ABSTRACT
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.
ABSTRACT
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.
ABSTRACT
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.
ABSTRACT
To elucidate antidiabetic effect and mechanism(s) of acarbose in a polygenic spontaneous hyperglycemic and hyperinsulinemic diabetic animal model, KKAy mice, acarbose was administered orally for 4 weeks and effects on body weight, plasma glucose and insulin levels, genetic expressions of intestinal sucrase-isomaltase (SI), sodium-glucose cotransporter (sGLT1) and glucose transporter in quadriceps muscle (GLUT4) were examined in this study. Although no differences in body weight were detected between control and acarbose-treated groups, plasma glucose level in acarbose-treated group was markedly reduced as compared to the control. In the mechanism study, acarbose downregulated the SI and SGLT1 gene expressions, and upregulated the GLUT4 mRNA and protein expressions when compared to the control group. In conclusion, the data obtained strongly implicate that acarbose can prevent the hyperglycemia in KKAy mice possibly through blocking intestinal glucose absorption by downregulations of SI and sGLT1 mRNA expressions, and upregulation of skeletal muscle GLUT4 mRNA and protein expressions.
Subject(s)
Animals , Mice , Absorption , Acarbose , Blood Glucose , Body Weight , Gene Expression , Glucose , Glucose Transport Proteins, Facilitative , Hyperglycemia , Insulin , Models, Animal , Muscle, Skeletal , Quadriceps Muscle , RNA, Messenger , Up-RegulationABSTRACT
Objective: To investigate the genes differently expressed in the livers of Kkay diabetic and normal mice, providing data to prevent human from diabetes and its chronic complications. Methods: cDNA microarray chips containing 8 192 cDNAs were used to investigate the gene expression pattern of the liver in Kkay mouse. Results: One hundred and fifty four genes were screened out, comprising 68 complete cDNAs and expressed sequence tags, among them 40 genes were up regulated and 114 genes were down regulated. Conclusion: The pathogenesis of type Ⅱ diabetes is complicated, including the disorder of the metabolism of carbohydrate, fat and protein, and many functional abnormalities of a number of vital proteins. [