五味子对糖尿病大鼠肾脏基质金属蛋白酶系统的影响

发布时间：2018-04-16 09:18

本文选题：五味子 + 糖尿病肾病　；参考：《吉林大学》2016年硕士论文

【摘要】：糖尿病肾病(diabetic nephropathy,DN)是由糖尿病引起的一种较为常见的慢性并发症,可导致终末期肾病,是糖尿病中危害性最大的一种并发症。DN时肾脏细胞外基质(extracellular matrixc,ECM)合成和降解失衡,导致ECM积聚,其主要与基质金属蛋白酶(Matrix metalloproteinases,MMPs)系统等有关。五味子是木兰科植物五味子或华中五味子的干燥成熟果实,属传统中药,但关于其单味药以及提取物对DN的作用尚无人研究。因此在本研究中,我们从多种机制研究了五味子提取物对DN大鼠的肾脏保护作用,分别检测了五味子提取物对DN大鼠的血糖水平、血脂代谢、氧化应激水平及肾功能变化等多方面的影响,并观察了五味子提取物对DN大鼠肾脏ECM成分(纤维连接蛋白FN、Ⅳ型胶原蛋白colⅣ)含量的影响,及MMP-2活性、TIMP-2表达情况,从基质降解的角度进行进一步的机制探讨,为五味子提取物治疗和改善DN提供理论基础。利用大鼠单次腹腔注射STZ(55 mg/kg)复制糖尿病动物模型,造模成功的糖尿病大鼠随机分为正常对照组(Control)、模型组(DN)、五味子提取物组(Sch)和贝那普利阳性药组(Bena),并于12周后观察五味子提取物对DN大鼠各项指标的作用。我们检测了各组大鼠常规血、尿生化指标,分别检测了血清、肾组织各项氧化应激水平;常规病理染色(HE、PAS)观察各组大鼠肾脏组织形态学改变;免疫组织化学法检测肾小球纤维连接蛋白(fibronectin,FN)、Ⅳ型胶原蛋白(CollagenⅣ,colⅣ)、TIMP-2在肾脏的表达;酶谱法检测MMP-2活性。研究结果表明,模型组大鼠肾脏部分肾小球出现血管管腔塌陷、系膜区细胞数增加、系膜区扩大等病理学改变,在给予五味子提取物治疗后,大鼠组织形态学改变明显改善。对血脂指标进行检测,模型组大鼠血T-CHO和LDL-C含量明显增多(P0.05,P0.01);与模型组比较,五味子提取物治疗组LDL-C明显降低(P0.05),T-CHO也有降低趋势。对氧化应激指标进行检测,模型大鼠血清GSH-PX活性降低、MDA增多,肾脏组织GSH-PX及CAT活性降低、MDA增多,均具统计学意义(P0.05),与模型组相比较,五味子组大鼠血清MDA减少(P0.05),肾脏组织CAT、SOD活性升高(P0.01),MDA减少(P0.05)。五味子组糖尿病大鼠肾小球病变有所缓解,系膜细胞增生减轻,基底膜变薄,ECM成分胶原蛋白减少,效果与贝那普利组相近。免疫组化显示给予五味子提取物治疗后肾小球内FN及ColⅣ表达明显减少,且抑制ECM降解的TIMP-2表达明显减少;酶谱法观察到糖尿病组大鼠MMP-2活性明显下降(P0.05),而与糖尿病组比较,五味子组大鼠MMP-2活性明显增强(P0.05)。结论:五味子提取物对STZ导致的实验性糖尿病大鼠具有肾脏保护作用,其机制可能与其降血脂、改善血清及肾组织氧化应激水平,提高糖尿病大鼠的肾脏MMP-2活性、抑制TIMP-2的表达、改善肾小球细胞外基质降解有关。创新点:本研究首次观察了五味子提取物对实验性糖尿病大鼠肾脏的干预作用,从ECM降解酶的角度探讨了其肾脏保护的可能作用机制。
[Abstract]:Diabetic nephropathy (diabetic nephropathy DN) is a common chronic complication caused by diabetes, which can lead to end-stage renal disease, is the most harmful as a complication of.DN extracellular matrix of kidney in diabetes (extracellular matrixc, ECM) synthesis and degradation of imbalance, leading to ECM accumulation, mainly with matrix metalloproteinase (Matrix metalloproteinases MMPs) system and so on. Chinensis is Magnoliaceae Schisandra sphenanthera or dried ripe fruit, is a traditional Chinese medicine, but the study on the effect of single herb extracts on DN and no one. In this study, we investigated the renal protective effect of Schisandra chinensis extract on rats from DN a variety of mechanisms, were detected in the blood glucose level, Schisandra chinensis extract on DN rats blood lipid metabolism, effect of stress level and renal oxidative changes in many aspects, and observe the The composition of Schisandra chinensis extract on renal ECM DN rats (fibronectin FN, type IV collagen col IV) were studied, and the activity of MMP-2, TIMP-2 expression, to further explore the mechanism of matrix degradation from the point of view, to provide the theoretical basis for improving the treatment of DN. And the extract of Schisandra chinensis by single intraperitoneal injection of STZ (rat 55 mg/kg) to replicate diabetic animal models and made diabetic rats were randomly divided into normal control group model (Control), model group (DN), Schisandra extract group (Sch) and Benner Pury positive drug group (Bena), and to observe the effect of extract of Schisandra chinensis on the indexes of DN rats after 12 weeks. Detection of the rats blood routine, urine biochemical indexes were detected in serum, the levels of oxidative stress in kidney tissue; pathological staining (HE, PAS) to observe the changes of renal morphology of rats; immunohistochemistry Glomerular fibronectin (fibronectin, FN), collagen type IV (Collagen IV, col IV), the expression of TIMP-2 in kidney; enzyme activity of MMP-2 was determined. The results show that the renal glomerular part of model rats, vascular lumen collapse, mesangial cell number, pathology department expanded film pathology. In the extract of Schisandra chinensis after treatment, morphology of rat tissue change significantly improved. To detect the serum lipids, the rats in the model group blood T-CHO and LDL-C content increased significantly (P0.05, P0.01); compared with the model group, the treatment group had significantly lower LDL-C extract of Schisandra chinensis (P0.05), T-CHO also had a decreasing tendency. To detect the oxidation stress index, decreased the activity of serum GSH-PX in a rat model of MDA increased, GSH-PX decreased, and the activity of CAT in kidney tissue increased MDA, were statistically significant (P0.05), compared with the model group, the serum of rats in MDA group decreased Schisandra (P0.05), renal tissue CAT, increased the activity of SOD (P0.01), MDA (P0.05). To reduce the glomerular lesions of Schisandra group of diabetic rats reduced, mesangial cell proliferation decreased, basement membrane thinning, ECM collagen decreased, the similar effect of benazepril group. Immunohistochemistry showed that the treated extract of Schisandra chinensis after treatment in glomeruli FN and Col IV expression was significantly reduced, and inhibits the degradation of ECM TIMP-2 expression significantly decreased; MMP-2 activity was observed in diabetic rats significantly decreased the enzyme spectrum (P0.05), and compared with the diabetic group, MMP-2 rats significantly increased activity of Schisandra chinensis (P0.05). Conclusion: the experimental diabetic rats of Schisandra chinensis extract on STZ to protect the function of kidney, the mechanism may be related to lowering blood lipid, improving oxidative stress level in serum and renal tissue of diabetic rats, improve the renal MMP-2 activity, expression of TIMP-2, improve glomerular cells Innovation: the first time we observed the intervention effect of Schisandra chinensis extract on the kidneys of experimental diabetic rats, and explored the possible mechanism of renal protection from the perspective of ECM degradation enzyme.

【学位授予单位】：吉林大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：R259

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