| 摘要: |
| 观察中药桃仁对2型糖尿病大血管纤维化大鼠模型中蛋白激酶B（AKT）信号通路的影响。方法：雄性SD大鼠200只，随机选取30只为空白对照组，余170只采用高脂高糖饲料喂养-腹腔注射链脲佐菌素（STZ）-高脂高糖饲料持续喂养的方法制备2型糖尿病大血管纤维化大鼠模型。造模成功后分为空白对照组（n=29）、模型对照组（n=34）、早期干预组（n=34）、低剂量组（n=32）和高剂量组（n=32）。空白对照组不予特殊处理，早期干预组和低剂量组予桃仁颗粒剂水溶液10 mL/(kg·d) 灌胃，高剂量组予桃仁颗粒剂水溶液20 mL/(kg·d) 灌胃，模型对照组给予10 mL/(kg·d) 生理盐水灌胃。干预7周后分别从各组随机选取5只大鼠处死、取材。实时荧光定量PCR（QPCR）检测AKT mRNA表达；免疫组织化学、Western blotting检测股动脉组织AKT信号通路关键信号分子AKT、磷酸化AKT（p-AKT）的表达。结果：免疫组化显示，空白对照组大鼠血管内皮细胞及中膜平滑肌细胞中有散在黄色物质，呈弱阳性改变，模型对照组呈强阳性反应，早期干预组和高剂量组呈弱阳性反应，低剂量组呈阳性反应。QPCR检测，与空白对照组（1.05±0.05）比较，模型对照组（5.68±0.61）、药物干预组（4.27±0.32、5.33±0.60、4.72±0.28）AKTmRNA表达上调（P<0.05或P<0.01）；与模型对照组比较，药物干预组AKT mRNA表达均上调，且以早期干预组（4.27±0.32）和高剂量组（4.72±0.28）为著（P<0.01）；Western blotting检测，与空白对照组（0.16±0.01、0.10±0.03）比较，模型对照组（0.38±0.03、0.21±0.02）、药物干预组（0.27±0.04、0.18±0.01；0.30±0.05、0.17±0.01；0.28±0.03、0.19±0.02）AKT、p-AKT表达均显著上调（P<0.01）；与模型对照组比较，药物干预组AKT、p-AKT表达均上调（P<0.01或P<0.05）；药物干预组两两比较显示，AKT、p-AKT表达无差异（P>0.05）。结论：中药桃仁可以抑制糖尿病大鼠大血管纤维化，其机制可能和抑制AKT信号通路有关。 |
| 关键词: 糖尿病大鼠 血管纤维化 蛋白激酶B 桃仁 |
| DOI：10.3969/j.issn.1007-6948.2019.01.013 |
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| 基金项目:国家自然科学基金资助项目（81473685） |
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| Effect of Peach Kernel on Protein Kinase B Signaling Pathway in Diabetic Macrovascular Fibrosis Rats |
| ZHOU Yu,LIU Guo-tao,LU Zeng-zhen,XU Yang,WANG Jun |
| Graduate School of Tianjin University of Traditional Chinese Medicine, Tianjin (300193), China |
| Abstract: |
| Objective To observe the effect of Peach Kernel Granule on protein kinase B (AKT) signaling pathway in type 2 diabetic rats with macrovascular fibrosis. Methods There were 200 male SD rats, 30 of which were randomly selected as blank control group. The remaining 170 rats were given high fat and sugar diet - intraperitoneal injection of streptozotocin (STZ) - continuous feeding with high fat and sugar diet to prepare macrovascular fibrosis model of type 2 diabetes mellitus. Thirty-nine rats were excluded from the loss of death during feeding and modeling, and were eventually divided into blank control group (n=29), model control group (n=34), early intervention group (n=34), low dose group (n=32) and high dose group (n=32). The blank control group was not given special treatment. The model control group was given 10 mL/(kg?d) normal saline by gavage, the early intervention group and the low dose group were given 10 mL/(kg?d) Peach kernel granule solution by gavage, and the high dose group was given 20 mL/(kg?d) Peach Kernel Granule solution by gavage. The early intervention group began to be intervened until the model of type 2 diabetes mellitus was established. The other groups began to be intervened after the model of type 2 diabetes mellitus macrovascular fibrosis rats was established. The intervention lasted for 7 weeks. At last, five rats were randomly selected from each group for experiment. Real-time fluorescence quantitative polymerase chain reaction (QPCR) was used to detect the expression of AKT. Immunohistochemistry and Western blotting were used to detect the expression of AKT and phosphorylated AKT in femoral artery tissue. Results Immunohistochemistry showed that yellow substances were scattered in vascular endothelial cells and smooth muscle cells of rats in blank control group, showing weak positive changes, strong positive reactions in model control group, weak positive reactions in early intervention group and high dose group, and positive reactions in low dose group. Compared with the blank control group (1.058687.png 0.05), the expression of mAKT in the model control group (5.688701.png0.61), and drug intervention groups (4.278714.png0.32, 5.338729.png0.60, 4.728743.png0.28) was up-regulated (P < 0.05 or P < 0.01) Compared with the model control group, the expression of mAKT in the drug intervention groups was up-regulated, especially in the early intervention group (4.278758.png0.32) and the high dose group (4.728771.png0.28) (P < 0.01). Compared with the blank control group (0.168784.png0.01, 0.108800.png0.03), the expression of AKT and p-AKT in the model control group (0.388811.png0.03, 0.218822.png0.02), and the drug intervention groups (0.278835.png0.04, 0.188849.png0.01; 0.308862.png0.05, 0.178877.png0.01; 0.288893.png0.03, 0.198908.png0.02) was significantly increased (P<0.01). Compared with the model control group, the expression of AKT and p-AKT in the drug intervention group was significantly increased (P < 0.01 or P < 0.05). The expression of AKT and p-AKT showed no difference between the two groups (P > 0.05). Conclusion Peach kernel can inhibit macrovascular fibrosis in diabetic rats, and its mechanism may be related to the inhibition of AKT signaling pathway. |
| Key words: Diabetic rats vascular fibrosis protein kinase B peach kernel |
