| Objective:Establish T2DM rat model by a high-fat diet combined with STZ.Make pharmacology study on Huanglian Wendan decoction(HLWDD)for treating T2DM rats in order to ascertain the optimal dose of Huanglian in this prescription.Analyze on the liver lipid metabolism signal pathway PPARa-LXRa-ABCA1 in this experiment,discuss the mechanism and effect target of HLWDD on basis of its improving T2DM lipid metabolism function,which can help provide scientific basis for this prescription’s further deep study and clinical rational use.Methods:100 male SD rats were randomly divided into normal control group(Normal)and model group were fed with normal diet and high fat diet for four weeks.Make intraperitoneal injection of STZ with 30 mg/kg dose to all the model group rats.After72 hours,if the rat’s FPG was greater than or equal to 11.7 mol/L,it would be involved in this trial.The successfully established model rats were randomly divided into model group(T2DM),Huanglian group(huanglian 10g,HL),WDD+huanglian5g(HLWDD-7.25),WDD+huanglianlOg(HLWDD-7.75),WDD+huanglianl5g(HLWDD-8.25),huanglian group(HL)and the positive control group(metformin hydrochloride,MHCL),the treated group rats were administrated with different doses as follows:7.25g·kg-1,7.75g·kg-1,8.25g·kg-1,1g·kg-1 and 0.05g·kg-1 by gavage.Rats in other groups adopted same volume water by the same way.All the rats continued to be fed with same kind of food as before.The therapy course were lasted for four weeks.Weight and FBG were measured once a week.After the last administration with fasting for 12h,made last measurements of body weight and glucose;detected TC,CHO,TG and HDL-C serum levels by automatic biochemical analyzer;The antioxidant ability indexes,GSH,MDA,SOD and GSH-PX in liver were also be determined.Observing the medical morphology of rat livers in normal group,T2DM group,HL group and the best compatibility proportion group with staining by HE.The expression of PPARa,LXRa and ABCA1 protein in rat liver in the above four groups were measured by RT-PCR and Western Blot method.Results:After administration,all the indexes in treated group rats were compared with T2DM group rats,the results were described as follows:the rats body weight in HL group and all dose HLWDD groups were increased significantly(P<0.01);Glucose in all treated group rats were significant decreased(P<0.01).;TC contents in serum were decreased significantly in all treated groups(P<0.01);CHO contents in serum were also significant decreased in MHCL group(P<0.01,P<0.05),HL group and HLWDD-8.25 group;HDL in serum were increased significantly in all treated groups rats except the MHCL group;LDL in serum were decreased significantly in all treated groups rats except the HLWDD-7.25 group(P<0.01).When compared with normal group,SOD,GSH-PX and GSH contents in T2DM group rats were decreased significantly(P<0.01,P<0.01,P<0.05),MDA was increased significantly(P<0.01);When compared with T2DM group,SOD and GSH-PX contents were increased significantly in all treated groups rats;MDA contents were decreased significantly in all treated groups rats except the HLWDD-7.25 group(P<0.01,P<0.05,P<0.05,P<0.05);GSH contents in liver were also significant increased in MHCL group,HL group and HLWDD-8.26 group(P<0.05);The HE staining results showed that,liver cell structures in Normal group were clear,uniform cytoplasm.Liver cell in T2DM group were arranged disorderly,some intracytoplasmic vacuoles were rounded by fat droplets.High,medium and low dose group and the positive control group of Liver cells in all treated group rats were arranged more neatly with less fat droplets vacuoles.The RT-PCR results showed that the protein expression of PPARa,LXRa and ABCA1 protein were all significantly down-regulated in T2DM rat livers when compared with Normal groups(p<0.05,p<0.05,p<0.05);The protein expression of PPARa,LXRa and ABCA1 protein were all significantly up-regulated in HL group and the best compatibility proportion group rat livers when compared with T2DM group.Western Blot results also showed that the protein relative expression of PPARa,LXRa were all significantly reduced in T2DM rat livers when compared with Normal groups(P<0.05);The protein relative expressions of PPARa,LXRa and ABCA1 were all significantly elevated in HL group and the best compatibility proportion group rat livers when compared with T2DM group(P<0.05).There showed no differences of the three target protein relative expressions between HL group and the best compatibility proportion group(P>0.05).Conclusion:HLWDD could help decrease the glucose levels,control weight descent,improve lipid metabolism,mitigate liver damage.WDD+huanglian10g(HLWDD-7.75)was the optimal dose group of Huanglian in HLWDD on basis of the pharmacology results.In conclusion,HLLWDD could make effective treatment on T2DM with lipid metabolism,mitigate liver damage by activating the liver lipid metabolism signal pathway PPARα-LXRα-ABCA1. |
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