The Effect And Mechanism Of Fasudil On Anti-inflammatory And Anti-oxidative Stress In Asthma

Objective: Recent studies show that fasudil plays animportant role in asthma, but the mechanism is still not very clear, so theexperimental study of the effect of Rho-1kinase inhibitor fasudil onanti-inflammatory and antioxidative in bronchial asthma (asthma),explore Rho kinase-1(ROCK-1) role in asthma. Methods: Thisexperiment is divided into three parts:1. animal experiment:24femalemice(5~6weeks) were randomly divided into control group.asthma groupand fasudil group, with8rats in each group. asthma group and fasudilgroup intraperitoneal injection and intranasal sensitization and challengedWith ovalbumin (OVA), fasudil group in nasal drops before usage offasudil intervention, control group was given the same dose of normalsaline. The groups were determination of airway responsiveness,bronchoalveolar lavage fluid (BALF) cell count, lung tissue HE stainingand PAS staining, Immunoblotting detection of NF-κB.PNF-κB.STAT6.PSTAT6.TNF-α and Rho kinase-1, the expression level of MUC5AC wasmeasured by immunofluorescence, expression of ROS in lung tissue offluorescence probe-DHE detection in ROS.2. Cell experiment: It wasdivided into control group, dust mite group and fasudil group, fasudilgroup was treated with fasudil (concentration of45umol/l) for an hour, dust mite group and fasudil group at the same time were given house dustmite (concentration of20ug/ml) for24h,Immunoblotting detection ofNF-κB.PNF-κB.STAT6.PSTAT6.TNF-α and Rho kinase-1, determinationof expression of MUC5AC and ROS fluorescence probe-DHE detectionof intracellular ROS immunofluorescence.3. Clinical trials:16cases ofpatients with collection of asthma, were divided into conventionaltreatment group and conventional+fasudil (intravenous,30mg/days)treatment group,8cases in each group, each group of treatment for1weeks, before and after the treatment, the serum collected a specimen,ELISA method was used to detect serum concentrations ofIL-3.IL-4.IL-17and TNF-α. Results:1. animal experiment:(1) Enhan-cement of forced expiratory airway (Penh) value change: fasudil groupcompared with asthma Penh value decreased, airway reactivity decreased(P<0.05).(2) Cytological changes in BALF: fasudil group compared withasthma in white blood cell counts, percentage of neutrophils andeosinophils (EOS) decreased (P<0.05).(3) Pathological changes in lungtissue of mice in each group: asthma group, inflammatory cell infiltrationin the bronchial and pulmonary tissue around, including neutrophil andlymphocyte in the most, with exudate bronchial and alveolar cavity,partial tracheal epithelial necrosis, and goblet cell and smooth muscle cellproliferation. Fasudil group compared with asthma airway inflammationwas alleviated, weaken the fluorescence intensity of MUC5AC (P<0.05), airway mucus secretion (P<0.05).(4) fasudil group compared withasthma group, decrease the expression of the concentration in lung tissueof ROS (P<0.05).(5) fasudil group compared with asthma group，NF-κB.PNF-κB.STAT6. PSTAT6.TNF-α and Rho kinase-1expressionlevels were significantly lower (P<0.05).2. Cell experiment:(1) fasudilgroup and dust mite group, expression of ROS in lung tissue decreased(P<0.05).(2) fasudil group compared with dust mite group， theMUC5AC fluorescence values decreased significantly (P<0.05).(3)fasudil group compared to dust mite group， NF-κB, PNF-κB,STAT6.PSTAT6. TNF-α and Rho kinase-1expression levels weresignificantly lower (P<0.05).3. Clinical trials: conventional treatmentgroup and conventional treatment+fasudil treatment group before andafter treatment, serum IL-3.IL-4.IL-17and TNF-α level differences werestatistically significant (P<0.05), after treatment than before treatmentdecreased, but regular asthma therapy+fasudil group before and aftertreatment of serum IL-3.IL-4.IL-17and TNF-α concentration differencebetween higher than regular asthma group, with statistical significance(P<0.05). Conclusion:1. animal experiment: asthma mice throughup-regulation of asthma related factors and protein expression andinduced airway hyperresponsiveness, lung tissue inflammation andoxidative stress, which causes a series of clinical asthma symptoms.Pretreatment with fasudil can mitigate this effect to a certain extent.2. cell experiment: dust mite induced16HBE oxidative stress, upregulationof the expression of inflammatory factors and protein, leading to celldamage. Fasudil early intervention can reduce the effect of, also provedthat fasudil have disease and antioxidant anti-inflammatory effect.3.Clinical trials: the concentration of fasudil can relieve asthma relatedcytokines in serum of patients with asthma, So once again proves thatfasudil can reduce inflammatory response of asthmatic patients.4. TheRho-1kinase may be involved in the process of airway inflammation inasthma allergen induced, Rho-1kinase inhibitor fasudil can significantlyreduce airway hyperresponsiveness in asthma, anti-inflammatory andanti-oxidative stress effect.