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400037 重庆,陆军军医大学(第三军医大学)第二附属医院眼科
[袁容娣,E-mail:1253450264@qq.comYUAN Rongdi, E-mail:1253450264@qq.com
收稿:2019-07-15,
修回:2019-08-28,
纸质出版:2020-01-15
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黄瑶, 范围, 杨梅, 等. Angptl2通过PirB激活视网膜Müller细胞参与眼内炎症[J]. 第三军医大学学报, 2020,42(1):39-44.
Yao HUANG, Wei FAN, Mei YANG, et al. Angptl2 mediates intraocular inflammation by activating Müller cells through PirB[J]. Journal of Third Military Medical University, 2020, 42(1): 39-44.
黄瑶, 范围, 杨梅, 等. Angptl2通过PirB激活视网膜Müller细胞参与眼内炎症[J]. 第三军医大学学报, 2020,42(1):39-44. DOI: 10.16016/j.1000-5404.201907167.
Yao HUANG, Wei FAN, Mei YANG, et al. Angptl2 mediates intraocular inflammation by activating Müller cells through PirB[J]. Journal of Third Military Medical University, 2020, 42(1): 39-44. DOI: 10.16016/j.1000-5404.201907167.
目的
2
探讨血管生成素样蛋白2(angiopoietin-like protein 2
Angptl2)在眼内促进炎症的作用和机制。
方法
2
实验组大鼠玻璃体腔注射0.2 mg LPS,对照组加入等量PBS,RT-PCR检测两组大鼠视网膜Angptl2的表达。以原代培养的SD大鼠视网膜Müller细胞为研究对象,加入外源性Angptl2后,细胞计数观察Müller细胞增殖变化,免疫荧光观察Müller细胞GFAP的表达以及RT-PCR检测促炎因子IL-1β、IL-6、TNF-α的表达变化。免疫荧光观察配对免疫球蛋白样受体B(paired immunoglobulin-like receptorB,PirB)在Müller细胞上的表达;Müller细胞外源性加入Angptl2后,再分别转染Ad-PirB-siRNA腺病毒载体和腺病毒对照Ad-C,分为PBS对照组、Angptl2组、Ad-C+Angptl2组、Ad-PirB-siRNA+Angptl2组,RT-PCR观察炎症因子的表达变化。
结果
2
LPS组Angptl2 mRNA表达较对照组明显增加(
P
<
0.05);Angptl2组较PBS对照组Müller细胞增殖增多,GFAP表达增加,IL-1β、IL-6、TNF-α表达也增加,差异具有统计学意义(
P
<
0.05);Müller细胞表达Angptl2的配体PirB,Ad-PirB-siRNA干扰Müller细胞PirB表达后,Ad-PirB-siRNA+Angptl2组较Angptl2组和Ad-C+Angptl2组炎症因子IL-6、TNF-α降低
差异具有统计学意义(
P
<
0.05)。
结论
2
Angptl2通过PirB激活Müller细胞分泌促炎因子IL-1β、IL-6、TNF-α参与眼内炎症。
Objective
2
To investigate the pro-inflammatory effects and mechanisms of angiopoietin-like protein 2 (Angptl2) in the eyes.
Methods
2
SD rats (8 weeks old) were given a single intravitreal injection of 0.2 mg LPS or PBS to establish intraocular inflammation model or sham operation. Then the Angptl2 expression in the retina was examined by real-time PCR (RT-PCR). Müller cells were isolated from the retina of newborn SD rats (with 24 h after born). After the obtained cells were purified and identified
the recombinant Angptl2 was added to the primarily cultured cells. Cell count
immunofluorescence assay and RT-PCR were used to assess the proliferation of Müller cells
the expression of GFAP
and levels of interleukin-1 beta (IL-1β)
interleukin-6 (IL-6) and tumor necrosis factor (TNF-α)
respectively. The expression of Angptl2 ligand
paired immunoglobulin-like receptor B (PirB)
in the Müller cells was observed by immunofluorescence assay. RT-PCR was employed to observed the expression of IL-6 and TNF-α in the Müller cells of PBS group
Angptl2 group
Ad-C (transfection of adenovirus negative control Ad-C)+Angptl2 group
and Ad-PirB-siRNA (transfection of adenovirus Ad-PirB-siRNA)+Angptl2 group.
Results
2
The mRNA level of Angptl2 was increased notably in the retina of the LPS-treated rats (intraocular inflammation model) than the control rats (
P
<
0.05). Exogenous Angptl2 treatment resulted in higher proliferation rate
expression of GFAP and the levels of IL-1β
IL-6 and TNF-α in Müller cells (
P
<
0.05). PirB was expressed in the retinal Müller cells. Interference of PirB by adenovirus Ad-PirB-siRNA significantly decreased the levels of IL-6 and TNF-α (
P
<
0.05).
Conclusion
2
Angptl2 mediates acute intraocular inflammation by activating Müller cells to secrets IL-1β
IL-6 and TNF-α though PirB.
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