清肺通络开玄法对呼吸道合胞病毒诱发幼龄大鼠肺炎性损伤肺组织HMGB1/RAGE轴的影响

Effect of therapy for clearing lung,unblocking collaterals and opening sweat pores on HMGB1/RAGE axis in lung tissue of young rats with pneumonia induced by respiratory syncytial virus

目的探讨清肺通络开玄法对呼吸道合胞病毒(RSV)诱发幼龄大鼠肺炎性损伤肺组织高迁移率族蛋白B1(HMGB1)/晚期糖基化终产物受体(RAGE)轴的影响。方法将24只3周龄雄性SD大鼠随机分为正常组、模型组、清肺通络开玄法组和清肺通络组,每组6只。除正常组外,其余各组采用RSV滴鼻法复制大鼠肺炎性损伤模型。第3天滴鼻接种病毒后,清肺通络开玄法组给予黄芩、大黄和大蒜贴膏贴敷于背部脊柱两侧,清肺通络组给予黄芩和大黄贴膏贴敷于背部脊柱两侧,1次/d,连续5 d;正常组和模型组不给予干预。干预5 d后,HE染色观察大鼠肺组织病理学改变,荧光定量PCR法检测肺组织中HMGB1和RAGE mRNA表达情况,免疫组化法检测肺组织中HMGB1和RAGE蛋白表达情况,酶联免疫吸附法检测血清中肿瘤坏死因子-α(TNF-α)、白细胞介素-6(IL-6)水平。结果干预5 d后,正常组大鼠未见肺组织病理损伤;模型组大鼠见明显肺炎性损伤;清肺通络开玄法组及清肺通络组大鼠肺炎性损伤程度较模型组轻,清肺通络开玄法组损伤更轻。模型组大鼠肺组织中HMGB1和RAGE mRNA相对表达量和蛋白表达平均光密度值均明显高于正常组(P均<0.05);清肺通络开玄法组及清肺通络组大鼠肺组织中RAGE mRNA相对表达量和HMGB1蛋白表达平均光密度值均明显低于模型组(P均<0.05),且清肺通络开玄法组RAGE mRNA相对表达量和HMGB1蛋白表达平均光密度值均明显低于清肺通络组(P均<0.05);清肺通络开玄法组大鼠肺组织中HMGB1 mRNA相对表达量和RAGE蛋白表达平均光密度值均明显低于模型组(P均<0.05),而清肺通络组HMGB1 mRNA相对表达量和RAGE蛋白表达平均光密度值与模型组比较差异均无统计学意义(P均>0.05)。模型组大鼠血清TNF-α、IL-6水平均明显高于正常组(P均<0.05);清肺通络开玄法组大鼠血清TNF-α、IL-6水平及清肺通络组大鼠血清IL-6水平均明显低于模型组(P均<0.05),且清肺通络开玄法组血清IL-6水平明显低于清肺通络组(P<0.05)。结论清肺通络开玄法能调控HMGB1/RAGE轴,抑制TNF-α、IL-6释放,减轻肺炎性组织损伤。

Objective It is to investigate the effect of therapy for clearing lung,unblocking collaterals(CLUC)plus opening sweat pores(OSP)on the high mobility group box-1 protein(HMGB1)/receptor for advanced glycoylation end-product(RAGE)axis in lung tissue of young rats with pneumonia induced by respiratory syncytial virus(RSV).Methods Twenty-four 3-week-old male SD rats were randomly divided into normal group,model group,CLUC+OSP group and CLUC group,with 6 rats in each group.Except for the normal group,the rats in the other groups were subjected to RSV nasal drops to establish the inflammatory lung injury models.On the 3rd day of intranasal inoculation of virus,the rats in the CLUC+OSP group were treated with scutellaria,rhubarb and garlic plaster on both sides of their back spine,and the rats in the CLUC group were treated with scutellaria and rhubarb plaster on both sides of their back spine,all once a day,continuously treated for5 days.The rats in the normal group and model group were not given intervention.After 5 days of treatment,the pathological changes of lung tissue were observed by HE staining,the expressions of HMGB1 and RAGE mRNA in lung tissue were detected by fluorescence quantitative PCR,and the expressions of HMGB1 and RAGE protein in lung tissue were detected by immunohistochemistry,the levels of tumor necrosis factor-α(TNF-α)and interleukin-6(IL-6)in serum were detected by enzyme-linked immunosorbent assay.Results After 5 days of treatment,there was no pathological injury in the lung tissue of the rats in the normal group;Obvious pneumonia injury was found in the rats in the model group.Compared with the model group,the degree of pneumonia injury in the CLUC+OSP group and CLUC group was lighter,and the injury in the CLUC group was more lighter.The relative expressions of HMGB1 and RAGE mRNA and the average optical density of protein expression in lung tissue of the rats in the model group were significantly higher than those of the rats in the normal group(all P<0.05).The relative expression of RAGE mRNA and the average optical density of HMGB1 protein in lung tissue of rats in the CLUC+OSP group and CLUC group were significantly lower than those of the rats in the model group(all P<0.05).The relative expression of RAGE mRNA and the average optical density of HMGB1 protein in the CLUC+OSP group were significantly lower than those in the CLUC group(both P<0.05).The relative expression of HMGB1 mRNA and the average optical density of RAGE protein in lung tissue of the rats in the CLUC+OSP group were significantly lower than those of rats in the model group(both P<0.05),but there were no significant differences in the relative expression of HMGB1 mRNA and the average optical density of RAGE protein between the OSP group and the model group(both P>0.05).The serum levels of TNF-αand IL-6 of the rats in the model group were significantly higher than those in the normal group(both P<0.05).The serum levels of TNF-αand IL-6 of the rats in the CLUC+OSP group and the serum level of IL-6 in the OSP group were significantly lower than those in the model group(all P<0.05),and the serum level of IL-6 in the CLUC+OSP group was significantly lower than that in the OSP group(P<0.05).Conclusion Therapy for clearing lung,unblocking collaterals and opening sweat pores can regulate HMGB1/RAGE axis,inhibit the release of TNF-αand IL-6,and reduce lung tissue injury induced by pneumonia.