缺血再灌注损伤后小鼠视网膜中 CXCR5 的抗炎作用。
The Anti-Inflammatory Effects of CXCR5 in the Mice Retina following Ischemia-Reperfusion Injury.
机构信息
Aier School of Ophthalmology, Central South University, Aier Eye Institute, Changsha, Hunan, China.
Department of Ophthalmology, Changsha Aier Eye Hospital, Changsha, Hunan, China.
出版信息
Biomed Res Int. 2019 Jul 4;2019:3487607. doi: 10.1155/2019/3487607. eCollection 2019.
OBJECT
Retinal ischemia-reperfusion (I/R) injury is a common pathological process in many ophthalmic diseases; there are no effective therapeutic approaches available currently. Increasing evidence indicates that microglia mediated neuroinflammation plays an important role in the retinal I/R injury. In this study, we aimed to investigate the roles of chemokine receptor CXCR5 in the pathological process of retinal I/R injury model.
METHOD
Retinal I/R injury model was established in CXCR5 knockout and wild mice by the acute elevation of intraocular pressure (AOH) for 60 minutes, and the eyes were harvested for further analyses. The cellular location of CXCR5 was detected by immunofluorescence staining; the expressions of CXCR5 and CXCL13 after I/R injury were analyzed by quantitative RT-PCR. The retinal microglia were detected as stained for Iba1 (+). Leakage of inflammatory cells was observed on the H&E stained cryosections. The protein expression and quantification of zonula occludens (ZO-1) were determined by Western blotting and densitometry. Capillary degeneration was identified on the intact retinal vasculatures prepared by trypsin digestion.
RESULTS
The number of activated microglia marked by Iba1 antibody in the retina was increased after retinal I/R injury in both KO and WT mice, more significant in KO mice. The leakage of inflammatory cells was observed largely at 2 days after injury, but there was no or little leakage at 7 days. The number of inflammatory cells (mainly neutrophils) was greater in CXCR5 KO mice than in WT mice, mainly located under internal limiting membrane. CXCR5 deficiency led to more ZO-1 degradation in CXCR5 KO mice compared to C57BL6 WT mice 2 days after reperfusion. The cellular capillaries were also significantly increased in the KO mice compared to the WT mice.
CONCLUSION
Our findings suggest that the chemokine receptor CXCR5 may protect retina from ischemia-reperfusion injury by its anti-inflammatory effects. Thus, CXCR5 may be a promising therapeutic target for the treatment of retinal I/R injury.
目的
视网膜缺血再灌注（I/R）损伤是许多眼科疾病中的一种常见病理过程；目前尚无有效的治疗方法。越来越多的证据表明，小胶质细胞介导的神经炎症在视网膜 I/R 损伤中起重要作用。在本研究中，我们旨在研究趋化因子受体 CXCR5 在视网膜 I/R 损伤模型中的病理过程中的作用。
方法
通过急性升高眼内压（AOH）60 分钟，在 CXCR5 敲除和野生型小鼠中建立视网膜 I/R 损伤模型，并对眼睛进行进一步分析。通过免疫荧光染色检测 CXCR5 的细胞位置；通过定量 RT-PCR 分析 I/R 损伤后 CXCR5 和 CXCL13 的表达。用 Iba1（+）染色检测视网膜小胶质细胞。用 H&E 染色的冷冻切片观察炎症细胞的渗漏。通过 Western blot 和密度测定法确定紧密连接蛋白（ZO-1）的蛋白表达和定量。通过胰蛋白酶消化制备完整的视网膜血管，鉴定毛细血管退化。
结果
在 KO 和 WT 小鼠的视网膜 I/R 损伤后，用 Iba1 抗体标记的激活小胶质细胞的数量增加，在 KO 小鼠中更为明显。损伤后 2 天观察到炎症细胞的渗漏较大，但在 7 天内无或很少有渗漏。在 CXCR5 KO 小鼠中，炎症细胞（主要是中性粒细胞）的数量比 WT 小鼠多，主要位于内界膜下。与 C57BL6 WT 小鼠相比，CXCR5 缺失导致再灌注后 2 天 CXCR5 KO 小鼠中 ZO-1 的降解更多。与 WT 小鼠相比，KO 小鼠的细胞毛细血管也明显增加。
结论
我们的研究结果表明，趋化因子受体 CXCR5 通过其抗炎作用可能保护视网膜免受缺血再灌注损伤。因此，CXCR5 可能是治疗视网膜 I/R 损伤的有前途的治疗靶点。
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