFA) levels of the mice were determined by a biochemical kit. The whole liver of mice was weighed and the morphological changes of the pancreas and liver in mice were observed by HE staining. The expression of AMP-activated protein kinase (AMPK), glucose transporter 4 (GLUT-4), and insulin receptor β (IRβ) protein in liver of mice was analyzed by Western blotting. Results Compared with the control group, the food intake, water intake, and urine volume of the model group were significantly increased (P < 0.001). Compared with the model group, each dose of artemether significantly reduced the water intake and urine volume of the mice (P < 0.01, 0.001); Artemether 400, 200, 100 mg/kg can significantly reduce the body weight and food intake in a dose-dependent manner (P < 0.05, 0.01); Artemether 400, 200, 100 mg/kg significantly reduced fasting blood glucose levels in mice, reduced the area under the curve of IPGTT (AUCs), and improved insulin resistance in mice (P < 0.01, 0.001). Compared with the control group, the TC, TG, and FFA levels of mice in the model group were significantly increased (P < 0.05). Compared with the model group, artemether significantly decreased the levels of TC, TG, and FFA in the serum of mice in a dose-dependent manner (P < 0.05), and significantly improved islet vacuolar degeneration and hepatic steatosis in db/db mice; The protein expression of AMPK, GLUT-4, and IRβ in the liver of mice was increased (P < 0.05). With the prolongation of the intervention time, the higher the dose of artemether, the higher the mortality rate and the incidence of adverse reactions in mice. Conclusion Artemether can significantly improve the high-fat state and insulin resistance of diabetic mice. It can treat fatty liver and may up-regulate the expression of GLUT-4 and IRβ protein through the AMPK pathway to exert its effects. It is expected to be used in the treatment of type 2 diabetes mellitus, which is mainly caused by metabolic syndrome. However, higher doses of artemether and longer-term application may lead to more adverse events.

Regulatory effect of mulberry leaf components on intestinal microflora in db/db mice / 药学学报

The 16S rDNA sequencing method was adopted to study the effects of mulberry leaf flavonoids, polysaccharides and alkaloids on intestinal microflora in db/db diabetic mice. The animal experiment was examined by the Ethics Committee of Nanjing University of Chinese Medicine. Ten db/m mice were control group and forty db/db mice were randomly divided into model group, metformin group, mulberry flavonoid (MF) group, mulberry polysaccharide (MP) group and mulberry alkaloid (MA) group. After intragastric administration for six weeks, fresh feces were collected for detection of intestinal microflora. There were Firmicutes, Bacteroidetes, Proteobacteria, Saccharibacteria, Tenericutes, Deferribacteres, Verrucomicrobia, Cyanobacteria in each group. The results showed that the intestinal microflora of db/db mice changed significantly from phylum level to genus level. The proportion of Firmicutes and Proteobacteria in model group decreased significantly, and the proportion of Bacteroidetes increased. The difference in species abundance distribution between model group and other groups was significant, which indicated that the community distribution was disordered in model group. After administration, the Bacteroidetes, Lachnospiraceae, Roseburia and Desulfovibrio were effectively regulated, especially in the alkaloid group. The difference in species abundance distribution between drug-treated group and blank group also became smaller. It is suggested that the active components of mulberry leaf have the effect of improving the intestinal microflora imbalance in db/db mice.

Protective effect and mechanism of Schisandra chinensis extract on oxidative stress in diabetic nephropathy mice / 中草药

Objective: To investigate the protective effect of Schisandra chinensis extract on oxidative stress in db/db mice, and to explore its possible mechanism. Methods: Ten male 9-week-old db/m mice and ten homologous male db/db mice of the same age were randomly divided into normal control group (C + V group), S. chinensis extract control group (C + SE group), model group (DN + V group), and S. chinensis extract-treated group (DN + SE group), with five mice in each group. The S. chinensis extract control group and S. chinensis extract-treated group were respectively ig administrated with 5 mg/(kg•d) of S. chinensis extract for 6 weeks. The body weight, blood glucose and 24 h urine micro-albumin were recorded at 0, 3, and 6 weeks. After 6 weeks, the mice were sacrificed and their kidney tissue specimens were collected. The oxidative stress index of malondialdehyde (MDA) were examined by lipid peroxidation kit. The pathological changes of kidney tissues were observed by PAS staining. The expression of antioxidant factor Nrf2 and HO-1 protein of kidney tissue was detected by Western Blotting. The expression of Nrf2 and its downstream target genes of kidney tissue were detected by qRT-PCR. Results: Compared with the normal control group, the body weight, blood glucose and 24h urine micro-albumin of the model group were significantly increased and the MDA content were increased. Therefore, the renal tissue pathological damage aggravated, and the expression of Nrf2 and HO-1, as well as its downstream target genes were all down- regulated. (P < 0.01 or P < 0.05). S. chinensis extract reduced blood glucose, 24 h urine micro-albumin and MDA level in diabetic nephropathy mice, improved renal pathological damage, and up-regulated Nrf2 and HO-1 and its downstream target gene expression. (P < 0.01 or P < 0.05). Conclusion: S. chinensis extract has protective effect on oxidative stress injury in db/db mice, and its mechanism may be related to up-regulation of antioxidant factor Nrf2 and its downstream genes.

Identification of a thermal stable allergen in yam (Dioscorea opposita) to cause anaphylaxis

Yam (Dioscorea opposita) is commonly consumed in East Asia, but allergic reaction to this plant food is rare. To date, there is no report of anaphylactic reaction after ingestion of cooked yam. We described 3 cases with anaphylaxis after eating boiled yam and 1 present with oral allergy syndrome as well. Basophil activation test in patients showed positive reactivity to boiled yam extract. In immunoblotting, a 30-kDa protein was recognized by all patients' sera and a 17-kDa band was detected by 1 patient. N-terminal amino acid revealed the 30-kDa IgE reacted band was DB3S, dioscorin in Dioscorea tuber. It promoted us that DB3S was a thermal stable oral allergen to trigger anaphylactic reaction and oral allergy syndrome in cooked yam (D. opposita) allergy. Patients with this plant food allergy should avoid both raw and well-cooked yam.

Resistant dextrin improves insulin resistance in db/db mice by enhancing insulin signaling pathway / 上海交通大学学报（医学版）

Objective: To investigate the effect of resistant dextrin on insulin resistance in db/db mice and preliminarily explore potential molecular mechanisms. Methods: Ten 8-week-old db/db mice were randomly assigned into two groups, i.e., control group and resistant dextrin group, which were administered with distilled water and resistant dextrin by gavage for 9 weeks, respectively. Fasting blood glucose (FBG) and body weight were measured every week. Intraperitoneal glucose tolerance test (IPGTT) and insulin tolerance test (ITT) were performed separately at the 8th and 9th week. Fasting serum insulin (FINS) was measured, homeostasis model assessment insulin resistance (HOMA-IR) index was calculated, and triacylglycerol (TAG), total cholesterol (TC), low density lipoprotein-cholesterol (LDL-Ch), high density lipoprotein-cholesterol (HDL-Ch) were detected after the mice were sacrificed. The relative expression of Ir, Irs-1, Pi3k, Akt, and Glut-2 in insulin signaling pathways in the liver was determined by real-time quantitative PCR. The expression of IRS-1, p-AKT, GLUT-2 in the liver were determined by Western blotting. Results: After 9 weeks of treatment with resistant dextrin, the body weight of the resistant dextrin group decreased, but the difference was not statistically significant (P=0.384); FINS decreased, and HOMA-IR index significantly decreased (P=0.032); insulin resistance was significantly improved (all P<0.05); the blood biochemical parameters, including TAG, TC, LDL-Ch, and HDL-Ch were lower, but only LDL-Ch was statistically different (P=0.034); the expression of Ir, Irs-1, Akt, and Glut-2 mRNA in the liver significantly increased (all P<0.05); the expression of IRS-1 and GLUT-2 proteins also significantly increased (P=0.026, P=0.039). Conclusion: Resistant dextrin may improve insulin resistance in db/db mice by enhancing insulin signaling pathway.

Global gene expression analysis in liver of db/db mice treated with catalpol / 中国天然药物

Catalpol, a major bioactive component from Rehmannia glutinosa, which has been used to treat diabetes. The present study was designed to elucidate the anti-diabetic effect and mechanism of action for catalpol in db/db mice. The db/db mice were randomly divided into six groups (10/group) according to their blood glucose levels: db/db control, metformin (positive control), and four dose levels of catalpol treatment (25, 50, 100, and 200 mg·kg), and 10 db/m mice were used as the normal control. All the groups were administered orally for 8 weeks. The levels of fasting blood glucose (FBG), random blood glucose (RBG), glucose tolerance, insulin tolerance, and glycated serum protein (GSP) and the globe gene expression in liver tissues were analyzed. Our results showed that catalpol treatment obviously reduced water intake and food intake in a dose-dependent manner. Catalpol treatment also remarkably reduce fasting blood glucose (FBG) and random blood glucose (RBG) in a dose-dependent manner. The RBG-lowering effect of catalpol was better than that of metformin. Furthermore, catalpol significantly improved glucose tolerance and insulin tolerance via increasing insulin sensitivity. Catalpol treatment significantly decreased GSP level. The comparisons of gene expression in liver tissues among normal control mice, db/db mice and catalpol treated mice (200 and 100 mg·kg) indicated that there were significant increases in the expressions of 287 genes, whichwere mainly involved in lipid metabolism, response to stress, energy metabolism, and cellular processes, and significant decreases in the expressions of 520 genes, which were mainly involved in cell growth, death, immune system, and response to stress. Four genes expressed differentially were linked to glucose metabolism or insulin signaling pathways, including Irs1 (insulin receptor substrate 1), Idh2 (isocitrate dehydrogenase 2 (NADP), mitochondrial), G6pd2 (glucose-6-phosphate dehydrogenase 2), and SOCS3 (suppressor of cytokine signaling 3). In conclusion, catalpol ecerted significant hypoglycemic effect and remarkable therapeutic effect in db/db mice via modulating various gene expressions.

Influence of extract from fermented buckwheat flower and leaf in kidney injury in type 2 diabetic db/db mice / 吉林大学学报(医学版)

Objective:To discuss the protective effects of extract from fermented buckwheat flower and leaf (EFBFL) on the kidney injury in the spontaneously obese type Ⅱ diabetic db/db mice,and to elucidate their possible action mechanisms from the protein expression levels of peroxisome proliferator-activated receptor γ(PPARγ) and nuclear factor-κB (NF-κB) in kidney tissue.Methods:The 8 weeks old male db/db mice were selected and the littermate db/m mice were used as normal control group;the db/db mice were randomly divided into model group,low dose of EFBFL group and high dose of EFBFL group,10 mice in each group.The mice in low and high doses of EFBFL groups were intragastrically given 50 and 100 mg · kg-1 EFBFL,and the mice in normal control group and model group were intragastrically given the equal amount of distilled water once daily for 8 weeks accordingly.The levels of fasting blood-glucose (FBG) of the mice in various groups were tested respectively before and after administration.Full automatic biochemical analyzer was used to detect the levels of serum creatinine (SCr) and blood urea nitrogen (BUN) of the mice,and the kidney index was calculated.The morphology of kidney tissue was observed by HE staining and Masson staining,and immunohistochemical method and Western blotting method were used to detect the expression levels of PPARγ and NF-κB protein in kidney tissue.Results:Compared with model group,the levels of FBG and the levels of serum SCr and BUN in EFBFL groups were significantly decreased (P＜0.05 or P＜0.01),and the kidney indexes were increased (P＜0.01).Compared with normal control group,the kidney glomerular of the mice in model group was weaked,glomerular basement membrane was diffusely thickened,the foot process was coalesced or disappeared,and the kidney tissue fibrosis was serious;compared with the model group,the above performance of the mice in EFBFL groups were improved in different degrees,especially in high dose of EFBFL group.Compared with model group,the expression levels of PPARγ in kidney tissue of the mice in EFBFL groups were increased (P＜0.01) and the expression levels of NF-κB in kidney tissue of the mice were decreased (P＜0.01).Conclusion:EFBFL can improve the kidney injury in the spontaneously obese type Ⅱ diabetic db/db mice,and its possible mechanism may be related to lowering the level of blood glucose,up-regulating the expression of PPARγ and down-regulating the expression of NF-κB in the kidney tissue of the mice.

Global gene expression analysis in liver of db/db mice treated with catalpol / 中国天然药物

Catalpol, a major bioactive component from Rehmannia glutinosa, which has been used to treat diabetes. The present study was designed to elucidate the anti-diabetic effect and mechanism of action for catalpol in db/db mice. The db/db mice were randomly divided into six groups (10/group) according to their blood glucose levels: db/db control, metformin (positive control), and four dose levels of catalpol treatment (25, 50, 100, and 200 mg·kg), and 10 db/m mice were used as the normal control. All the groups were administered orally for 8 weeks. The levels of fasting blood glucose (FBG), random blood glucose (RBG), glucose tolerance, insulin tolerance, and glycated serum protein (GSP) and the globe gene expression in liver tissues were analyzed. Our results showed that catalpol treatment obviously reduced water intake and food intake in a dose-dependent manner. Catalpol treatment also remarkably reduce fasting blood glucose (FBG) and random blood glucose (RBG) in a dose-dependent manner. The RBG-lowering effect of catalpol was better than that of metformin. Furthermore, catalpol significantly improved glucose tolerance and insulin tolerance via increasing insulin sensitivity. Catalpol treatment significantly decreased GSP level. The comparisons of gene expression in liver tissues among normal control mice, db/db mice and catalpol treated mice (200 and 100 mg·kg) indicated that there were significant increases in the expressions of 287 genes, whichwere mainly involved in lipid metabolism, response to stress, energy metabolism, and cellular processes, and significant decreases in the expressions of 520 genes, which were mainly involved in cell growth, death, immune system, and response to stress. Four genes expressed differentially were linked to glucose metabolism or insulin signaling pathways, including Irs1 (insulin receptor substrate 1), Idh2 (isocitrate dehydrogenase 2 (NADP), mitochondrial), G6pd2 (glucose-6-phosphate dehydrogenase 2), and SOCS3 (suppressor of cytokine signaling 3). In conclusion, catalpol ecerted significant hypoglycemic effect and remarkable therapeutic effect in db/db mice via modulating various gene expressions.

Influence of extract from fermented buckwheat flower and leaf in kidney injury in type 2 diabetic db/db mice / 吉林大学学报(医学版)

Objective: To discuss the protective effects of extract from fermented buckwheat flower and leaf (EFBFL) on the kidney injury in the spontaneously obese type II diabetic db/db mice, and to elucidate their possible action mechanisms from the protein expression levels of peroxisome proliferator-activated receptor y (PPARy) and nuclear factor-κB (NF-κB) in kidney tissue. Methods: The 8 weeks old male db/db mice were selected and the littermate db/m mice were used as normal control group; the db/db mice were randomly divided into model group, low dose of EFBFL group and high dose of EFBFL group, 10 mice in each group. The mice in low and high doses of EFBFL groups were intragastrically given 50 and 100 mg · kg: EFBFL, and the mice in normal control group and model group were intragastrically given the equal amount of distilled water once daily for 8 weeks accordingly. The levels of fasting blood-glucose (FBG) of the mice in various groups were tested respectively before and after administration. Full automatic biochemical analyzer was used to detect the levels of serum creatinine (SCr) and blood urea nitrogen (BUN) of the mice, and the kidney index was calculated. The morphology of kidney tissue was observed by HE staining and Masson staining, and immunohistochemical method and Western blotting method were used to detect the expression levels of PPARγ and NF-κB protein in kidney tissue. Results: Compared with model group, the levels of FBG and the levels of serum SCr and BUN in EFBFL groups were significantly decreased (P<0. 05 or P<0. 01), and the kidney indexes were increased (P<0. 01). Compared with normal control group, the kidney glomerular of the mice in model group was weaked, glomerular basement membrane was diffusely thickened, the foot process was coalesced or disappeared, and the kidney tissue fibrosis was serious; compared with the model group, the above performance of the mice in EFBFL groups were improved in different degrees, especially in high dose of EFBFL group. Compared with model group, the expression levels of PPARγ in kidney tissue of the mice in EFBFL groups were increased (P<0. 01) and the expression levels of NF-κB in kidney tissue of the mice were decreased (P<0. 01). Conclusion: EFBFL can improve the kidney injury in the spontaneously obese type II diabetic db/db mice, and its possible mechanism may be related to lowering the level of blood glucose, up-regulating the expression of PPARγ and down-regulating the expression of NF-κB in the kidney tissue of the mice.

Dynamic pathological characteristics of kidney in db/db mice with type 2 diabetes mellitus / 吉林大学学报(医学版)

Objective; To investigate the dynamic pathological characteristics of kidney in the db / db mice with type 2 diabetes mellitus and to clarify the pathogenesis and development mechanism of diabetic nephropathy, and to provide the experimental evidence for the further study on diabetic nephropathy. Methods: The male SPF db/db mice aged 7 - 8 weeks were selected as model group (n=16) and the db/m mice with the same ages were selected as control group (n=16). The body weights and levels of fasting blood glucose (FBG) of the mice were detected after 8, 16, and 32 weeks. Then eight mice in each group were sacrificed and the kidney tissue was dissected at 8, 16 and 32 weeks. HE and Masson staining were used to observe the pathomorphology of the kidney tissue; the ultrastructures of kidney tissue were observed under electron microscope. Results: Compared with normal group, the body weights of the mice in model group at 8, 16 and 32 weeks were significantly increased (P<0. 01), the FBG levels were significantly increased (P<0. 01), and the double kidney indexes were significantly decreased (P<0. 01); the double kidney index of the mice in model group at 32 weeks was lower than that at 16 weeks (P< 0. 05). The HE and Masson staining results showed that the morphological changes of kidney tissue of the mice at 16 weeks were significant, the glomerular volume was expansion and the kidney tubular epithelial cells were edema, which turned serious at 32 weeks. A large number of blue dye substances in the kidney tissue was found. Under electron microscope, the glomerular basement membranes got thickened, the foot processes got fused and the number of mitochondria in kidney tubular epithelial cells was reduced and swell was found in the mice in model group at 16 weeks; the lesions in the kidney tissue were serious at 32 weeks, and the collagenous fiber was visible at 32 weeks. Conclusion: The pathological changes in the kidney tissue of the db/db mice at 16 weeks are significant, such as glomerular basement thickening and footwork fusion. The kidney tissue of the db/db mice at 32 weeks shows prominent fibrosis.

Alpinia oxyphylla Miq. extract changes miRNA expression profiles in db-/db- mouse kidney

BACKGROUND: A number of dysregulated miRNAs have been identified and are proposed to have significant roles in the pathogenesis of type 2 diabetes mellitus or renal pathology. Alpinia oxyphylla has shown significant anti-inflammatory properties and play an anti-diabetes role. The objective of this study was to detect the alteration of miRNAs underlying the anti-diabetes effects of A. oxyphylla extract (AOE) in a type II diabetic animal model (C57BIKsj db-/db-). RESULTS: Treatment with AOE for 8 weeks led to lower concentrations of blood glucose, urine albumin, and urine creatinine. 17 and 13 miRNAs were statistically identified as differentially regulated in the DB/DB and db-/db- AOE mice, respectively, compared to the untreated db-/db- mice. Of these, 7 miRNAs were identified in both comparison groups, and these 7 miRNAs were verified by quantitative real-time PCR. Functional bioinformatics showed that the putative target genes of 7 miRNAs were associated with several diabetes effects and signaling pathways. CONCLUSIONS: These founding suggest that the potential of AOE as a medicinal anti-diabetes treatment through changes in the expressions of specific miRNAs. The results provide a useful resource for future investigation of the role of AOE-regulated miRNAs in diabetes mellitus.

Biological characteristics of a db/db mouse model of type 2 diabetes / 中国比较医学杂志

Objective To observe the biological characteristics of db/db mice, and to provide the basis for application of db/db mice in experimental research.Methods Spontaneous type 2 diabetes BKS.Cg-Dock7m +/+ Leprdb/JNju mice and wild type mice of the same age were used in this study.Their fasting blood glucose was determined at 8,12,16, 20 and 24 weeks of age, the body weight was recorded at 10, 12,16, 20 and 24 weeks of age, the levels of serum insulin, total cholesterol and triglyceride were measured, the organ weight and liver coefficient were determined, and the liver and pancreas were taken for pathological examination at 24 weeks of age.Results The db/db mice maintained a high level of fasting blood glucose and body weight.Levels of serum insulin, total cholesterol, and triglyceride were significantly higher than the wild type mice.Compared with wild type mice, liver weight and kidney weight were also significantly increased.Obvious pathological changes of liver and pancreas were observed in 24-week old db/db mice.Conclusions db/db mouse has obvious characteristics of type 2 diabetes, such as hyperglycemia, hyperlipidemia, and hyperinsulinemia, can maintain stable levels of high blood glucose,and is an ideal animal model for experimental study of type 2 diabetes mellitus.