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目的观察黑逍遥散对Aβ25-35诱导阿尔茨海默病(Alzheimer’s disease,AD)大鼠模型海马基因表达谱的影响。方法选择雌性SD大鼠,以海马注射Aβ25-35淀粉样蛋白建立AD模型,并设立假手术组、模型组、西药组及中药高、中、低剂量组,每组14只。大鼠造模7天后连续灌胃[3 m L/(kg·d)]28天,假手术组与模型组给予生理盐水,西药组给予石杉碱甲片水溶液[0.02 mg/(kg·d)],中药高、中、低剂量组分别给予黑逍遥散水煎液[17.00、8.50、4.25 g/(kg·d)]。灌胃结束后处死大鼠解剖取得大鼠海马组织,并提取组织RNA,利用大鼠基因表达谱芯片筛选差异表达的基因,而后对获得的部分剂量依赖性变化的差异表达基因进行qRT-PCR验证。结果与假手术组比较,模型组583个基因表达上调,579个下调,wisp1、crebbp、igfbp-1、znf483、zfp37及zic4 mRNA表达升高,casq2、bcl-2 mRNA表达降低(P<0.05)。与模型组比较,中药高、中、低剂量组276个基因表达上调,170个下调,其中71个基因剂量依赖性上调表达,70个基因剂量依赖性下调表达,西药组igfbp-1、znf483、zfp37及zic4 mRNA降低,casq2、bcl-2 mRNA升高(P<0.01);中药高剂量组wisp1、crebbp、igfbp-1、znf483、zfp37及zic4降低(P<0.01),casq2、bcl-2 mRNA升高(P<0.01);中药中剂量组crebbp、igfbp-1、znf483、zfp37及zic4降低(P<0.01,P<0.05),casq2、bcl-2 mRNA升高(P<0.01,P<0.05);中药低剂量组igfbp-1、znf483、zfp37及zic4 mRNA降低(P<0.01)。与中药高剂量组比较,中药中剂量组crebbp、zfp37、zic4 mRNA升高(P<0.01),igfbp-1、bcl-2 mRNA降低(P<0.01,P<0.05);中药低剂量组crebbp、znf483、zfp37 mRNA升高(P<0.01,P<0.05),igfbp-1、bcl-2、zic4 mRNA降低(P<0.01)。与中药中剂量组比较,中药低剂量组casq2、bcl-2、zic4 mRNA降低(P<0.01,P<0.05)。结论黑逍遥散可通过调控zfp37、znf483、zic4表达影响AD发生;通过抑制wnt信号通路相关基因wisp-1、crebbp、igfbp-1、casq2的表达而影响Aβ代谢和Tau蛋白异常磷酸化。
Objective To observe the effects of Heixiaoyao powder on the gene expression profiles of hippocampus in Alzheimer’s disease (AD) -induced Aβ25-35 induced AD rats. Methods Female SD rats were selected, AD model was established by injecting Aβ25-35 amyloid into the hippocampus, and 14 rats in each group were established as sham operation group, model group, western medicine group and Chinese medicine high, medium and low dose groups. The rats were given intragastric administration of [3 m L / (kg · d)] for 7 days and the rats in the sham-operation group and the model group were given normal saline. The rats in the western medicine group were treated with huperzine A solution [0.02 mg / (kg · d )], Chinese medicine high, medium and low dose groups were given Xiaoyaoyao powder decoction [17.00,8.50,4.25 g / (kg · d)]. The rats were killed after gavage, the hippocampus was dissected and the tissue RNA was extracted. The differentially expressed genes were screened by rat gene expression microarray. Then the differentially expressed genes that were obtained in some dose-dependent manner were verified by qRT-PCR . Results Compared with the sham operation group, 583 genes were up-regulated and 579 were down-regulated in the model group. The expressions of wisp1, crebbp, igfbp-1, znf483, zfp37 and zic4 mRNA and casq2 and bcl- . Compared with model group, 276 genes were up-regulated and 170 were down-regulated in high, medium and low dose groups of Chinese medicine group, of which 71 genes were up-regulated in a dose-dependent manner and 70 genes were down-regulated in a dose-dependent manner. Western medicine group igfbp-1, znf483, zfp37, zfp37 and zic4were decreased (P <0.01), while the expression of casq2 and bcl-2 mRNA was decreased (P <0.01) (P <0.01, P <0.01). The levels of crebbp, igfbp-1, znf483, zfp37 and zic4 in middle-dose group decreased ); Low dose Chinese medicine group igfbp-1, znf483, zfp37 and zic4 mRNA decreased (P <0.01). The crebbp, zfp37 and zic4 mRNA in middle-dose group were significantly increased (P <0.01), and the mRNA expressions of igfbp-1 and bcl-2 in the middle-dose group were significantly decreased znf483 and zfp37 mRNA (P <0.01, P <0.05), and the mRNA expressions of igfbp-1, bcl-2 and zic4 decreased (P <0.01). Compared with the medium dose group, the levels of casq2, bcl-2 and zic4 mRNA in the low dose group decreased (P <0.01, P <0.05). Conclusion Heixiaoyao San can affect the occurrence of AD by regulating the expression of zfp37, znf483 and zic4, and affect the metabolism of Aβ and abnormal phosphorylation of Tau by inhibiting the expression of wnt-1, crebbp, igfbp-1 and casq2 in wnt signaling pathway.