Effects of Angelica sinensis polysaccharide on the apoptosis of cardiomyocytes in diabetic KK-Ay mice / 中国药房

OBJECTIVE To investigate the effects of Angelica sinensis polysaccharide on the apoptosis of cardiomyocytes in diabetic KK-Ay mice. METHODS KK-Ay mice were randomly divided into model group， metformin group （200 mg/kg） and A. sinensis polysaccharide high-dose， medium-dose and low-dose groups （400， 200 and 100 mg/kg）； C57BL/6J mice were included in blank group， with 8 mice in each group. Each group was given relevant medicine intragastrically or normal saline， once a day， for consecutive 4 weeks. After the final administration， the levels of fasting glucose， total cholesterol （TC）， low-density lipoprotein cholesterol （LDL-C） and insulin （INS） were detected； the protein expressions of B-cell lymphoma 2 （Bcl-2）， cleaved- caspase-3， apoptosis signal-regulated kinase 1 （ASK1）， phosphorylated c-Jun N-terminal kinase （p-JNK）， phosphorylated inositol- requiring enzyme 1α （p-IRE1α） in myocardium， and apoptosis in cardiomyocytes were also detected. RESULTS Compared with model group， the fasting glucose， TC and LDL-C content， apoptotic rate of cardiomyocyte， protein expressions of p-JNK and p- IRE1α， ASK1， cleaved-caspase-3 were significantly lower in the metformin group and A. sinensis polysaccharide medium-dose， high-dose groups； INS level and relative expression of Bcl-2 protein were significantly increased （P＜0.05 or P＜0.01）. CONCLUSIONS A. sinensis polysaccharide can improve the levels of blood glucose and blood lipid and inhibit cardiomyocyte apoptosis in diabetic KK-Ay mice， and the mechanism may be related to the inhibition of IRE1/ASK1/JNK signaling pathway.

Effect of Tibetan medicine E-se on insulin resistance in KK-Ay mice with spontaneous type 2 diabetes / 中国药理学通报

Aim To evaluate the effect of E-se extract on insulin resistance in KK-Ay mice with spontaneous type 2 diabetes anrl explore its mechanism.Methods Ten C57/6J mice were assigned to a normal control group.Fifty KK-Ay model mice were randomly divided into model group, positive control group ( rosiglita- zone, 2.67 mg • kg 1 ), and low- ( 0.75 g • kg 1 ) , medium- ( 1.50 g • kg 1 ) , and high-dose ( 3.00 g • kg ') E-se groups, with 10 mice in each group.All mice were measured for body weight and fasting blood glucose weekly, insulin tolerance on the 32nd day, and insulin after the last administration on the 35th day, and the insulin resistance/sensitivity indexes were calculated.The pancreas was stained by hematoxylin- eosin ( HE ).Islet cell apoptosis was detected by TUNEL staining.Glucagon-like peptide-1 ( GLP-1 ) was detected by immunohistochemistry.Results j j Compared with the model group, the E-se groups showed reduced body weight, fasting blood glucose, serum insulin concentration, and insulin resistance in¬dex, elevated insulin sensitivity index, decreased le¬sion grading score of pancreatic tissues and apoptosis percentage of islet cells, and increased content of GLP- 1 protein in pancreatic tissues.Conclusions E-se ex¬tract can improve insulin resistance by reducing serum insulin level, inhibiting islet cell apoptosis, and in¬creasing the sensitivity of the body to insulin.

Effect of Gegen Qinliantang on TLR4/IKKβ/NF-κB Pathway in KKAy Mice with Diabetes / 中国实验方剂学杂志

ObjectiveTo observe the effect of classical prescription Gegen Qinliantang（GGQLT） on inflammatory factors and key targets in the inflammatory pathways mediated by lipopolysaccharide in KKAy mice and explore its mechanism in improving spontaneous type 2 diabetes mellitus （T2DM）. MethodSixty-five SPF KKAy mice with spontaneous T2DM and 13 C57BL/6J mice （control） were selected in the barrier system and fed on a high-fat diet. The model was properly induced in 44 mice in the context of random blood glucose exceeding or equal to 13.9 mmol·L-1. Then the mice were assigned into a normal group （20 mL∙kg-1 normal saline）， a model group （20 mL∙kg-1 normal saline）， an acarbose group （3.9 mg∙kg-1）， and high- and low-dose GGQLT groups （1.82 and 0.45 g∙kg-1）， with 11 mice in each group. The mice in each group were treated correspondingly by gavage for eight weeks， once per day. Blood glucose and body weight were systematically evaluated. Twelve hours after the last administration， blood samples were collected from the eyes， and the serum and muscle and liver tissues were extracted. The levels of tumor necrosis factor-α （TNF-α）， interleukin-6 （IL-6）， and glucose transporter type 4 （GluT4） were detected by semi-quantitative enzyme-linked immunosorbent assay （ELISA）. The protein expression of IκB kinase β （IKKβ） and nuclear factor-κB （NF-κB） in muscle tissues and Toll-like receptor 4 （TLR4） in liver tissues was detected by Western blot. ResultCompared with the normal group， the model group showed increased body weight and blood glucose （P<0.01）. Compared with the model group， the acarbose group and the GGQLT groups showed reduced body weight and blood glucose （P<0.05， P<0.01）. As revealed by ELISA results， compared with the normal group， the model group showed increased levels of TNF-α and IL-6 （P<0.01） and deceased GluT4 level （P<0.05）. Compared with the model group， the groups with drug treatment showed reduced levels of TNF-α and IL-6 （P<0.05， P<0.01）， and the acarbose group and the high-dose GGQLT group showed increased GluT4 level （P<0.05， P<0.01）. As displayed by Western blot results， compared with the normal group， the model group showed increased protein expression of IKKβ， NF-κB， and TLR4 （P<0.01）. Compared with the model group， the acarbose group and the GGQLT groups showed reduced protein expression of IKKβ， NF-κB， and TLR4 （P<0.05， P<0.01）. ConclusionGGQLT can inhibit the inflammatory cascade effect and improve T2DM by down-regulating the levels of key inflammatory factors in the TLR4 pathway， inhibiting their activation， and increasing the translocation and activity of GluT4 on the basis of the regulation of intestinal flora.

Renal Protective Effect of Tangshiping Capsule on Diabetic Nephropathy Kkay Mice and Its Effect on Wnt/β-catenin Signal Transduction Pathway / 中国实验方剂学杂志

Objective:To investigate the renal protective effect of Tangshenping capsule (Tangshenping) on diabetic nephropathy (DN) KKAy mice and its effect on Wnt/β-catenin signaling pathway. Method:Sixty female Sprague-Dawley KKAy mice aged 10 weeks old were induced with KKAy rat feed for 10 weeks. The DN animal model was successfully determined with blood glucose (>16.7 mmol·L-1) and 24 hour urine protein (>0.4 mg). The model mice were randomly divided into a model group, an irbesartan group, and low, medium and high-dose Tangshenping group, with 10 female C57BL/6J mice as a control group. The treatment groups were given the corresponding drugs by gavage. The normal group and the model group were given an equal volume of deionized water by gavage. The intragastric dose was 0.01 mL·g-1 body weight coefficient once a day. The general conditions of the mice were observed, the body mass was weighed every 4 weeks, and 24 h urine protein was quantified. At the 26th week, the blood was collected from eyeballs, and the mice were put to death. The quality of the kidneys, serum blood urea nitrogen (BUN), serum creatinine (SCr), triglyceride (TG), malondialdehyde (MDA), nitric oxide (NO) and superoxide dismutase (SOD) content were measured. In situ hybridization and immunohistochemistry were used to detect the expressions of Wnt4, glycogen synthase kinase 3β(GSK3β) and β-catenin in kidney tissues. Result:Compared with model group, body mass, kidney mass/body mass, and 24 h urine protein were significantly lower in high-dose Tangshenping group (PPPβ and β-catenin were decreased (PConclusion:Tangshenping may inhibit the activation of Wnt/β-catenin signaling pathway, reverse the transdifferentiation of renal tubular epithelial cells in DN KKAy mice, delay the progression of renal interstitial fibrosis, and then exert renal protection.

Experimental study of bicyclol for type 2 diabetic treatment using KKAy mice / 药学学报

To investigate the effects of small molecule compound bicyclol on type 2 diabetes mellitus (T2DM) and its mechanism of action, KKAy mice were treated with various doses of bicyclol (100, 200, and 400 mg·kg-1·d-1) with metformin (200 mg·kg-1·d-1) as a positive control, respectively. Age-matched C57BL/6J mice were used as the non-diabetic control (Con). The effect on hyperglycemia was evaluated by the levels of no-fasting blood glucose, fasting blood glucose (FPG), and glucose tolerance. Whole body insulin sensitivity was evaluated by fasting plasma insulin (FPI) and homeostasis model assessment-insulin resistance (HOMA-IR). The hepatic response to insulin was evaluated by insulin-induced activation of insulin signaling pathway. Western blot was performed to detect hepatic protein expressions. All animal experimental procedures were approved by the Animal Ethics Committee of Chinese Academy of Medical Sciences. KKAy mice showed T2DM characteristics such as hyperglycemia and insulin resistance, including attenuated response to insulin in the liver. A 28-day treatment of bicyclol suppressed both FPG and no-fasting blood glucose, in a dose- and time-dependent manner. Moreover, FPI and HOMA-IR values were both significantly decreased, and hepatic insulin-induced-phosphorylation of IRβ and Akt were up-regulated in KKAy mice after bicyclol treatment. Phosphorylation of FoxO1, the key transcription factor for regulating gluconeogenesis, was also significantly elevated by bicyclol treatment. These results suggested that bicyclol has some therapeutic effects on hyperglycemia in a time- and dose-dependent manner in KKAy mice. Its mechanism might be attributed to improving insulin resistance, enhancing hepatic insulin signaling pathway, and inhibiting gluconeogenesis. No significant interference on the hypoglycemic effect of metformin by bicyclol was observed in this study.

Experimental study of anti-diabetic effects of gastrodin in KK-Ay mice / 中国药理学通报

Aim To study the anti-diabetic effects of natural product gastrodin ( GSTD ) in KK-Ay mice. Methods C57BL/6J mice were used as normal con-trol, while KK-Ay diabetic mice were divided into five groups, namely the untreated group, GSTD 10 mg· kg-1, 20 mg·kg-1, 50 mg·kg-1 groups, and the metformin ( Met) 200 mg·kg-1 group, respectively, with 10 mice in each group. GSTD and Met were ad-ministered intragastrically for eight weeks. Before ex-periment and once a week during the experiment, the fasting blood glucose ( FBG) levels were determined. During the 7th week of drug treatment, oral glucose tolerance test ( OGTT ) and insulin tolerance test ( ITT) were conducted. Before the end of experiment, 24 h urine samples were collected for the assay of rela-tive parameters. At the end of experiment, blood sam-ples were collected for the assay of glycosylated hemo-globin ( GHb) ; serums were isolated for the determina-tion of insulin concentration and other biochemical in-dexes. After sacrifice, the livers, kidneys, and pan-creases of the mice were harvested for pathological ex-amination; the contents of advanced glycation end product ( AGE) and triglyceride ( TG) in renal tissues were assayed by kits. Results GSTD at all doses sig-nificantly reduced FBG, urine glucose, GHb, serum insulin level, and the insulin resistance index in KK-Ay diabetic mice. In addition, GSTD greatly inhibited body weight gain and improved glucose tolerance and insulin tolerance ( P <0.05 or P <0.01 vs untreated group ) . The pathological examination showed that GSTD significantly increased the glycogen content of liver tissues, reduced islet volume and improved its pathological changes. In addition, the glomerulosclero-sis induced by diabetes was greatly ameliorated by GSTD. Meanwhile, GSTD greatly reduced serum crea-tine ( Scr) , 24 h urine amount, 24 h urine total pro-tein and microalbumin ( mAlb) , as well as renal AGE and TG contents in KK-Ay mice ( P <0.05 or P <0.01 vs untreated group) . The anti-diabetic effect of GSTD at 50 mg·kg-1 was comparable to that of 200 mg·kg-1 of Met. Conclusions When used to treat KK-Ay diabetic mice, GSTD has potent activities in lowering blood glucose, improving insulin resistance and ameliorating diabetic nephropathy. However, the detailed mechanisms of GSTD in modulating glucose metabolism and increasing insulin sensitivity still need further investigation.

Xiao Ke Qing improves glycometabolism and ameliorates insulin resistance by regulating the PI3K/Akt pathway in KKAy mice / 医学前沿

Xiao Ke Qing (XKQ) granule has been clinically used to treat type 2 diabetes mellitus (T2DM) for 10 years in Chinese traditional medication. However, its mechanisms against hyperglycemia remain poorly understood. This study aims to investigate XKQ mechanisms on diabetes and diabetic liver disease by using the KKAy mice model. Our results indicate that XKQ can significantly reduce food and water intake. XKQ treatment also remarkably decreases both the fasting blood glucose and blood glucose in the oral glucose tolerance test. Additionally, XKQ can significantly decrease the serum alanine aminotransferase level and liver index and can alleviate the fat degeneration in liver tissues. Moreover, XKQ can ameliorate insulin resistance and upregulate the expression of IRS-1, PI3K (p85), p-Akt, and GLUT4 in the skeletal muscle of KKAy mice. XKQ is an effective drug for T2DM by ameliorating insulin resistance and regulating the PI3K/Akt signaling pathway in the skeletal muscle.

Effect of Compound Tangshenning on Proliferation and RhoA/ROCK Signaling Pathway of High Glucose-induced Epithelial Cells of Renal Tubules / 世界科学技术-中医药现代化

Objective:To study the effect of the prescription Tangshenning on the proliferation of the high glucoseinduced cells of near-end kidney tubules.Method:To prepare durg-contained serum from mice to enter into in vitor reaction system,the cells were randomly divided into 4 groups:the normal group,the model group,the,the irbesartan group,the tangshenning group and then detect the effects of all serum sections on the proliferation of high glucoseinduced epithelial cells of kidney tubules by the MTT colorimetric method,and the expression of RhoA,ROCK 1,Ecadherin and α-SMA in each group were detected by Western blotting.Result:The high glucose group of renal tubular epithelial cells form from the flat irregular polygon into long fusiform;After adding the drug-containing serum corresponding intervention into the cells into a flat in irregular polygon.The high glucose group of renal tubular epithelial cells show obvious proliferation condition,In the condition of 24 h,48 h,The high glucose group proliferation is better than the normal group (P＜0.01),in 60 h,The high glucose group proliferation is better than the normal group (P＜0.05);In 24 h,compared with the irbesartan,Tangshenning has better effect of inhibiting the cell proliferation(P＜0.05);In 48 h,Tangshenning has the better effect than irbesartan and Y27632 (P＜0.01);In 60 h,Tangshenning has the better effect than irbesartan and Y27632 (P＜0.05);Western blotting:Western blotting analysis showed that compared with the normal group,the expression of RhoA protein in high glucose group and Y27632 decreased (P＜0.01);The high glucose group and Tangshenning have significant difference (P＜0.01);Y27632 and Tangshenning have difference (P＜0.05).compared with the normal group,the expression of ROCK1 protein in high glucose group decreased (P＜0.01);The high glucose group,Tangshenning,the irbesartan and Y27632 have difference (P＜0.05).Compared with the normal group,the expression of a-SMA protein in high glucose group decreased (P＜0.01);The high glucose group,Tangshenning,the irbesartan and Y27632 have difference (P＜0.05).Compared with the normal group,the expression of E-Cadherin protein in high glucose group increased (P＜0.05).Y27632 and Tangshenning have difference (P＜0.05).The high glucose group,Tangshenning,the irbesartan and Y27632 have difference (P＜0.05).Conclusion:The prescription Tangshenning is able to inhibit the proliferation of high glucose-induced epithelial cells of kidney tubules and and can reverse renal tubularepithelial cell transdifferentiation via regulating RhoA/ROCK signaling pathway,and restrain renal interstitial fibrosis,thereby delaying the pathogenesis of diabetic kidney disease.

Renoprotective Effect of Tang-Shen-Ning and Its Influence on Notch/Snail1 Pathway in Diabetic Nephropathy KKAy Mice / 世界科学技术-中医药现代化

This paper was aimed to study the renal protective effect of Tang-Shen-Ning (TSN) on diabetic nephropathy (DN) KKAy mice by inhibiting the Notch/snail1 signal transduction pathway.A total of 30 KKAy mice,which were fed with mice-dedicated food for 10 weeks and with the blood glucose over 16.7 mmol· L-1,24-hour urinary albumin larger than 0.4 mg,were made into the DN model.The DN mice were randomly divided into the model group,irbesartan group and TSN group according to their blood glucose and weight.Intragastric administration of medication was given.A total of 10 female C57BL/6J mice were selected as the control group.The general condition,body weight and 24-hour urinary protein quantitation were detected.After 16-week intervention,mice were sacrificed.Levels of blood glucose,blood urea nitrogen (BUN) and serum creatinine (Scr) were detected.HE and Mallory staining were applied to renal tissues.In situ hybridization (ISH) and western blotting were used to detect the Notch/snail 1 pathway,α-SMA,E-Cadherin protein and mRNA expression in renal tissues.Statistical analysis was made by SPSS20.0 software.The results showed that compared with the model group,the rats' general conditions were improved;body weight and 24-hour urinary protein quantitation were significantly decreased (P＜0.01);contents of BUN and Scr were reduced (P＜0.01,P＜0.05).The pathological staining showed significantly reduction on renal interstitial fibrosis.The Notch/snail1 pathway,protein and mRNA expression of α-SMA were significant reduced with statistical significance (P＜0.01);protein and mRNA expression of E-Cad protein were significant increased with statistical significance (P＜0.01).It was concluded that TSN can protect the renal function of DN,delay the disease progression of DN,and inhibit epithelial-mesenchymal transdifferentiation (EMT) of renal tubular epithelial cells and renal interstitial fibrosis.Furthermore,the inhibition on EMT may be through the regulation of the Notch/snail1 pathway.

Effect of Tangshenping Medicated Serum on Proliferationand RhoA / ROCK Signaling Pathway of High Glucose-induced Epithelial Cells of Renal Tubules / 世界科学技术-中医药现代化

To study the effect of the Tangshenping containing serum on the proliferation of the high glucose-induced epithelial cells of renal tubules.To prepare durg-contained serum from rats to enter into in vitor reaction system,the cellscultured via 10％ FBS-RPMI 1640 were randomly divided into 7groups:the normal group,themodel group,the Y27632 group,theirbesartangroup,the small dose Tangshenping group,the medium dose Tangshenping group and the high dose Tangshenping group and The cells were cultured in 3000 cells/well and grown in 96-well plates.Each group had 8 wells,then detect the effects of all serum sections on the proliferation of high glucose-induced epithelial cells of kidney tubules by the MTT colorimetric method after cultured for 12 h,24 h,48 h,and 60 h.Based on the results above,cell protein were extracted from each group at 24 h,and the expression of RhoA,ROCK1,α-SMA and E-cadherin in each group were detected by Western blotting.After high glucose stimulation,the shape of cell was shuttle-like or irregular triangle,the way it grew was radial;after the intervention of the corresponding serum,the shape of the cell was fiat and irregular polygonal.Started with 12h,compared with the normal group,OD value of other groups increased;at the 24h、48hand 60h,compared with the normal group,OD value of high glucose groupincreased significantly (P＜0.01);compared with the high glucose group,OD value of treatment groups decreased (P＜0.05);and 48 h,compared with the Y27632group,irbesartan groupand Tangshenping high dose group,OD value of Tangshenping low and medium dose groups decreased (P＜0.05);60 h,compared with Y27632 group,OD value of Tangshenping medium dose groups decreased;compared with irbesartan group,OD value of o Tangshenpinggroupsdecreased (P＜0.05);compared with Tangshenping high dose group,OD value of Tangshenpinglow groupsdecreased (P＜0.05) Western blotting analysis showed that compared with normal group,the expression of E-Cadherin protein in high glucose group reduced,and the expression of RhoA,ROCK1 and α-SMA protein increased;compared with high glucose group,the expression of E-Cadherin protein in each treating group increased,and the Tangshenping large dose group wassignificantly different (P＜0.01);the expression of RhoA,ROCK1 and or-SMA protein reduced,Tangshenping,the large dose group was significantlydifferent (P＜0.01);Compared with the Y27632 group,the expression of E-cadherin,ROCK1 and α-SMA protein in Tangshenping large dose group had no significant difference,while the expression of RhoAproteinreduced (P ＜0.01).Compared with theirbesartan group,the expression of E-cadherin,RhoA,ROCK1 and α-SMA protein in Tangshenping large dose group had no significant difference (P＞0.05).The Tangshenping containing serum is abletoinhibit the proliferation of high glucose-induced epithelial cells of kidney tubules,and can reverse renal tubular-epithelial cell transdifferentiation via regulating RhoA/ROCK signaling pathway,and restrain renal interstitial fibrosis,thereby delaying the pathogenesis of diabetic kidney disease.

Renoprotective Effects of Tang-Shen-Ping on RhoA/ROCK Signaling Pathway in Diabetic Kidney Disease KKAy Mice / 世界科学技术-中医药现代化

This study was aimed to explore the renoprotective effects of Tang-Shen-Ping (TSP) on RhoA/ROCK signaling pathway in KKAy mice with diabetic kidney disease (DKD).A total of 60 female 10-week SPF degree KKAy mice,which were fed with KK special food for 10 weeks,were made into DKD model.Mice were randomly divided in the model group,irbesartan group,low-,medium-and high-dose TSP group (0.525 g· kg-1,1.05 g· kg-1,and 2.1 g· kg-1).Ten female C57BL/6J mice were used as the normal control group.Mice of each group were intragastrically administered with corresponding medicine,respectively,while mice of the control group and the model group were given deionized water of the equal volume.The body weight was measured and the 24-hour urine protein quantification was detected every 4 weeks.At the end of the 26th week,all mice were sacrificed and the biochemical indicators,such as fasting blood glucose (FBG),serum blood urea nitrogen (BUN),serum creatinine (Scr),and triglyceride (TG) were measured.HE staining,Mallory staining and PAS staining were used to observe the pathological morphology of kidney tissues.Immunohistochemistry (IHC) and in situ hybridization (ISH) were used in the detection of transforming growth factor-β1 (TGF-β1),Ras homolog gene family member A (RhoA),Rho-associated coiled-coil-containing protein kinase 1 (ROCK1),α-smooth muscle actin (α-SMA),E-Cadherin (E-Cad) mRNA and protein expression.The results showed that compared with the model group,there were significant differences on body weight,the ratio of kidney weight to body weight,and urinary protein in the middle-and high-dose TSP group (P ＜ 0.01);the renal pathological damage was obviously decreased;contents of FBG,BUN,Scr and TG decreased (P ＜ 0.01);mRNA and protein expression of E-Cadherin increased;mRNA and protein expression of TGF-β1,RhoA,ROCK1 and α-SMA decreased with significant difference in the middle-and high-dosc TSP group (P ＜ 0.01).It was concluded that the renoprotective effects and epithelial-mesenchymal transdifferentiation (EMT) of renal tubular epithelial cells of TSP on DKD KKAy mice may be related to the regulation of RhoA/ROCK signaling pathway.

Effect of Gegen Qinlian Decoction on LPS, TNF-α, IL-6, and intestinal flora in diabetic KK-Ay mice / 中草药

Objective: To explore the effect of Gegen Qinlian Decoction (GGQLD) on LPS, TNF-α, IL-6, and intestinal flora in diabetic KK-Ay mice. Methods: C57BL/6J mice with ordinary feed were taken as the normal control group and orally administrated with equal distilled water. The KK-Ay mice fed with high-fat diet were divided into five groups: pioglitazone group, blank group (model group), high, medium, and low dose GGQLD group, and orally administrated with pioglitazone hydrochloride (5 mg/kg), distilled water, and GGQLD (crude drug 40, 13.3, and 4.44 g/kg), respectively. The oral administration for six groups lasted for four weeks. Tumor necrosis factor (TNF-α), interleukin 6 (IL-6), and endotoxin (LPS) levels in the plasma were determined by enzyme-linked immunosorbent assay (ELISA); Gut microbial communities were assayed by polymerase chain reaction (PCR) and PCR-denaturing gradient gel electrophoresis (PCR-DGGE) methods. Results: Compared with the model group, the LPS levels in the plasma of mice were significantly reduced by 15.61% and 14.48% respectively in the Gegenqinlian Decoction of high and medium dose group (P < 0.05), the IL-6 levels in plasma of mice were significantly reduced by 56.86%, 37.12% and 30.21% respectively in high, medium, and low dose GGQLD group (P < 0.05), and the TNF-α levels in plasma of mice were significantly reduced by 28.32%, 30.70%, and 23.42% respectively in high, medium, and low dose GGQLD group (P < 0.05). The number of DGGE bands in high dose group significantly increased, and by cloning, sequencing, and Blast analysis, Lactobacillus johnsonii only existed in the high dose group; The results showed that GGQLD could regulate the structure of intestinal flora in KK-Ay mice. Conclusion: The mechanisms of anti-diabetic effects of GGQLD in type 2 diabetic KK-Ay mice are probably related with the anti-inflammation and regulation of intestinal flora.

Effects of astragalus injection combined with puerarin injection on process of endoplasmic reticulum stress through PERK pathway in dabe-tic nephropathy mice / 中国病理生理杂志

AIM: To investigate the effects of astragalus injection combined with puerarin injection on endo-plasmic reticulum stress through PERK pathway in diabetic nephropathy mice .METHODS: Male KKAy mice were ran-domly divided into model group ( injected with normal saline ) and treatment group ( injected with astragalus and puerarin ) . The male C57BL/6J mice served as normal group .The mice were sacrificed 4 weeks after treatments for observing morpho-logical changes under electron microscope .The renal tissues were collected to determine the expression of protein kinase R-like endoplasmic reticulum kinase ( PERK ) , eukaryotic initiation factor 2α( eIF2α) and glucose-regulated protein 78 (GRP78) at mRNA and protein levels by real-time PCR and Western blot.RESULTS: Under electron microscope, the renal tubular epithelial cells in model group and treatment group showed the swelling of the nucleus , endoplasmic reticulum and mitochondria .The results of real-time PCR and Western blot showed that the expression of PERK , eIF2αand GRP78 at mRNA and protein levels in model group was higher than that in normal group (P<0.05), while that in treatment group was lower than that in model group .CONCLUSION: Astragalus injection combined with puerarin injection reduces the mRNA and protein expression of PERK , eIF2αand GRP78, thus inhibiting the endoplasmic reticulum stress in type 2 dia-betic mice to protect the kidney function .

Inhibition of Water extract from Jiangtang Decoction on inflammation in KK-Ay mice by PI3K/Akt/NF-κB signaling pathway / 药物评价研究

Objective To study the effect and mechanism of Water extract from Jiangtang Decoction (WEJTD) on diabetes mellitus and diabetic nephropathy (DN) in spontaneous type 2 diabetes mellitus model KK-Ay mice.Methods Totally 50 KK-Ay mice were randomly divided into five groups:model group,metformin (positive drug,250 mg/kg) group,WEJTD low,medium and high dose (2,4,and 8 g/kg) group,with 10 C57BL/6J mice as normal group.The relative drugs were ig administered once a day for 12 weeks,and mice in control group and model group were perfused with distilled water of equal volume.After 12 weeks' oral administration,mice were put into metabolism cages,and the food-intake,water-intake and urine volume were calculated and collected.Blood were collected to detect the concentration of IL-6,ICAM-1 and TNF-α.Then mice were executed,and HE staining and PASM staining were used to check the effect of WEJTD on kidney.Western blotting and qRT-PCR were used to detect the concentration of PI3K,Akt,NF-κB,IL-6,ICAM-1 and TNF-α in kidney.Results WEJTD can alleviate the symptoms of diabetes,such as food ration,polydipsia and polyuria (P ＜ 0.05,0.01,and 0.001);Relief the pathological changes of kidney and significantly decreased glycogen deposition (P ＜ 0.001),down-regulate the increase of IL-6,ICAM-1 and TNF-α in serum and kidney (P ＜ 0.05,0.01 and 0.001),up-regulate the phosphorylation of PI3K and Akt (P ＜ 0.05,0.01,and 0.001),and inhibit the phosphorylation of NF-κB (P ＜ 0.001).Conclusion WEJTD had positive effects on kidney morphology of KK-Ay,and the underlying mechanism might be related to the regulation of PI3K-Akt and NF-κB-mediated inflammation.

Effect ofQiwei Granules on Renal Podocyte Injury in Diabetic Nephropathy Mice / 世界科学技术-中医药现代化

This article was aimed to study the effect ofQiwei granules on the podocyte in KK-Ay mice kidney. The 28 8-week-old male KK-Ay mice were randomly divided into the model group, low-dosage, middle-dosage and high-dosageQiwei granule group. Eight C57BL/6J mice were used as the normal control. The general conditions, blood glucose and 24 h albuminuria were recorded in the experiment. After 10-week treatment, renal indexes including serum creatinine and urea nitrogen were measured. The kidneys of mice were collected and measured. The hematoxylin and eosin (HE), Masson’s Trichrome, and periodic acid-Schiff (PAS) were used on renal tissues of mice. The immunohistochemical staining for WT-1 was made. Software analysis was combined in the calculation of renal podocyte amount. Western blot was used in the detection of nephrin protein expressions in the kidney of mice. RT-PCR was used in the detection of nephrin mRNA expression. The results showed that compared with the model group, the body weight, blood glucose, 24 h albuminuria and the serum creatinine were obviously decreased after 10-week treatment ofQiwei granules. It can effectively improve the glomerular mesangial proliferation and preserve the podocyte number. Meanwhile, after the treatment ofQiwei granules, the nephrin protein expression and mRNA expression were obviously higher than the model group. It was concluded thatQiwei granules probably managed nephrin expression to improve the podocyte injury in the diabetic nephropathy of KK-Ay mice.

Pharmacology and pharmacokinetics studies of MBX-2982, a novel GPR119 agonists / 中国药学杂志

OBJECTIVE: To observe the glucose tolerance and evaluate the hypoglycemic effect of MBX-2982 respectively in the normal mice and in the KK-Ay mice. And further pharmacokinetics investigation was experimented in rats. METHODS: The influence of glucose tolerance was evaluated in KM mice which underwent a single dose of MBX-2982. Body weight, fasting blood glucose, glucose tolerance, triglyceride, serum insulin were all tested in order to evaluate the hypoglycemic effect on KK-Ay mice. The pharmacokinetics of MBX-2982 suspension and solution were investigated in rats to calculate the oral bioavailability. RESULTS: The blood glucose of each test point were all reduced after MBX-2982 (3, 10, 30 mg · kg-1) were administered orally to KM mice. After MBX-2982 (10, 30 mg · kg-1) treatment to KK-Ay mice for 4 weeks, the fasting blood glucose and triglyceride were significantly reduced and the serum insulin was remarkably increased. Meanwhile the area under glucose curve was significant reduced of 30 mg · kg-1. After oral administration of MBX-2982 (4 mg · kg-1) in rats, the oral bioavailability of suspension (0.4% CMC) and solution (DMSO-Cremopor EL-NS = 1:1:8) were 35.2% and 98.2% receptively. CONCLUSION: Animal experiment results show that the MBX-2982 as a GPR119 agonists had a good hypoglycemic effect. The absolute bioavailability of the solution is closed to 100%, which is higher than that of suspension.

Preliminary evaluation of antihyperglycemic effect of Rhizoma Coptidis alkaloids and their structure-activity relationships / 中国药理学通报

Aim To find the material foundation of treatment for diabetes in Coptidis Rhizoma ( RC ) . Methods The antihyperglycemic effect of RC alka-loids ( berberine, coptisine, palmatine, epiberberine, and jatrorrhizine) was evaluated in spontaneity diabe-tes KK-Ay mice. Results After 40 days′ oral admin-istration ( 225 mg · kg-1 · d-1 , ig ) , berberine and coptisine significantly suppressed the elevated fasting blood glucose level and ameliorated the glucose toler-ance . Body weight gain of KK-Ay mice was significant-ly decreased in the epiberberine-treated group. Berber-ine improved insulin resistance and jatrorrhizine in-creased the SOD activity, decreased the MDA level. Conclusions These results indicate that the main an-tihypoglycemic effect constituents are berberine and coptisine, while they show different mechanisms. Pal-matine, epiberberine and jatrorrhizine display different potential roles in the treatment of diabetes. The meth-ylene-dioxy groups at the C-2 , C-3 , C-9 and C-10 po-sitions are indispensable for antihyperglycemic effect of RC alkaloids.

Effect tetrahydrocurcumin-solid dispersion on KK-Ay mice / 中国药学杂志

OBJECTIVE: To study the effects tetrahydrocurcumin-solid dispersion on KK-AY mice. METHODS: C 57/6 J mice were used as controls, KK-Ay mice were randomly divided into model group, tetrahydrocurcumin-solid dispersion groups(100, 50, 15 mg · kg-1) and rosiglitazone group(2.67 mg · kg-1), gavage for 35 d, mouse weight, fasting blood glucose, oral glucose tolerance, serum insulin and blood lipid indexes were detected. RESULTS: The weight mice tetrahydrocurcumin-solid dispersion group (100 mg · kg-1) on twenty-first days administration began to decrease, and maintained at a low level during the administration; tetrahydrocurcumin-solid dispersion in each dose group showed impaired fasting blood glucose lowering from the fourteenth day after the administration began, and maintained at a low level during the administration, tetrahydrocurcumin-solid dispersion in each dose group can decrease postprandial blood glucose and AUC value; tetrahydrocurcumin-solid dispersion in each dose group can decrease the value TC, LDL-C, increase the insulin sensitivity index, tetrahydrocurcumin-solid dispersion in each dose group glycosylated serum protein values were decreased, while only in the 100 mg · kg-1 group was statistical significant differences compared with the model group. CONCLUSION: Tetrahydrocurcumin-solid dispersion, which can effectively reduce the blood glucose levels KK-AY mice, and has good effect on glucose metabolism, lipid metabolism. The mechanism action may be related to the increase insulin sensitivity.

Effect of Tang-Nai-Kang on Trans-differentiation of Renal Tubular Epithelial Cell in KKAy Mice / 世界科学技术-中医药现代化

This study was aimed to explore the effect of Tang-Nai-Kang (TNK) on trans-differentiation of renal tubular epithelial cell in KKAy mice in order to discuss the possible mechanism. Fifty 12-week-old male KKAy mice were randomly divided into the model group, valsartan group, TNK high-dose, middle-dose and low-dose group, with 10 rats in each group. Ten C57BL/6J mice were used in the normal group. Rats in the model group and normal group were given 0.9% sodium chloride solution. Rats in other groups were given the corresponding drugs. After 8 weeks of gavage administration, kidneys of all mice were sampled and given Mosson and PAS dyeing. Expression distribution of α-smooth muscle actin (α-SMA) and E-cadherin in kidney tissues were observed under immunohistochemical staining. Expression of transforming growth factor-β1 (TGF-β1) was measured by western blot. The results showed that compared with the normal group, the area of renal fibrosis in the model group was significantly increased (P < 0.01); the expression of α-SMA was stronger; and the expression of E-cadherin was weaker. Compared with the model group, the area of renal fibrosis in the valsartan group, TNK high-dose, middle-dose and low-dose groups were significantly decreased (P< 0.01); the expression of α-SMA was weaker (P< 0.01);and the expression of E-cadherin was obviously increased (P < 0.05). The TGF-β1 expression in the model group was significantly higher than that in the normal group (P < 0.01). Compared with the model group, the TGF-β1 expression in the valsartan group, TNK low-dose, middle-dose and high-dose groups were significantly lowered (P<0.01). And the TGF-β1 expression in the TNK high-dose group was even lower than that in the valsartan group. It was concluded that TNK was able to suppress the epithelial-mesenchymal transition (EMT) of renal tubular epithelial cell, and lessen the renal tubule interstitial fibrosis, in order to protect the kidney.

Effect of Gymnema Sylvestre on Protein Kinase B in Adipose Tissues of Insulin Resistance Mice / 世界科学技术-中医药现代化

This article was aimed to study effects and mechanisms of Gymnema sylvestre on protein kinase B (PKB) and its phosphorylation in adipose tissues of KKAy mice which were mainly characterized by insulin resistance (IR). A total of 18 KKAy mice were randomly divided into the diabetes model (DM) group and Gymnema sylvestre (GS) group according to body weight levels. And 9 normal C57BL/6J mice were used as the normal control (NC) group. Intragastric administration of medication was given to mice for 8 weeks. At the end of the experiment, all animals were tested for fasting plasma glucose (FPG) and fasting insulin level (Fins) for evaluation of insulin sensitivity index (ISI). Expressions of phosphoinositide-dependent kinase-1 (PDK1), PKB, P-PKB (Ser473), P-PKB (Thr 308) in adi-pose tissues of epididymis were determined. The expression of phosphatase and tensin homolog (PTEN) mRNA was also determined. The results showed that compared with the DM group, the GS group showed lower FPG and Fins, higher ISI. The expression of P-PKB (Ser473) phosphorylation and P-PKB (Thr 308) were increased, and the PDK1 and PTEN mRNA were decreased. It was concluded that GS can improve insulin sensitivity of KKAy mice through activating PKB by up-regulate the expression of P- PKB (Ser473) and its phosphorylation ratio and P- PKB (Thr 308) in adipose tissues.