五子衍宗丸提高大鼠睾丸支持细胞抵抗热应激能力的基因表达谱分析

Effects of Wuzi Yanzong Wan on improving heat stress resistance of sertoli cell in rats based on gene expression profile

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 study the mechanism of Wuzi Yanzong Wan （Five-Seed Progeny Pill） on improving heat stress resistance of sertoli cells in rats based on data from the whole genome microarray.Methods Sertoli cells in primary culture were randomly divided into Wuzi Yanzong Wan （WZYZW） group, normal group and model group.The cells in three groups were given WZYZW solution and equal volume of PBS respectively.After 24 hours, rats in WZYZW group and model group were placed in water bath at 43 ℃ for 15 minutes, while rats in normal group were put in water bath at 37 ℃ for 15 minutes, then total RNA of three groups were extracted respectively and data was analyzed with gene chip technology.Results Compared with the model group, there were totally 1,203 differently genes in WZYZW group based on Sertoli cell gene expression profiles, in which 697 were down regulated and 506 were up-regulated.There were totally 473 differently expressed genes in the normal group, including 168 up-regulated genes and 305 down regulated genes.Among these down-regulated gene expressions, RGD1305298, Trpt1, Dll3, Abi3 and Ppp1r12b were significant in the WZYZW group and normal group, while Dusp7 was significant among up-regulated gene expressions.The genes mentioned above were related to cell cycle, apoptosis, cell metabolism, cell adhesion and Notch signal pathway, regulation of actin cytoskeleton, MAPK signaling pathway and tight junction pathway, etc.Conclusion The mechanism of Wuzi Yanzong pill in improving heat stress resistance of sertoli cells may involve maintenance of tight junctions between sertoli cells, inhibition of sertoli cells apoptosis and promotion of cell differentiation and maturation.