Pretreatment with recombinant human Slit2 protein relieves endotoxin-induced uveitis in rats

Yong DU; Shaoqiu JIANG; Xiyuan ZHOU

doi:10.16016/j.1000-5404.201712095

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Pretreatment with recombinant human Slit2 protein relieves endotoxin-induced uveitis in rats

Basic Medicine | 更新时间：2026-01-09

- Pretreatment with recombinant human Slit2 protein relieves endotoxin-induced uveitis in rats
- Journal of Third Military Medical University Vol. 40, Issue 14, Pages: 1263-1270(2018)
- 作者机构：
  
  400016 重庆，重庆医科大学附属第二医院眼科，眼科学重庆市重点实验室
- 作者简介：
- 基金信息：
  
  the General Program of National Natural Science Foundation of China(81170858)
- DOI：10.16016/j.1000-5404.201712095
  CLC： R773.9; R967; R977.6
- Received：11 December 2017，
  
  Revised：2018-04-15，
  
  Published：30 July 2018
- 稿件说明：
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Yong DU, Shaoqiu JIANG, Xiyuan ZHOU. Pretreatment with recombinant human Slit2 protein relieves endotoxin-induced uveitis in rats[J]. Journal of Third Military Medical University, 2018, 40(14): 1263-1270.
DOI：

Yong DU, Shaoqiu JIANG, Xiyuan ZHOU. Pretreatment with recombinant human Slit2 protein relieves endotoxin-induced uveitis in rats[J]. Journal of Third Military Medical University, 2018, 40(14): 1263-1270. DOI： 10.16016/j.1000-5404.201712095.

摘要

目的

探讨重组人Slit2蛋白(recombinant human slit2

rhSlit2)对内毒素诱导的Sprague-Dawley(SD)大鼠葡萄膜炎(endotoxin-induced uveitis

EIU)的作用。

方法

给予SD大鼠玻璃体腔内分别注射rhSlit2(10、30、100 ng/眼)。24 h后，将2 g/L的脂多糖(lipopolysaccharides

LPS)注射到大鼠双侧后足底肉垫内(200 μg/鼠)。LPS注射24 h后观察大鼠前房炎症并进行临床评分；取大鼠房水进行房水蛋白浓度测定和房水细胞计数；LPS注射24 h后摘除大鼠眼球，制作石蜡切片HE染色后观察大鼠眼部形态学变化；采用Real-time PCR检测大鼠眼部炎症因子白介素-6(interleukin-6

IL-6)、肿瘤坏死因子-α(tumor necrosis factor-α

TNF-α)mRNA水平的表达；Western blot检测大鼠眼部IL-6、TNF-α和PI3K/Akt信号通路蛋白的表达。

结果

在LPS注射24 h后，rhSlit2对前房炎症有明显的抑制作用，且具有剂量依赖性。LPS+rhSlit2(100 ng/眼)组临床评分明显低于LPS+PBS组[(1.40±0.84)

(5.60±0.97)分，

＜0.01

]

；HE染色可见各组大鼠眼内结构层次完好。与LPS+PBS组比较，LPS+rhSlit2(100 ng/眼)组眼内炎症细胞渗出数量明显减少[(65.21±18.38)×10

/mL

(319.60±28.88)×10

/mL，

＜0.01

]

；房水蛋白浓度明显降低[(12.59±3.04)

(31.50±3.09) mg/mL，

＜0.01

]

；眼内炎症因子IL-6 (19.69±5.28

77.09±12.61)、TNF-α(1.78±0.33

4.06±0.58)的mRNA表达降低(

＜0.01)；大鼠眼内IL-6(1.92±0.60

4.42±0.11)、TNF-α(1.33±0.11

1.69±0.21)和P-Akt(0.18±0.37

0.55±0.34)蛋白水平也降低(

＜0.01)。

结论

大鼠玻璃体腔RhSlit2预注射对内毒素诱导的SD大鼠葡萄膜炎具有抑制作用。RhSlit2可通过抑制PI3K/Akt信号通路的活化以抑制LPS诱导炎症反应。

Abstract

Objective

To investigate the effect of recombinant human Slit2 protein (rhSlit2) on endotoxin-induced uveitis in rats.

Methods

Adult SD rats were subjected to injection of 2 μL PBS or rhSlit2 (10

or 100 ng/eye) into the vitreous cavity

followed 24 h later by injection of 200 μg lipopolysaccharides (LPS) into the posterior foot pads. Twenty-four hours after LPS injection

the clinical scores of the anterior chamber inflammation of the rats were assessed

and the protein concentrations and the total number of cells in the aqueous humor were determined. The changes in ocular morphologies were observed using HE staining 24 h after LPS injection. Real-time PCR was used to detect the expression of interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) in the eyes

and the protein levels of IL-6

TNF-α

Akt and P-Akt were measured using Western blotting.

Results

Pretreatment with rhSlit2 24 h before LPS injection significantly inhibited anterior chamber inflammation in a dose-dependent manner. Compared with PBS

pretreatment with rhSlit2 at 100 ng/eye resulted in a significantly lower mean clinical score of anterior chamber inflammation (5.60±0.97

1.40±0.84

0.01) and protected the integrity of the ocular structure in LPS-challenged rats. Compared with those with PBS+LPS treatment

the rats pretreated with rhSlit2 (100 ng/eye) before LPS challenge exhibited a significantly lower number of intraocular inflammatory cells [(319.60±28.88)×10

/mL

(65.21±18.38)×10

/mL

0.01

]

lower protein concentrations in the aqueous humor (

0.01)

lower mRNA levels of IL-6 (77.09±12.61

19.69±5.28

0.01) and TNF-α (4.06±0.58

1.78±0.33

0.01)

and lower protein levels of IL-6 (4.42±0.11

1.92±0.60

0.01)

TNF-α (1.69±0.21

1.33±0.11

0.01) and P-Akt (0.55±0.34

0.18±0.37

0.01) in the eye tissues.

Conclusion

Intravitreal injection of rhSlit2 can ameliorate the inflammation in SD rats with endotoxin-induced uveitis possibly by inhibiting the activation of the PI3K/Akt signaling pathway.

关键词

Keywords

references

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