沙参麦冬汤对新生大鼠支气管肺发育不良的保护作用及其机制

Protective effect of Radix Salviae Miltiorrhizae Decoction on bronchopulmonary dysplasia in neonatal rats and its mechanism

目的:观察沙参麦冬汤作用下慢性支气管肺发育不良(BPD)新生大鼠肺的发育情况,阐明沙参麦冬汤对高体积分数氧(高氧)所致新生大鼠慢性BPD的影响及其作用机制。方法:200只新生大鼠分为对照组,模型组,地塞米松组,低和高剂量沙参麦冬汤组,每组40只。除对照组外,其余4组大鼠采用持续吸入高氧造模,沙参麦冬汤组和地塞米松组大鼠灌胃给予6.00和24.0 mg·kg-1沙参麦冬汤及0.75μg·kg-1地塞米松注射液,对照组大鼠给予等量生理盐水。给药21 d后检测各组大鼠肺组织湿质量/干质量(W/D)值,HE染色法观察各组大鼠肺组织病理形态表观,计算各组大鼠肺组织中辐射状肺泡计数(RAC),检测各组大鼠肺组织中丙二醛(MDA)、超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-Px)水平,ELISA方法检测各组大鼠肺组织中细胞炎性因子白细胞间介素1(IL-1)、白细胞间介素10(IL-10)、肿瘤坏死因子α(TNF-α)水平,Western blotting法检测各组大鼠肺组织中促凋亡蛋白Bax、抗凋亡蛋白Bcl-2和caspase-3蛋白表达水平。结果:与对照组比较,模型组大鼠肺组织W/D值明显升高(t=3.144,P<0.05),可见明显肺水肿;氧暴露21 d后肺泡腔扩大、结构简化,间隔增厚,RAC明显减少(t=3.989,P<0.05),炎症细胞浸润,肺组织中MDA、SOD、GSH-Px、IL-1和TNF-α水平以及Bax和caspase-3蛋白表达水平明显升高(t=6.562,t=1.971,t=1.972,t=20.241,t=32.808,t=22.663,t=19.234,P<0.05),IL-10水平明显降低(t=7.857,P<0.05);与模型组比较,地塞米松组和不同剂量沙参麦冬汤组大鼠肺水肿程度改善,肺组织W/D值明显降低(t=1.018,t=2.691,t=0.760,P<0.05),肺泡结构紊乱情况减轻,肺组织中MDA、SOD、GSH-Px、IL-1和TNF-α水平以及Bax和caspase-3蛋白表达水平降低(t=6.562,t=1.971,t=1.972,t=20.241,t=32.808,t=22.663,t=19.234,P<0.05),IL-10水平和Bcl-2蛋白表达水平升高(t=7.857,t=7.743,P<0.05),且以高剂量沙参麦冬汤组效果为佳。结论:沙参麦冬汤能有效保护由高氧造成的新生大鼠支气管和肺的损伤。

Objective:To observe the developmental situation of the lung of the neonatal rats with chronic bronchopulmonary dysplasia(BPD)after treated with Radix Salviae Miltiorrhizae Decoction,and to clarify the effect of Radix Salviae Miltiorrhizae Decoction on the BPD in the neonatal rats induced by high-volume fractinal oxygen(hyperoxia)and its mechanism.Methods:A total of 200 neonatal rats were divided into control group,model group,dexamethasone group,low and high doses of Radix Salviae Miltiorrhizae Decoction groups,and there were 40 rats in each group.Except for the rats in control group,the rats in the other four groups were induced by continuous inhalation of hyperoxia.The rats in dexamethasone group and Radix Salviae Miltiorrhizae Decoction group were treated with dexamethasone solutions(0.75μg·kg-1)and 6.00 amd 24.00 mg·kg-1 Radix Salviae Miltiorrhizae Decoction,respectively,and the rats in control group were given the equal volume of normal saline.After 21 d of administration,the wet mass/dry mass(W/D)values of lung tissue of the rats in various groups were measured,thePathomorphology of lung tissue of the rats in various groups were observed by HE staining,and the number of radical alveolar counts(RAC)in lung tissue of the rats in various groups was calculated;the levels of malondialdehyde(MDA),superoxidedismutase(SOD)and glutathionePeroxidase(GSH-Px)in lung tissue of the rats in various groups were detected,the levels of interleukin-1(IL-1),interleukin-10(IL-10)and tumor necrosis factor-α(TNF-α)in lung tissue of the rats in various groups were determined by ELISA method,the expression levels of Bax,Bcl-2 and caspase-3Proteins in lung tissue of the rats in various groups were measured by Western blotting method.Results:Compared with control group,the W/D value of lung tissue of the rats in model group was increased significantly(t=3.144,P<0.05),and the obviousPulmonary edema was found.After 21 d of oxygen exposure,the lung tissue of the rats in model group was shown as enlarged alveolar cavity,simplified structure,and thickened interval,and the number of RAC was decreased significantly(t=3.989,P<0.05);the inflammatory cell infiltration and the levels of MDA,SOD,GSH-Px,IL-1β,and TNF-αand the expression levels of Bax and caspase-3Proteins in lung tissue of the rats in model group were increased significantly(t=6.562,t=1.971,t=1.972,t=20.241,t=32.808,t=22.663,t=19.234,P<0.05),and the level of IL-10 in lung tissue of the rats was decreased significantly(t=7.857,P<0.05).Compared with model group,the degrees ofPulmonary edema of the rats in dexamethasone and Radix Salviae Miltiorrhizae Decoction groups were improved,and the W/D values of lung tissue were significantly decreased(t=1.018,t=2.691,t=0.760,P<0.05),and the alveolar structural disorder was alleviated;the levels of MDA,SOD,GSH-Px,IL-1,and TNF-αand the expression levels of Bax and caspase-3Proteins in lung tissue of the rats were decreased(t=6.562,t=1.971,t=1.972,t=20.241,t=32.808,t=22.663,t=19.234,P<0.05),the levels of IL-10 and the expression levels of Bcl-2Protein were increased(t=7.857,t=7.743,P<0.05),expecially in high dose of Radix Salviae Miltiorrhizae Decoction group.Conclusion:Radix Salviae Miltiorrhizae Decoction can effectivelyProtect the bronchus and lung injuries induced by hyperoxia in the neonatal rats.