海黄补肺方对矽肺大鼠炎症因子和T淋巴细胞亚群的影响

Effects of Haihuang Bufei Formula on inflammatory factors and T lymphocyte subsets in silicotic rats

目的探讨海黄补肺方对矽肺大鼠炎症因子和T淋巴细胞亚群的影响,进一步探讨海黄补肺方的治疗作用。方法采用气管滴注二氧化硅(silicon dioxide,SiO2)悬液的方法建立大鼠矽肺模型,造模后24 h随机分成正常对照组、模型组、阳性药对照组及海黄补肺方低、中、高剂量组,每组10只,每天给药1次,连续给药1个月。末次给药1 h后处理大鼠,测定大鼠肺指数,血清肿瘤坏死因子α(tumor necrosis factorα,TNF-α)、白细胞介素1β(interleukin 1β,IL-1β),肺组织羟脯氨酸(hydroxyproline,HYP)、超氧化物歧化酶(superoxide dismutase,SOD)及丙二醛(malondialdehyde,MDA)含量,苏木精-伊红(hematoxylin-eosin,HE)、Masson染色法观察大鼠肺组织病理变化,流式细胞仪检测大鼠外周血T淋巴细胞亚群。结果与模型组比较,海黄补肺方中、高剂量组大鼠的肺指数,血清IL-1β,肺组织HYP、MDA含量降低(P<0.05),海黄补肺方高剂量组血清TNF-α含量降低[(238.64±51.80)ng/L]、肺组织SOD含量升高[(55.30±13.77)pg/mL](P<0.05)。HE、Masson染色结果显示海黄补肺方中、高剂量能减轻肺组织病理损伤及纤维化。流式细胞仪检测显示海黄补肺方高剂量组大鼠外周血分化簇(cluster of differentiation,CD)8^(+)T细胞百分含量降低(P<0.05),CD4^(+)/CD8^(+)比值升高(1.44±0.19)(P<0.05),CD4^(+)T细胞百分含量变化无统计学意义(P>0.05)。结论海黄补肺方对矽肺大鼠具有保护作用,可降低胶原蛋白含量,减轻肺组织病理损伤及纤维化,可能与其抑制炎症因子表达、减少氧化应激、调节机体免疫系统有关。

Objective To investigate the effects of Haihuang Bufei Formula on inflammatory factors and T lymphocyte subsets in silicotic rats,and to further explore the therapeutic effects of Haihuang Bufei Formula.Methods The silicotic rat models were constructed by intratracheal instillation of silicon dioxide(SiO2)suspension.After 24 hours of modeling,the rats were randomly divided into normal control group,model group,positive drug control group,and low,medium,high dose group of Haihuang Bufei Formula.Each group contained 10 rats,which were administered once daily for one month.One hour after the last administration,the rats were sacrificed.Lung index,serum tumor necrosis factorα(TNF-α),interleukin 1β(IL-1β),hydroxyproline(HYP),superoxide dismutase(SOD),and malondialdehyde(MDA)contents in lung tissue were measured.Hematoxylin-eosin(HE)and Masson staining methods were used to observe the pathological changes in lung tissues.Flow cytometry was used to detect the T lymphocyte subsets in the peripheral blood of rats.Results Compared with the model group,the lung index,serum IL-1β,lung tissue HYP,and MDA contents in the middle and high dose groups of Haihuang Bufei Formula treatment were significantly reduced(P<0.05).In the high-dose group,the serum TNF-αcontent decreased to(238.64±51.80)ng/L,and the lung tissue SOD content increased to(55.30±13.77)pg/mL(P<0.05).HE and Masson's staining results indicated that the medium and high doses of Haihuang Bufei Formula could alleviate pathological damage of silicosis and fibrosis in lung tissue.Flow cytometer analysis showed that the percentage of CD8^(+)T cells in the high-dose group of Haihuang Bufei Formula was significantly decreased(P<0.05).The ratio of CD4^(+)/CD8^(+)increased to(1.44±0.19)(P<0.01),and the percentage of CD4^(+)T cells showed no statistically significant change among groups(P>0.05).Conclusions Haihuang Bufei Formula has a protective effect on silicotic rats by reducing collagen content,alleviating pathological damage and fibrosis of lung tissue,which may be related to inhibiting the expression of inflammatory factors,reducing oxidative stress,and regulating the immune system.