[关键词]
[摘要]
目的 运用基因芯片技术筛选补阳还五汤延缓大鼠胫前肌失神经肌萎缩的靶基因,并对变化显著的PI3K基因进行验证。方法 建立大鼠腓总神经夹伤模型,将造模的20只SPF级SD大鼠随机分为补阳还五汤组与模型组,术后ig给药,18 d后芯片筛选差异表达基因。并运用RT-PCR与Western blotting法验证PI3K基因与蛋白的差异表达。结果 与模型组相比,补阳还五汤组基因表达谱有显著性差异,其中14条基因表达上调,10条基因表达下调,其中Angptl4与Pik3c2g基因表达变化较明显(上调5倍);同时基因验证表明:与模型组相比,补阳还五汤中、高剂量(生药12.96、25.92 g/kg)组PI3K mRNA与蛋白表达均显著增加(P<0.05、0.01)。结论 补阳还五汤对失神经肌萎缩的防治作用与其对多种相关基因的调控有关,其机制可能是通过促进能量合成与血管新生,保护神经,抑制细胞凋亡与胶原合成等方面来防治失神经肌萎缩的发生;补阳还五汤可能通过增加PI3K基因与蛋白表达,从而延缓肌萎缩的发生;PI3K/Akt信号转导通路的激活可能在补阳还五汤延缓失神经肌萎缩中发挥重要作用。
[Key word]
[Abstract]
Objective To screen the target gene of Buyang Huanwu Decoction (BHD) used for delaying denervated tibial muscle atrophy of rats by genechip technique and to verify the differentially expressed gene PI3K. Methods After 20 Sprague-Dawley (SD) rats were subjected to common peroneal nerve crush model of 5 mm injury, they were randomly divided into BHD and model groups, for daily ig administration of drugs after operation. The drug administration lasted for 18 d, the genechip analysis was performed to screen the differentially expressed gene. Then, RT-PCR and Western-blotting were used to detect the differential expression of PI3K and protein. Results Compared with the model group, there was a significant difference of gene expression profile in BHD group. Among them, the expression of 14 genes had up-regulation and 10 genes had down-regulation. In the differential expression genes, Angptl4 and Pik3c2g had the more obvious change (up-regulation for 5-fold). Simultaneously, the validation tests showed: Compared with the model group, the PI3K mRNA and protein expression in the BHD mid- and high-dose (crude drug 12.96 and 25.92 g/kg) groups increased significantly (P < 0.05, 0.01). Conclusion The preventive and therapeutic effects of BHD on denervated tibial muscle atrophy of rats are relevant with the regulation of many related genes. The mechanism may be as follows: promoting energy synthesis and blood neogenesis, neuroprotection, anti-apoptosis, and inhibition of collage synthesis; BHD could delay the denervated tibial muscle atrophy of rats by increasing the expression of PI3K and protein; The activation of PI3K/Akt signal transduction pathway probably play an important role in delaying the denervated muscle atropy by BHD.
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[基金项目]
国家自然科学基金(青年科学基金)资助项目(81302890);江苏省高校自然科学基金面上项目(13KJB360003);江苏省自然科学基金面上项目(BK2011816);高等学校博士学科点专项科研基金面上项目(新教师类)(20123237120004);南京中医药大学青年自然基金项目(12XZR09)