The Effects Of Uygur Herb Hyssopus Officinalis L. On The Process Of Chronic Asthma And The Change Of Tfh Cells In Asthmatic Mice

Objective: The Uygur Herb Hyssopus officinalis L. was used to treat early inchronic asthmatic mice. We observed the effect of Hyssopus officinalis L. on thedevelopment of airway hyperreactivity；we compared the relationship between MMP-9、TIMP-1and T cells subsets, and revealed that Hyssopus officinalis L. could improved theairway remodling by regulating the balance of MMP-9/TIMP-1;meanwhile, we detectedthe change of phenotype and content of the Tfh cell in different groups in order to knowwhether Tfh cells involved in the pathogenesis of chronic asthma and whether Hyssopusofficinalis L. regulated the expression of Tfh cells. It included three parts in this study:①To replicate the chronic asthmatic mice model and observe the airway reactivity, theratio of eosinophils in BALF, the change of structure in different groups. To investigatethe effect of Hyssopus officinalis L. on the development of airway hyperreactivity.②According to compare thecorrelation of MMP-9, TIMP-1and main cytokines of Tcells, it revealed the relationship among them in the process of chronic asthma;at thesame time, we observed that Hyssopus officinalis L. could improved the airwayremodling by regulating the balance of MMP-9/TIMP-1.③We observed the change ofrelating markers (such as BCL-6、CXCR5、ICOS、ICOSL) of the Tfh cells and cytokines(IL-21). It may improve the reaserch on the pathogenesis of immune in chronic asthma;To investigate if the change of the phenotype and content of Tfh cells can be regulated byHyssopus officinalis L so it can further reveal the mechanism of this drug. Methods:Part one: The Preparation of water extract of Hyssopus officinalis L. Thirty-two femaleBalb/c mice were randomly divided into four groups: control group, asthma group,Dexamethasone group and Hyssopus offcinalis L. group. The indexes were detected after the last challenge, as follows: airway reactivity was tested in the context of sobriety andnon-wound; bronchoalveolar lavage fluid was extracted, WBC was classified andcounted; we abserved the structure change of collagen fiber in the different lung tissuesand the deposition, serection of mucus. Part two: The Preparation of water extract ofHyssopus officinalis L. Thirty-two female Balb/c mice were randomly divided into fourgroups: control group, asthma group, Dexamethasone group and Hyssopus offcinalis L.group. The indexes were detected after the last challenge, Those indexes (includingMMP-9、TIMP-1、IFN-γ、IL-4、IL-17、TGF-β) were be measured by Real-time PCR inthe level of mRNA;and be compared the correlation among them. Besides, the expressionof MMP-9、TIMP-1in the level of protein were be detected. Part three: The Preparationof Hyssopus offcinalis L water extract. Thirty-two female Balb/c mice were randomlydivided into four groups: control group, asthma group, Dexamethasone group andHyssopus offcinalis L. group. The indexes were detected after the last challenge. Theratio of Tfh cells in the spleen and the peripheral blood were measured by flow cytometry;the mRNA and protin expession of BCL-6、CXCR5、ICOS、ICOSL and IL-21weredetected and comparied by RT-PCR, Western Blot and immunohistochemistry. Results:Part one: Comparied with control group, Pench increased obviously when inhalated withMch in asthma group (P＜0.05), this difference caused by the different concentration; thechanges of Pench in Dexamethasone group and Hyssopus offcinalis L. group were greaterthan control group, but lower than that of asthma group. There were statistical differences(P<0.05). The content of WBC and the ratio of EOS were increased in asthmatic BALF(P<0.05), while declined in Dexamethasone group and Hyssopus offcinalis L. group(P<0.05). The serum concentration of IgE increased (P＜0.05), but it decreased afterintervented by Hyssopus offcinalis L.. Besides, The outcome of Masson staning and PASstaning indicated that there were massive deposition of collagen fibers and secretion ofmucus in asthmatic lung, but the deposition and secretion were relieved inDexamethasone group and Hyssopus offcinalis L. group. Part two: The expession ofMMP-9and TIMP-1were raised in asthmatic lung tissues, especially TIMP-1, there werestatistical differences in asthma group and control group (P<0.05). wih treatment theexpession of MMP-9and TIMP-1reduced in Dexamethasone group and Hyssopusoffcinalis L. group and were stronger than that of asthma group (P<0.05). The expessionof IL-4, IL-17and TGF-βmRNA were decreased after intervented by Dexamethasone andHyssopus offcinalis L (P＜0.05), while, The expession of IFN-γmRNA were increased (P＜0.05). Correlation analysis revealed that MMP-9and IL-4, IL-17were directly related respectively, but MMP-9and IFN-γ were negative correlation; the expession of TIMP-1is closely related to other indexes. In addtion, the expession of MMP-9and TIMP-1wereincreased in asthma group (P＜0.05), but decreased in Dexamethasone group andHyssopus offcinalis L. group (P＜0.05). Part three: The ratio of CD4+CXCR5+/CD4+and CD4+CXCR5+ICOS+/CD4+CXCR5+in normal spleen and the peripheral blood lowerthan that in asthma group, there were statistical differences (P＜0.05). Comparied withcontrol group, the mRNA and protein expression of BCL-6, CXCR5, ICOS, ICOSL andIL-21were incerseed in asthma group (P＜0.05). The ratio of CD4+CXCR5+inDexamethasone group and Hyssopus offcinalis L. were less than that in asthma group,especially in Hyssopus offcinalis L. group (P＜0.05). the mRNA and protein expressionof BCL-6, CXCR5, ICOS, ICOSL and IL-21by different methods has the same change,the indexes in Hyssopus offcinalis L. group were lower than that in asthma group (P＜0.05). Conelusions:①There were some changes after treated by Hyssopus offcinalis L.,including responsiveness of airway, infiltration of inflammatory cells, deposition ofcollagen fibers, secretion of mucus, proliferation of smooth muscle all relieved. Itsuggested that Hyssopus offcinalis L. could decreased the inflammatory cells andcytokines, relieved the airway hyperreactivity and tissue damage by effect the expessionof Th cells.②There were the relativity between MMP-9, TIMP-1and main cytokines ofTh cells; Hyssopus offcinalis L. could decrease the expession of IL-4, IL-17and TGF-β,but increase the the expession of IFN-γ, and correct the imbalance of MMP-9/TIMP-1; Itsuggested that the abnormal expession of T cells could effect the secrection of MMP-9and TIMP-1. While, Hyssopus offcinalis L. might relieve the airway remodeling byregulating the expession of T cells subsets and correct the imbalance of MMP-9/TIMP-1.③The ratio of Tfh cells increased in asthmatic spleen and peripheral blood, and themRNA and protin expression BCL-6, CXCR5, ICOS, ICOSL and IL-21were increased.It indicated that Tfh cells may take part in the process of chronic asthma. Compared withasthma group, the ratio of CD4+CXCR5+/CD4+和CD4+CXCR5+ICOS+/CD4+CXCR5+were lower than that in Hyssopus offcinalis L. group; the mRNA and protein expressionof BCL-6, CXCR5, ICOS, ICOSL, IL-21were also decreased. It indicated that Hyssopusoffcinalis L. might play a part role in the therapy of asthma by regulating the content andphenotype of Tfh cell.