| ObjectiveTo investigate the expression of cold-induced RNA binding protein(CIRP),transforming growth factor β1(TGF-β1 and matrix metalloproteinase-9(MMP-9)in different types of chronic rhinosinusitis(CRS),and to analyze their correlation between tissue remodeling in CRS and these three factors CIRP TGF-β、MMP-9.MethodsSelected patients in hospital are divided into the following groups:individuals with chronic rhinosinusitis without nasal polyps(CRSsNP,20 cases)and individuals with chronic rhinosinusitis with nasal polyps(CRSwNP,20 cases)as the case groups,individuals with maxillary sinus cyst as the control group(20 cases).All the patients in this study underwent endoscopic sinus surgery in the First Affiliated Hospital of Zhengzhou University from 2020.09 to 2021.01.The mucosal tissues of the ethmoid sinus of patients with CRSsNP,mucosal tissues of nasal polyps of CRSwNP and uncinate process mucosa tissues of patients with maxillary sinus cyst were collected.HE staining and MT staining were used to observe the remodeling of mucosal tissues.The expressions of CIRP,TGF-β1 and MMP-9 in tissues were detected by immunohistochemistry and real-time quantitative polymerase chain reaction(qRTPCR).Kruskal-Wallis(K-W)test was used for comparison between multiple groups,Pearson correlation analysis was used for correlation between variables,one-way ANOVA was used for comparison of age distribution between groups,and chi-square test was used for comparison of gender distribution,P<0.05 was considered statistically significant.Results1 HE staining resultsThere were obvious tissue remodeling changes in.CRS,and the pathological changes of the mucosal tissues in the CRSwNP group and the CRSsNP group were different.2 Masson staining resultsCollagen fibers are stained green and are deposited in the basement membrane region and near vascular glands.The difference between the three groups was statistically significant(H=31.740,P<0.001).Pair comparison showed that collagen volume fraction in CRSsNP group was significantly higher than that in CRSwNP group(significance value was adjusted by Bonferroni correction method,both P<0.001 after adjustment)and control group(P=0.037 after adjustment),and collagen volume fraction in CRSwNP group was significantly lower than that in control group(P=0.005 after adjustment).3 Immunohistochemical results3.1 Expression of CIRPThe expression of CIRP was observed in different types of CRS and the control group,the expression of CIRP in sinus mucosa was mainly located in the nucleus and cytoplasm.The expression level of CIRP in the CRSwNP group was higher than that in the control group and the CRSsNP group.The difference among the three groups was statistically significant(H=26.943,P<0.001).Pair comparison showed that the expression of CIRP in the CRSwNP group was significantly higher than that in the CRSsNP group and the control group(the significance value was adjusted by Bonferroni correction method,and the adjusted P values were both<0.001).The expression of CIRP in the CRSsNP group was not significantly different from that in the control group(the adjusted P=1.000).3.2 Expression of TGF-β1TGF-β1 was expressed in CRSsNP group,CRSwNP group and control group,and the expression of TGFβ1 in rhinosinus mucosa tissue was mainly located in the cytoplasm.The difference between the three groups was statistically significant(H=39.004,P<0.001).Pairwise comparison showed that the expression of TGF-β1 in CRSsNP group was significantly higher than that in CRSwNP group(the significance values was adjusted by Bonferroni correction method,and the adjusted P values<0.001)and control group(the adjusted P=0.041),and the expression of TGF-β1 in CRSwNP group was significantly lower than that in control group(the adjusted P=0.001).3.3 Expression of MMP-9The expression of MMP-9 can be seen in different types of CRS and the control group.The positive expression of MMP-9 is mainly located in the cytoplasm,mainly expressed in the epithelium,glands,vascular endothelial cells and inflammatory cells infiltrating the submucosal lamina propria.The difference between the three groups was statistically significant(H=16.975,P<0.001).Pairwise comparisons show:the expression of MMP-9 in CRSwNP group is significantly higher than that in the control group(adjust the significant value by Bonferroni correction method,adjusted P<0.001),the expression of MMP-9 in CRSsNP group is higher than that in the control group,but there was no statistically significant difference(P=0.100 after adjustment),the expression of MMP-9 in CRSwNP group is higher than that in CRSsNP group,but there was no statistically significant difference(P=0.139 after adjustment).4 qRT-PCR results4.1 Relative expression level of CIRPThere were significant differences among the three groups(H=94.800,P<0.001).After pairwise comparison,the relative mRNA expression of CIRP in the CRSwNP group was significantly higher than that in the control group and the CRSsNP group,(significance value was adjusted by Bonferroni correction method,the adjusted P values were both<0.001),while the difference between the CRSsNP group and the control group was not statistically significant(P=1.000 after adjustment).4.2 Relative expression level of TGF-β1The difference among the three groups was statistically significant(H=138.591,P<0.001).The pairwise comparison showed that the mRNA expression of TGF-β1 in the CRsNP group was significantly higher than that in the control group and the CRSwNP group,and the mRNA expression of TGF-β1 in the CRSwNP group was significantly lower than that in the control group and the CRSsNP group(the significance value was adjusted by Bonferroni correction method,and the adjusted P values were all<0.001).4.3 Relative expression level of MMP-9The difference among the three groups was statistically significant(H=100.800,P<0.001).Pairwise comparison showed that the mRNA expression of MMP-9 in the CRSwNP group was significantly higher than that in the CRSsNP group and the control group(the significance value was adjusted by Bonferroni correction method,and the adjusted P values were all<0.001),while the difference between the CRSsNP group and the control group was not statistically significant(P=0.604 after adjustment).5 Correlations between the expression of CIRP,TGF-β1 and MMP-9 proteinPair correlation analysis of the expression of CIRP,TGF-β1 and MMP-9 protein in each tissue showed that there was a significant positive correlation between CIRP and MMP-9 protein expression(r=0.481,P<0.001).CIRP was negatively correlated with TGF-β1 protein expression(r=-0.338,P=0.008).MMP-9 was negatively correlated with the protein expression of TGF-β1(r=-0.291,P=0.024).6 Correlation between protein expression of CIRP,TGF-β1,MMP-9 and collagen depositionThe protein expression of CIRP,TGF-β1 and MMP-9 in each tissue was respectively correlated with CVF.The results showed that the protein expression of CIRP was negatively correlated with CVF(r=-0.283,P=0.029).TGF-β1 protein expression was positively correlated with CVF(r=0.437,P<0.001).There was no significant correlation between MMP-9 protein expression and CVF(r=-0.196,P=0.134).Conclusion1.The expression of CIRP was up-regulated in CRSwNP tissues,but had no significant change in CRSsNP tissues,which was involved in the tissue remodeling of CRSwNP.2.TGF-β1 expression was down-regulated in CRSwNP tissues and up-regulated in CRSsNP tissues,which promoted collagen deposition and was involved in tissue remodeling of CRS.3.The expression of MMP-9 was up-regulated in CRSwNP,but there was no significant change in CRSsNP tissues,which promoted the degradation of extracellular matrix and was involved in the tissue remodeling of CRSwNP.4.There was a significant positive correlation between CIRP and MMP-9,CIRP was negatively correlated with TGF-β1,MMP-9 was negatively correlated with TGFβ1.5.CIRP,TGF-β1 and MMP-9 are involved in the pathogenesis of CRS and are expected to be new targets for the treatment of CRS. |
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