橙皮素对矽尘暴露大鼠肺组织中细胞因子的影响

Effects of hesperetin on cytokines in lung tissue of rats exposed to silica

目的探讨橙皮素干预对二氧化硅染尘大鼠肺组织损伤的拮抗作用及其对细胞因子的影响。方法将54只健康SPF级Wistar雄性大鼠随机分为6组,分别为阴性对照(生理盐水)组、二氧化硅(50 mg/ml)模型组、吡非尼酮(100 mg/kg)对照组和低(100 mg/kg)、中(200 mg/kg)、高剂量(400 mg/kg)橙皮素治疗组,每组9只。二氧化硅模型组、吡非尼酮对照组和不同剂量橙皮素治疗组均给予1 ml二氧化硅溶液(50 mg/ml)一次性经气管灌注造模;24 h后,吡非尼酮对照组和各剂量橙皮素治疗组采用灌胃方式进行干预,染毒容量为10 ml/kg,每天1次,连续28 d;阴性对照组和二氧化硅模型组均给予等容积生理盐水。检测大鼠肺组织中转化生长因子-β1(TGF-β1)、肿瘤坏死因子-α(TNF-α)、白细胞介素-1β(IL-1β)、IL-4、IL-10、干扰素-γ(IFN-γ)的水平,并进行肺组织肺泡炎及纤维化评分。结果与阴性对照组比较,二氧化硅模型组、吡非尼酮对照组和各剂量橙皮素治疗组大鼠的肺泡炎和肺纤维化评分均较高,差异有统计学意义(P<0.05);且随着橙皮素染毒剂量的升高,大鼠的肺泡炎和肺纤维化评分均呈下降趋势。与二氧化硅模型组比较,吡非尼酮对照组和各剂量橙皮素治疗组大鼠的肺泡炎和肺纤维化评分均较低,差异有统计学意义(P<0.05)。与阴性对照组比较,二氧化硅模型组、吡非尼酮对照组和各剂量橙皮素治疗组大鼠肺组织中TGF-β1、IL-1β、TNF-α、IL-4、IL-10和IFN-γ的水平均较高,差异有统计学意义(P<0.05);且随着橙皮素染毒剂量的升高,大鼠肺组织中TGF-β1、IL-1β的水平均呈下降趋势,TNF-α、IL-4、IL-10的水平均呈先上升后下降的趋势,而IFN-γ的水平呈上升趋势。与二氧化硅模型组比较,吡非尼酮对照组和各剂量橙皮素治疗组大鼠肺组织中TGF-β1、IL-1β、TNF-α、IL-4的水平均降低,除低剂量橙皮素治疗组IL-1β和吡非尼酮对照组IL-4外,差异均有统计学意义(P<0.05);而中、高剂量橙皮素治疗组大鼠肺组织中IL-10的水平和高剂量橙皮素治疗组大鼠肺组织中的IFN-γ水平均较高,差异有统计学意义(P<0.05)。结论橙皮素可能通过抑制促炎因子的释放、促进抗炎因子的分泌从而减轻二氧化硅对肺组织的损伤作用,进而抑制矽肺纤维化的形成和发展。

Objective To understand the antagonism of hesperetin on rat lung damage and cytokines induced by silica.Methods A total of 54 healthy SPF male Wistar rats were randomly divided into six groups:negative control group(normal saline),silica model group(50 mg/ml),pirfenidone control group(100 mg/kg),and low(100 mg/kg),moderate(200 mg/kg)and high dose(400 mg/kg)hesperetin treatment group,nine rats in each group.The silica model group,the pirfenidone control group and treatment groups of hesperetin with different doses were all given 1 ml of silica solution(50 mg/ml)once through tracheal infusion to develop the damage models;Then 24 hours later,the rats in pirfenidone control group and each hesperetin treatment group were treated with pirfenidone or hesperetin through gavage at 10 ml/kg once a day for 28 consecutive days.Both the negative control group and the silica model group were given equal volume of normal saline.The levels of transforming growth factor-β1(TGF-β1),tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β),IL-4,IL-10 and interferon-γ(IFN-γ)in lung tissue of rats were determined,and the pulmonary alveolitis and fibrosis scores were evaluated.Results Compared with the negative control group,the alveolitis and pulmonary fibrosis scores of the silica model group,the pirfenidone control group and the various hesperetin treatment groups were significantly higher(P<0.05);With the increase of the dose of hesperetin,the alveolitis and pulmonary fibrosis scores of rats showed a downward trend.Compared with the silica model group,the alveolitis and pulmonary fibrosis scores of the pirfenidone control group and the various doses of hesperetin treatment group were significantly lower(P<0.05).Compared with negative control group,the levels of TGF-β1,IL-1β,TNF-α,IL-4,IL-10 and IFN-γin lung tissue of rats in silica model group,pifenidonecontrol group and hesperetin treatment groups were significantly higher(P<0.05);With the increase of the dosage of hesperetin,the levels of TGF-β1 and IL-1βin the lung tissue of rats showed a downward trend,and the levels of TNF-α,IL-4 and IL-10 all showed an increasing trend first and then decreasing,while the levels of IFN-γshowed an increasing trend.Compared with the silica model group,the levels of TGF-β1,IL-1β,TNF-α,and IL-4 in the lung tissue of the pirfenidone control group and each hesperetin-treated group were decreased,except the levels of IL-1βin the low-dose hesperetin treatment group and IL-4 in the pirfenidone control group,the difference was statistically significant(P<0.05);And significant higher levels of IL-10 were observed in lung tissue in the moderate and high dose groups of hesperetin and significant higher levels of IFN-γwere observed in lung tissue in the high dose group of hesperetin(P<0.05).Conclusion Hesperetin may inhibit the lung injury and fibrosis in rats induced by silica by inhibiting the release of pro-inflammatory factors and promoting the secretion of anti-inflammatory factors.