Effects Of The Alcoholic Extracts Of Ligustrum Officinalis And Radix Caulis Lonicerae On The Expressions Of NF-κB Inflammatory Signaling Pathway-related Molecules In Asthmatic Mice

Objective: To investigate the effects of alcohol extracts of the alcoholic extracts of Ligustrum officinalis and Radix caulis lonicerae on the expression of NFkappa B related molecules in inflammatory signaling pathway of asthmatic mice with Cold Fluid in the Lung Syndrome and the mechanism of the alcoholic extracts of Ligustrum officinalis and Radix caulis lonicerae on alleviating airway inflammation and remodeling in asthmatic mice with Cold Fluid in the Lung Syndrome was explored.Methods: 120 female Kunming mice of SPF grade were randomly divided into blank group with 15 mice,Model group、Xiaoqinglong Decoction group、the alcoholic extracts of Ligustrum officinalis and Radix caulis lonicerae were divided into three dosage groups: low,medium and high,with 21 rats in each group.The blank group was sensitized by intraperitoneal injection of normal saline and inhalation by atomization.The remaining models were sensitized by intraperitoneal injection of fresh Ovalbumin(OVA)sensitizing solution and stimulated by ultrasonic atomization.The mice were exposed to cold stimulation at-4-4 C and were fed with ice water for 14 days to establish the model of asthma with Cold Fluid in the Lung Syndrome.After successful establishment of the model,the blank group and the model group were given normal saline by gastric lavage,while the Xiaoqinglong Decoction group was given Xiaoqinglong decoction,lignum ginger and Radix caulis lonicerae alcohol extract of corresponding concentration,once a day,for four weeks.Four weeks later,according to the requirements,locate the samples and test the indicators.The pathological changes of mice were observed by HE staining.Reverse transcription polymerase chain reaction(RT-qPCR)was used to detect the m RNA expression levels of NF-κB p65 and ECM in mice.Western blotting(WB)with the expression levels of NF-κB p65 and ECM protein in mice were detected,and the contents of TGF-β1 and VEGF were detected by enzyme-linked immunosordental assay(ELISA).Results:(1)HE staining test: Compared with the blank group,airway remodeling was observed in the model group.Compared with the model group,airway remodeling in each treatment group was controlled to varying degrees.The alcoholic extracts of Ligustrum officinalis and Caulis lonicerae in high and medium dose groups and Xiaoqinglong Decoction group could obviously relieve inflammatory infiltration of lung tissue cells and improve airway remodeling and other pathological symptoms.(2)RT-qPCR detection: Compared with the blank group,the NF-κ B p65 and ECM genes in the lung tissue of the model group were highly expressed,and the difference was statistically significant(P<0.05).Compared with the model group,NF-κ B p65 and ECM in all treatment groups showed a downward trend(P<0.05),among which the high dose group of Ligustrum officinalis and Caulis lonicerae and Xiaoqinglong decoction group decreased most(P<0.01).(3)Westernblot test: Compared with the blank group,the expression of NF-κ B p65 and ECM protein in the lung tissue of the model group was significantly different(P<0.05).Compared with the model group,the levels of NF-κ B p65 and ECM protein in each treatment group showed a downward trend(P<0.05),especially in the high-dose group of Ligustrum officinalis,Caulis lonicerae and Xiaoqinglong decoction group(P<0.01).(4)ELISA test: Compared with the blank group,the levels of TGF-β 1 and VEGF in the lung tissue of the model group increased significantly,and the difference was statistically significant(P<0.05).Compared with the model group,the TGF-β 1 and VEGF in the lung tissue of each treatment group decreased(P<0.05);especially the high dose group of Ligustrum officinalis and Caulis lonicerae and Xiaoqinglong decoction group were the most significant(P<0.01).Conclusion: Ethanol extract of Ligustrum officinalis and Caulis lonicerae can down-regulate the expression or level of NF-kappa B p65,ECM,TGF-beta 1 and VEGF in the inflammatory signaling pathway of mice with asthma Cold Fluid in the Lung Syndrome,reduce the infiltration of inflammatory cells,slow down the inflammatory response,inhibit the proliferation of airway smooth muscle cells(ASMCs),and interfere with airway remodeling is its effective mechanism in treating asthma.