The protective effect of Ulinastatin on sepsis through miR-21/p38MAPK pathway
10.3760/cma.j.issn.1671-0282.2018.11.012
- VernacularTitle:乌司他丁通过miR-21/p38MAPK通路对脓毒症保护作用的研究
- Author:
Yang TAO
1
;
Ling XIAO
;
Jian HUANG
;
Yu MA
Author Information
1. 重庆市急救医疗中心重症医学科
- Keywords:
Ulinastatin;
Sepsis;
miR-21;
PTEN;
p38MAPK
- From:
Chinese Journal of Emergency Medicine
2018;27(11):1242-1247
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the effect of ulinastatin on miR-21/p38 mitogen-activated protein kinase (p38MAPK) signaling pathway in sepsis. Methods SPF BALB/c mice (n=30) were randomly divided into 3 groups (n=10, each group): LPS group (LPS 10 mg/kg disposable intraperitoneal injection), UTI group (Ulinastatin 105 U/kg was pretreated by intraperitoneal injection for 2 hours, and LPS was administered intraperitoneally 10 mg/kg), and the normal group (same volume of normal saline was injected at the same time point). After 24 h, the mice were sacrificed and the pathological results of lung tissue were observed. The expression of miR-21 in lung tissue was detected by real-time quantitative PCR. The expression of PTEN and p-p38MAPK in lung tissues of each group were detected by immunohistochemistry. In addition, mouse macrophage RAW264.7 cells were treated with LipofectamineTM 2000 transfection reagent, miR-21 mimic agent (100 nmol/L) and inhibitor (100 nmol/L) for 6 h, then the supernatant was discarded and Ulinastatin (1000 U/mL) was added for 2 h treatment, and finally 500 ng/mL LPS were added. After 24 h, the cells were collected. The expression of miR-21 in lung tissue was determined by PCR, and the expression of PTEN and p-p38MAPK protein were detected by Western blot. Results In animal experiments, the expression of miR-21(2.73±0.17) and p-p38MAPK was up-regulated in the LPS group, and the expression of PTEN protein was down-regulated in the downstream target gene of miR-21. Whereas the expression levels of miR-21(2.14±0.13) and p-p38MAPK in the UTI group were decreased, and the expression of PTEN protein was up-regulated. In the cell experiment, compared with the LPS group(1.93±0.21), the expression of p-p38MAPK protein in the LPS+miR-21 inhibitor group(1.51±0.06) decreased, while the expression of p-p38MAPK protein in the LPS+miR-21 mimic group increased(2.52±0.07). After UTI intervention, the expression of p-p38MAPK protein in the LPS+UTI group(0.61±0.04) was lower than that in the LPS group. Compared with the LPS+UTI group, the expression of p-p38MAPK protein in the LPS+UTI+miR-21 inhibitor group was increased(2.36±0.15). The expression of p-p38MAPK protein was decreased in the LPS +UTI+miR-21 mimc group(0.22±0.05). Conclusions Ulinastatin plays a protective role in sepsis, and its mechanism is related to the regulation of miR-21/p38MAPK pathway.
