滋阴潜阳法对肾性高血压大鼠AT1受体介导的信号通路的影响

发布时间：2018-03-10 19:07

本文选题：高血压大鼠　切入点：滋阴潜阳法　出处：《皖南医学院》2016年硕士论文　论文类型：学位论文

【摘要】：目的:实验通过对二肾一夹肾性高血压大鼠(2K1C-RHR)模型的复制,观察滋阴潜阳方药对模型大鼠的降压作用,并观察胸腺与脾脏中AT1R、PKA、CREB蛋白表达和血清中IL-2含量的变化。通过研究滋阴潜阳方药对高血压大鼠AT1受体介导的信号通路的影响,探讨神经、免疫、内分泌三大系统在高血压病的病理过程中发挥着整体作用及滋阴潜阳方药在2H1C-RHR中的降压机制,进一步阐述滋阴潜阳方药降压、调节神经免疫的作用靶点。方法:筛选复制成功的模型大鼠60只,随机分为模型组、氯沙坦钾组、天麻钩藤组和滋阴潜阳低、中、高组,以及正常对照组与假手术组共8组,每组10只大鼠。分别对各组大鼠进行相应处理后,观察检测各项的客观指标。采用动物无创血压测量分析仪测量用药前后大鼠的血压,观察各组大鼠的血压变化情况;AT1R、CREB、PKA采用免疫组织化学法测定,IL-2采用ELISA法测定,HE染色观察各组大鼠右肾的形态变化。数据统计方法:实验数据均以均数±标准差((?)±s)表示,多组间数据比较采用单因素方差分析,两组间数据比较采用t检验,并且用SPSS18.0软件包进行处理,P0.05、P0.01为差异显著,有统计学意义。结果:1、滋阴潜阳法对2K1C-RHR收缩压的影响:造模后2周,模型组及各用药组大鼠的收缩压均明显升高,与假手术组、正常组比较,有统计学意义(P0.01);且SBP值≥160mm Hg,造模前后收缩压差值绝对值≥20mm Hg,表明模型复制成功;用药4周后,滋阴潜阳各剂量组(32、16、8g/kg)、氯沙坦钾组和天麻钩藤组大鼠SBP均明显降低,与模型组比较,有统计学意义(P0.01)。2、滋阴潜阳法对2K1C-RHR肾组织病理学的影响:从肾脏的形态上观察到,模型组右侧受夹肾脏,与左侧肾脏相比明显萎缩,表面凹凸不平,坏死区域质地较硬为灰黄色,未坏死区为暗红色;左侧肾脏表面光滑,呈暗红色,体积较假手组明显增大。从HE染色病理切片上显示,模型组大鼠的肾小球萎缩趋势明显,肾小管发生变性并且排列紊乱;而滋阴潜阳方(低、中、高)组大鼠均有不同程度的改善。因而表明滋阴潜阳方各剂量组对肾脏具有一定的保护作用。3、滋阴潜阳法对2K1C-RHR血清IL-2的影响:模型组大鼠血清中IL-2含量明显降低(P0.01);与模型组相比,各用药组均能明显升高血清中IL-2含量,差异有统计学意义(P0.01)。4、滋阴潜阳法对2K1C-RHR胸腺、脾脏中AT1R、PKA和CREB表达的影响:(1)AT1R表达情况:模型组大鼠脾脏、胸腺中AT1R表达明显升高(P0.01);与模型组相比,各用药组均能降低脾脏、胸腺中AT1R表达,差异有统计学意义(P0.01)。(2)PKA表达情况:模型组大鼠脾脏、胸腺中PKA表达明显升高(P0.01);与模型组相比,各用药组均能降低脾脏、胸腺中PKA表达,差异有统计学意义(P0.01)。(3)CREB表达情况:模型组大鼠脾脏、胸腺中CREB表达明显升高(P0.01);与模型组相比,各用药组均能降低脾脏、胸腺中CREB表达,差异有统计学意(P0.01)。结论:1、滋阴潜阳方具有较为显著的降压效果。2、滋阴潜阳方能够保护靶器官肾脏。3、2K1C-RHR存在免疫系统调节的紊乱,滋阴潜阳方能够提高外周血中炎症因子IL-2水平,结合其对RAS系统的影响,表明滋阴潜阳方药可能通过神经免疫内分泌系统的角度发挥其对机体的整体调节作用。4、滋阴潜阳方药同时降低胸腺和脾脏AT1R、CREB、PKA的表达,提示对淋巴细胞AT1受体介导的G蛋白-cAMP-PKA信号通路的影响可能是其对高血压大鼠机体整体调节作用机制之一。
[Abstract]:Objective: through the experiment on two kidney one clip renal hypertensive rats (2K1C-RHR) model, observe the antihypertensive effect of Ziyin Qianyang Decoction on the rat model, and observe the AT1R, thymus and spleen in PKA, change of IL-2 content and expression of CREB protein in the serum. Through the influence of yin and Yang. The medicine on signal pathway of AT1 receptor induced hypertensive rats by the nerve, immunity, play the overall effect and mechanism of antihypertensive prescription of nourishing yin and Yang in the 2H1C-RHR three system in the pathological process of endocrine hypertension, further elaborated the Yin and Yang recipe buck, targets of regulation of neural immune. Copy the successful model: screening of 60 rats were randomly divided into model group, losartan group, Tianmagoutengyin group and Yin and Yang, low, high group and normal control group and sham operation group 8 group, 10 rats in each group. The rats were respectively. Should be treated objectively. The detection indexes were observed by animal noninvasive blood pressure of rats before and after blood pressure analyzer medication, blood pressure changes were observed; AT1R, CREB, PKA by immunohistochemistry assay, IL-2 was determined by ELISA method, to observe the morphological changes of the right kidney of the rats with HE staining methods: statistical data. The experimental data are mean standard deviation ((?) + s) said that the data were compared by single factor variance analysis, the data of the two groups were compared with t test, and SPSS18.0 software was used for processing, P0.05, P0.01 are significantly different, with statistical significance. Results: 1, Ziyinqianyang systolic blood pressure effects on 2K1C-RHR: 2 weeks after modeling, the systolic pressure of model group and each treatment group rats were significantly increased, and the sham operation group, normal group, with statistical significance (P0.01); and the SBP value is more than 160mm Hg, systolic blood pressure difference before and after modeling The absolute value is more than 20mm Hg, showed that the model was established successfully; after 4 weeks of medication, the Yin and Yang of each dose group (32,16,8g/kg), losartan group and Tianmagoutengyin rats in group SBP were significantly decreased, compared with the model group, there was statistical significance (.2, P0.01) Ziyinqianyang effect on 2K1C-RHR in renal tissue pathology study: from the kidney morphology observed in model group, the right kidney by the clip, compared with left renal atrophy, uneven surface, hard texture and gray areas of necrosis, no necrosis and dark red; the left kidney surface smooth, dark red, the volume was significantly higher than control group. HE staining showed that the pathological increase from section, glomerular atrophy trend of rats in model group obviously, renal tubular degeneration and arranged in disorder; and Ziyin Qianyang recipe (low, high) improved in different degree. So that group to have the Yin and yang side groups have on the kidney The protective effect of.3, effect of Ziyinqianyang on serum 2K1C-RHR IL-2: significantly decreased the serum levels of IL-2 rats in the model group (P0.01); compared with the model group, the treatment group can significantly increase the content of IL-2 in serum, the difference was statistically significant (.4, P0.01) Ziyinqianyang in 2K1C-RHR thymus. In the spleen of AT1R, expression of PKA and CREB: (1) the expression of AT1R in spleen of rats in the model group, the expression of AT1R was significantly increased in the thymus (P0.01); compared with the model group, the treatment group can reduce the expression of AT1R in spleen, thymus, the difference was statistically significant (P0.01) (2). The expression of PKA in spleen of rats in the model group, the expression of PKA was significantly increased in the thymus (P0.01); compared with the model group, the treatment group can reduce the expression of PKA in spleen, thymus, the difference was statistically significant (P0.01). (3) the expression of CREB in spleen model group rats significantly increased the expression of CREB in the thymus (P0.01) and model; Group, the treatment group can reduce the expression of CREB in spleen, thymus, there was statistically significant difference (P0.01). Conclusion: 1, yin and Yang has.2 significant antihypertensive effect, nourishing yin and yang can regulate the immune system disorders exist to protect the target organ of.3,2K1C-RHR in kidney, yin and yang to inflammatory factors the IL-2 level increased in the peripheral blood, combined with its impact on the RAS system, that Ziyin Qianyang decoction may regulate.4 by neuroimmunoendocrine system angle play an important role in the whole body, nourishing yin and Yang prescription while reducing the thymus and spleen AT1R, CREB, PKA expression, suggesting that effects on G protein -cAMP-PKA lymphocyte signaling pathway mediated by AT1 receptor may be the body of hypertensive rats overall adjustment mechanism.

【学位授予单位】：皖南医学院
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：R259

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