JP5685751B2 - Subconjunctival fibroblast proliferation inhibitor and method for inhibiting subconjunctival fibroblast proliferation - Google Patents
Subconjunctival fibroblast proliferation inhibitor and method for inhibiting subconjunctival fibroblast proliferation Download PDFInfo
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Description
本発明は結膜下線維芽細胞増殖抑制剤および結膜下線維芽細胞増殖抑制方法に係り、特に安全かつ効果的に、眼科手術後における結膜癒着や結膜下組織瘢痕形成を抑制することの可能な、結膜下線維芽細胞増殖抑制剤および結膜下線維芽細胞増殖抑制方法に関する。
The present invention relates to a subconjunctival fibroblast growth inhibitor and the subconjunctival fibroblast growth inhibition method, particularly safe and effective, it possible to suppress the conjunctiva adhesion and subconjunctival tissue scarring after ophthalmic surgery, subconjunctival fibroblast growth inhibitor and to subconjunctival fibroblast growth suppression method.
眼科領域の手術では、手術後に、結膜下の線維芽細胞増殖による瘢痕形成によって、術後成績が左右されることがある。緑内障に対して行う線維柱帯切除は、その代表的なものである。従来、線維柱帯切除手術術後の結膜下組織瘢痕形成を予防する目的で代謝阻害薬マイトマイシンＣが術中に使用されている。 In surgery in the ophthalmic field, postoperative results may be affected by scar formation due to subconjunctival fibroblast proliferation after surgery. Trabeculectomy for glaucoma is a typical example. Conventionally, the metabolic inhibitor mitomycin C has been used intraoperatively for the purpose of preventing subconjunctival tissue scar formation after trabeculectomy.
一方、ドライアイに対する有効性を有するものとしてトレハロース点眼液（１００ｍＭ，２００ｍＭ）が既に報告されているなど、トレハロースを眼科領域で応用する技術的提案がなされている（後掲特許文献）。 On the other hand, trehalose ophthalmic solution (100 mM, 200 mM) has already been reported as having an effect on dry eye, and technical proposals for applying trehalose in the ophthalmic field have been made (patent document).
このうち特許文献１は、シェーグレン症候群における眼の臨床症状の治療・予防剤として、トレハロースを有効成分として含有する眼科用医薬組成物を技術開示するものである。また特許文献２は、角膜保護作用を有する安全な眼科用医薬組成物として、トレハロースを用いる医薬組成物を開示するものである。 Among them, Patent Document 1 discloses a technical ophthalmic pharmaceutical composition containing trehalose as an active ingredient as a therapeutic / preventive agent for clinical symptoms of eyes in Sjogren's syndrome. Patent Document 2 discloses a pharmaceutical composition using trehalose as a safe ophthalmic pharmaceutical composition having a cornea protecting action.
なお、トレハロースは、２つのα−グルコース（ブドウ糖）が１，１−グリコシド結合してできた二糖類であり、以下の化学的特徴を有している。
ＩＵＰＡＣ名：２−（ヒドロキシメチル）−６−［３，４，５−トリヒドロキシ−６−（ヒドロキシメチル）テトラヒドロピラン−２−イル］オキシ−テトラヒドロピラン−３，４，５−トリオール
別名：α−Ｄ−グルコピラノシル−α−Ｄ−グルコピラノシド（α，α‐トレハロース）
分子式：Ｃ１２Ｈ２２Ｏ１１（無水物）、Ｃ１２Ｈ２２Ｏ１１・２Ｈ２Ｏ（二水和物）
分子量：３４２．２９（無水物）、３７８．３３（二水和物） ｇ／ｍｏｌ
Trehalose is a disaccharide formed by linking two α-glucoses (glucose) with a 1,1-glycoside, and has the following chemical characteristics.
IUPAC name: 2- (hydroxymethyl) -6- [3,4,5-trihydroxy-6- (hydroxymethyl) tetrahydropyran-2-yl] oxy-tetrahydropyran-3,4,5-triol -D-glucopyranosyl-α-D-glucopyranoside (α, α-trehalose)
Molecular formula: C 12 H 22 O 11 (anhydride), C 12 H 22 O 11 · 2H 2 O (dihydrate)
Molecular weight: 342.29 (anhydride), 378.33 (dihydrate) g / mol
さて上述のように従来、線維柱帯切除手術術後の結膜下組織瘢痕形成予防の目的で、代謝阻害薬マイトマイシンＣが術中に使用されているが、この薬剤は正常細胞に対する増殖抑制も強いため、瘢痕形成抑制のみならず正常結膜上皮細胞にも傷害を及ぼすことがあり、術後晩期合併症が頻発し、臨床上問題となっている。 As described above, conventionally, the metabolic inhibitor mitomycin C has been used during the operation for the purpose of preventing subconjunctival tissue scar formation after trabeculectomy, but this agent also has a strong inhibition of proliferation against normal cells. In addition to the suppression of scar formation, normal conjunctival epithelial cells may be damaged, and post-operative late complications frequently occur, causing clinical problems.
また従来、上述のようにトレハロースが眼科領域において用いられているとはいえ、線維柱帯切除手術術後の結膜下組織瘢痕形成との関連においては、これまで研究を行った者もなく、また何らの報告もなされていない状況である。 In addition, although trehalose has been used in the ophthalmic field as described above, no research has been conducted so far in relation to subconjunctival tissue scar formation after trabeculectomy. There are no reports.
そこで本発明が解決しようとする課題は、かかる従来技術の問題点を踏まえ、安全かつ効果的に、眼科手術後における結膜癒着や結膜下組織瘢痕形成を抑制することの可能な、結膜下線維芽細胞増殖抑制剤および結膜下線維芽細胞増殖抑制方法を提供することである。
Therefore object of the present invention is to provide, such conventional light of problems of the art, safe and effective, capable of suppressing the conjunctiva adhesion and subconjunctival tissue scarring after ophthalmic surgery, subconjunctival fibroblasts cytostatic agents and to provide a subconjunctival fibroblast growth suppression method.
さて本願発明者らは、食品や化粧品で頻用されているトレハロースの瘢痕抑制作用に注目し、トレハロース点眼薬を緑内障に対する線維柱帯切除手術術後などの結膜下組織瘢痕形成抑制の目的で使用可能か否かの研究を行ってきた。
参考文献
Matsuo T, Tsuchida Y, Morimoto N. Trehalose eye drops in the treatment of dry eye syndrome. Ophthalmology 109 (11), 2024-2029, 2002
Matsuo T. Trehalose versus hyaluronan or cellulose in eyedrops for the treatment of dry eye. Jpn J Ophthalmol 48 (4), 321-327, 2004
The inventors of the present application pay attention to the scar suppression effect of trehalose frequently used in foods and cosmetics, and can use trehalose eye drops for the purpose of suppressing subconjunctival tissue scar formation after trabeculectomy for glaucoma. I have been studying whether or not.
References
Matsuo T, Tsuchida Y, Morimoto N. Trehalose eye drops in the treatment of dry eye syndrome. Ophthalmology 109 (11), 2024-2029, 2002
Matsuo T. Trehalose versus hyaluronan or cellulose in eyedrops for the treatment of dry eye.Jpn J Ophthalmol 48 (4), 321-327, 2004
上述の通り、トレハロース点眼液のドライアイその他に対する有効性が報告されてはいるが、結膜下組織における線維芽細胞増殖抑制効果に対してトレハロースが有効であることについては、これまで研究を行った者もなく、何らの報告もなされていない。 As described above, the effectiveness of trehalose ophthalmic solution against dry eye and others has been reported, but we have studied so far that trehalose is effective in suppressing fibroblast proliferation in subconjunctival tissue. There was no report and no reports were made.
本願発明者は、かかる課題について検討した結果、トレハロースが結膜下線維芽細胞の抑制作用および創傷治癒遅延作用を備えていることを見出し、本発明の完成に至った。すなわち、上記課題を解決するための手段として本願で特許請求される発明、もしくは少なくとも開示される発明は、以下の通りである。 As a result of studying such problems, the inventor of the present application has found that trehalose has an inhibitory effect on subconjunctival fibroblasts and an effect of delaying wound healing, thereby completing the present invention. That is, the invention claimed in the present application, or at least the disclosed invention, as means for solving the above-described problems is as follows.
〔１〕 正常結膜上皮細胞に対して何ら傷害を及ぼすことなく眼科手術後の結膜癒着や結膜下組織瘢痕形成を抑制するための、有効成分としてトレハロースのみを用いた点眼薬形態の結膜下線維芽細胞増殖抑制剤。
〔２〕 正常結膜上皮細胞に対して何ら傷害を及ぼすことなく眼科手術後の結膜癒着や結膜下組織瘢痕形成を抑制するための、有効成分としてトレハロースのみを用いた点眼薬形態の結膜癒着抑制剤。
〔３〕 正常結膜上皮細胞に対して何ら傷害を及ぼすことなく眼科手術後の結膜癒着や結膜下組織瘢痕形成を抑制するための、有効成分としてトレハロースのみを用いた点眼薬形態の結膜下組織瘢痕形成抑制剤。
〔４〕 正常結膜上皮細胞に対して何ら傷害を及ぼすことなく眼科手術後の結膜癒着や結膜下組織瘢痕形成を抑制するための、有効成分としてトレハロースのみを用いた、結膜下線維芽細胞増殖抑制用トレハロース点眼薬。
〔５〕 トレハロース濃度が５％以上であることを特徴とする、〔４〕に記載のトレハロース点眼薬。
[1] Subconjunctival fibroblasts in the form of eye drops using only trehalose as an active ingredient to suppress conjunctival adhesion and subconjunctival tissue scar formation after ophthalmic surgery without causing any damage to normal conjunctival epithelial cells Cell growth inhibitor.
[2] A conjunctival adhesion inhibitor in the form of eye drops using only trehalose as an active ingredient to suppress conjunctival adhesion and subconjunctival tissue scar formation after ophthalmic surgery without causing any damage to normal conjunctival epithelial cells .
[3] Subconjunctival tissue scar in the form of eye drops using only trehalose as an active ingredient to suppress conjunctival adhesion and subconjunctival tissue scar formation after ophthalmic surgery without causing any damage to normal conjunctival epithelial cells Formation inhibitor.
[4] Inhibition of subconjunctival fibroblast proliferation using only trehalose as an active ingredient to suppress conjunctival adhesion and subconjunctival tissue scar formation after ophthalmic surgery without causing any damage to normal conjunctival epithelial cells Trehalose eye drops.
[5] The trehalose ophthalmic solution according to [4], wherein the trehalose concentration is 5% or more.
〔６〕 眼科手術の術中に供給し、また術後においては日単位で１日以上の期間に亘り供給することにより、術後晩期合併症の頻発を解消できることを特徴とする、〔４〕 または〔５〕 に記載のトレハロース点眼薬。
〔７〕 人間以外の動物の眼科手術において、術中に手術対象に対して〔４〕ないし〔６〕のいずれかに記載のトレハロース点眼薬を供給し、術後においては日単位で１日以上の期間に亘り手術対象に対して該トレハロース点眼薬を供給する、結膜下線維芽細胞増殖抑制方法。
[6] The frequent occurrence of postoperative late complications can be eliminated by supplying it during ophthalmic surgery and supplying it for a period of 1 day or more after surgery. [4] or [5] The trehalose eye drop according to [5].
[7] In ophthalmic surgery for animals other than humans, the trehalose eye drop according to any one of [4] to [6] is supplied to the surgical subject during the surgery, and after surgery, the trehalose eye drops for 1 day or more per day A method for suppressing subconjunctival fibroblast proliferation, which supplies the trehalose eye drop to a surgical subject over a period of time.
本発明の結膜下線維芽細胞増殖抑制剤および結膜下線維芽細胞増殖抑制方法は上述のように構成されるため、これによれば、安全かつ効果的に、眼科手術後における結膜癒着や結膜下組織瘢痕形成を抑制することができる。
For subconjunctival fibroblasts cytostatic agents and methods subconjunctival fibroblast growth inhibition of the present invention is constructed as described above, according to this, safely and effectively, conjunctival adhesion and subconjunctival after ophthalmic surgery Tissue scar formation can be suppressed.
つまり本発明完成の過程において発明者は、トレハロースが、結膜切除および線維柱帯切除手術後に起こる結膜下組織の線維芽細胞増殖を選択的に抑制し、その結果、結膜下組織の瘢痕形成を特異的に抑制する作用を明らかにしたものである。かかる作用は結膜上皮細胞にはみられない。すなわち、正常細胞に対する増殖抑制が発生することはない。 In other words, in the process of completing the present invention, the inventor found that trehalose selectively suppressed fibroblast proliferation in the subconjunctival tissue that occurred after conjunctival and trabeculectomy surgery, resulting in specific scar formation in the subconjunctival tissue. This clearly shows the inhibitory action. Such an effect is not seen in conjunctival epithelial cells. That is, growth suppression for normal cells does not occur.
つまり本発明によれば、眼最表面の結膜上皮の創傷治癒機転には何ら影響を与えることなく（正常結膜上皮細胞に対しては何ら傷害を及ぼすことなく）、結膜下組織の瘢痕形成や結膜癒着のみを抑制することができる。したがって、術後晩期合併症が頻発するという従来技術の欠点も、問題なく解消される。 In other words, according to the present invention, there is no effect on the wound healing mechanism of the conjunctival epithelium on the outermost surface of the eye (without any damage to normal conjunctival epithelial cells), and scar formation and conjunctiva in the subconjunctival tissue. Only adhesions can be suppressed. Therefore, the disadvantage of the prior art that postoperative late complications occur frequently can be solved without any problems.
本発明について、さらに詳細に説明する。
本発明は、トレハロースを用いた結膜下線維芽細胞増殖抑制剤、結膜癒着抑制剤、あるいは結膜下組織瘢痕形成抑制剤である。その限りでは剤形が限定されるものではないが、点眼薬の形態が主であるといえる。したがって、かかる剤形をとる本発明は、結膜下線維芽細胞増殖抑制のためのトレハロース点眼薬であるといえる。
The present invention will be described in further detail.
The present invention, subconjunctival fibroblast growth inhibitor using trehalose, conjunctival adhesion inhibitor, or a subconjunctival tissue scarring inhibitor. To that extent, the dosage form is not limited, but it can be said that the form of eye drops is the main. Accordingly, the present invention takes such a dosage form can be said to be trehalose eye drops for subconjunctival fibroblasts growth inhibition.
実施例に後述するように、本発明トレハロース点眼薬は、特にトレハロース濃度が５％以上であるように調製されたものを用いることが、その作用効果を充分に発揮する上で望ましい。 As will be described later in Examples, it is desirable that the trehalose ophthalmic solution of the present invention be prepared so that the trehalose concentration is 5% or more, in order to sufficiently exhibit its effects.
そして特に、本発明トレハロース点眼薬は、緑内障またはその他の眼科手術の術後に発生する結膜癒着を抑制するため、または結膜下組織瘢痕形成を抑制するために用いて、優れた抑制効果を得ることができる。 In particular, the trehalose ophthalmic solution of the present invention is used for suppressing conjunctival adhesion occurring after glaucoma or other ophthalmic surgery, or for suppressing subconjunctival tissue scar formation, to obtain an excellent suppressing effect. Can do.
主な使用方法を述べる。
眼科手術において、術中に、手術対象と連続する空間に対して本発明トレハロース点眼薬を供給する。すなわち、該空間を介して手術対象に対してトレハロース点眼薬は供給されて、結膜癒着抑制作用、あるいは結膜下組織瘢痕形成抑制作用がなされる。このとき、かかるトレハロース点眼薬の増殖抑制作用は、結膜上皮細胞に対しては及ばない。すなわち、眼最表面の結膜上皮の創傷治癒機転には何ら影響を与えることなく、結膜癒着や結膜下組織の瘢痕形成のみを抑制することができる。
The main usage is described.
In ophthalmic surgery, the trehalose eye drop of the present invention is supplied to a space continuous with a surgical object during the operation. That is, trehalose eye drops are supplied to the surgical subject through the space, and a conjunctival adhesion suppressing action or a subconjunctival tissue scar forming suppressing action is performed. At this time, the growth inhibitory action of such trehalose eye drops does not reach conjunctival epithelial cells. That is, only the conjunctival adhesion and the scar formation of the subconjunctival tissue can be suppressed without affecting the wound healing mechanism of the conjunctival epithelium on the outermost surface of the eye.
術後においては、日単位で１日以上の期間に亘り、手術対象と連続する空間に対してトレハロース点眼薬を供給する。すなわち、該空間を介して手術対象に対してトレハロース点眼薬は供給されて、さらに結膜癒着抑制作用、あるいは結膜下組織瘢痕形成抑制作用がなされる。このとき、かかるトレハロース点眼薬の増殖抑制作用は、結膜上皮細胞に対しては及ばない。すなわち、眼最表面の結膜上皮の創傷治癒機転には何ら影響を与えることなく、結膜癒着や結膜下組織の瘢痕形成のみを抑制することができる。 After the operation, trehalose ophthalmic solution is supplied to the space continuous with the operation target for a period of one day or more in daily units. That is, trehalose eye drops are supplied to the surgical subject through the space, and further, a conjunctival adhesion inhibitory action or a subconjunctival tissue scar formation inhibitory action is performed. At this time, the growth inhibitory action of such trehalose eye drops does not reach conjunctival epithelial cells. That is, only the conjunctival adhesion and the scar formation of the subconjunctival tissue can be suppressed without affecting the wound healing mechanism of the conjunctival epithelium on the outermost surface of the eye.
なお本発明は、主として人間の眼科手術と術後における利用を想定したものであるが、人間以外の動物、たとえば犬・猫といった愛玩動物の眼科手術においても同様に、術中に手術対象に対してトレハロース点眼薬を供給し、術後においては日単位で１日以上の期間に亘り手術対象に対してトレハロース点眼薬を供給するという方法で、使用することは可能である。 The present invention is mainly intended for human ophthalmic surgery and postoperative use, but in the case of ophthalmic surgery for non-human animals, for example, pets such as dogs and cats, It can be used by supplying trehalose eye drops and supplying trehalose eye drops to the surgical subject over a period of one day or more after surgery.
以下、本発明を実施例によってさらに詳細に説明するが、本発明がかかる実施例に限定されるものではない。なお実施例は、本願発明者による研究過程の概要を記すものである。
＜１ 試験方法＞
１） 試験管内（ｉｎ ｖｉｔｒｏ）実験
Ｉｎ ｖｉｔｒｏにおいて、正常ヒト皮膚由来線維芽細胞およびヒト表皮角化細胞の培養を行い、トレハロース各濃度下にてその増殖能につき対照群と比較した（なお、濃度は重量％。以下も同様である。）。
EXAMPLES Hereinafter, although an Example demonstrates this invention further in detail, this invention is not limited to this Example. In addition, an Example describes the outline | summary of the research process by this inventor.
<1 test method>
1) In vitro experiment In vitro, normal human skin-derived fibroblasts and human epidermal keratinocytes were cultured, and compared with the control group for their ability to proliferate under each concentration of trehalose. % By weight, and so on.)
２） 動物 （ｉｎ ｖｉｖｏ） 実験
白色家兎に対し結膜単純切除後、５％トレハロース点眼（１４７ｍＭ）を連日行い、術後結膜癒着の程度を評価した。別の白色家兎群に線維柱帯切除を行い、術中トレハロースにて洗眼し、かつ術後連日点眼を行いながら、経時的に眼圧を測定後に眼球摘出し評価した。組織評価として、Ｍａｓｓｏｎｎ−ｔｒｉｃｈｒｏｍｅ染色および免疫染色を行い、対照群と比較検討した。なお免疫染色は、抗ｖｉｍｅｎｔｉｎ抗体による免疫染色、抗α−ＳＭＡ抗体（α−ｓｍｏｏｔｈ ｍｕｓｃｌｅ ａｃｔｉｎ の特異モノクローナル抗体）による免疫染色、およびこれら二種の免疫染色を重複して行う二重免疫染色を行った。
2) Animals (in vivo) After simple conjunctival resection of white rabbits, 5% trehalose instillation (147 mM) was performed every day to evaluate the degree of postoperative conjunctival adhesion. A trabeculectomy was performed on another white rabbit group, the eyes were washed with trehalose during the operation, and the eyeballs were measured over time while performing eye drops every day after the operation. As tissue evaluation, Massonn-trichrome staining and immunostaining were performed and compared with the control group. The immunostaining is performed by immunostaining with an anti-vimentin antibody, immunostaining with an anti-α-SMA antibody (specific monoclonal antibody of α-smooth muscle actin), and double immunostaining that duplicates these two types of immunostaining. It was.
＜２ 結果と考察＞
図１は、各種濃度のトレハロース存在下での培養線維芽細胞の形態を示す顕微鏡写真図である。試験したトレハロース濃度は、０％（Ｃｏｎｔｒｏｌ）、０．６２５％、１．２５％、２．５％、５．０％、１０．０％、２０．０％ である。図示されるように、５％以上の濃度のトレハロース存在下では、濃度依存性に（濃度が高くなるほど）線維芽細胞の増殖を抑制している。ただし濃度を高くしていっても、上皮細胞の増殖にはまったく影響を与えなかった。すなわちｉｎ ｖｉｔｒｏの実験結果では、線維芽細胞はトレハロース５％以上で濃度依存性に増殖が抑制される結果であった。
<2 Results and discussion>
FIG. 1 is a photomicrograph showing the morphology of cultured fibroblasts in the presence of various concentrations of trehalose. The trehalose concentrations tested are 0% (Control), 0.625%, 1.25%, 2.5%, 5.0%, 10.0%, 20.0%. As shown in the figure, in the presence of trehalose at a concentration of 5% or more, the proliferation of fibroblasts is suppressed in a concentration-dependent manner (as the concentration increases). However, increasing the concentration had no effect on the proliferation of epithelial cells. That is, in the in vitro experimental results, the growth of fibroblasts was suppressed in a concentration-dependent manner when trehalose was 5% or more.
Ｉｎ ｖｉｖｏの実験結果を説明する。
図２は、白色家兎における結膜単純切除術後の組織所見（Ｍａｓｓｏｎ−ｔｒｉｃｈｒｏｍｅ染色）を示す顕微鏡写真図である。図示されるように５％トレハロース点眼群（上段）では、明らかな癒着が認められず、結膜組織の瘢痕形成が抑制されていることが認められた。しかし、結膜上皮細胞による上皮創傷治癒機転には、まったく影響を与えなかった。すなわちｉｎ ｖｉｖｏの実験では、結膜単純切除後では、瘢痕状組織が認められた対照群（Ｃｏｎｔｒｏｌ 下段）に比較して明らかな結膜の癒着抑制が認められた。
In vivo experimental results will be described.
FIG. 2 is a photomicrograph showing the histological findings (Masson-trichrome staining) after conjunctival simple resection in white rabbits. As shown in the figure, in the 5% trehalose ophthalmic group (upper), no obvious adhesion was observed, and it was confirmed that the scar formation of the conjunctival tissue was suppressed. However, it did not affect the mechanism of epithelial wound healing by conjunctival epithelial cells. That is, in an in vivo experiment, after conjunctival simple excision, clear suppression of conjunctival adhesion was recognized as compared with the control group in which scar-like tissue was observed (Control lower stage).
図３は、白色家兎線維柱帯切除術後の眼圧経過を示すグラフである。図示されるように、５％または１０％トレハロース使用群では、不使用群（Ｃｏｎｔｒｏｌ）と比較して眼圧の下降効果が維持されている結果であった。またその眼圧下降効果は、従来用いられているマイトマイシンＣ（ＭＭＣ）ともさほど差異のないものだった。すなわち線維柱帯切除モデルでは、観察期間内において、手術効果を裏付ける眼圧の下降を確認することができた。 FIG. 3 is a graph showing the course of intraocular pressure after white rabbit trabeculectomy. As shown in the figure, the 5% or 10% trehalose use group maintained the lowering effect of intraocular pressure compared to the non-use group (Control). Further, the intraocular pressure lowering effect was not so different from the conventionally used mitomycin C (MMC). That is, in the trabeculectomy model, it was possible to confirm a decrease in intraocular pressure that supported the surgical effect during the observation period.
図４は、白色家兎線維柱帯切除術後９０日間トレハロース点眼後の組織所見（Ｍａｓｓｏｎ−ｔｒｉｃｈｒｏｍｅ染色）を示す顕微鏡写真図である。図では、左上が５％トレハロース点眼、右上が１０％トレハロース点眼、中央左が対照、下が０．０４％ＭＭＣ による各組織所見を示す。図示されるように、各トレハロース点眼群ではいずれも、結膜−ｆｌａｐ−ｂｌｅｂ間にスペースが認められた（図中の矢印の示す位置）。すなわち、トレハロース使用群で明らかに線維形成が抑制されており、５％または１０％トレハロース点眼を９０日間継続した後では、線維芽細胞が充分に抑制されていることが認められた。 FIG. 4 is a photomicrograph showing tissue findings (masson-trichrome staining) after instillation of trehalose for 90 days after white rabbit trabeculectomy. In the figure, the upper left is 5% trehalose instillation, the upper right is 10% trehalose instillation, the center left is the control, and the lower is each tissue finding by 0.04% MMC. As shown in the figure, in each trehalose ophthalmic group, a space was observed between the conjunctiva-flap-bleb (position indicated by the arrow in the figure). That is, the formation of fibers was clearly suppressed in the trehalose use group, and it was confirmed that fibroblasts were sufficiently suppressed after 5 days or 10% trehalose instillation was continued for 90 days.
図５は、白色家兎線維柱帯切除術後９０日間トレハロース点眼後の免疫組織化学所見を示す顕微鏡写真図である。図では上段が対照群、中段が５％トレハロース点眼群、下段が１０％トレハロース点眼群である。また、左列が抗ｖｉｍｅｎｔｉｎ抗体による、中央列が抗α−ＳＭＡ抗体による、そして右列が両者を用いた二重免疫染色による、各免疫組織化学所見である。 FIG. 5 is a photomicrograph showing the immunohistochemical findings after 90 days of trehalose instillation after white rabbit trabeculectomy. In the figure, the upper row is the control group, the middle row is the 5% trehalose eye drop group, and the lower row is the 10% trehalose eye drop group. The left column shows anti-vimentin antibodies, the center column shows anti-α-SMA antibodies, and the right column shows immunohistochemical findings by double immunostaining using both.
図示されるように、５％、１０％の各トレハロース点眼群では明らかに、ｖｉｍｅｎｔｉｎ／α−ＳＭＡの発現の低下を認めた。つまり、抗ｖｉｍｅｎｔｉｎ抗体による免疫組織化学所見、抗α−ＳＭＡ抗体による免疫組織化学所見、および二重免疫染色による免疫組織化学所見のいずれにおいても、線維芽細胞の増殖が抑制されていることが認められた。 As shown in the figure, a decrease in the expression of vimentin / α-SMA was clearly observed in the 5% and 10% trehalose ophthalmic groups. That is, it is recognized that the proliferation of fibroblasts is suppressed in any of the immunohistochemical findings by anti-vimentin antibody, the immunohistochemical findings by anti-α-SMA antibody, and the immunohistochemical findings by double immunostaining. It was.
本発明の結膜下線維芽細胞増殖抑制剤および結膜下線維芽細胞増殖抑制方法によれば、安全かつ効果的に、眼科手術後における結膜癒着や結膜下組織瘢痕形成を抑制することができる。つまり本発明によれば、手術後に起こる結膜下組織の線維芽細胞増殖を選択的に抑制し、しかも結膜上皮の創傷治癒機転には何ら影響を及ぼさない。
According to subconjunctival fibroblast growth inhibitor and the subconjunctival fibroblast growth inhibiting method of the present invention, it is possible to suppress a safe and effective, conjunctival adhesion and subconjunctival tissue scarring after ocular surgery. That is, according to the present invention, the fibroblast proliferation in the subconjunctival tissue that occurs after surgery is selectively suppressed, and the conjunctival epithelial wound healing mechanism is not affected at all.
この点をさらに応用すれば、翼状片に行う翼状片切除手術の術後に再発予防薬としてトレハロースを使用できる可能性や、斜視手術術後にも瘢痕形成抑制の目的として使用できる可能性も認められ、産業上利用性も高い発明である。
If this point is further applied, there is a possibility that trehalose can be used as a preventive agent for recurrence after pterygium resection surgery for pterygium, and that it can also be used for the purpose of suppressing scar formation after strabismus surgery. It is an invention with high industrial applicability.
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