Effents Of Different Consentrations Of Moxa Smoke On OMP、iNOS Protein Expression In Rat Nasal Mucosaand Olfactory Function

Objective:To explore the effects of different concentrations of moxa smoke on the olfactory function of the body,so as to provide theoretical basis for the safety of moxa smoke produced in clinical moxibustion.Methods:1:According to the moxa smoke concentration in the simulated acupuncture clinic previously measured by our group,the moxa smoke concentration was reasonably divided according to the content of PM2.5 in moxa smoke,so as to divide the environment of high,middle and low moxa smoke concentration.2:After stratification with body mass,adult SD rats were randomly divided into normal control group(normal group),low concentration moxa smoking group(low smoking group),medium concentration moxa smoking group(middle smoking group)and high concentration moxa smoking group(high smoking group).Then the rats in each concentration of moxa smoke group were carried out the moxa smoking experiment for 90 days,twice a day and 4 hours each time.There was no intervention in the normal control group.Smoking lasted for 90 days.At the end of the experiment,the behavior of rats was observed by double bottle test and food globule test.3: At the end of the experiment,rat serum and nasal mucosal tissue were taken to detect the pathological damage of nasal mucosa by HE staining and TUNEL.The contents of IL-1 β,IL-6 and TNF-α in serum and nasal mucosa were detected by ELISA and the correlation among them was counted.The expression of OMP positive cells was detected by immunohistochemical method.The expressions of OMP,iNOS mRNA and protein were detected by Real-time PCR and immunoblotting respectively.Results:By comparing the behavioral changes of rats in each group,we found that there was no significant difference between normal group and low smoking group(P > 0.05),but there were significant differences between middle smoking group,high smoking group,normal group and low smoking group(P < 0.01,P < 0.05).The results of HE staining showed that the structure of nasal mucosa in normal group was clear,neatly arranged,ciliated on the surface,no inflammatory cells were found in submucosa,a small number of inflammatory cells could be seen in low smoking group compared with normal group,the structure of nasal mucosa in middle smoking group was unclear and disordered,and there were more inflammatory cells in submucosa.In the high smoke group,the structure of nasal mucosa was unclear,disordered,necrotic,and there were a large number of submucosal infiltration of acting cells.TUNEL detection showed that the apoptotic cells in nasal mucosa of rats in all groups increased significantly with the increase of moxa smoke concentration.The apoptotic cells in nasal mucosa of rats in middle and high smoking groups were significantly higher than those in normal group and low smoking group(P < 0.01),and there was significant difference between high smoking group and middle smoking group(P < 0.01).ELISA detection showed that the contents of IL-1 β,IL-6 and TNF-α in serum and nasal mucosa of rats in each moxa smoke group were significantly higher than those in the normal group(P < 0.001).The content of inflammatory factors increased with the increase of moxa smoke concentration(P < 0.01),and there was a significant correlation between them(P < 0.05).The expression of OMP positive cells in nasal mucosa of rats in each group was detected by immunohistochemical method.The results showed that the expression of OMP in low smoking group was lower than that in normal group(P >0.05).The levels in middle smoking group and high smoking group were also significantly lower than those in low smoking group(P < 0.01).The expression of OMP and iNOS mRNA was detected by Real-time PCR and Western blotting.It was found that the expression of OMP mRNA and protein decreased with the increase of moxa smoke concentration in normal grou(P < 0.01),low smoking group,middle smoking group and high smoking group,while the expression of iNOS mRNA and protein increased with the increase of moxa smoke concentration(P < 0.01).Conclusion:1 The long-term low concentration moxa smoke environment will not cause obvious pathological changes of nasal mucosa,while medium and high concentration moxa smoke environment will cause nasal mucosal pathological changes.2 The long-term low concentration of moxa smoke environment will not have a significant impact on the olfactory sensitivity of the body,medium and high concentration of moxa smoke environment will damage the olfactory sensitivity of the body.