[1]王应璇,罗鑫,罗泓杨,等.甲状腺相关眼病患者眼外肌中胶原蛋白和HSP47的表达[J].眼科新进展,2023,43(11):873-876.[doi:10.13389/j.cnki.rao.2023.0175]
　WANG Yingxuan,LUO Xin,LUO Hongyang,et al.Expression of collagen and heat shock protein 47 in extraocular muscle tissues of patients with thyroid-associated ophthalmopathy[J].Recent Advances in Ophthalmology,2023,43(11):873-876.[doi:10.13389/j.cnki.rao.2023.0175]
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《眼科新进展》[ISSN:1003-5141/CN:41-1105/R]

卷:
43卷
期数:
2023年11期
页码:
873-876
栏目:
应用研究
出版日期:
2023-11-05

文章信息/Info

Title:
Expression of collagen and heat shock protein 47 in extraocular muscle tissues of patients with thyroid-associated ophthalmopathy
作者:
王应璇; 罗鑫; 罗泓杨; 刘太祥
563000　贵州省遵义市，遵义医科大学附属医院眼科临床技能实验室（王应璇，罗泓杨）；563000 贵州省遵义市，遵义医科大学附属医院眼科（罗鑫，刘太祥）
Author(s):
WANG Yingxuan1; LUO Xin2; LUO Hongyang1; LIU Taixiang2
1.Ophthalmic Clinical Skills Laboratory，Affiliated Hospital of Zunyi Medical University，Zunyi 563000，Guizhou Province，China
2.Department of Ophthalmology,Affiliated Hospital of Zunyi Medical University,Zunyi 563000,Guizhou Province,China
关键词:
甲状腺相关眼病; 纤维化; 眼外肌; 胶原 ; 热休克蛋白47
Keywords:
thyroid-associated ophthalmopathy;  fibrosis;  extraocular muscles;  collagen;  heat shock protein 47
分类号:
R777.5
DOI:
10.13389/j.cnki.rao.2023.0175
文献标志码:
A
摘要:
目的　探讨甲状腺相关眼病（TAO）患者眼外肌中胶原蛋白（Ⅰ、Ⅲ、Ⅴ型）和热休克蛋白47（HSP47）的表达特点。
方法　收集2019年5月至2019年9月在中山大学眼科中心明确诊断为静止期 TAO 患者4例右眼眼外肌（TAO 组），同期收集于我院行斜视矫正手术的共同性斜视患者4例右眼眼外肌（对照组）。对两组眼外肌分别进行HE染色、Masson染色分析样本中胶原蛋白（Ⅰ、Ⅲ、Ⅴ型）相对表达和纤维化的程度。实时荧光定量聚合酶链反应（qRT-PCR）检测两组眼外肌中胶原蛋白 （Ⅰ、Ⅲ、Ⅴ 型）和HSP47的mRNA 相对表达量；Western blot检测两组眼外肌中胶原蛋白（Ⅰ、Ⅲ、Ⅴ型）和HSP47表达情况。
结果　Masson染色结果显示，TAO组和对照组眼外肌中均显示大量胶原纤维，但对照组的眼外肌的形态模糊，胶原纤维分布较散乱；而TAO组眼外肌形态较清晰，胶原纤维排列较密集。HE染色结果显示，对照组眼外肌形态不清，肌纤维少且排列紊乱；TAO组眼外肌形态较清晰，肌纤维分布较集中，排列比较规则，横截面积大小不一。Western blot检测结果显示，TAO组和对照组眼外肌中胶原蛋白（Ⅰ、Ⅲ、Ⅴ型）和HSP47均有表达，但对照组眼外肌中胶原蛋白（Ⅰ、Ⅲ、Ⅴ型）相对表达量均高于 TAO 组（均为P<0.05）。Pearson相关分析结果显示，TAO组与对照组眼外肌中HSP47分别与两组眼外肌中的胶原蛋白（Ⅰ、Ⅲ、Ⅴ型）表达均呈正相关（均为P<0.01）。qRT-PCR检测结果显示，TAO组与对照组眼外肌中胶原蛋白（Ⅰ、Ⅲ、Ⅴ型）和HSP47的mRNA相对表达量差异均无统计学意义(均为P>0.05）。
结论　HSP47可在TAO患者眼外肌中表达且眼外肌的纤维化可能与HSP47蛋白表达有关。
Abstract:
Objective　To discover the expression characteristics of collagen (types Ⅰ, Ⅲ and Ⅴ) and heat shock protein 47 (HSP47) in extraocular muscle tissues of patients with thyroid-associated ophthalmopathy (TAO).
Methods　Extraocular muscle tissues of four right eyes were collected, respectively, from patients with resting TAO diagnosed at the Eye Center of Sun Yat-sen University between May 2019 and September 2019 (TAO group) and from patients who underwent strabismus correction surgery at our hospital for concomitant strabismus during the same period (control group). The hematoxylin-eosin (HE) staining and Masson staining were performed on the extraocular muscle tissues in the two groups to assess the collagen (types I, Ⅲ and V) expression and fibrosis. The relative mRNA expression levels of collagen (types I, Ⅲ and V) and HSP47 in the extraocular muscle tissues were measured by quantitative real-time polymerase chain reaction (qRT-PCR). The expression levels of collagen (types Ⅰ, Ⅲ and Ⅴ) and HSP47 in the extraocular muscle tissues were measured by Western blot.
Results　Masson staining revealed many collagen fibers in the extraocular muscle tissues in both the TAO and control groups. The morphology of extraocular muscle tissues in the control group was vague, and collagen fibers were distributed diffusely; however, in the TAO group, the morphology of extraocular muscle tissues was clear, and collagen fibers were distributed densely. HE staining showed that the extraocular muscle tissues in the control group were unclear in morphology, and muscle fibers were few and irregularly distributed; however, in the TAO group, the extraocular muscle tissues were clear in morphology, and muscle fibers varied in the cross-sectional area but were distributed densely and regularly. Western blot showed that collagen (types Ⅰ, Ⅲ and Ⅴ) and HSP47 were expressed in the extraocular muscle tissues in both the TAO and control groups, but the expression levels of collagen (types Ⅰ, Ⅲ and Ⅴ) in the control group were higher than those in the TAO group (all P<0.05). Pearson analysis showed that the expression of HSP47 was positively correlated with that of collagen (types Ⅰ, Ⅲ and Ⅴ) in both the TAO and control groups, respectively (all P<0.01). The qRT-PCR data showed that there was no statistically significant difference in the relative mRNA expression levels of collagen (types Ⅰ, Ⅲ and Ⅴ) and HSP47 between the two groups (all P>0.05).
Conclusion　HSP47 is expressed in the extraocular muscle tissues of TAO patients. The fibrosis of the extraocular muscle tissues may be related to the HSP47 protein expression.

参考文献/References:

［1］　DEBNAM J M,KOKA K,ESMAELI B.Extrathyroidal manifestations of thyroid disease:graves eye disease ［J］.Neuroimaging Clin N Am,2021,31(3):367-378.
［2］　MEN C J,KOSSLER A L,WESTER S T.Updates on the understanding and management of thyroid eye disease ［J］.Ther Adv Ophthalmol,2021,13:25158414211027760.
［3］　HU C,YANG J,QI Z,WU H,WANG B,ZOU F,et al.Heat shock proteins:biological functions,pathological roles,and therapeutic opportunities ［J］.MedComm 2022,3(3):e161.
［4］　ABRAHAM E T,OECAL S,MORGELIN M,SCHMID P W N,BUCHNER J,BAUMANN U,et al.Collagen’s primary structure determines collagen:HSP47 complex stoichiometry ［J］.J Biol Chem,2021,297(6):101169.
［5］　KOHLER A,MORGELIN M,GEBAUER J M,OCAL S,IMHOF T,KOCH M,et al.New specific HSP47 functions in collagen subfamily chaperoning ［J］.FASEB J,2020,34(9):12040-12052.
［6］　ITO S,NAGATA K.Roles of the endoplasmic reticulum-resident,collagen-specific molecular chaperone Hsp47 in vertebrate cells and human disease ［J］.J Biol Chem,2019,294(6):2133-2141.
［7］　BELLAYE P S,BURGY O,BONNIAUD P,KOLB M.HSP47:a potential target for fibrotic diseases and implications for therapy ［J］.Expert Opin Ther Targets,2021,25(1):49-62.
［8］　GUO Q,TIAN Q,TIAN X,LIU T.Effect of regulating the expression of HSP47 on collagen metabolism in scleral fibroblasts ［J］.Curr Eye Res,2021,46(3):408-416.
［9］　LUO H,LIU T,YANG H,YE H,LUO X.Expression of collagen (types Ⅰ,Ⅲ,and Ⅴ),HSP47,MMP-2,and TIMP-1 in retrobulbar adipose tissue of patients with thyroid-associated orbitopathy ［J］.J Ophthalmol,2020,4(21):4929634.
［10］　NABI T,RAFIQ N.Factors associated with severity of orbitopathy in patients with Graves’ disease ［J］.Taiwan J Ophthalmol,2020,10(3):197-202.
［11］　HATTON M P,RUBIN P A.The pathophysiology of thyroid-associated ophthalmopathy ［J］.Ophthalmol Clin North Am,2002,15(1):113-119.
［12］　LUN A Y,LE BRAS M,SCHARBARG E,MORCEL P,HADJADJ S,LEBRANCHU P,et al.Graves’ orbitopathy:diagnosis,epidemiology and treatments ［J］.Rev Med Interne,2022,43(4):242-251.
［13］　YUE Z,MOU P,CHEN S,TONG F,WEI R.A novel competing endogenous rna network associated with the pathogenesis of graves’ ophthalmopathy ［J］.Front Genet,2021,12:795546.
［14］　HU H,LIANG L,GE Q,JIANG X,FU Z,LIU C,et al.Correlation between peripheral T cell subsets and the activity of thyroid-associated ophthalmopathy ［J］.Int J Endocrinol,2022:3(8):2705650.
［15］　SMITH T J.Insights into the role of fibroblasts in human autoimmune diseases ［J］.Clin Exp Immunol,2005,141(3):388-397.
［16］　SZELOG J,SWANSON H,SNIEGOWSKI M C,LYON D B.Thyroid eye disease ［J］.Mo Med,2022,119(4):343-350.
［17］　VIEIRA I H,RODRIGUES D,PAIVA I.The mysterious universe of the tsh receptor ［J］.Front Endocrinol (Lausanne),2022,13:944715.
［18］　YOON J S,KIKKAWA D O.Thyroid eye disease:from pathogenesis to targeted therapies ［J］.Taiwan J Ophthalmol,2022,12(1):3-11.
［19］　WANG X,YE H,YANG S,SHA X,WANG X,ZHANG T,et al.Disulfiram exerts antifibrotic and anti-inflammatory therapeutic effects on perimysial orbital fibroblasts in Graves’ orbitopathy ［J］.Int J Mol Sci,2022,23(9)：5261.
［20］　CAI H,SASIKUMAR P,LITTLE G,BIHAN D,HAMAIA W,ZHOU A,et al.Identification of HSP47 binding site on native collagen and its implications for the development of HSP47 inhibitors ［J］.Biomolecules,2021,11(7)：983.
［21］　NAITOH M,HOSOKAWA N,KUBOTA H,TANAKA T,SHIRANE H,SAWADA M,et al.Upregulation of HSP47 and collagen type Ⅲ in the dermal fibrotic disease,keloid ［J］.Biochem Biophys Res Commun,2001,280(5):1316-1322.
［22］　郭立云,孙恒.间歇性外斜视患者眼外肌及其本体感受器的病理学改变 ［J］.眼科新进展,2012,32(8):729-732.
GUO L Y,SUN H.Pathological changes of extraocular muscles and proprioceptors in intermittent exotropia ［J］.Rec Adv Ophthalmol，2012，32(8):729-732.
［23］　KANEHIRA C,YAMAMOTO M,HIROUCHI H,ISHIZUKA S,SAKIYAMA K,HIGA K,et al.Tendinous annulus of Zinn for a common origin of the extraocular rectus muscles:a histological study of the orbital apex from donated elderly cadavers ［J］.Anat Sci Int,2022,97(4):369-379.
［24］　余新平,麦光焕,余焕云,陈静嫦,邓大明,林小铭.正常人眼外肌组织学初步研究 ［J］.眼科研究,2008,26(7):494-497.
YU X P,MAI G H,YU H Y,CHEN J E,DENG D M,LIN X M.Histiocytic study of extraocular muscles of normal human ［J］.Chin Ophthal Res,2008,26(7):494-497.
［25］　栾瑛,王淑霞,刘斌,周博研,周丽娜.共同性斜视眼外肌病理改变的研究 ［J］.国际眼科杂志,2008,8(1):94-96.
LUAN Y,WANG S X,LIU B,ZHOU B Y ,ZHOU L N.Pathological changes of extraocular muscles in concomitant strabismus ［J］.Int Eye Sci,2008,8(1):94-96.
［26］　胡依博,艾立坤,吴怡,顼晓琳,陈雪.甲状腺相关性眼病眶内组织的病理改变 ［J］.眼科,2014,23(2):130-134.
HU Y B,AI L K,WU Y,XU X L,CHEN X.Pathological changes of intraorbital tissues of thyroid associated ophthalmopathy ［J］.Ophthalmol CHN,2014,(2):130-134.
［27］　PAN H,HALPER J.Regulation of heat shock protein 47 and type I procollagen expression in avian tendon cells ［J］.Cell Tissue Res,2003,311(3):373-382.
［28］　侯洋,史建刚,袁文.缺氧状态对骨形态发生蛋白-2诱导分化体外复合富血小板血浆凝胶的骨髓间充质干细胞的影响［J］.脊柱外科杂志,2016,14 (2):101-105.
HOU Y,SHI J G,YUAN W.Effect of hypoxia on bone morphogenetic protein-2 induced differentiation of bone marrow stromal stem cells combined with platelet-rich plasma gel in vitro ［J］.J Spinal Surg,2016,14(2):101-105.
［29］　DENG G,DAI C,CHEN J,JI A,ZHAO J,ZHAI Y,et al.Porous Se@SiO(2) nanocomposites protect the femoral head from methylprednisolone-induced osteonecrosis ［J］.Int J Nanomedicine,2018,13:1809-1818.
［30］　GHELLIONI G V,DA SILVA L S,PIOVEZAN A P,MARTINS R O.Effect of methylprednisolone use on the rotator cuff in rats:biomechanical and histological study ［J］.Rev Bras Ortop,2015,50(3):260-265.
［31］　GU L,HAN Y,LIU W,MAO Y,LI J,WANG H.The expression of IL-2 and IL-4 in CD4(+) T cells from mouse lymph nodes and spleen during HSV-1-induced facial palsy ［J］.Inflamm Res,2014,63(2):117-125.
［32］　KHOSHSIRAT S,ABBASZADEH H A,AHRABI B,BAHRAMI M,ABDOLLAHI M A,KHORAMGAH M S,et al.Evaluation of the effect of BMSCs condition media and methylprednisolone in TGF-beta expression and functional recovery after an acute spinal cord injury ［J］.Bratisl Lek Listy,2018,119(11):684-691.
［33］　BAI X J,CHU X J,YANG T,TONG B D,QI X,PENG Y Y,et al.Changes of mRNA expression underlying orbital adipogenesis in thyroid-associated orbitopathy ［J］.Arch Med Sci,2022,18(6):1708-1715.

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备注/Memo

备注/Memo:
国家自然科学基金地区项目（编号：81860181，81960183）
更新日期/Last Update: 2023-11-05