轻度低温联合芦丁通过抑制 TGF-β/smad 通路对脊髓损伤及炎症因子的治疗作用。

The effect of mild hypothermia plus rutin on the treatment of spinal cord injury and inflammatory factors by repressing TGF-β/smad pathway.

机构信息

MM. Department of Spine Surgery - Guizhou Orthopedic Hospital - Guizhou Province, China.

PhD. Department of Spine Surgery - Guizhou Orthopedic Hospital - Guizhou Province, China.

出版信息

Acta Cir Bras. 2021 May 7;36(3):e360307. doi: 10.1590/ACB360307. eCollection 2021.

Abstract

PURPOSE

To probe the mechanism of mild hypothermia combined with rutin in the treatment of spinal cord injury (SCI).

METHODS

Thirty rats were randomized into the following groups: control, sham, model, mild hypothermia (MH), and mild hypothermia plus rutin (MH+Rutin). We used modified Allen's method to injure the spinal cord (T10) in rats, and then treated it with MH or/and rutin immediately. BBB scores were performed on all rats. We used HE staining for observing the injured spinal cord tissue; ELISA for assaying TNF-α, IL-1β, IL-8, Myeloperoxidase (MPO), and Malondialdehyde (MDA) contents; Dihydroethidium (DHE) for measuring the reactive oxygen species (ROS) content; flow cytometry for detecting apoptosis; and both RT-qPCR and Western blot for determining the expression levels of TGF-β/Smad pathway related proteins (TGF-β, Smad2, and Smad3).

RESULTS

In comparison with model group, the BBB score of MH increased to a certain extent and MH+Rutin group increased more than MH group (p < 0.05). After treatment with MH and MH+Rutin, the inflammatory infiltration diminished. MH and MH+Rutin tellingly dwindled TNF-β, MDA and ROS contents (p < 0.01), and minified spinal cord cell apoptosis. MH and MH+Rutin could patently diminished TGF-β1, Smad2, and Smad3 expression (p < 0.01).

CONCLUSIONS

MH+Rutin can suppress the activation of TGF-β/Smad pathway, hence repressing the cellular inflammatory response after SCI.

摘要

目的

探讨亚低温联合芦丁治疗脊髓损伤(SCI)的机制。

方法

将 30 只大鼠随机分为对照组、假手术组、模型组、亚低温组(MH)和亚低温联合芦丁组(MH+Rutin)。采用改良 Allen 法损伤大鼠脊髓(T10),伤后立即给予 MH 或/和芦丁治疗。对所有大鼠进行 BBB 评分。采用 HE 染色观察损伤脊髓组织;ELISA 法检测 TNF-α、IL-1β、IL-8、髓过氧化物酶(MPO)和丙二醛(MDA)含量;二氢乙啶(DHE)法测定活性氧(ROS)含量;流式细胞术检测细胞凋亡;以及 RT-qPCR 和 Western blot 测定 TGF-β/Smad 通路相关蛋白(TGF-β、Smad2 和 Smad3)的表达水平。

结果

与模型组相比,MH 组 BBB 评分有所升高,MH+Rutin 组升高更为明显(p<0.05)。MH 和 MH+Rutin 治疗后,炎症浸润减轻。MH 和 MH+Rutin 显著降低 TNF-β、MDA 和 ROS 含量(p<0.01),减少脊髓细胞凋亡。MH 和 MH+Rutin 明显降低 TGF-β1、Smad2 和 Smad3 的表达(p<0.01)。

结论

MH+Rutin 可抑制 TGF-β/Smad 通路的激活,从而抑制 SCI 后细胞炎症反应。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6151/8112105/f68e05e58261/1678-2674-acb-36-3-e360307-gf01.jpg

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