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目的探讨肿瘤坏死因子受体1(TNFR-1)在毛囊周期中表达的规律及意义。方法采用石蜡切片免疫荧光、Western blot、RT-PCR检测小鼠毛囊生长期(出生后8、15 d,P8d、P15d)、退化期(出生后18 d,P18d)和静止期(出生后24 d,P24d)各时期TNFR-1的表达情况;免疫荧光检测毛囊细胞分化标记物AE13的周期性表达;TUNEL检测毛囊周期中的细胞凋亡情况。结果在4个时相点的皮肤中TNFR-1都存在不同强度的表达,RT-PCR与Western blot检测结果呈现出一致的规律,即从生长期中期(P8d)到生长期末期(P15d)呈现高表达趋势,退化期(P18d)和静止期(P24d)表达减弱并维持较低水平表达。免疫荧光的结果显示:TNFR-1表达在生长期主要表达于表皮层,毛囊的毛干和内根鞘以及毛母质,生长末期在外根鞘还出现明显表达;退化期在外根鞘、毛干及表皮层有表达,在次级毛芽出现很强的特异性表达;在静止期表皮层及杵状毛中呈现弱表达。AE13在生长期主要表达于表皮层以及毛干的皮质和毛小皮;退化期在毛干尤其是毛干基部端(即角质化区域和杵状毛形成区)强表达;在静止期弱表达于表皮层及杵状毛中。TUNEL检测凋亡结果显示:在生长期TUNEL阳性染色主要出现在表皮层,毛囊的毛干、内根鞘以及毛母质,生长末期还出现在外根鞘;在退化期出现在外根鞘、表皮层以及毛干基部端;在静止期P24d出现在表皮层、杵状毛、bulge区和皮脂腺。结论 TNFR-1表达于毛囊的各个时期,生长期中期到生长期末期呈现高表达趋势,退化期和静止期表达减弱并维持较低水平表达,其表达与毛囊细胞终末分化和凋亡有关。
Objective To investigate the expression and significance of tumor necrosis factor receptor 1 (TNFR-1) in hair follicle cycle. Methods Paraffin sections were used for immunofluorescence. The growth of mouse hair follicles (8, 15 d, P8 d, P15 d), degeneration period (18 d, 18 d) and resting period (24 d after birth) , P24d) TNFR-1 expression in each period; immunofluorescence staining of hair follicle cell differentiation marker AE13 of the periodic expression; TUNEL detection of hair follicle cycle apoptosis. Results The expression of TNFR-1 in the skin at four time points showed different intensity. The results of RT-PCR and Western blot showed the same pattern, that is, from the mid-growth period (P8d) to the end of the growth period (P15d) The trend of high expression, degeneration (P18d) and quiescent (P24d) expression decreased and maintained a low level of expression. The results of immunofluorescence showed that TNFR-1 was mainly expressed in the epidermis, hair follicles and inner root sheaths as well as hair and females in the growth phase, and was also expressed in the outer root sheath at the end of growth. The epidermis was expressed in the secondary buds showed a strong specific expression; in the quiescent layer and clubbed hair showed weak expression. AE13 mainly expressed in the epidermis and the hair follicles cortical and epidermal cells during the growth phase. The degenerative phase was strongly expressed in the hair shaft, especially in the basal part of the hair shaft (keratinized area and clubbed hairy area) In the epidermis and clubbing hair. The results of TUNEL assay showed that TUNEL positive staining mainly occurred in the epidermis, hair follicle, root sheath and hair plume during the growth phase, and appeared in the outer root sheath at the end of growth; Hairy stem base end; in the quiescent period P24d appeared in the epidermis, clubbed hair, bulge area and sebaceous glands. Conclusion The expression of TNFR-1 in the hair follicle at each stage, from the mid-growth period to the end of the growing period showed a high expression, decreased and quiescent expression decreased and maintained at a low level, the expression of hair follicle terminal differentiation and apoptosis.