Wu Qi, Zhou Yao, Zhou Xian-Mei
Physiology Department, Xuzhou Medical University, Xuzhou 221009, China.
Pathophysiology Department, Xuzhou Medical University, Xuzhou 221009, China.
Evid Based Complement Alternat Med. 2019 Feb 4;2019:1528586. doi: 10.1155/2019/1528586. eCollection 2019.
To investigate the intervention effect and functioning mechanism of citrus alkaline extract (CAE) on bleomycin- (BLM-) induced pulmonary fibrosis in mice.
42 C57BL/6 male mice were assigned randomly to the normal group, model group, low (16mg/kg), medial (32mg/kg) and high (64mg/kg) CAE dosage groups, prednisone group (6mg/kg), and pirfenidone group (100mg/kg), respectively. One day after model construction, intragastric administration was applied to the mice once a day for 28 days and then killed. Body weights of mice were recorded. Their pulmonary tissues were subjected to HE staining and Masson's staining and then their degree of pulmonary alveolitis as well as pulmonary fibrosis was scored. Contents of hydroxyproline (HYP) and prostaglandin E2 (PGE2) in pulmonary tissues and levels of interleukin-17 (IL-17) in serum and bronchoalveolar lavage fluid (BALF) were determined by ELISA method. Expression of collagen I, collagen III, and Prosurfactant protein C (Pro-SPC) proteins in pulmonary tissue were measured immunohistochemically and that of nuclear transcription factor B (NF-B) and vimentin was determined by the immunofluorescence method. Apoptosis of pulmonary tissue was tested by the Tunel staining method, while the expression of MAPK-related protein was recorded by Western Blot assay.
After CAE treatment, the body weight, PGE2 level, and Pro-SPC protein expression of pulmonary fibrosis mice were increased, while the score of pulmonary alveolitis and pulmonary fibrosis, levels of HYP and cell apoptosis, IL-17 contents of serum and BALF in pulmonary tissues, and expression of collagen I, collagen III, vimentin, NF-B, and p-p38 were reduced.
CAE effectively delayed the progression of BLM-induced pulmonary fibrosis in pulmonary fibrosis mice and a possible mechanism is the inhibition of cell apoptosis of NF-B/p38-mediated signaling pathway.
探讨柑橘碱性提取物（CAE）对博来霉素（BLM）诱导的小鼠肺纤维化的干预作用及作用机制。
将42只C57BL/6雄性小鼠随机分为正常组、模型组、低剂量（16mg/kg）CAE组、中剂量（32mg/kg）CAE组、高剂量（64mg/kg）CAE组、泼尼松组（6mg/kg）和吡非尼酮组（100mg/kg）。造模后1天开始对小鼠进行灌胃给药，每天1次，共28天，然后处死。记录小鼠体重。对其肺组织进行HE染色和Masson染色，然后对肺泡炎和肺纤维化程度进行评分。采用ELISA法测定肺组织中羟脯氨酸（HYP）和前列腺素E2（PGE2）含量以及血清和支气管肺泡灌洗液（BALF）中白细胞介素-17（IL-17）水平。采用免疫组织化学法检测肺组织中Ⅰ型胶原、Ⅲ型胶原和表面活性蛋白C（Pro-SPC）蛋白的表达，采用免疫荧光法检测核转录因子B（NF-κB）和波形蛋白的表达。采用Tunel染色法检测肺组织细胞凋亡情况，采用Western Blot法检测MAPK相关蛋白的表达。
CAE治疗后，肺纤维化小鼠的体重、PGE2水平和Pro-SPC蛋白表达增加，而肺泡炎和肺纤维化评分、HYP水平、细胞凋亡、肺组织血清和BALF中IL-17含量以及Ⅰ型胶原、Ⅲ型胶原、波形蛋白、NF-κB和p-p38的表达降低。
CAE有效延缓了肺纤维化小鼠BLM诱导的肺纤维化进展，其可能机制是抑制NF-κB/p38介导的信号通路细胞凋亡。
