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目的采用全基因组表达谱芯片数据分析五子衍宗丸提高大鼠睾丸支持细胞抵抗热应激能力的作用机制。方法将原代培养的睾丸支持细胞分为五子衍宗丸组、正常组和模型组,分别给予五子衍宗丸溶液和等体积的PBS,培养24 h后,五子衍宗丸组和模型组给予43℃水浴15 min,正常组给予37℃水浴15 min处理。分别提取3组细胞总RNA,通过基因芯片技术进行数据分析。结果在支持细胞全基因组表达谱中,五子衍宗丸组与模型组比较共有1 203个差异表达基因,其中下调基因有697个,上调基因506个;正常组与模型组比较共有473个差异表达基因,其中上调基因168个,下调基因305个;与模型组相比,五子衍宗丸组和正常组共同下调的基因中RGD1305298、Trpt1、Dll3、Abi3和Ppp1r12b共5个基因表达差异明显,共同上调的基因中Dusp7表达差异明显,这些差异表达基因涉及到细胞周期、细胞凋亡、细胞代谢、细胞黏附等分子功能及Notch信号通路、细胞骨架调节、MAPK信号通路和紧密连接等通路。结论五子衍宗丸可能是通过维持支持细胞间紧密连接,抑制支持细胞凋亡,促进支持细胞分化成熟来提高睾丸支持细胞热应激的能力。
Objective To analyze the mechanism of Wuzi Yanzong Pills in improving the ability of testicular sertoli cells to resist heat stress by using genome-wide expression microarray data. Methods Primary cultured testicular sertoli cells were divided into five sub-Yanzong pill group, normal group and model group, and were given Wuzi Yanzong pill solution and an equal volume of PBS. After culturing for 24 hours, Wuzi Yanzong pill group and model group were given 43 ℃ water bath 15 min, normal group given 37 ℃ water bath for 15 min treatment. Three groups of total RNA were extracted and analyzed by gene chip technology. Results There were 1 203 differentially expressed genes in Wuziyanzong pill group compared with model group, of which 697 genes were down-regulated and 506 genes were up-regulated in the whole-genome expression profiles of supporting cells. There were 473 differentially expressed genes in normal group and model group , Of which 168 genes were up-regulated and 305 genes were down-regulated. Compared with the model group, the expression of RGD1305298, Trpt1, Dll3, Abi3 and Ppp1r12b in the genes down-regulated by Wuzi Yanzong pill and normal group were significantly different Gene Dusp7 expression differences significantly, these differentially expressed genes involved in cell cycle, apoptosis, cell metabolism, cell adhesion and other molecular functions and Notch signaling pathway, cytoskeleton regulation, MAPK signaling pathway and tight junctions and other pathways. Conclusion Wuzi Yanzong Pill may enhance the heat stress of testicular sertoli cells by maintaining the tight junctions between supporting cells, inhibiting the apoptosis of supporting cells and promoting the differentiation and maturation of supporting cells.