[1]王婷,李轩,王玉川,等.microRNA146a在干眼中的表达及作用[J].眼科新进展,2018,38(10):930-934.[doi:10.13389/j.cnki.rao.2018.0220]
　WANG Ting,LI Xuan,WANG Yu-Chuan,et al.The expression and role of microRNA146a in patients with dry eye[J].Recent Advances in Ophthalmology,2018,38(10):930-934.[doi:10.13389/j.cnki.rao.2018.0220]
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microRNA146a在干眼中的表达及作用/HTML
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《眼科新进展》[ISSN:1003-5141/CN:41-1105/R]

卷:
38卷
期数:
2018年10期
页码:
930-934
栏目:
实验研究
出版日期:
2018-10-05

文章信息/Info

Title:
The expression and role of microRNA146a in patients with dry eye
作者:
王婷; 李轩; 王玉川; 汤欣
300020　天津市，天津市眼科医院，天津市眼科研究所，天津医科大学眼科临床学院，天津市眼科学与视觉科学重点实验室
Author(s):
WANG Ting; LI Xuan; WANG Yu-Chuan; TANG Xin
Tianjin Eye Hospital and Institute，Clinical College of Ophthalmology of Tianjin Medical University，Tianjin Key Laboratory of Ophthalmology and Vision Science，Tianjin 300020，China
关键词:
干眼; 炎症; microRNA146a; 白细胞介素受体相关激酶1; 肿瘤坏死因子受体相关因子6
Keywords:
dry eye; inflammation; microRNA146a; interleukin-1 receptor-associated kinase 1; tumor necrosis factor receptor associated factor-6
分类号:
R777
DOI:
10.13389/j.cnki.rao.2018.0220
文献标志码:
A
摘要:
目的　研究microRNA146a在干眼小鼠模型中的表达及作用。方法　5周龄BALB/c雄性小鼠20只，左眼为正常对照组（20眼），右眼为干眼组（20眼）。用2 g·L-1苯扎氯铵诱导小鼠建立中重度干眼模型。造模完成3 d后，采用泪膜破裂时间（break-up time，BUT）和角膜荧光素钠染色（fluorescein staining，FLS）对小鼠进行眼表相关检查；随后随机选取10只小鼠，采用HE染色观察正常对照组和干眼组角膜和结膜的组织结构差异，结膜PAS染色后计数杯状细胞的量并对比；剩余10只小鼠分别提取角膜与结膜组织的总RNA，采用实时荧光定量PCR检测microRNA146a、白细胞介素（interleukin，IL）-1、IL-6、IL-8以及IL-1受体相关激酶1（IL-1 receptor-associated kinase 1，IRAK1）和肿瘤坏死因子受体相关因子6（tumor necrosis factor receptor associated factor-6，TRAF6）的相对表达量。结果　完成干眼造模3 d后，干眼组BUT为 （3.640±0.493）s，明显低于正常对照组的（10.645±0.583）s，差异有统计学意义（P<0.05）；干眼组角膜FLS评分为（14.650±0.860）分，明显高于正常对照组的（1.233±0.927）分，差异有统计学意义（P<0.05）。小鼠角膜和结膜组织HE染色结果显示：与正常对照组相比，干眼组角膜上皮欠平整，细胞数量增多，角膜基质层胶原纤维排列紊乱，成纤维细胞活化；干眼组小鼠结膜上皮细胞数量增多，排列欠规整，并伴有缺损。小鼠结膜PAS染色结果显示：干眼组结膜杯状细胞数量为（9.500±4.506）个，相较于正常对照组的［（29.667±8.756）个］明显降低，差异有统计学意义（P<0.05）。实时荧光定量PCR结果显示：干眼组角膜和结膜中microRNA146a、IL-1、IL-6、IL-8的相对表达量相较于正常对照组均升高，差异均有统计学意义（均为P<0.05）。相较于正常对照组，干眼组角膜中IRAK1的相对表达量降低，TRAF6的相对表达量升高，而干眼组结膜中IRAK1的相对表达量升高，TRAF6的相对表达量降低，差异均有统计学意义（均为P<0.05）。结论　干眼时机体可通过增加microRNA146a的表达，以及其对炎症中靶点的调控作用，从而形成对干眼炎症反应的负反馈调节机制。
Abstract:
Objective　To investigate the expression and role of microRNA146a in patients with dry eye.Methods　The left eyes of twenty five-week-old BALB/c mice were categorized into normal control group （20 eyes）；the right eyes of these mice were categorized into dry eye group （20 eyes）.Right eyes of these mice were topically treated with 2 g·L-1 benzalkonium chloride for fourteen days to establish moderate or severe dry eye mouse model.Three days after establishment，dry eye-related clinical tests were performed，including BUT （tear film break-up time） and FLS （fluorescein staining） for the cornea.Then ten mice were chosen from these twenty mice randomly，HE （hematoxylin eosin staining） and PAS （periodic acid-Schiff staining） were performed to observe the differences of cornea and conjunctiva between the normal control group and the dry eye group.The number of goblet cells in conjunctiva were counted after PAS （periodic acid-schiff staining），and the difference of them in two groups was compared.As for the remaining ten mice，total RNA of the cornea and conjunctiva were extracted，and the expression of microRNA146a，interleukin-1 （IL-1），interleukin-6 （IL-6），interleukin-8 （IL-8），interleukin-1 receptor associated kinase 1 （IRAK1） and tumor necrosis factor receptor associated factor 6 （TRAF6） were detected through real-time fluorescent quantitative PCR.Results　Three days after establishment of dry eye mouse model，BUT in the dry eye group was （3.640±0.493） s，which was significantly declined in comparison with in the normal control group［ （10.645±0.583）s］ （P<0.05）.FLS scores in dry eye group ［（14.650±0.860） scores］ was significantly higher than that in the normal control group ［（1.233±0.927） scores］ （P<0.05）.Compared to the normal control group，HE of the dry eye group showed that the corneal epithelial cell layer was uneven，and the number of cells increased.Collagenous fibers in the corneal stromal layer were irregular and accompanied with fibroblast activation.The number of conjunctival epithelial cells was also increased and uneven and accompanied with the defect.PAS showed that the number of goblet cells in dry eye group ［（9.500±4.506） cells］ was significantly lower than that in the normal control group ［（29.667±8.756） cells］ （P<0.05）.Real-time fluorescent quantitative PCR showed that the expression levels of microRNA146a，IL-1，IL-6，IL-8 were increased in the cornea and conjunctiva in dry eye group in comparison with the normal control group （P<0.05）.Compared to the cornea in the normal control group，the expression of IRAK1 was down-regulated，and TRAF6 was up-regulated in the cornea of the dry eye group （P<0.05）.However，compared to the conjunctiva in the normal control group，the expression of IRAK1 was up-regulated，and TRAF6 was down-regulated in the conjunctiva of the dry eye group （P<0.05）.Conclusion　In patients with dry eye，owning to the onset of inflammation，the organism can build negative feedback regulation which is dependent on the up-regulation expression of microRNA146a and the effect of microRNA146a on the target genes in inflammation.

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备注/Memo

备注/Memo:
国家自然科学基金资助项目（编号：81170828、81670837）；天津市应用基础与前沿技术研究计划项目 （编号：15JCZDJC35300）；天津市卫生行业重点攻关项目（编号：14KGl33）
更新日期/Last Update: 2018-09-28