| At present,the diabetes has become a chronic disease seriously threating our physical and mental health. The high incidence of diabetes is considered closely related to modern people’s production and life style. Overweight and obesity caused by continuous high intake of the fat have become the key pathogenic factor. Studies have confirmed that the diabetes especially the insulin resistance(IR) of type II diabetes has much to do with the disorder of fat metabolism.High FFA level caused by the disorder of fat metabolism, can result in excessive deposition of lipid in adipose tissue, there by reducing the sensitivity of insulin. FFA can reduce the release of tumor necrosis factorα( TNF2α),plasminogen activator inhibitor(PAI-1), interleukin-6(IL-6),C- reactive protein and other cytokines and secretion of bioactive polypeptides adiponectin, resulting in IR. It a necessary to develop the drugs which have the effects of hypoglycemic as well as hypolipidemic.In the recent years, the Chinese herbal hypoglycemic medicine has been widely used in clinics. Many studies have confirmed that, hypoglycemic, lipid-lowering effects are obvious in patients treated with JLD. But it is unclear at present for the specific mechanisms and targets.Objective : Toestablish a high-fatinduced insulinresistance mode and observe the effects of JLD on insulin resistance and the change of fatty tissue metablish in Apo E- /- mice, the expression analysis of proteins involved in the transfer of glucose-lipid metabolism was alanlysed to determine the underlying mechanisms.Methods:Glucose tolerance tests were done firstly in the animals after the end of the experiments, and then the areas under the curve of blood glucose(AUCglu) vs. time were Calculated. 48 hours later afterthe stress reaction disappeared in mice, Fasting venous blood of the mice following a 14-16 over-night fast was get by remove the eyeball, the blood was centrifuged for detecting the serum FBG,FFA,TC,TG,HDL-C,and LDL-C levels by automatic biochemical analyzer, and the fasting insulin level was tested with the radiation immunity analysis method, then HOMA-LR was calculated as follows:FBG×FINS/22.5. The mice were executed by cervical dislocation, and the pancreas and part of Abdominal subcutaneous tissue were fixed for HE staining, some of the adipose tissue was collected and homogenized for detecting the contents of MDA and SOD in adipose tissue by ELISA test, and the protein expressions of PPARγ,IRS-1,IRS-2,GLUT-4,SREBP-1c,LDLR,FAT,CD36,and CPT1 in adipose tissue were detected using western blotmethod.Results: Insulin sensitivity evaluation. The levels of Fins, FBG and HOMA-IR were increased in HF group compared with that of NF group(P<0.01 respectively). Those values were decreased in different Jinlida groups compared with HF group(P<0.05 or 0.01respectively), JLDH group showed an efficacy similar to that of LGLT group. After The mouse were administrated with a large amount of glucose, blood sugar levels rose to the highest levels in all groups at 30 minute,then begin to decline. The peak values of blood glucosein JLDH and JLDM groups decreased obviously compared with those of HF group(P<0.05).Biochemical alterations in plasma: The levels of TC, TG, FFA and LDLC were increased while the level of HDL-C lowered significantly in HF group compared with those of NF group(P<0.01 respectively),TG values were decreased in all Jinlida groups compared with those in HF group(P<0.05), the levels of TC,FFA and LDL-C were decreased while HDL-C level was increased in both JLDH group and JLDM group( P<0.01or<0.05 respectively).The contents of SOD and MDA in adipose tissue: The SOD contents were decreased while the MDA contents were increased in HF group compared with those in NF group(P<0.05 respectively). MDA levels were significantly decreased in all Jinlida groups compared with those of HF group(P<0.05).SOD levels were significantly increased in JLDH group and JLDM goup compared with those of HF group.HE staining of adipose tissue and pancreatic tissue: Compared with NF group, the volume of adipose tissue cell in HF group was increased, and the cell number was reduced. The cell volume was decreased in LGLT and all JLDH groups compared with that in HF group by different degree.Compared with HF group, in the cells were reduced in JLDM group and JLDL group. The pancreas after HE staining showed that unclear islet structure and incomplete cell were found in HF group. Compared with HF group, the injuries in pancreas tissue in LGLT group and different JLD groups were reduced in different degree.Results of Western Blot: The expressions of FAT and SREBP-1C were increased while the expressions of CPT1, GLUT-4, IRS-1, IRS-2, and PPARγ were decreased in adipose tissue in HF group compared with those in NF group. The decreased IRS-2 expression in HF group was attenuated in all JLD groups, and the decreased expressions of CPT1, IRS-1, IRS-2 and increased expression of SREBP-1C were attenuated in JLDM group and JLDH group, the expressions of GLUT-4 and PPARγwere increased in JLDH group compared with HF group. The expression changes of GLUT-4, IRS-1, IRS-2, and SREBP-1C in HF group were attenuated in LGLT group.Conclusions:1 Insulin resistance and lipid metabolism disorderswere found in APOE- /- mice with high fat diet for months,which may be caused by severe oxidative stress,or by the changes of protein expression involved in metabolism of glucose and lipid in the adipose tissue.2 Insulin resistance and lipid metabolism disorders in APOE- /- mice with high fat dietwere attanuated by the treatment of JLD, which may be caused by reduaction of oxidative stress, or by promoting the use of blood sugar and lipid transformation. |
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