NLRP6基因敲除对小鼠生长繁育和实质器官的影响

Effect of NLRP6 gene knockout on growth and reproduction and parenchymal organs in mice

摘要:

目的 NOD样受体热蛋白结构域相关蛋白6（NOD-like receptor thermal protein domain associated protein 6，NLRP6）是新发现的寡聚化核苷酸结合结构域样受体家族成员，广泛表达于肠道、肝脏、肾脏、脾脏和肌肉等组织器官，在炎症、焦亡和自噬等多种生物过程发挥广泛的调节作用。近期研究表明，NLRP6在应激条件下对多种组织器官的疾病表型具有重要影响。然而，NLRP6对组织器官生长发育的影响尚未可知。方法 采用CRISPR/Cas9基因编辑技术构建NLRP6基因敲除小鼠，饲养并记录小鼠的生长和繁殖情况。将小鼠的脾、肝、心、肾、脑、四肢和后背皮肤等组织器官进行解剖和切片染色，以评估NLRP6基因敲除对实质器官的宏观发育影响和对组织结构的微观影响。结果 在自然状态下，NLRP6基因敲除缩短了雄鼠的性成熟期并使成年雄鼠的睾丸发生不可逆的破溃与萎缩。在四肢发育上，NLRP6基因敲除诱导成年雄鼠后肢横纹肌断裂，导致后肢明显萎缩。在脾脏发育上，NLRP6基因敲除不仅显著增加了雄鼠的脾脏体积（P<0.01）还诱导雄鼠脾脏出现炎性细胞浸润。在后背皮肤上，NLRP6基因敲除引发雄鼠后背皮肤出现明显的溃疡损伤、胶原纤维增生和炎性细胞浸润。结论 在自然生长发育条件下，NLRP6基因敲除选择性影响小鼠的生殖器发育与性成熟期、后肢肌肉发育、脾脏大小及其炎症免疫状态以及后背皮肤的结构完整性，而且这一作用具有明显的雄激素依赖性。

Abstract:

Objective NLRP6（NOD-like receptor thermal protein domain associated protein 6），a recently identified member of the nucleotide-binding oligomerization domain（NOD）-like receptors family，is abundantly expressed in the intestine，liver，kidney，spleen，muscle，and other tissues or organs，playing a regulatory role in various biological processes such as inflammation，pyroptosis，and autophagy. Recently，NLRP6 was reported to exert a significant impact on the disease phenotypes of various tissues and organs under stress conditions. However，the role of NLRP6 in the growth and development of tissues and organs in a natural state remains unclear.Methods A mouse model of NLRP6 gene knockout was established using CRISPR/Cas9 gene editing technique. The mice were raised and observed for their growth and reproduction. The spleen，liver，heart，kidney，brain，limbs，and dorsal skin of the mice were dissected，sectioned，and stained to evaluate the effect of NLRP6 gene knockout on the macroscopic development of parenchymal organs and microscopic tissue structure.Results In the natural state，NLRP6 knockout shortened the sexual maturity in male mice，resulting in irreversible ulceration and atrophy of the testicles in adult male mice. NLRP6 gene knockout led to the rupture of striated muscle in the hindlimbs in adult male mice，resulting in obvious atrophy of the hindlimbs. NLRP6 gene knockout not only significantly increased the volume of the spleen（P<0.01）but also induced inflammatory cell infiltration in male mice. NLRP6 gene knockout caused significant ulcerous damage，collagen fiber proliferation，and inflammatory cell infiltration in the dorsal skin in male mice.Conclusion In the natural condition of growth and development，NLRP6 gene knockout selectively affects genital development and sexual maturity，hindlimb muscle development，the size and immune response of the spleen，and the structural integrity of the dorsal skin in mice. This effect is significantly androgen-dependent.