益气养阴，化瘀祛浊中药抗糖尿病大鼠大血管病变及作用机理研究
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摘要
目的:探讨益气养阴,化瘀祛浊法抗糖尿病(DM)大鼠低度炎症及大血管病变的作用机理,为临床中医药的推广提供有效数据及理论依据。方法:1、建立糖尿病大鼠大血管病的实验模型。通过高糖、高脂、胆固醇、胆酸盐等膳食方法喂饲一个月及腹腔注射链脲佐菌素(STZ)45mg/kg,复制糖尿病大鼠大血管病的病理模型。并设正常组、空白模型对造组、AS对照组、阳性药对照组、中药预防、治疗组。预防组在给予高糖高脂膳食的同时立即给予中药。AS对照组不给高糖、不注射链脲佐菌素。各组成模后治疗8周。并与福辛普利进行对照比较。2、各组大鼠实验12周后处死,提取血清、主动脉、通过生化、放免、免疫组化、RT—PCR法及蛋白印迹杂交法进行研究。观察本实验药对血糖、血脂、IL—6、TNF-α的影响。并通过免疫组化观察PAI—1的表达。光镜观察动脉血管内皮,血管平滑肌增生情况。蛋白印迹杂交及RT—PCR检测:TGF—β、PAI—1、PPARr蛋白及基因表达。结果:益气养阴,化瘀祛浊中药能降低早期DM大鼠血糖;降低TG、TC、LDL-C;预防组能抑制IL—6、TNF-α的表达,免疫组化观察:预防组能抑制PAI—1的表达。光镜观察:中药组可保护动脉血管内皮的完整,抑制血管平滑肌增生;RT-PCR显示:能抑制TGF—β、PAI—1蛋白及基因表达。增强PPARr蛋白及基因表达。均P<0.01。结论:1、调节血糖、脂代谢的紊乱;2、降低PAI-1,改善循环;3、抑制炎性因子IL-6、TNF-α的表达;4、抑制血管TGF—β、PAI—1蛋白及基因表达。增强PPARr蛋白及基因表达。5、保护动脉血管内皮的完整,抑制血管平滑肌增生。6、该中药对早期DM大鼠的大血管有保护作用。防治DM大鼠大血管病。
Object:
To investigate the pharamacodynamic mechanism of boost Qi nourishing Yi dissipating blood stasis removing turbid urine recipe to the low inflammation and macrovascular disease in diabetes mellitus rats,so as to provide effective data and theoretical evidence for spreading use of the Chinese clinic medicine.
Methods:
1. The macrovascular disease models of diabetes mellitus rats were set up. The rats were fed with the diets: high sucrose and fat, cholesterol and cholate in a month, getting intra-abdominal injection of STZ 45mg/kg to make the models of DM macrovascular disease established. The experimental rats were randomly divided into six groups: normal group,blank pattern contrast group, AS contrast group, positive medicine contrast group, prevention group and cure group. Prevention group was fed with fat diet, meanwhile, taking Chinese herbal medicine. AS contrast group wasn’t given high sucrose Intra-abdominal injection of STZ. After each group models were set up, they were treated for eight weeks, taking Fosinopril as the comparative drug.
2. After sacrificing the rats in twelve weeks, serum and aorta were studied through the techniques: biochemistry, radioimmunoassay, immuno-histochemistry (IHC), reverse transcriptase PCR(RT-PCR) and western blot. Observing the experimental medicine effect on blood fat, blood glucose, inflammatory factor IL-6 and TNF-α, researching the expression of PAI-1by immuno-histochemistry further, observing artery vascular endothelial and the hyperplasia of vascular smooth muscle by light microscope and studying the expression of protein and gene of TGF-β,PAI-1and PPARr by western blot and RT-PCR.
Results:
Boost Qi nourishing Yi, dissipating blood stasis and removing turbid urine recipe lowered blood glucose, TG, TC and LDL-C in early diabetes mellitus rats. In prvention group,the expression of IL-6 and TNF-αwas reduced. The results showed that : the expression of PAI-1 was inhibited in prevention group by immuno-histochemistry, the Chinese herbal medicine protected integral artery vascular endothelial and inhibited the hyperplasia of vascular smooth muscle observed by light microscope, and by RT-PCR the expression protein and gene of TGF-βand PAI-1was inhibited, while that of PPARr was strengthened (p<0.01) .
Conclusion:
1. Regulating glycometabolism,lipid metabolism;
2. Decreasing PAI-1,and improving microcirculation;
3. Inhibiting the expression of inflammatory factor IL-6 and TNF-α;
4. Inhabiting the expression of protein and gene of vascular TGF-βand PAI-1, and strengthening that of PPARr.
5. Protecting artery vascular endothelial integrity and inhabiting the hyperplasia of vascular smooth muscle ;
6. It was indicated that the medicine prevented and treated the macrovascularity in early diabetes mellitus rats.
Object:
To investigate the pharamacodynamic mechanism of boost Qi nourishing Yi dissipating blood stasis removing turbid urine recipe to the low inflammation and macrovascular disease in diabetes mellitus rats,so as to provide effective data and theoretical evidence for spreading use of the Chinese clinic medicine.
Methods:
1. The macrovascular disease models of diabetes mellitus rats were set up. The rats were fed with the diets: high sucrose and fat, cholesterol and cholate in a month, getting intra-abdominal injection of STZ 45mg/kg to make the models of DM macrovascular disease established. The experimental rats were randomly divided into six groups: normal group,blank pattern contrast group, AS contrast group, positive medicine contrast group, prevention group and cure group. Prevention group was fed with fat diet, meanwhile, taking Chinese herbal medicine. AS contrast group wasn’t given high sucrose Intra-abdominal injection of STZ. After each group models were set up, they were treated for eight weeks, taking Fosinopril as the comparative drug.
2. After sacrificing the rats in twelve weeks, serum and aorta were studied through the techniques: biochemistry, radioimmunoassay, immuno-histochemistry (IHC), reverse transcriptase PCR(RT-PCR) and western blot. Observing the experimental medicine effect on blood fat, blood glucose, inflammatory factor IL-6 and TNF-α, researching the expression of PAI-1by immuno-histochemistry further, observing artery vascular endothelial and the hyperplasia of vascular smooth muscle by light microscope and studying the expression of protein and gene of TGF-β,PAI-1and PPARr by western blot and RT-PCR.
Results:
Boost Qi nourishing Yi, dissipating blood stasis and removing turbid urine recipe lowered blood glucose, TG, TC and LDL-C in early diabetes mellitus rats. In prvention group,the expression of IL-6 and TNF-αwas reduced. The results showed that : the expression of PAI-1 was inhibited in prevention group by immuno-histochemistry, the Chinese herbal medicine protected integral artery vascular endothelial and inhibited the hyperplasia of vascular smooth muscle observed by light microscope, and by RT-PCR the expression protein and gene of TGF-βand PAI-1was inhibited, while that of PPARr was strengthened (p<0.01) .
Conclusion:
1. Regulating glycometabolism,lipid metabolism;
2. Decreasing PAI-1,and improving microcirculation;
3. Inhibiting the expression of inflammatory factor IL-6 and TNF-α;
4. Inhabiting the expression of protein and gene of vascular TGF-βand PAI-1, and strengthening that of PPARr.
5. Protecting artery vascular endothelial integrity and inhabiting the hyperplasia of vascular smooth muscle ;
6. It was indicated that the medicine prevented and treated the macrovascularity in early diabetes mellitus rats.
引文
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