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血清半胱氨酸蛋白酶-1、Gasdermin家族蛋白D、Gasdermin家族蛋白D-N端在银屑病患者血清及皮损中的表达和意义

Expression and significance of Caspase-1, Gasdermin D and N-terminal fragment of GSDMD in serum and lesions of patients with psoriasis vulgaris

    摘要
  • 摘要:
    目的 检测寻常型银屑病患者血清半胱氨酸蛋白酶-1(Caspase-1)及皮损Caspase-1、Gasdermin家族蛋白D(GSDMD)、Gasdermin家族蛋白D-N端(GSDMD-N)的表达水平, 进一步探索细胞焦亡在银屑病中的作用。
    方法 将进行期寻常型银屑病的30例患者纳入银屑病组(PSO组)，收集30例患者的血清样本和其中20例患者的皮损样本，并收集同时期在苏北人民医院体检中心体检的30例健康者的血清和整形外科20例健康求美者的皮肤组织(皮损)纳入健康对照组(CON组)。采用实时荧光定量聚合酶链反应(PCR)法(qRT-PCR)检测2组血清Caspase-1 mRNA的表达水平; 采用酶联免疫吸附法(ELISA)检测2组血清Caspase-1的含量; 采用蛋白免疫印迹法检测2组皮肤组织中Caspase-1蛋白的表达水平; 采用免疫组织化学(IHC)法检测2组皮肤组织中GSDMD、GSDMD-N的表达及分布情况。
    结果 PSO组患者血清中Caspase-1 mRNA表达水平高于CON组，差异有统计学意义(P<0.01)。PSO组血清中Caspase-1蛋白表达水平为(1 047.74±276.20) pg/mL, 高于CON组的(371.20±160.84) pg/mL, 差异有统计学意义(P<0.01)。PSO组与CON组的Caspase-1总体蛋白表达水平比较，差异无统计学意义(P>0.05); PSO组皮损中的Pro-Caspase-1蛋白表达水平低于CON组, Cleaved Caspase-1蛋白表达水平和GSDMD蛋白表达阳性率高于CON组，差异有统计学意义(P<0.01)。PSO组皮损中GSDMD-N蛋白表达阳性率高于CON组，差异有统计学意义(P<0.01)。
    结论 进行期寻常型银屑病患者血清及皮损中Caspase-1和皮损中GSDMD、GSDMD-N表达增加，提示细胞焦亡通路可能参与寻常型银屑病的发生和发展。

     

    Abstract:
    Objective To detect the expressions of Caspase-1 in serum and Caspase-1, Gasdermin D (GSDMD) as well as N-terminal fragment of GSDMD(GSDMD-N) in lesions of patients with psoriasis vulgaris, and further explore the role of pyroptosis in psoriasis.
    Methods A total of 30 patients with progressive psoriasis vulgaris were enrolled into psoriasis group (PSO group). Serum samples from 30 healthy subjects who underwent physical examinations in the Physical Examination Center of Northern Jiangsu People's Hospital and skin tissue (skin lesions) from 20 healthy aesthetic seekers in Department of Plastic Surgery were collected and included in healthy control group (CON group). The expression of Caspase-1 mRNA in the serum was detected by Quantitative Real Time-Polymerase Chain Reaction (qRT-PCR); the levels of serum Caspase-1 of two groups were detected by enzyme linked immunosorbent assay (ELISA); the expression of proteins of Caspase-1 in samples of skin tissues were detected by western blot; the expression and distribution of GSDMD and GSDMD-N in samples of skin tissues were detected by immunohistochemical method(IHC).
    Results The expression level of Caspase-1 mRNA in serum of the PSO group was significantly higher than that of in the CON group (P<0.01). The level of serum Caspase-1 protein expression in the PSO group was (1 047.74±276.20) pg/mL, which was significantly higher than (371.20±160.84) pg/mL in the CON group (P<0.01). The expression level of total Caspase-1 protein between PSO group and CON group showed no significant difference(P>0.05). The expression level of Pro-Caspase-1 protein in skin lesions of the PSO group was significantly lower than that in the CON group, and the positive rates of expression of Cleaved Caspase-1 protein and GSDMD protein of the PSO group were significantly higher than those in the CON group (P<0.01). The positive rates of expression of GSDMD-N protein in skin lesions of the PSO group was significantly higher than that in the CON group (P<0.01).
    Conclusion The expressions of Caspase-1 in serum and skin lesions, GSDMD and GSDMD-N in skin lesions are increased in patients with psoriasis vulgaris in active stage, suggesting that the pyroptosis pathway may be involved in the occurrence and development of psoriasis vulgaris.

     

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