PURPOSE High myopia can lead to blindness. Genipin is a collagen cross-linking agent that may be used to treat myopia. However, the mechanism of action of genipin for the treatment of myopia is unclear. This study investigated the effect of genipin on the scleral expression of the miR-29 cluster, matrix metalloproteinase 2 (MMP2), and collagen alpha1 chain of type I (COL1A1) in a guinea pig model of myopia. METHODS The model of myopia was established by treating guinea pigs with a - 8D lens on both eyes for 21 days, and eyes with a refractive error of - 6D or greater were included. Quantitative reverse transcription-polymerase chain reaction (RT-PCR) and western blot were used to examine the mRNA and protein expression, respectively. A dual-luciferase assay was used to determine the direct targeting of the miR-29 cluster on the 3'-untranslated region (UTR) of the COL1A1 gene. RESULTS The scleral expression of miR-29a, miR-29b, and miR-29c as well as MMP2 was significantly increased, and the scleral expression of COL1A1 was significantly decreased in the myopia group. Genipin treatment reversed these effects in myopic eyes. The dual-luciferase assay showed that the luciferase activities were significantly decreased in human embryonic kidney (HEK) cells transfected with miR-29a and miR-29b, but not miR-29c, compared with those transfected with control miRNAs. CONCLUSIONS Genipin inhibits the scleral expression of the miR-29 cluster and MMP2 and promotes COL1A1 expression in a guinea pig model of myopia. Thus, genipin may promote COL1A1 expression by reducing the expression of the miR-29 cluster.

中文翻译：

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Genipin抑制豚鼠近视眼的巩膜表达miR-29和MMP2，并促进COL1A1表达。

目的高度近视会导致失明。Genipin是一种胶原蛋白交联剂，可用于治疗近视。但是，Genipin治疗近视的作用机理尚不清楚。这项研究调查了吉尼平对近视豚鼠模型中miR-29簇，基质金属蛋白酶2（MMP2）和I型胶原alpha1链（COL1A1）的巩膜表达的影响。方法通过用双眼-8D晶状体治疗豚鼠21天来建立近视模型，并包括屈光不正-6D或更大的眼睛。定量逆转录-聚合酶链反应（RT-PCR）和蛋白质印迹分别用于检测mRNA和蛋白质表达。使用双重荧光素酶测定法确定miR-29簇直接靶向3' -COL1A1基因的非翻译区（UTR）。结果在近视组中，miR-29a，miR-29b和miR-29c以及MMP2的巩膜表达显着增加，而COL1A1的巩膜表达显着下降。Genipin治疗可逆转近视眼中的这些作用。双重荧光素酶测定法显示，与转染了对照miRNA的人相比，用miR-29a和miR-29b而不是miR-29c转染的人胚肾（HEK）细胞中荧光素酶活性显着降低。结论Genipin可抑制近视豚鼠模型中miR-29簇和MMP2的巩膜表达，并促进COL1A1表达。因此，京尼平可通过减少miR-29簇的表达来促进COL1A1表达。近视组中miR-29b和miR-29c以及MMP2显着增加，而COL1A1的巩膜表达显着降低。Genipin治疗可逆转近视眼中的这些作用。双重荧光素酶测定法显示，与转染了对照miRNA的人相比，转染miR-29a和miR-29b而不是miR-29c的人胚肾（HEK）细胞的荧光素酶活性显着降低。结论Genipin抑制近视豚鼠模型中miR-29簇和MMP2的巩膜表达，并促进COL1A1表达。因此，genipin可能通过减少miR-29簇的表达来促进COL1A1表达。近视组中miR-29b和miR-29c以及MMP2显着增加，而COL1A1的巩膜表达显着降低。Genipin治疗可逆转近视眼中的这些作用。双重荧光素酶测定法显示，与转染了对照miRNA的人相比，用miR-29a和miR-29b而不是miR-29c转染的人胚肾（HEK）细胞中荧光素酶活性显着降低。结论Genipin可抑制近视豚鼠模型中miR-29簇和MMP2的巩膜表达，并促进COL1A1表达。因此，京尼平可通过减少miR-29簇的表达来促进COL1A1表达。Genipin治疗可逆转近视眼中的这些作用。双重荧光素酶测定法显示，与转染了对照miRNA的人相比，用miR-29a和miR-29b而不是miR-29c转染的人胚肾（HEK）细胞中荧光素酶活性显着降低。结论Genipin可抑制近视豚鼠模型中miR-29簇和MMP2的巩膜表达，并促进COL1A1表达。因此，genipin可能通过减少miR-29簇的表达来促进COL1A1表达。Genipin治疗可逆转近视眼中的这些作用。双重荧光素酶测定法显示，与转染了对照miRNA的人相比，用miR-29a和miR-29b而不是miR-29c转染的人胚肾（HEK）细胞中荧光素酶活性显着降低。结论Genipin可抑制近视豚鼠模型中miR-29簇和MMP2的巩膜表达，并促进COL1A1表达。因此，genipin可能通过减少miR-29簇的表达来促进COL1A1表达。结论Genipin可抑制近视豚鼠模型中miR-29簇和MMP2的巩膜表达，并促进COL1A1表达。因此，genipin可能通过减少miR-29簇的表达来促进COL1A1表达。结论Genipin可抑制近视豚鼠模型中miR-29簇和MMP2的巩膜表达，并促进COL1A1表达。因此，genipin可能通过减少miR-29簇的表达来促进COL1A1表达。