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Objective To clear, amplify and detect the activity of the recombinant adeno-assoeiated virus vector with adiponectin( rAAV2/1-Aerp30 ). Methods Recombinant plasmid pSNAV2.0-Acrp30 was obtained. The recombinant plasmid was then transfected into BHK21 cells using LipofectAMINETM 2000. The G418 resistant cells were obtained consequently. These cells were infected with HSVI-rc/△UL2 which has the function of packaging and copying recombinant AAV. After purification, the construction of recombinant rAAV2/1-Aerp30 was collected. Results The construction of recombinant pSNAV2.0-Acrp30 was confirmed by PCR electrophoresis and digestion with restriction enzyme. The gene sequencing showed that the recombinant pSNAV2.0-Acrp30 was correct. The virus titer was about 1.0×1012 μg/ml. The purity f the recombinant AAV2/1 was fairly high using the SDS-PAGE method. Conclusion With this method, rAAV2/-Aerp30 with high virus titers and purity can be acquired successfully and it can meet the demands of the experimental study of Acrp30 gene therapy of GK rats.
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0.05). The blood glucose, glycosylated hemoglobin and oxidized low density lipoprotein degrade, insulin resistance were improved in probucol group after treatment, while the adiponectin level was increased(P
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Objective To observe the effects of blocking renin-angiotensin system with losartan or perindopril on renal angiotensinⅡand its type 1 receptor （AT1R） expression in STZ-induced （diabetic） rats. Methods Male SD rats were randomly divided into 2 groups: normal control group （NC） and （diabetic） group. Diabetes was induced by STZ （65 mg/kg） abdominal injection. Four weeks later, diabetic rats were further divided into 4 groups: diabetic rats treated with losartan (DL, 20 mg·kg-1·d-1),diabetic rats treated with perindopril (DP,2 mg·kg-1·d-1) and diabetic untreated control group（DC）. Radioimmunoassay was used to measure plasma and renal angiotensin （Ⅱ.）Expression of AT1R was detected by RT-PCR in each group before and after 12 week′s treatment. Results At the 4th week, renal AT1R expression decreased. At the 16th week, plasma angiotensinⅡ,renal angiotensinⅡand AT1R mRNA in diabetic untreated group were significantly higher than those in normal rats. （Renal） angiotensinⅡand AT1R in losartan or perindopril-treatmented group were significantly lower than those in DC group. Conclusion Intrarenal RAS is abnormal in STZ-induced diabetic rats. Renoprotective effects of losartan and perindopril may partly induced by inhibiting renal angiotensinⅡ and AT1R （expression.）
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AIM: To observe the effects of puerarin on blood lipid and expression of aorta laminin B_1 mRNA in streptozotocin-induced diabetic rats. METHODS: Diabetic nephropathy rats were induced by intraperitoneal injection of STZ, and the experimental rats were divided into normal control group, model group, and puerarin group. During and after the treatment for 12 weeks, the general state, blood suger(BS), triglyceride(TC), cholesterol, low density lipoprotein(LDL-C), high density lipoprotein(HDL-C), glycosylated hemoglobin(HbA1c) and glycosylated low density lipoprotein(G-LDL) were detected. Aorta alteration of tissue morphology was observed by H.E staining, and the expressions of laminin B1 mRNA were determined by in situ hybridization analysis. RESULTS: Diabetes mellitus and aorta lesion occurred in the two model groups. Puerarin could improve the general state, decrease the level of triglyceride(P
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Objective To observe the short term therapeutic effects of continuous subcutaneous insulin infusion (CSII) in treatment of type 1 diabetic and type 2 diabetic patients,and compared with intensive therapy of insulin multiple subcutaneous injection by conventional scheme.Methods 48 diabetic patients including 14 cases of type 1 and 34 cases of type 2,were randomly divided into CSII group and control group. In CSII group,the short acting Novolin was subcutaneously infused using insulin pump continuously. In control group,the short acting Novolin was subcutaneously injected before three meals a day and medium acting Novolin was subcutaneously injected before sleep at night.Results Hyperglycemia of the CSII group was depressed,and decreasing degree was similar to that of the control group.However,therapeutic time,insulin dosage and insulin induced hypoglycemia reaction incidence in the CSII group were obviously lower than thoses in control group(P
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Objective To study the renal protective effect of perindopril in diabetic rats. Methods The following groups of rats were studied: normal control rats ,diabetic control rats and diabetic rats treated with perindopril (2mg?kg -1 ?d -1 ) . After 4 weeks or 12 weeks treatment , alterations of renal ultrastructure were observed in each group, Urinary albumin excretion was observed every 4 weeks.Results Urinary albumin excretion of diabetic rats treated with perindopril were significantly lower than that of diabetic untreated group(P
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Objective To observe the effect of losartan on urinary albumin excretion in streptozotocin diabetic rats.Methods The following groups of rats were studied:normal control rats(NC),diabetic control rats (DC),diabetic rats treated with losartan 〔20mg/(kg*d)〕(DL) and diabetic rats treated with perindopril （2mg/(kg*d)〕(DP).Urinary albumin was observed at the 4th、8th、12th and 16th week.Results Urinary albumin excretion of diabetic rats treated with losartan or perindopril were significantly lower than that of diabetic untreated group (P＜0.01).The effect was not different between losartan treated and perindopril treated rats.Conclusion The results suggested that losartan can reduce urinary albumin excretion in diabetic rats.
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Objective To observe the effect of puerarin on the ICAM-1 gene expression in streptozotocin-Induced diabetic nephropathy rats. Methods Diabetic nephropathy rats were induced by intraperitoneal injection of STZ.Rats were allocated into normal control group,model group,and puerarin group.The puerarin treatment lasted for 16 weeks.During and after the treatment,the general health conditions,blood glucose levels,blood urea nitrogen,serum creatinine,urinary albumin excretion rate of 24-hours,and clearance rate of creatinine of the rats were observed.The ICAM-1mRNA expressions were also determined by in situ hybridization analysis in the kidney tissue of the rats.Results Diabetes mellitus and renal function lesion occurred in the model group and puerarin group.Puerarin could improve the general health of the rats,decrease blood urea nitrogen,serum creatinine,and urinary albumin excretion rate of 24-hours;and increase the clearance rate of creatinine.The ICAM-1mRNA expressions in the kidney were significantly inhibited by puerarin treatment.Conclusion Puerarin has a function of renal protection.
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Objective To study the effects of rAAV2/1-Acrp30 on sICAM-1 and sVCAM-1 level as well as NF-?B expression in GK rats with diabetic arteriosclerosis so as to explore the effect of adiponectin on diabetic macroangiopathy. Methods A total number of 30 atherosclerotic GK rat models were randomly divided into three groups:① Model group one (M1):hind limb intramuscular injection of normal saline; ② Model group two (M2):hind limb intramuscular injection of vacuity virus rAAV2/1; ③ Treatment group (T):hind limb intramuscular injection of rAAV2/1-Acrp30 at a dose of 1?1012mg/L. After 8 weeks' treatment,the rats were killed,and serum sVCAM-1 and sICAM-1 level as well as aortic NF-?Bp65mRNA expression were measured in each group. Aortic NF-?Bp65mRNA expression was measured by RT-PCR.Results Compared with those in control model groups (M1 and M2),sVCAM-1 and sICAM-1 levels were decreased significantly in the treatment (T) group (P