The Therapeutic Effects Of UTI On Allergic Rhinitis And The Relationship With Heme Oxygenase-1

Objective: Allergic rhinitis is a clinical common chronic disease, studieshave shown that the onset of allergic rhinitis is not only a traditional conceptconsiders IgE-mediated typeⅠallergic nasal mucosa, and is susceptible toindividual who exposed allergen and lead to IgE-mediated inflammatorytransmitter release and a variety of immune active cells, Th1/Th2cytokinenetwork imbalance are caused chronic inflammatory diseases at the nasalmucosa and submucosal vessels. The global incidence of Allergic rhinitis isabout10%25%, the prevalence of allergic rhinitis is about9%-24.6%inChina, and with the development of national industrial, there is an upwardtrend in incidence, seriously affect the quality of people’s lives.Heme oxygenase (Heme oxygenase, HO) is a kind of microsomalenzymes, HO is the starting and the rate-limiting enzyme of heme degradation,HO-1for its induction type. Recent research data show that HO-1gene as aprotective gene, its overexpression have important biological effects such asanti-oxidative stress, anti-inflammation, resisting proliferation of malignantcell, regulating cell cycle activity as well as to the immunoregulatory effectsof Treg cells.HO-1has protective effect on allergic rhinitis.Nowadays, the treatment of allergic rhinitis is mostly used hormonedrugs, which have lots of adverse drug reactions. And ulinastatin results fromthe human body,which has less adverse drug reactions. In clinic, Ulinastatinhas been widely used in the treatment such as pancreatitis, shock, systemicinflammatory response, and the effect of ulinastatin is remarkable. Ulinastatincan inhibit excessive release of inflammatory mediators, improvemicrocirculation and perfusion of tissue, restrain cell apoptosis. At the sametime, Ulinastatin can stabilize the cell membrane, inhibit proteindecomposition, eliminate oxygen free radicals and inhibit the expression of IL -6, IL–8and ICAM-1, and so on.But ulinastatin has not been applied in thetreatment of allergic rhinitis, the specific role of ulinastatin in mechanism oftreatment allergic inflammation remains to be further discussed.Because of both HO-1and ulinastatin has immunoregulatory andanti-inflammatory effects, So we are studied as follows,in vitro and in vivoexperiments to explore the expression of HO-1protein levels which impactedby ulinastatin in mice and mast cells in the pathogenesis of allergic rhinitis,and compared with hormone drugs. Whether the ulinastatin through raised theexpression of HO-1to achieve a therapeutic effect. To provide a theoreticalbasis for mechanism of ulinastatin and HO-1in the treatment of allergicrhinitisMethods:40FVB mice were randomly divided into control group (con),allergic rhinitis group (OVA), dexamethasone intervention group (OVA+dex)and ulinastatin intervention group (OVA+UTI).AR group conducted thesensitization with10%ovalbumin (OVA). Dexamethasone and ulinastatinintervention group conducted intraperitoneal injection of dexamethasone andulinastatin before local excitation. Behavioral points are evaluated animalmodel. Behavior observation was used to evaluate the model. The mice wereanaesthetize after the last challenge, then took the heart blood, using forsurvey the level of IL-4、INF-γ、OVA and specific IgE content with ELISA.Preparing the Nasal lavage fluid（Nasal lavage fluid, NLF）, which was countfor inflammatory cell. Lung tissue specimen was homogenized to determinethe protein content of HO-1.The nasal tissue were fixed with paraform,embedded with paraffin and pathological section was made, then stained withhematoxlin-eosin for histological analyze and immunohistochemical stainingfor the expression of HO-1protein. Meanwhile, the mouse mast cellscultured in vitro were randomly divided into control group (con),Inflammation model group (TNF-α), dexamethasone intervention group(TNF-α+Dex) and ulinastatin intervention group (TNF-α+UTI). Thesupernatant was collected to detect the tryptase activity. Cells were collectedrespectively to performed immunofluorescence, ROS detection, Western blot, and RT-PCR to compare the effect of ulinastatin in mast cells in inflammationmodel and the expression of HO-1protein in mast cells in each group, andto explore whether there is time and dose dependence between the HO-1andulinastatin in mast cell.Results：1Ulinastatin can obviously improve nasal symptoms of allergic rhinitis inmiceAR models were successfully established in10animals, which hadtypical nasal symptoms, and all of their behavioral stack scores were morethan5points; Compared with AR group, symptoms of dexamethasone andulinastatin intervention group were significantly relieved, stack scores wereless than5points; control group almost had no nasal symptoms of allergicrhinitis.2Ulinastatin has obvious protective effect to airway mucosa in mice withallergic rhinitisThe nasal mucosa of control group was well arranged has no obviousinflammatory changes; The AR group showed different degrees of edema, afew epithelial exfoliation, the lager size of goblet cell, gland hyperplasia, theinfiltration of submucosal eosinophils, lymphocytes and monocytes. Nasalinflammation in dexamethasone and ulinastatin intervention group weresignificantly reduced.3Ulinastatin can significantly inhibit number of eosinophils in nasal lavagefluid in mice with allergic rhinitisIn AR model group, the total number of EOS were increased significantly,the differences were significant (P<0.05). Compared the dexamethasone andulinastatin intervention group with control group, the amount of EOS wassignificantly decreased, but the difference was statistically significant（P<0.05）.4Ulinastatin can reverse the imbalance of Th1/Th2cytokines in mice allergicrhinitis In AR mice, IL-4and OVA specific IgE in serum were significantlyhigher than the other there groups, and the differences were significant (P<0.05). The control group, dexamethasone intervention group and ulinastatinintervention group had no significant difference（P>0.05）.5Ulinastatin can significantly inhibit mast cell degranulation that stimulatedby TNF-αTryptase activity in mast cells was enhanced after TNF-α stimulation,significantly higher than that in control group, ulinastatin intervention groupand dexamethasone intervention group, the differences were significant(P<0.05). The control group, dexamethasone intervention group andulinastatin intervention group had no significant difference（P>0.05）.6Ulinastatin could inhibit the ROS expression by TNF-α challenged in mastcellsROS was low expression in normal group of mast cells, the expression ofROS in mast cells was significantly increased after TNF-α stimulation,andwas significantly decreased after dexamethasone and ulinastatin intervention,but still higher than control group.7Ulinastatin can induce expression of HO-1in nasal mucosa and lungtissue of allergic rhinitis mice, and mast cellsCompared the control group with the other three groups, the expressionof HO-1protein was significantly higher（P<0.05）,他he AR group,dexamethasone intervention group and ulinastatin intervention group had nosignificant difference（P>0.05）.8The expression of HO-1as a dose and time dependence in UTI treated mastcells both in protein and geneThe expression of HO-1in mast cells gradually increased with the timeincreased under ulinastatin intervention,significantly at4hours. After4hoursof Ulinastatin intervention, the expression of HO-1increased gradually withthe increase of ulinastatin concentration.Conclusion:1Ulinastatin inhibit eosinophilic granulocyte increased at mice airway, regulating Th1/Th2cytokine imbalance,and compared withdexamethasone UTI also have immunomodulatory effect, but theanti-inflammatory was superior than dexamethasone.2Protective effects of ulinastatin on mast cell inflammatory models caninhibit the degranulation of mast cells and oxidative stress.3Ulinastatin can up-regulate the expression of HO-1protein in allergicrhinitis mouse model.Ulinastatin can up-regulate the expression of HO-1protein in mast cells inflammatory model，the expression of HO-1has timeand dose dependence with ulinastatin.4The immunoregulatory and anti-inflammatory effects of ulinastatin onallergic rhinitis may be associated with the up-regulation of HO-1expression.