Geniposide Inhibited Lipopolysaccharide-induced Apoptosis by Modulating TLR4 and Apoptosis-related Factors in Mouse Mammary Glands
Geniposide Inhibited Lipopolysaccharide-induced Apoptosis by Modulating TLR4 and Apoptosis-related Factors in Mouse Mammary Glands
复制标题
京尼平苷通过调节 TLR4 和凋亡相关因子抑制脂多糖诱导的小鼠乳腺细胞凋亡
DOI：
10.1016/j.lfs.2014.10.006
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发表时间：
2014-12-05
期刊：
影响因子：
6.1
通讯作者：
Zhang, Naisheng
中科院分区：
文献类型：
--
作者：
Song, Xiaojing;Guo, Mengyao;Zhang, Naisheng
目的：京尼平苷是栀子果实中发现的一种主要环烯醚萜苷，因其抗炎、抗肿瘤和抗细胞凋亡活性而在亚洲国家广泛使用。尽管京尼平苷的抗炎作用已被广泛报道，但其在乳腺炎中的抗细胞凋亡作用仍不清楚。本研究旨在探讨京尼平苷在脂多糖（LPS）诱导的小鼠乳腺中是否具有抗凋亡活性。主要方法：建立LPS诱导的小鼠乳腺炎模型和LPS刺激的原代小鼠乳腺上皮细胞（mMECs）模型，探讨京尼平苷的抗凋亡作用及其潜在作用机制。在体内研究中，通过TUNEL检测乳腺细胞凋亡。采用实时定量聚合酶链反应（qRT-PCR）和Western blot分析Bax、Bcl-2、Caspase-3和p53的表达。在体外研究中，通过活死染色测量乳腺上皮细胞的凋亡。采用Western blot和qRT-PCR分析Bax、Bcl-2、Caspase-3、p53和TLR4的表达。主要发现：京尼平苷在体内减轻乳腺细胞凋亡，下调Bax表达，抑制Caspase-3裂解和p53磷酸化，上调Bcl-2表达。在体外，京尼平苷以剂量依赖性方式降低死细胞比例。京尼平苷抑制 Bax 表达和 Caspase-3 裂解，并上调 Bcl-2 表达。此外，京尼平苷还能下调 TLR4 的表达并抑制 p53 的磷酸化。 意义：这些结果表明京尼平苷的抗凋亡特性是由于其在 LPS 诱导的小鼠乳腺炎中对 TLR4 和凋亡相关因子（p53、Bax、Bcl-2 和 Caspase-3）的调节。 (C) 2014 Elsevier Inc. 保留所有权利。
Aims: Geniposide, a major iridoid glycoside found in gardenia fruit, is widely used in Asian countries for its anti-inflammatory, anti-tumor and anti-apoptotic activities. Although the anti-inflammatory effect of geniposide has been widely reported, its anti-apoptotic role in mastitis remains unclear. In the present study, we investigated whether geniposide exerts anti-apoptotic activity in lipopolysaccharide (LPS)-induced mouse mammary glands.Main methods: We established a LPS-induced mouse mastitis model and LPS-stimulated primary mouse mammary epithelial cells (mMECs) model to investigate the anti-apoptotic effect of geniposide and the underlying mechanism of action. In the in vivo studies, apoptosis in mammary glands was detected by TUNEL. Quantitative real-time polymerase chain reaction (qRT-PCR) and Western blot were used to analyze the expression of Bax, BcI-2, Caspase-3 and p53. In the in vitro study, the apoptosis in mammary epithelial cells was measured by Live-Dead staining. Western blot and qRT-PCR analysis were used to analyze the expression of Bax, BcI-2, Caspase-3, p53 and TLR4.Key findings: Geniposide alleviated mammary gland apoptosis, down-regulated Bax expression, inhibited Caspase-3 cleavage and p53 phosphorylation and up-regulated BcI-2 expression in vivo. In vitro, geniposide decreased the ratio of dead cells in a dose-dependent manner. Geniposide inhibited Bax expression and Caspase-3 cleavage, and up-regulated the expression of BcI-2. Moreover, geniposide down-regulated the expression of TLR4 and repressed the phosphorylation of p53.Significance: These results demonstrate that the anti-apoptotic property of geniposide is due to its modulation of TLR4 and apoptosis-related factors (p53, Bax, BcI-2 and Caspase-3) in LPS-induced mouse mastitis. (C) 2014 Elsevier Inc. All rights reserved.