真菌性鼻窦炎大鼠鼻窦黏膜组织Maspin、IKKα表达水平及意义

Expression and significance of Maspin and IKKα in sinusoidal mucosa of rats with fungal rhinosinusitis

目的探讨真菌性鼻窦炎(FRS)大鼠鼻窦黏膜组织Maspin、IKKα表达水平及意义。方法 40只SD级大鼠建立真菌性鼻窦炎模型,按照随机数字表法分为鼻塞组、FRS组、免疫抑制剂组、侵袭性FRS组,每组各10只,另选择10只健康大鼠作为空白对照组;对照组正常喂养;鼻塞组仅鼻腔塞入止血棉,腹腔与鼻腔内给予等体积0.9%Na Cl注射液;FRS组腹腔注射等量0.9%Na Cl注射液,鼻腔注射烟曲霉菌孢子悬液;免疫抑制剂组腹腔注射环磷酰胺,鼻腔内注射等量0.9%Na Cl注射液;侵袭性FRS组腹腔注射环磷酰胺、鼻腔注射烟曲霉菌孢子悬液建立侵袭性FRS大鼠模型。采用酶联免疫吸附试验检测血清白细胞介素6(IL-6)、肿瘤坏死因子α(TNF-α)表达水平;免疫组织化学染色法检测各组大鼠鼻腔组织Maspin、IKKα蛋白表达水平,荧光定量PCR法检测各组大鼠鼻腔组织Maspin mRNA、IKKαmRNA表达水平。结果各组大鼠血清IL-6、TNF-α表达水平比较差异有显著性(P<0.05),其中FRS组血清IL-6、TNF-α显著高于对照组、鼻塞组、免疫抑制剂组和侵袭性FRS组【(69.3±10.9)ng/L vs(45.2±7.1)ng/L,(46.4±6.7)ng/L,(21.3±4.5)ng/L,(20.9±4.3)ng/L;(30.4±4.8)ng/L vs(14.8±2.7)ng/L,(13.9±1.4)ng/L,(7.9±0.6)ng/L,(7.8±0.4)ng/L】(P<0.05),对照组血清IL-6、TNF-α又显著高于免疫抑制剂组和侵袭性FRS组(P<0.05),免疫抑制剂组、侵袭性FRS组间血清IL-6、TNF-α表达水平比较差异无显著性(P>0.05)。免疫组织化学染色结果显示FRS组、侵袭性FRS组Maspin蛋白表达水平显著低于对照组、鼻塞组、免疫抑制剂组,IKKα蛋白表达水平显著高于对照组、鼻塞组、免疫抑制剂组(P<0.05);其中侵袭性FRS组Maspin蛋白表达显著低于FRS组,IKKα蛋白表达则显著高于FRS组(P<0.05)。FRS组、侵袭性FRS组Maspin mRNA表达水平显著低于对照组、鼻塞组、免疫抑制剂组,IKKαmRNA表达水平显著高于对照组、鼻塞组、免疫抑制剂组(P<0.05);其中侵袭性FRS组Maspin mRNA表达显著低于FRS组,IKKαmRNA表达显著高于FRS组(P<0.05)。结论 IKKα活化后下调Maspin表达是导致FRS发病的主要原因,这一过程可能也是侵袭性FRS分子作用机制之一。

Objective To investigate the expression and significance of M aspin and IKKα in nasosinusoidal mucosa of rats with fungal rhinosinusitis （ FRS）. Methods A total of 40 SD rats were used to establish the FRS model, and randomly divided into nasal obstruction group, FRS group, immunosuppressive group and invasive FRS group, 10 rats in each group. Another 10 normal rats were used as control group. Mice in the control group were fed with normal diet. In the nasal obstruction group, the mice had only hemostatic cotton stuffed in the nasal cavity and injection of 0. 9% NaCl in the abdominal and nasal cavities. In the FRS group, the mice were injected Aspergillus fumigatus spore suspension into the nasal cavity and 0. 9% NaCl i.p. The mice of the immunosuppressive group were given cyclophusphamide i.p. and 0. 9% NaCl injection into the nasal cavity, The invasive FRS group was injected with cyclophosphamide i. p. and Aspergillus fumigatus spore suspension into the nasal cavity. The serum levels of interleukin-6 （IL-6） and tumor necrosis factor-α （TNF-α） were measured by enzyme-linked immunosorbent assay （ELISA）. The expression of Maspin and IKKα in nasosinusoidal mucosa was detected by immunohistochemical staining. The expression of Maspin mRNA and IKKα mRNA in the nasosinusoidal mucosa was detected by fluorescence quantitative PCR. Results The serum levels of IL-6 and TNF- α in different groups were significantly different （P 〈 0. 05）. The level of IL-6 in the FRS group was （69.3 ±10.9） ng/ L, significantly higher than those in the control group, nasal obstruction group, immunosuppressive group and invasive FRS group [ （45.2 ± 7. 1）ng/L, （46.4 ±6.7） ng/L, （21.3 ± 4.5） ng/L, and （20.9 ±4.3 ng/L）] （P 〈 0.05）. The level of TNF-α in the FRS group was （30.4±4.8） ng/L, significantly higher than those in the control group, nasal obstruction group, immunosuppressive group and invasive FRS group [ （ 14.8 ± 2.7） ng/L, （ 13.9± 1.4） ng/L, （7.9 ± 0.6） ng/L, and （7.8 ± 0.4 ng/L）] （P 〈 0.05）. The levels of IL-6 and TNF-α in the control group were significantly higher than those in the immunosuppressive group and invasive FRS group （P 〈 0.05）. There was no significant difference between the immunosuppressive group and the invasive group （P 〉 0.05 ）. Theresuh of immunohistochemical staining showed that the protein expression of Maspin in the FRS group and invasive FRS group was significantly lower than that in the control group, nasal obstruction group and immunosuppressive group, while the expression of IKKα protein was significantly higher than that of control group, nasal obstruction group and immunosuppressive group （P 〈 0. 05）. The protein expression of Maspin in the invasive FRS group was significantly lower than that in the FRS group, by contrast, the expression of IKKα protein was significantly higher （ P 〈 0.05）. The PCRresuh revealed that the expression levels of Maspin and IKKα mRNA were （0. 217 ± 0. 013 ） and （0. 193± 0. 012）, significantly lower than that in the control, obstruction and immunosuppressive groups [ （0. 309± 0. 021 ）, （0. 302±0. 017） , and （0. 293 ±0. 02） ] （P 〈 0.05） , while the expressions level of IKKα mRNA were significantly higher [（0.319 ± 0.043）, （0.384±0.048） vs （0.169± 0.015）, （0.171± 0.018）, and （0. 175 ± 0.019）] （P 〈 0. 05）. Conclusions Down-regulation of Maspin expression after IKKα activation is the main cause of the onset of FRS, which may also be one of the mechanisms of invasive FRS.