Dangua Fang(丹瓜方)regulating tricarboxylic acid cycle and respiratory chain and its mechanism in diabetic rats

OBJECTIVE:To investigate the influence and possible targets of Dangua Fang(丹瓜方)on tricarboxylic acid(TCA)cycle and respiratory chain to enrich the prescription’s mechanism of effective intervention on glycolipid metabolic diseases such as type 2 diabetes.METHODS:After interventional rats were fed with high glucose and high fat diet ad libitum for 4 weeks,intraperitoneally injected streptozotocin to induce diabetic model.According to blood glucose level,28 diabetic rats were selected and continued to be fed with high glucose and high fat diet,were stratified by body weight,and divided randomly by blood glucose into Model group(was given sterile water by gastric perfusion and injected aquae pro injection intraperitoneally),Dangua group[Dangua liquor(丹瓜方液)20.5 g·kg^(-1)·d^(-1) by perfusion and aquae pro injection intraperitoneally],Inhibitor group[sterile water by perfusion and nicotinamide phosphoribosyl transferase(Nampt)specific blocker GEN-6171.25 mg/kg intraperitoneally],DanInhit group(Dangua liquor and GEN-617 synchronously).Control group were continuously fed with ordinary diet.The intervention was last for 10 weeks.Body weight(BW),liver index(LI),glycosylated hemoglobin(HbA1c),TC,TG,free fatty acids(FFA),creatinine(Cr),and A-ketoglutarate(α-KG),Iso-citric acid(ICA),oxaloacetic acid(OAA)were tested.The cytochrome C oxidase(COX)and Succinate dehydrogenase(SDH)were evaluated by Colorimetry;Nampt protein,Adenosine triphosphate(ATP)synthase(ATPs),Nicotinamide adenine dinucleotide(NAD^(+))and its reduced(NADH)in liver were measured by enzyme linked immunosorbent assay.The expressions of Nampt and mitochondrialnadhdehydrogenase-1(mt-ND1)gene in liver was assessed by real-time polymerase chain reaction.Hepatic tissue staining was also completed.RESULTS:The levels of BW,ICA,α-KG and Nampt-mRNA in the Model group are lower than that in the Normal group(P<0.05),conversely,liver weight,LI,TC,HbA1c,SDH and ATPs,mt-ND1-mRNA,and Nampt protein in the Model group are higher(P<0.01,P<0.05).Compared with Model group,the levels of ICA,Nampt-mRNA and Nampt in Dangua group are significantly increased,and FFA obviously raised(P<0.01 and P<0.05);liver weight,BW,SDH are obviously lower,and HbA1c decreased significantly(P<0.01,P<0.05).TG,FFA and Nampt protein increased in the DanInhit group,TC,TG,BW obviously increased in the Inhibitor group,but SDH is decreased in both the two groups(P<0.05,P<0.01).Compared with Dangua group,DanInhib group has the lower levels of ICA,mt-ND1-mRNA,Nampt-mRNA,and the higher level of BW,LI and HbA1c.In the Inhibitor group,ICA and Nampt protein decreased,BW and LI,HbA1c and TG increased(P<0.01 or P<0.05).Tissue staining display that,in the model group there is obvious pathologic changes ie:fibrosis,steatosis and inflammatory cell infiltration.Lesions in the Dangua group are mild,and those of Inhibitor group are more obvious than the Model group,and DanInhit group is intermediately affected compared to Dangua group and Inhibitor group.CONCLUSION:Dangua Fang increases the metabolic flux of TCA cycle and optimizes respiratory chain function by up-regulating Nampt expression.

Dingxin recipe alleviates atherosclerosis injury in Apo E-knockout mice via downregulation of visfatin expression and inhibition of the visfatin-induced inflammatory response

OBJECTIVE:To further elucidate the mechanism underlying the anti-atherosclerotic effect of Dingxin recipe(DXR).METHODS:Fifty 6-week-old male Apo E^-/-mice were randomly divided into the following groups:model,simvastatin(5 mg·kg^-1·d^-1),DXR low-dose(9.30 g·kg^-1·d^-1),DXR middle-dose(18.59 g·kg^-1·d^-1)and DXR high-dose(37.18 g·kg^-1·d^-1)(n=10).Ten male C57BL/6J mice were used as the control group.All Apo E^-/-mice were fed a high-fat diet(HFD)and the control mice received a common diet.After HFD for 12 weeks,the mice were treated with DXR or simvastatin for another 12 weeks.The expression of inflammatory cytokines and visfatin was determined in serum and atherosclerotic lesions by enzyme-linked immunosorbent assay.Visfatin expression was also assessed in aortic atherosclerotic plaques.Cultured vessel endothelial cells(VECs)were pretreated with DXR sera prior to visfatin.The effects of DXR were analyzed to elucidate its protective mechanism against visfatin-induced inflammation in VECs.RESULTS:DXR regulated blood lipids and reduced tumor necrosis factor-α(TNF-α),interleukin-6(IL-6),intercellular adhesion molecules-1(ICAM-1),vascular cell adhesion molecule-1(VCAM-1)and visfatin expression in Apo E^-/-mice,particularly at the higher doses.The areas of atherosclerotic lesions in the DXR groups were significantly smaller than those in the model group.DXR alleviated visfatin-induced VEC injury via downregulation of TNF-α,IL-6,ICAM-1 and VCAM-1 through mitogen-activated protein kinase pathways.CONCLUSION:DXR alleviated atherosclerosis injury via downregulation of visfatin expression and inhibition of the visfatin-induced inflammatory response in VECs.