NLRP3在介导小鼠皮肤伤口修复过程中的作用

Roles of NLRP3 in mediating skin wound repair in mice

本试验进行了小鼠肌成纤维细胞及骨髓源巨噬细胞的体外培养,随后使用野生型C57BL/6J小鼠或NLRP3基因敲除鼠耳皮肤肌成纤维细胞培养上清液(myofibroblast conditioned medium,MFbCM)处理M1和M2型巨噬细胞,分别检测巨噬细胞组织修复相关极化调节分子精氨酸酶(Arginase)的活性、NO的表达量、IL-10的分泌量以及几丁质酶3样分子(Ym1)的表达。同时,通过野生型C57BL/6J小鼠和NLRP3基因敲除小鼠背部创伤模型的建立,分析皮肤伤口愈合情况。通过免疫荧光检测小鼠皮肤创面组织中Ym1和Arginase表达量。结果显示,在MFbCM诱导的条件下,骨髓源巨噬细胞的精氨酸酶活性、NO的表达、IL-10的分泌和Ym1的表达均依赖于小鼠肌成纤维细胞NLRP3的表达;当肌成纤维细胞NLRP3表达缺失后,巨噬细胞上述组织修复相关极化调节因子均出现下调现象,进而可能对伤口愈合产生不利影响。此外,与之相一致的是,NLRP3基因敲除小鼠的伤口愈合速度相比于野生型C57BL/6J小鼠显著缓慢。在创面组织中,NLRP3缺失后Ym1和Arginase表达显著下调。结果表明,NLRP3对小鼠肌成纤维细胞培养上清液诱导的骨髓源巨噬细胞组织修复相关极化调节分子和伤口修复可能具有重要的影响。

Mice bone marrow-derived macrophages and myofibroblasts were cultured in vitro.Subsequent,M1 and M2 macrophages were treated with myofibroblast conditioned medium(MFbCM)of the wild-type C57 BL/6 J or NLRP3mice.The activity of arginase,the expression of NO,the secretion of IL-10 and the expression of Ym1,which are related to the polarization regulator molecules related to the tissue repair of macrophages,were detected respectively.At the same time,wild-type C57 BL/6 J and NLRP3mice were wounded on the back,and the wound healing was analyzed.The expression of Ym1 and Arginase in mouse skin wound tissue was detected by immunofluorescence.The results showed that under the conditions induced by MFbCM,the Arginase activity,NO expression,IL-10 secretion and Ym1 expression of bone marrow-derived macrophages all depended on the expression of NLRP3 in mice myofibroblasts.After knockout of NLRP3 receptor,the above-mentioned tissue repair-related polarization regulators of macrophages were down-regulated,which would adversely affect wound healing.In addition,the wound healing speed of NLRP3 knockout mice was significantly slower than that of the wild-type C57 BL/6 J mice.The expression of Ym1 and Arginase was significantly down-regulated after NLRP3 deletion in wound tissues.In summary,NLRP3 play key roles in the expression of tissue repair-related polarization regulators of bone marrow-derived macrophages induced by MFbCM and wound repair.