[尾加压素受体抑制剂尿降压肽对小鼠急性肝细胞凋亡的影响]

[Effects of urantide, a urotensin receptor inhibitor, on acute hepatocyte apoptosis in mice].

作者信息

Yu Fang-ping, Zhao Liang, Liu Liang-ming, Liang Dong-yu, Yang Dao-hua, Zhang Fang-fang, Ye Chang-gen

机构信息

Cadres Ward, Shanghai Songjiang Central Hospital Affiliated to Nanjing Medical University, Songjiang Hospital Affiliated to First People's Hospital, Shanghai Jiaotong University, Shanghai 201600, China.

出版信息

Zhonghua Yi Xue Za Zhi. 2011 Dec 20;91(47):3358-62.

PMID:22333205

Abstract

OBJECTIVE

To explore the effects of urantide, a urotensin II receptor (UT) inhibitor, on lipopolysaccharide (LPS)/D-galactosamine (D-GalN)-induced acute hepatocyte apoptosis in mice.

METHODS

Male BALB/c mice were randomly divided into 4 groups (n = 6 each): normal control, pre-treatment control, model and pre-treatment model. The pre-treatment control and pre-treatment model groups received urantide (0.6 mg/kg body weight) by a caudal vein injection. At 30 minutes post-injection, the model and pre-treatment model groups were treated with LPS/D-GalN to induce acute hepatocyte apoptosis via an intraperitoneal injection. Hepatocyte apoptosis was examined by terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling (TUNEL) and caspase-3 colorimetric assay. The expressions of proinflammatory cytokines, such as tumor necrosis factor alpha (TNF-α), interferon-γ (IFN-γ) and interleukin-1 beta (IL-1β), were detected by reverse transcription-polymerase chain reaction (RT-PCR) and enzyme-linked immunosorbent assay.

RESULTS

Massive hepatocyte apoptosis were detected in the model group. The apoptotic index was clearly reduced in the pre-treatment model group [(26 ± 11)% vs (77 ± 20)%, P < 0.01]. And the activity of caspase-3 was also lower in the pre-treatment model group than that in the model group [(2.50 ± 0.83) pmol · min(-1) · mg(-1) vs (3.76 ± 0.42) pmol · min(-1) · mg(-1), P < 0.01]. In addition, the serum and liver tissue levels of TNF-α, IL-1β and IFN-γ in the pre-treatment model group were significantly lower than those in the model group[1.69 ± 0.47 vs 3.57 ± 0.79, 0.31 ± 0.02 vs 0.46 ± 0.06, 2.81 ± 0.72 vs 3.35 ± 0.84, (233 ± 36) pg/ml vs (441 ± 157) pg/ml, (228 ± 21) pg/ml vs (364 ± 20) pg/ml, (93.8 ± 5.2) pg/ml vs (180.3 ± 4.3) pg/ml, all P < 0.01].

CONCLUSION

LPS/D-GalN-induced acute hepatocyte apoptosis can be inhibited by a pretreatment of urantide through an inhibition of expression and secretion of proinflammatory cytokines. The UII/UT receptor system plays a pivotal role in the liver immuno-inflammatory injury of acute liver failure (ALF). And it may become a new drug target of ALF therapy.

摘要

目的

探讨尾加压素Ⅱ受体（UT）抑制剂尿抑胃素对脂多糖（LPS）/D-氨基半乳糖（D-GalN）诱导的小鼠急性肝细胞凋亡的影响。

方法

雄性BALB/c小鼠随机分为4组（每组n = 6）：正常对照组、预处理对照组、模型组和预处理模型组。预处理对照组和预处理模型组通过尾静脉注射给予尿抑胃素（0.6 mg/kg体重）。注射后30分钟，模型组和预处理模型组通过腹腔注射LPS/D-GalN诱导急性肝细胞凋亡。通过末端脱氧核苷酸转移酶介导的dUTP-生物素缺口末端标记法（TUNEL）和半胱天冬酶-3比色法检测肝细胞凋亡。通过逆转录-聚合酶链反应（RT-PCR）和酶联免疫吸附测定法检测促炎细胞因子如肿瘤坏死因子α（TNF-α）、干扰素-γ（IFN-γ）和白细胞介素-1β（IL-1β）的表达。

结果

模型组检测到大量肝细胞凋亡。预处理模型组的凋亡指数明显降低[（26±11）%比（77±20）%，P < 0.01]。预处理模型组半胱天冬酶-3的活性也低于模型组[（2.50±0.83）pmol·min⁻¹·mg⁻¹比（3.76±0.42）pmol·min⁻¹·mg⁻¹，P < 0.01]。此外，预处理模型组血清和肝组织中TNF-α、IL-1β和IFN-γ的水平明显低于模型组[1.69±0.47比3.57±0.79，0.31±0.02比0.46±0.06，2.81±0.72比3.35±0.84，（233±36）pg/ml比（441±157）pg/ml，（228±21）pg/ml比（364±20）pg/ml，（93.8±5.2）pg/ml比（180.3±4.3）pg/ml，均P < 0.01]。