孢子丝菌病患者皮损中Toll样受体2、4及髓样分化因子88的表达

Expressions of Toll-like receptor 2, Toll-like receptor 4 and myeloid differentiation factor 88 in skin lesions of patients with sporotrichosis

目的通过观察孢子丝菌病患者皮损中Toll样受体2、4（TLR2、TLR4）以及髓样分化因子88（MyD88）的表达情况，探讨其在孢子丝菌病免疫识别及免疫介导中的作用。方法选择孢子丝菌病患者19例及健康人对照12例，应用免疫组化法分别检测患者组皮损及对照组皮肤组织中TLR4及MyD88的表达情况，同时应用实时荧光定量PCR技术检测TLR2及MyD88mRNA的表达情况。所有结果数据以虿±s表示，采用SPSS17．0统计软件进行数据分析，两组间比较采用独立样本t检验，P〈0．05认为差异有统计学意义。结果免疫组化分析：孢子丝菌病患者皮损中TLR4、MyD88的表达部位主要是除角质层外的表皮全层和真皮及皮下组织中的浆细胞和淋巴细胞。对照组皮肤几乎不表达TLR4，MyD88于真皮及皮下组织呈弱表达。孢子丝菌病组TLR4表达水平（63．767±3．829）高于对照组（5．167±3．246），差异有统计学意义（t=4．82，P〈0．05）；MyD88表达水平（57．236±4．744）亦高于对照组（10．588±1．640），差异有统计学意义（t=3．30，P〈0．05）。实时荧光定量PCR分析：孢子丝菌病患者皮损TLR2mRNA和MyD88mRNA的相对表达水平分别为1．974±1．452和2．028±2．061，均显著高于对照组（分别为1．430±1．073和0．688±0．422），均P〈0．05。结论孢子丝菌病发病可能与真菌通过Toll样受体途径作用于机体免疫系统有关。

Objective To evaluate the roles of Toll-like receptor 2 （TLR2）, TLR4 and myeloid differentiation factor 88 （MyD88） in immune recognition and immune mediation in sporotrichosis by detecting their expressions in skin lesions of sporotrichosis. Methods Biopsy specimens were obtained from the skin lesions of 19 patients with sporotrichosis and normal skin of 12 healthy human controls. Immunohistochemistry was performed to observe the expressions of TLR4 and MyD88, and real-time fluorescence-based quantitative PCR to quantify the expressions of TLR2 and MyD88 mRNAs. Data were expressed as mean ＋ standard error. Statistical analysis was done with the SPSS17.0 software. Independent samples t-test was conducted to compare the parameters between the lesional and control specimens. A p-value of less than 0.05 was considered to be statistically significant. Results TLR4 and MyD88 were mainly observed in the whole epidermis except the stratum corneum as well as plasma ceils and lymphocytes in the dermis and subcutaneous tissue in lesional skin. However, TLR4 was nearly absent and MyD88 was weakly expressed in the dermis and subcutaneous tissue in normal skin. The expression levels of TLR4 and MyD88 were significantly higher in the lesional skin than in the control skin （TLR4, 63.767 ± 3.829 vs. 5.167 ± 3.246, t = 4.82, P 〈 0.05; MyD88, 57.236 ± 4.744 vs. 10.588 ± 1.640, t = 3.30, P 〈 0.05）. Similarly, the lesional skin showed significantly stronger expressions of TLR2 and MyD88 mRNAs compared with the normal skin（TLR2, 1.974 ± 1.452 vs. 1.430 ± 1.073, P 〈 0.05; MyD88, 2.028 ± 2.061 vs. 0.688 ± 0.422, P 〈 0.05）. Conclusion Sporothrix may induce the development of sporotrichosis by interacting with host immune system via TLR signaling pathways.