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[低聚原花青素对百草枯染毒小鼠的保护作用]

[Protective effects of oligomeric proanthocyanidins in mice exposed to paraquat].

作者信息

Liu P, Zhou Y S, Li L, Qin Y L, Mu J S

机构信息

Department of Emergency Medicine, , Intensive Care Medicine Center, The Fifth Medical Center of PLA General Hospital, Beijing 100039, China.

出版信息

Zhonghua Lao Dong Wei Sheng Zhi Ye Bing Za Zhi. 2019 Aug 20;37(8):571-575. doi: 10.3760/cma.j.issn.1001-9391.2019.08.003.

Abstract

To investigate the protective effect of oligomeric proanthocyanidins (OPCs) in paraquat-exposed mice. An acute lung injury model was established by a single intraperitoneal injection of paraquat (PQ) in BALB/c mice. The mice were randomized into control group, paraquat-exposed group (PQ group) , oligomeric proanthocyanidins group (OPCs group) , and paraquat and oligomeric proanthocyanidins-exposed group (PQ+OPCs group) , with 10 mice in each group. Only normal saline was intraperitoneally injected into the mice in the control group. The mice in the PQ group were divided into 8 subgroups according to the dose of poison administered, i.e., 0, 25, 50, 75, 100, 150, 200, and 300 mg/kg; the mice in each subgroup were given a single intraperitoneal injection of PQ and were observed and recorded for death at 3, 6, 12, 24, 36, 48, 60, 84, and 96 hours after PQ injection. Origin 8.0 was used to calculate the median lethal dose (LD(50)) of the mice at 24, 36, 48, and 60 hours after PQ injection, and the PQ dose (100 mg/kg, ip) was chosen based on the accumulated mortality rate. An OPCs-treated experimental model was established by an intraperitoneal injection of OPCs followed by a single PQ injection (100 mg/kg, ip) 1 hour later to observe the effects of OPCs on the apparent poisoning effect and fatality rate in PQ-induced mice. Immunohistochemistry was used to determine the effect of OPCs on PQ-induced lung tissue lesions. The peripheral blood samples of the mice were collected to determine the effects of OPCs on PQ-induced inflammatory factors such as tumor necrosis factor-α (TNF-α) , interleukine-1β (IL-1β) , and transforming growth factor-β1 (TGF-β1) using enzyme-linked immunosorbent assay. The mortality rate was significantly correlated with the dose and exposure time in PQ-exposed mice; the mortality rate gradually increased with increasing dose and exposure time of the poison (<0.05) . The LD(50) values for the mice were 216.67, 124.11, and 71.24 mg/kg at 24, 48, and 72 hours after PQ exposure, respectively. PQ could induce animal death at 12 hours after injection, and the mortality rate of the animals was 40% (4/10) at 48 hours after PQ exposure. The PQ-induced mortality rate of the mice in the PQ+OPCs group was reduced, and the mortality rate of the animals was 10% (1/10) at 48 hours after PQ exposure. Compared with treatment in the control group, OPCs exposure alone had no significant effect on the expression of TNF-α and TGF-β1 in the peripheral blood (>0.05) , but it significantly inhibited the expression of IL-1β (<0.05) . After 48 hours, the expression of TNF-α, TGF-β1, and IL-1β in peripheral blood significantly increased by 39%, 45%, and 38%, respectively, in the PQ group (<0.05) , but they significantly decreased by 31%, 13%, and 22%, respectively, in the OPCs+PQ group as compared with the PQ group (<0.05) . OPCs pretreatment can significantly alleviate PQ-induced poisoning effect.

摘要

探讨寡聚原花青素(OPCs)对百草枯染毒小鼠的保护作用。通过对BALB/c小鼠单次腹腔注射百草枯(PQ)建立急性肺损伤模型。将小鼠随机分为对照组、百草枯染毒组(PQ组)、寡聚原花青素组(OPCs组)和百草枯与寡聚原花青素染毒组(PQ+OPCs组),每组10只小鼠。对照组小鼠仅腹腔注射生理盐水。PQ组小鼠根据染毒剂量分为8个亚组,即0、25、50、75、100、150、200和300 mg/kg;每个亚组的小鼠单次腹腔注射PQ,并在注射PQ后3、6、12、24、36、48、60、84和96小时观察并记录死亡情况。使用Origin 8.0计算PQ注射后24、36、48和60小时小鼠的半数致死剂量(LD(50)),并根据累积死亡率选择PQ剂量(100 mg/kg,腹腔注射)。通过腹腔注射OPCs,1小时后再单次注射PQ(100 mg/kg,腹腔注射)建立OPCs处理的实验模型,观察OPCs对PQ诱导小鼠明显中毒效应和死亡率的影响。采用免疫组织化学法测定OPCs对PQ诱导的肺组织损伤的影响。采集小鼠外周血样本,使用酶联免疫吸附测定法测定OPCs对PQ诱导的炎症因子如肿瘤坏死因子-α(TNF-α)、白细胞介素-1β(IL-1β)和转化生长因子-β1(TGF-β1)的影响。PQ染毒小鼠的死亡率与剂量和暴露时间显著相关;死亡率随毒物剂量和暴露时间的增加而逐渐升高(<0.05)。PQ暴露后24、48和72小时小鼠的LD(50)值分别为216.67、124.11和71.24 mg/kg。PQ注射后12小时可导致动物死亡,PQ暴露后48小时动物死亡率为40%(4/10)。PQ+OPCs组小鼠的PQ诱导死亡率降低,PQ暴露后48小时动物死亡率为10%(1/10)。与对照组处理相比,单独暴露OPCs对外周血中TNF-α和TGF-β1的表达无显著影响(>0.05),但显著抑制IL-1β的表达(<0.05)。48小时后,PQ组外周血中TNF-α、TGF-β1和IL-1β的表达分别显著增加39%、45%和38%(<0.05),但与PQ组相比,OPCs+PQ组分别显著降低31%、13%和22%(<0.05)。OPCs预处理可显著减轻PQ诱导的中毒效应。

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