补肾通窍方对Ⅰ型糖尿病大鼠下丘脑前区及隔外侧核神经元Bax与Bcl-2蛋白表达的影响

【作者】 蔚芳；

【导师】 张跃明；

【作者基本信息】 浙江中医药大学 ， 中医学， 2010， 硕士

【摘要】 目的观察补肾通窍方对Ⅰ型糖尿病大鼠下丘脑前区和隔外侧核神经元Bax与Bcl-2蛋白表达的影响,研究该方抗糖尿病大鼠下丘脑前区及隔外侧核神经元退行性变的作用方法将40只清洁级雄性大鼠随机分为四组：正常对照组(NC),糖尿病模型组(DM),中药防治组(ZY)和胰岛素治疗组(XY)。通过腹腔注射链脲佐菌素(STZ)55mg/kg建立Ⅰ型糖尿病大鼠模型。其中中药防治组自造模前一周起,每天予补肾通窍方煎剂(4g生药/ml药液)1ml灌胃治疗；胰岛素治疗组在造模成功后每天皮下注射精蛋白锌重组人胰岛素注射液2u；其余二组灌服等量的生理盐水。观察各组大鼠的形态、精神等状况,并且每周定时测定各组大鼠的体重、空腹血糖1次。4周后麻醉大鼠行灌注固定,取脑组织行连续冠状石蜡切片,用免疫组化SABC法检测下丘脑神经元Bax和Bcl-2蛋白的表达水平。运用显微镜目镜网格计数单位面积内阳性神经元数目作为阳性细胞密度。结果(1)糖尿病模型组与正常对照组相比,大鼠反应行动迟缓,体重减轻(P<0.01),空腹血糖增高(P<0.01),下丘脑前区、隔外侧核Bax阳性细胞密度增加(P<0.01)；Bcl-2阳性细胞密度减少(P<0.01)。(2)中药防治组与糖尿病模型组相比,大鼠反应行动灵活,体重增加(P<0.01),空腹血糖降低(P<0.01),下丘脑前区及隔外侧核Bax阳性细胞密度均减少(P<0.01)；Bcl-2阳性细胞密度均增加(P<0.01)。(3)中药防治组与胰岛素治疗组相比,下丘脑前区、隔外侧核Bax阳性细胞密度均减少(P<0.05)；Bcl-2阳性细胞密度均增加(P<0.05)。结论补肾通窍方对链脲佐菌素诱导的Ⅰ型糖尿病大鼠下丘脑前区及隔外侧核神经元退行性变有保护和治疗作用。更多还原

【Abstract】 Objective To investigate the effects of the Bushen Tongqiao Recipe (BS TQR) on expressions of Bax and Bcl-2 in the hypothalamus neurons of I T diabetic rats and to research the therapeutic effect of Bushen Tongqiao Recipe on the degeneration of the hypothalamus neurons of diabetic rats.Methods 40 SD male rats were randomly divided into 4 groups:normal control group were without any treatment (NC); diabetes model control group(DM); traditional Chinese medicine prevention and cure group(ZY) and the insulin control group(XY). Except the normal control group, the other three groups were treated with intraperitoneal infection 55mg/kg of streptozocin (STZ). The rats in traditional Chinese medicine prevention and cure group were treated with Bushen Tongqiao Recipe lml pre-treating 1 week before the day STZ was injected for five weeks continuously. The insulin control group was cured by insulin 2u since the day the model was established while the rats in other two groups under the amount of saline filling. Observe the spirit form each rat related symptoms etc. At the same time of every week, the weight and fasting blood-glucose of the rats were detected.40 rats were killed four weeks later, fixative was injected intravascularly into the brain and tissues were picked up to make into continual paraffin was sections. The Bax and the Bcl-2 expression level was elaluated by immuno-histochemistry SABC. Totaling the number of the positive expressions as their positive cell’s density with ocular reseau of optical microscope.Results (1) Compared with NC group, the rat’s reaction and action of DM group were stupid, the weight was lower (P<0.01), the fasting blood-glucose of the rat was higher than that of the DM group (P<0.01), the positive cell’s density of the expressions of Bax increased in Hypothalamus Anterior(A) and Lareral septal nucleus (L)areas of the DM group (P<0.05) while the positive cell’s density of the expressions of Bcl-2 decreased (P<0.05). (2)Compared with the DM group, the ZY group’s reaction and action were smart and the weight was higher(P<0.01)the fasting blood-glucose was lower of rats in the DM group(P<0.01), the positive cell’s density of the expressions of Bax decreased in Hypothalamus A and L areas of ZY group (P<0.01)while the positive cell’s density of the expressions of Bcl-2 increased(P<0.01). (3)Compared with the XY group, the positive cell’s density of the expressions of Bax decreased in Hypothalamus A and L areas of ZY group(P<0.05) while the positive cell’s density of the expressions of Bcl-2 increased<0.05).Conclusions It was suggested that modified Bushen Tongqiao Recipe was effective in preventing and treating the degeneration of the hypothalamus neuron of IT diabetic rats induced by STZ.更多还原