ROCK inhibitors beneficially alter the spatial configuration of TGFβ2-treated 3D organoids from a human trabecular meshwork (HTM).
ROCK inhibitors beneficially alter the spatial configuration of TGFβ2-treated 3D organoids from a human trabecular meshwork (HTM).
复制标题
岩石抑制剂有益地改变了人类小梁网（HTM）的TGFβ2处理的3D器官的空间构型。
DOI：
10.1038/s41598-020-77302-9
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发表时间：
2020-11-20
影响因子：
4.6
通讯作者：
Hikage F
中科院分区：
文献类型：
--
作者：
Ota C;Ida Y;Ohguro H;Hikage F
为了阐明rho相关的含有蛋白激酶抑制剂（ROCK-i， Ripasudil和Y27632）的分子药理学对其水流出效率的影响，在TGFβ2存在下制备了人小梁网（HTM）的2D或3D培养物。通过跨内皮电阻（TEER, 2D），电子显微镜（EM， 2D和3D），细胞外基质（ECM）的表达，包括胶原1 (COL1)， COL4和COL6，以及通过免疫标记和/或定量PCR （3D）检测纤维连接蛋白（FN），以及通过微挤压器检测3D类器官的固体性。tgf - β2在二维培养中显著增加TEER值，ECM表达表明三维类器官呈现更密集的排列形状。ROCK-i可显著降低tgf - β2诱导的三维类器官TEER和免疫标记ECM表达的增强。相比之下，COL1的mRNA表达增加，COL4和FN的mRNA表达不变。电镜显示tgf - β2使HTM细胞变得更加致密，并在三维类器官内观察到丰富的ECM沉积。ROCK-i显著抑制了这些反应。由tgf - β2引起的致密固体被ROCK-i显著抑制。目前的研究表明，ROCK-i对tgf - β2诱导的HTM 3D类器官的空间构型有有益的影响。
To elucidate molecular pharmacology of Rho-associated coiled-coil containing protein kinase inhibitors (ROCK-i, Ripasudil and Y27632) on their efficiency for aqueous outflow, 2D or 3D cultures of a human trabecular meshwork (HTM) were prepared in the presence of TGFβ2. Those were examined by transendothelial electrical resistance (TEER, 2D), electronic microscopy (EM, 2D and 3D), expression of the extracellular matrix (ECM) including collagen1 (COL1), COL4 and COL6, and fibronectin (FN) by immunolabeling and/or quantitative PCR (3D), and solidity of 3D organoids by a micro-squeezer. TGFβ2 significantly increased the TEER values in 2D cultures, and the ECM expression indicated that the 3D organoids assumed a more densely packed shape. ROCK-i greatly reduced the TGFβ2-induced enhancement of TEER and the immunolabeled ECM expression of the 3D organoids. In contrast, the mRNA expression of COL1 was increased, and those of COL4 and FN were unchanged. EM revealed that TGFβ2 caused the HTM cells to become more compact and abundant ECM deposits within the 3D organoids were observed. These were significantly inhibited by ROCK-i. The dense solids caused by the presence of TGFβ2 were significantly suppressed by ROCK-i. Current study indicates that ROCK-i cause beneficial effects toward the spatial configuration of TGFβ2-induced HTM 3D organoids.
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影响因子：
19
作者：
Huh, Dongeun;Hamilton, Geraldine A.;Ingber, Donald E.
通讯作者：
Ingber, Donald E.
影响因子：
4.6
作者：
Kaneko Y;Ohta M;Inoue T;Mizuno K;Isobe T;Tanabe S;Tanihara H
通讯作者：
Tanihara H
影响因子：
3.9
作者：
Cho, Hee Jun;Yoo, Jiyun
通讯作者：
Yoo, Jiyun
影响因子：
7.5
作者：
Kadler KE;Hill A;Canty-Laird EG
通讯作者：
Canty-Laird EG
影响因子：
3.5
作者：
Nakagawa, O;Fujisawa, K;Narumiya, S
通讯作者：
Narumiya, S