肾阳虚与肾阴虚模型小鼠睾丸生殖相关差异蛋白的比较研究

Reproduction-related proteins differentially expressed in the testes of the mice with kidney-yang or kidney-yin deficiency

目的:比较氢化可的松诱导的肾阳虚和肾阴虚模型小鼠睾丸表达的差异蛋白,探讨两种肾虚导致雄性不育共同和特异的物质基础。方法:将30只昆明小鼠随机分为对照组、肾阳虚组和肾阴虚组,对照组:腹腔注射生理盐水0.2 ml,1次/d,连续7 d;肾阳虚组:腹腔注射氢化可的松25 mg/(kg·d),连续10 d;肾阴虚组:腹腔注射氢化可的松50 mg/(kg·d),连续7 d。HE染色分析睾丸组织病理变化,同位素标记相对和绝对定量(iTRAQ)技术及生物信息学方法比较两种肾虚小鼠睾丸表达的差异蛋白。结果:与对照组相比,Sod1为两种肾虚小鼠睾丸表达的生殖相关节点差异蛋白,且在肾阴虚组表达显著高于肾阳虚组(P<0.05);两种肾虚小鼠睾丸共同表达的生殖相关节点蛋白有5个,上调的为Rps28,下调的为Rpl11、Rplp2、Svs2和Svs3a(P<0.05)。结论:Sod1可能是区别两种肾虚不育小鼠的关键物质基础之一,Rps28、Rpl11、Rplp2、Svs2和Svs3a则可能是两种肾虚不育小鼠共同的重要的物质基础。

Objective: To compare the differentially expressed proteins in mice with kidney-yang deficiency and those with kidney-yin deficiency induced by hydrocortisone, and explore the similar and different material bases of male infertility caused by the two types of kidney deficiency. Methods: Thirty Kunming mice were equally randomized into a normal control, a kidney-yang deficiency and a kidney-yin deficiency group. The animals of the normal control group were injected intraperitoneally with normal saline at 0.2 ml qd for 7 days, while those of the latter two groups with hydrocortisone at 25 mg/kg/d for 10 days and 50 mg/kg/d for 7 days, respectively, for establishment of kidney-yang deficiency and kidney-yin deficiency models. Then the pathological changes in the testicular tissue of the mice were observed by HE staining and the differentially expressed proteins were compared among different groups using isobaric tags for relative and absolute quantitation(iTRAQ) and the bioinformatics method. Results: Sod1 was found to be a reproduction-related node protein differentially expressed in the testis tissues of the two types of kidney-deficiency mice, more highly expressed in the kidney-yin than in the kidney-yang deficiency group(P < 0.05). Five reproduction-associated node proteins were co-expressed in the testes of the two groups of kidney-deficiency mice, with significantly up-regulated expression of Rps28 and down-regulated expressions of Rpl11, Rplp2, Svs2 and Svs3 a(P < 0.01). Conclusion: Sod1 may be one of the key material bases for the differentiation of male infertility caused by kidney-yang deficiency from that induced by kidney-yin deficiency, while Rps28, Rpl11, Rplp2, Svs2 and Svs3 a may be the common material bases of male infertility caused by the two types of kidney deficiency.