Department of Critical Care Medicine, Hunan Cancer Hospital, No.283 of Tongzipo Road, Changsha, Hunan, China.
Department of Thoracic Radiotherapy, Hunan Cancer Hospital, No.283 of Tongzipo Road, Changsha, Hunan, China.
Apoptosis. 2017 Sep;22(9):1147-1156. doi: 10.1007/s10495-017-1393-z.
To investigate the inhibitory effects of chlorogenic acid on pulmonary fibrosis and the internal mechanisms in vivo and in vitro. 30 male BALB/C mice were randomized into 5 groups: control group, pulmonary fibrosis model group, low, middle and high dose of chlorogenic acid groups. Mice in pulmonary fibrosis model group were administered 5.0 mg/kg bleomycin with intracheal instillation and mice in 3 chlorogenic acid groups were treated with chlorogenic acid every day for 28 days after bleomycin administration. Lung tissue histology was observed using HE staining. Primary pulmonary fibroblasts were isolated and cultured. The expressions of fibrosis related factors (α-SMA and collagen I), as well as ER stress markers (CHOP and GRP78) were determined by both real-time PCR assay and Western blotting, while the expressions of other ER stress signaling pathway factors PERK, IRE-1, ATF-6 and protein levels of caspase-12, caspase-9, caspase-3, PARP were determined by Western blotting. RLE-6TN cell line induced by TGF-β1 was also used to verify the amelioration effects in vitro study. In both in vivo and in vitro studies, TUNEL staining was used to evaluate cell apoptosis. Expressions of collagen I, α-SMA, GRP78, and CHOP were significantly inhibited by chlorogenic acid in dose-dependent manner. Similarly, decreasing levels of cleaved caspase-12, caspase-9, caspase-3 and increasing level of uncleaved PARP were observed in chlorogenic acid groups compared with those in the fibrosis group both in vivo and in vitro. Chlorogenic acid could also significantly down-regulate the level of phosphorylation of PERK and cleaved ATF-6 in vivo study. Moreover, MTT assay demonstrated chlorogenic acid could enhance proliferation of RLE-6TN cells induced by TGFβ1 in vitro. And the apoptosis assays indicated that chlorogenic acid could significantly inhibit cell apoptosis both in vivo and in vitro studies. Chlorogenic acid could inhibit the pulmonary fibrosis through endoplasmic reticulum stress inhibition in vivo and in vitro.
研究绿原酸对肺纤维化的抑制作用及其体内和体外的作用机制。
30 只雄性 BALB/C 小鼠随机分为 5 组：对照组、肺纤维化模型组、低、中、高剂量绿原酸组。肺纤维化模型组小鼠经气管内滴注 5.0mg/kg 博来霉素，绿原酸组小鼠在博来霉素给药后每天给予绿原酸治疗 28 天。采用 HE 染色观察肺组织病理学变化。分离培养原代肺成纤维细胞。采用实时 PCR 法和 Western blot 法检测纤维化相关因子（α-SMA 和胶原 I）以及内质网应激标志物（CHOP 和 GRP78）的表达，Western blot 法检测其他内质网应激信号通路因子 PERK、IRE-1、ATF-6 以及 caspase-12、caspase-9、caspase-3、PARP 的蛋白水平。还使用 TGF-β1 诱导的 RLE-6TN 细胞系验证体外研究的改善作用。在体内和体外研究中，均采用 TUNEL 染色评估细胞凋亡。
绿原酸呈剂量依赖性抑制体内和体外的胶原 I、α-SMA、GRP78 和 CHOP 的表达。同样，与纤维化组相比，体内和体外的绿原酸组均观察到 cleaved caspase-12、caspase-9、caspase-3 的水平降低和未切割的 PARP 水平升高。体内研究还表明，绿原酸还可以显著下调 PERK 的磷酸化水平和 cleaved ATF-6 的水平。此外，MTT 试验表明，绿原酸可增强 TGFβ1 诱导的 RLE-6TN 细胞的增殖。凋亡试验表明，绿原酸可显著抑制体内和体外的细胞凋亡。
绿原酸可通过抑制内质网应激在体内和体外抑制肺纤维化。
