CD31 induces inflammatory response by promoting hepatic inflammatory response and cell apoptosis
CD31 induces inflammatory response by promoting hepatic inflammatory response and cell apoptosis
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DOI：
10.26355/eurrev_201811_16296
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发表时间：
2018-11-01
影响因子：
3.3
通讯作者：
Li, Y-Y
中科院分区：
文献类型：
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作者：
Cheng, G-Y;Jiang, Q.;Li, Y-Y
目的：目的：探讨CD 31对扑热息痛诱导的肝损伤的调节作用，为药物性肝炎的防治提供新的方向。材料与方法：野生型（WT）小鼠分别给予对乙酰氨基酚（APAP）（250 mg/kg）或等剂量的磷酸盐缓冲液（PBS）。分别于给药后1、3、6、12 h，采用实时荧光定量逆转录聚合酶链反应（RT-PCR）和免疫印迹法（Western blotting）检测小鼠肝脏CD 31 mRNA和蛋白表达水平。一旦证实CD 31参与APAP诱导的肝损伤，则建立APAP诱导的WT小鼠和CD 31基因缺陷（CD 31-/-）小鼠的急性肝损伤模型。分别于注射APAP（250 mg/kg）后8 h和24 h采集血清，测定血清丙氨酸氨基转移酶（ALT）活性。取小鼠肝组织，HE染色观察肝组织病理变化。同时，从小鼠肝组织中分离单个核细胞（MNCs）。流式细胞仪检测浸润的巨噬细胞和中性粒细胞的数量，并分析这些细胞的活化水平。RT-PCR检测肝组织TNF-α、IL-1 β、KC、MCP-1、IL-6等促炎细胞因子的表达水平。采用酶联免疫吸附试验（ELISA）检测血清中细胞因子的表达水平。Western blotting检测肝组织JNK、Caspase-3和细胞色素P450 2 E1（CYP 2 E1）蛋白表达水平。WT小鼠ALT水平明显高于CD 31-/-小鼠，同时WT小鼠肝细胞坏死或凋亡明显增多。结果还表明，炎症细胞因子，包括KC，IL-1 β，MCP-1和IL-6的表达水平在WT小鼠中显著较高。WT小鼠肝组织中浸润的巨噬细胞和中性粒细胞数量明显多于CD 31-/-小鼠。结论：APAP处理的CD 31-/-小鼠肝损伤较WT小鼠轻。我们还证实了CD 31通过促进肝脏炎症反应和细胞凋亡参与了APAP诱导的炎症反应，这可能为药物性肝炎的防治提供了新的策略。
OBJECTIVE: To investigate whether CD31 could regulate paracetamol-induced liver injury, thereby providing a new direction for the prevention and treatment of drug-induced hepatitis.MATERIALS AND METHODS: Wild-type (WT) mice were treated with acetaminophen (APAP) (250 mg/kg) or isodose of phosphate-buffered saline (PBS). 1, 3, 6 and 12 h after the treatment, the messenger RNA (mRNA) and protein expression level of CD31 in the liver of mice were determined by Real-time reverse transcription polymerase chain reaction (RT-PCR) and Western blotting, respectively. Once CD31 was confirmed to be involved in APAP-induced liver injury, the acute liver injury model in WT mice and CD31 gene deficient (CD31-/-) mice induced by APAP was established. Serum samples were collected at 8 and 24 h after APAP injection (250 mg/kg), and the activity of serum alanine aminotransferase (ALT) was measured. The liver tissues of mice were isolated and analyzed by hematoxylin and eosin (HE) staining. Meanwhile, mononuclear cells (MNCs) were isolated from the liver tissues of mice. The number of infiltrating macrophages and neutrophils was detected by flow cytometry, and the activation level of these cells was analyzed. The expression levels of proinflammatory cytokines in liver tis- sues, such as TNF-alpha, IL-1 beta, keratinocyte chemo- attractant (KC), MCP-1 and IL-6, were determined by RT-PCR. The expression levels of cytokines in serum were detected by enzyme-linked immunosorbent assay (ELISA). Moreover, the protein expression levels of JNK, Caspase-3, and cytochrome P450 2E1 (CYP2E1) in liver tissues were detected by Western blotting.RESULTS: After APAP treatment, we found that WT mice were more sensitive to APAP-induced liver injury. The level of ALT in WT mice was significantly higher than that of CD31-/- mice, meanwhile, more necrotic or apoptotic cells were found in WT mice. Results also indicated that the expression levels of inflammatory cytokines, including KC, IL-1 beta, MCP-1 and IL-6, were significantly higher in WT mice. Meanwhile, the number of infiltrating macrophages and neutrophils in the liver tissues of WT mice were much more than that of CD31-/- mice.CONCLUSIONS: APAP-treated CD31-/- mice exhibited less liver injury when compared with WT mice. We also confirmed that CD31 was greatly involved in APAP-induced inflammatory response by promoting hepatic inflammatory and cell apoptosis, which might provide a new strategy for the prevention and treatment of drug-induced hepatitis.