ORM1通过SPTLC1对干眼症鞘脂代谢的影响

ORM1 Effect on Sphingolipid Metabolism in Dry Eye via SPTLC1

目的研究人源血清类粘蛋白1(orosomucoid 1,ORM1)通过丝氨酸棕榈酰转移酶长链碱基亚基1(serine palmitoyltransferase long chain base subunit 1,SPTLC1)对干眼症鞘脂代谢的影响,为干眼症的发病机制提供新的研究方向。方法通过皮下注射氢溴酸东莨菪碱构建干眼症模型,通过晶状体注射腺相关病毒过表达ORM1。行泪液分泌量检测,杯状细胞检测,角膜荧光素染色,泪膜破裂时间检测,HE检测评估角膜结膜损伤,泪液质量,以及组织病理变化。通过生化试剂盒检测神经酰胺和鞘磷脂含量,通过qPCR和WB检测ORM1和SPTLC1的表达。结果(1)ORM1可增加干眼症模型的泪液分泌(P<0.0001);(2)ORM1可增加干眼症模型的眼表泪膜稳定性(P<0.001);(3)ORM1可改善干眼症模型的角膜上皮损伤(P<0.0001);(4)ORM1可增加干眼症模型的角膜组织中杯状细胞数量(P<0.001);(5)过表达ORM1后,角膜组织上皮层变厚,基底层细胞排列较为紧密,细胞层数变多,空泡减少;(6)ORM1可抑制干眼症模型的炎症基因表达(P<0.01);(7)ORM1可促进总神经酰胺和鞘磷脂含量(P<0.01);(8)ORM1可促进SPTLC1的mRNA和蛋白表达(P<0.001)。结论ORM1可通过调控SPTLC1参与调控干眼症鞘脂代谢,为探讨干眼症的发生发展机制和寻找有效且安全的治疗方案提供新的视角。

Objective To study the effect of ORM1 on sphingolipid metabolism in dry eye via SPTLC1 so as to provide a new research direction for the pathogenesis of dry eye.Methods By the subcutaneous injection of scopolamine hydrobromide(SCOP),a dry eye model was constructed,and adeno-associated virus overexpressing ORM1 was injected through the lens.Tear secretion assay,goblet number,corneal fluorescein staining,lacrimal gland rupture time,and HE assay were performed to assess the corneal conjunctival damage,tear quality,and histopathological changes.Ceramide and sphingomyelin content were detected by biochemical kits,and the expression of ORM1 and SPTLC1 was detected by qPCR and WB.Results(1)ORM1 increased tear secretion in the dry eye model(P<0.0001);(2)ORM1 increased the stability of the lacrimal gland on the ocular surface in the dry eye model(P<0.0001);(3)ORM1 improved the corneal epithelial damage in the dry eye model(P<0.0001);(4)ORM1 increased the number of goblet in the corneal tissue in the dry eye model(P<0.0001);(5)After the overexpression of ORM1,the epithelium of the corneal tissue became thicker and the cells of the basal lamina were more closely arranged.cell layers became more numerous and vacuoles were reduced;(6)ORM1 inhibited the inflammatory gene expression in a dry eye model(P<0.01);(7)ORM1 promoted the total ceramide and sphingomyelin content(P<0.01);(8)ORM1 promoted the rise of mRNA and protein expression of SPTLC1(P<0.001).Conclusion ORM1 can participate in the regulation of sphingolipid metabolism in dry eye disease through the regulation of SPTLC1,which provides new perspectives for exploring the mechanism of the development of dry eye disease and searching for effective and safe therapeutic options.