清肺通络方对肺炎支原体肺炎小鼠体内氧化应激的影响

目的探讨清肺通络方对肺炎支原体肺炎模型小鼠体内氧化应激的影响。方法将100只BALB/c小鼠随机分为正常组、模型组、清肺通络方组、阿奇霉素组、中西医结合组,每组20只。正常组给予0.9%氯化钠50μL/次早晚滴鼻,其余组给予支原体国际标准株50μL/次早晚缓慢滴鼻腔造模。造模第1天开始,正常组和模型组给予生理盐水连续灌胃10 d;清肺通络方组给予清肺通络方连续灌胃10 d;阿奇霉素组给予阿奇霉素药液灌胃,第1~3天和第8~10天;中西医结合组给予阿奇霉素药液灌胃,第1~3天和第8~10天,同时给予清肺通络方连续灌胃10 d。实验第6天、第11天每组每次各处死小鼠10只,HE染色观察肺组织病理生理形态,PCR法检测肺组织中肺炎支原体DNA含量,生化法检测肺泡灌洗液、肺组织匀浆、血清标本中一氧化氮合酶(NOS)、诱导型一氧化氮合酶(iNOS)、一氧化氮(NO)、丙二醛(MDA)、超氧化物歧化酶(SOD)表达水平。结果清肺通络方组、阿奇霉素组、中西医结合组小鼠肺组织病理改变较模型组有不同程度改善,其中中西医结合组改善最明显。实验第6天和第11天,模型组小鼠肺组织中肺炎支原体DNA含量均显著增高(P均<0.05),清肺通络方组、阿奇霉素组和中西医结合组均明显低于同期模型组(P均<0.05),且中西医结合组明显低于同期清肺通络方组和阿奇霉素组(P均<0.05)。实验第6天和第11天,模型组小鼠肺泡灌洗液、肺组织、血清中NOS、iNOS、NO、MDA表达水平均明显高于正常组(P均<0.05),SOD表达水平均明显低于正常组(P均<0.05);清肺通络方组、阿奇霉素组和中西医结合组NOS、iNOS、NO、MDA表达水平均明显低于模型组(P均<0.05),SOD表达水平均明显高于模型组(P均<0.05)。实验第6天,阿奇霉素组肺泡灌洗液中NOS、NO、SOD,肺组织中iNOS、NO、MDA,血清中NOS、MDA表达水平改善情况均明显优于清肺通络方组(P均<0.05);中西医结合组肺泡灌洗液中NOS、iNOS、NO表达水平,肺组织中iNOS表达水平,血清中NO、MDA表达水平均明显低于清肺通络方组和阿奇霉素组(P均<0.05),肺泡灌洗液中MDA表达水平,肺组织中NO、MDA表达水平,血清中NOS、iNOS表达水平均明显低于清肺通络方组(P均<0.05),而各样本中SOD表达水平均明显高于清肺通络方组和阿奇霉素组(P均<0.05)。实验第11天,阿奇霉素组与清肺通络方组上述各指标比较差异均无统计学意义(P均>0.05);中西医结合组肺泡灌洗液、肺组织中SOD表达水平均明显高于清肺通络方组(P均<0.05)。结论清肺通络方可通过抑制NOS、iNOS活力,降低NO、MDA含量,提高SOD活力发挥体内抗氧化作用,从而减轻肺部炎症损伤,清肺通络方与阿奇霉素联用作用更为显著。

Effect of Qingfei Tongluo Decoction on oxidative stress in mice with mycoplasma pneumoniae pneumonia

Objective It is to investigate the effect of Qingfei Tongluo Decoction(QTD)on oxidative stress in mice with mycoplasma pneumoniae pneumonia.Methods 100 BALB/c mice were randomly divided into normal group,model group,QTD group,azithromycin group,and integrated traditional Chinese and western medicine group(combination group),with 20 mice in each group.The normal group was given 0.9% sodium chloride 50μL by nasal drip every morning and night,and the other groups were given 50μL international standard strain of mycoplasma by slowly instilling the nasal cavity every morning and night for modeling.From the first day of modeling,the normal group and model group were treated with normal saline by gavage for 10 days;the QTD group was treated with QTD by gavage for 10 days;the azithromycin group was given azithromycin solution by gavage,from 1st-3rd day and 8th to 10th day;the combination group was given azithromycin liquid by gavage from the 1st to 3rd and 8th to 10th days,QTD by gavage for 10 consecutive days.On the 6th and 11th day of the experiment,10 mice were killed each time in each group.The pathophysiological morphology of lung tissue was observed by HE staining.The DNA content of mycoplasma pneumoniae in lung tissue was detected by PCR method.The expression levels of nitric oxide synthase(NOS),inducible nitric oxide synthase(iNOS),nitric oxide(NO),malondialdehyde(MDA),and superoxide dismutase(SOD)in alveolar lavage fluid,homogenate of lung tissue and serum samples were detected by biochemical method.Results The pathological changes of lung tissue of mice in the QTD group,azithromycin group,and combination group were improved compared with the model group,and the improvement was the most significant in the combination group.On the 6th and 11th days of the experiment,the DNA content of mycoplasma pneumoniae in the lung tissues of mice in the model group was significantly increased(all P<0.05),and the content in the QTD group,azithromycin group and combination group were significantly lower than those in the model group at the same period(all P<0.05),and the combination group was significantly lower than those in the QTD group and azithromycin group at the same period(P<0.05).On the 6th and 11th days of the experiment,the expression levels of NOS,iNOS,NO,and MDA in the alveolar lavage fluid,homogenate of lung tissue and serum samples of the model group were significantly higher(all P<0.05),and the SOD expression level was significant lower than those in the normal group(all P<0.05);the expression levels of NOS,iNOS,NO,MDA in the QTD group,azithromycin group,and combination group were significantly lower(all P<0.05),and SOD expression levels were significantly higher than those in the model group(all P<0.05).On the 6th day of the experiment,the expression levels of NOS,NO,SOD in alveolar lavage fluid,iNOS,NO,MDA in lung tissue,and NOS and MDA in serum were improved more significantly in the azithromycin group than those in the QTD group(all P<0.05);the expression levels of NOS,iNOS,and NO in the alveolar lavage fluid,iNOS in the lung tissue,and NO and MDA in the serum in the combination group were significantly lower than those in the QTD group and azithromycin group(all P<0.05),the expression level of MDA in the alveolar lavage fluid,NO and MDA in the lung tissue,and NOS and iNOS in the serum in were significantly lower than those in the QTD group(all P<0.05),while the expression level of SOD in each sample was significantly higher than that of QTD group and azithromycin group(all P<0.05).On the 11th day of the experiment,there was no significant difference in the above indicators between the azithromycin group and the QTD group(P>0.05);the expression of SOD in the alveolar lavage fluid and lung tissue of the combination group was significantly higher than that in the QTD group(all P<0.05).Conclusion QTD can inhibit the activity of NOS and iNOS,reduce the content of NO and MDA,and increase the activity of SOD to play an anti-oxidant effect in the body,thereby reducing lung inflammation.The combined effect of QTD plus azithromycin is more significant.