Juming Zhu, Dongmei Ding, Tao Sun, Yuting Zhang, Huizi Miao, Yunjie Gu, Ming Dai, Manhui Zhu
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IF on mouse choroid flat mounts was done to confirm the effect of ORI on BRB integrity. Indocyanine green angiography and IF on mouse retina-RPE-choroid flat mounts were performed to determine the effect of ORI on CNV formation and retinal function. Hematoxylin and eosin staining and TUNEL staining were carried out to appraise ocular and systemic cytotoxicity caused by ORI.</p><p><strong>Results: </strong>ORI protected ARPE-19 cells from hypoxia-induced destruction of barrier properties and promoted the expression of ZO-1 and occludin by the TGFβR/SUV39H1/KLF11 axis, maintaining barrier properties of ARPE-19 cells with hypoxia. ORI improved BRB integrity during laser-induced CNV in mice and mitigated laser-induced CNV formation in mice without any ocular or systemic cytotoxicity (n = 4-5 in each group).</p><p><strong>Conclusions: </strong>ORI ameliorates BRB integrity and subsequent formation of CNV via regulating the TGFβR/SUV39H1/KLF11 pathway in RPE cells.</p>","PeriodicalId":14620,"journal":{"name":"Investigative ophthalmology & visual science","volume":"66 2","pages":"56"},"PeriodicalIF":5.0000,"publicationDate":"2025-02-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11855140/pdf/","citationCount":"0","resultStr":"{\"title\":\"Oridonin Preserves Retinal Pigmented Epithelial Cell Tight Junctions and Ameliorates Choroidal Neovascularization.\",\"authors\":\"Juming Zhu, Dongmei Ding, Tao Sun, Yuting Zhang, Huizi Miao, Yunjie Gu, Ming Dai, Manhui Zhu\",\"doi\":\"10.1167/iovs.66.2.56\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><strong>Purpose: </strong>To investigate the role and mechanism of oridonin (ORI), a bioactive diterpenoid extracted from the Chinese herbal medicine Rabdosia rubescens, on the integrity of outer blood-retinal barrier (oBRB) during choroidal neovascularization (CNV).</p><p><strong>Methods: </strong>ARPE-19 cells were exposed to hypoxia and treated with ORI. 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引用次数: 0
摘要
目的：研究从中草药冬凌草中提取的活性二萜类化合物冬凌草苷（ORI）对脉络膜新生血管（CNV）过程中血视网膜外屏障（oBRB）完整性的影响及其机制。方法：对ARPE-19细胞进行缺氧处理和ORI处理。采用WB、染色质免疫沉淀法、荧光素报告活性法和免疫荧光法检测tgf - β r /SUV39H1/KLF11轴ZO-1和occludin的表达，并研究ORI对ARPE-19细胞屏障特性的影响。建立激光诱导小鼠CNV模型，灌胃给药。在小鼠脉络膜平载上进行IF实验，以证实ORI对BRB完整性的影响。对小鼠视网膜- rpe -脉络膜平载进行吲哚菁绿血管造影和IF观察，观察ORI对CNV形成和视网膜功能的影响。采用苏木精和伊红染色及TUNEL染色评价ORI引起的眼部和全身细胞毒性。结果：ORI保护ARPE-19细胞免受缺氧对屏障特性的破坏，促进tgf - β r /SUV39H1/KLF11轴ZO-1和occludin的表达，维持缺氧状态下ARPE-19细胞的屏障特性。ORI改善了激光诱导小鼠CNV过程中BRB的完整性，减轻了激光诱导小鼠CNV的形成，无任何眼部或全身细胞毒性（每组n = 4-5）。结论：ORI通过调控RPE细胞中tgf - β r /SUV39H1/KLF11通路改善BRB的完整性和随后CNV的形成。
Purpose: To investigate the role and mechanism of oridonin (ORI), a bioactive diterpenoid extracted from the Chinese herbal medicine Rabdosia rubescens, on the integrity of outer blood-retinal barrier (oBRB) during choroidal neovascularization (CNV).
Methods: ARPE-19 cells were exposed to hypoxia and treated with ORI. The expression of ZO-1 and occludin in the axis of TGFβR/SUV39H1/KLF11 was detected by WB, chromatin immunoprecipitation, luciferin report activity assay, and immunofluorescence assay (IF), and the effect of ORI on the barrier properties of ARPE-19 cells was studied. A laser-induced mouse CNV model was constructed, and ORI was administrated by oral gavage. IF on mouse choroid flat mounts was done to confirm the effect of ORI on BRB integrity. Indocyanine green angiography and IF on mouse retina-RPE-choroid flat mounts were performed to determine the effect of ORI on CNV formation and retinal function. Hematoxylin and eosin staining and TUNEL staining were carried out to appraise ocular and systemic cytotoxicity caused by ORI.
Results: ORI protected ARPE-19 cells from hypoxia-induced destruction of barrier properties and promoted the expression of ZO-1 and occludin by the TGFβR/SUV39H1/KLF11 axis, maintaining barrier properties of ARPE-19 cells with hypoxia. ORI improved BRB integrity during laser-induced CNV in mice and mitigated laser-induced CNV formation in mice without any ocular or systemic cytotoxicity (n = 4-5 in each group).
Conclusions: ORI ameliorates BRB integrity and subsequent formation of CNV via regulating the TGFβR/SUV39H1/KLF11 pathway in RPE cells.
期刊介绍：
Investigative Ophthalmology & Visual Science (IOVS), published as ready online, is a peer-reviewed academic journal of the Association for Research in Vision and Ophthalmology (ARVO). IOVS features original research, mostly pertaining to clinical and laboratory ophthalmology and vision research in general.
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