银屑1号对Trappin-2刺激下肿瘤坏死因子α干预造模的HaCaT细胞分泌NE、Trappin-2的影响

Effects of Yinxie Yihao Prescription(银屑 1 号) on Neutrophil Elastase and Trappin-2 Secreted by HaCaT Cells Interfered and Modeled by Tumor Necrosis Factor α Stimulated by Trappin-2

目的探讨银屑1号治疗银屑病的可能作用机制。方法 24只大鼠随机分为银屑1号组、雷公藤组、空白组,每组8只。银屑1号组大鼠给予银屑1号煎煮浓缩液196 g/(kg·d)灌胃,雷公藤组给予雷公藤多甙片混悬液6 mg/(kg·d)灌胃,空白组给予生理盐水灌胃,均每日2次,每次5 ml,连续5天,制备含药血清。体外培养Ha Ca T细胞,以肿瘤坏死因子α(TNF-α)刺激Ha Ca T细胞建立银屑病细胞模型。实验设置空白组、雷公藤组、抑制剂组、银屑1号组、抑制剂+银屑1号组。空白组、雷公藤组、银屑1号组分别加入基础培养基3.8 ml与相应含药血清,抑制剂组、抑制剂+银屑1号组分别加入空白含药血清和银屑1号含药血清,同时均加入Trappin-2(0.3μg/ml)培养基3.8 ml。体外培养48 h,检测Ha Ca T细胞中中性粒细胞弹性蛋白酶(NE)、Trappin-2含量及NE、Trappin-2 mRNA表达。结果与空白组比较,雷公藤组、抑制剂组、银屑1号组、抑制剂+银屑1号组NE、Trapppin-2含量及mRNA含量明显降低(P<0.05或P<0.01)。与雷公藤组比较,抑制剂组、银屑1号组、抑制剂+银屑1号组NE、Trapppin-2含量及mRNA表达明显升高(P<0.01)。与抑制剂组比较,银屑1号组、抑制剂+银屑1号组NE、Trapppin-2表达明显降低(P<0.05或P<0.01);银屑1号组NE mRNA表达明显升高(P<0.01);抑制剂+银屑1号组Trapppin-2mRNA表达降低(P<0.05)。与银屑1号组比较,抑制剂+银屑1号组NE、Trapppin-2含量及mRNA表达明显降低(P<0.05或P<0.01)。结论银屑1号可降低Ha Ca T细胞中NE、Trappin-2含量及基因表达水平,从而减轻炎症反应,可能是其治疗银屑病的作用机制之一。

Objective To explore the possible mechanism of Yinxie Yihao Prescription（YXYHP）（银屑 1 号） on psoriasis. Methods A total of 24 rats were randomly divided into YXYHP group,Leigongteng group and blank group,with 8 rats in each group. The rats in YXYHP group were given concentrate decoction of YXYHP 196 g/（kg·d） in gavage. Rats in Leigongteng group were given suspension liquid of Triperygium wilfordii multiglucoside tablet6 mg/（kg·d） in gavage. Rats in the blank group were given normal saline in gavage,twice a day,5 ml each time for continuous 5 days. The serum containing medicine from each group was prepared. The psoriatic cell model was established stimulating HaCaT cell cultivated in vitro with tumor necrosis factor α（TNF-α）. The experiment set blank group,Leigongteng group,inhibitor group,Chinese medicine group,and inhibitor ＋ Chinese medicine group. The blank group,Leigongteng group,and Chinese medicine group were added basal culture medium 3. 8 ml,Trappin-2 culture medium and serum containing Chinese medicine respectively. The inhibitor group,and inhibitor ＋ Chinese medicine group were added Trappin-2（0. 3 μg/ml） culture medium 3. 8 ml and Trappin-2 culture medium serum containing related medicine. After culturing in vitro for 48 h,the neutrophil elastase（NE）,Trapinin-2 and expression of NE,Trappin-2 mRNA in HaCaT cells were detected. Results Compared with the control group,the contents of NE,Trappin-2 and mRNA expression in Leigongteng group,inhibitor group,Chinese medicine group,and inhibitor ＋Chinese medicine group were significantly decreased（P〈0. 05 or P〈0. 01）. Compared with Leigongteng group,the contents of NE,Trappin-2 and mRNA expression in inhibitor group,Chinese medicine group,and inhibitor ＋ Chinese medicine group were significantly increased（P〈0. 01）. Compared with the inhibitor group,the expression of NE and Trappin-2 in the Chinese medicine group,inhibitor ＋ Chinese medicine group was significantly decreased（P〈0. 05 or P〈0. 01）. The NE and mRNA expression in the Chinese medicine group was significantly increased（P〈0. 01）. The expression of Trappin-2 mRNA was decreased in inhibitor ＋ Chinese medicine group（P〈0. 05）. Compared with the Chinese medicine group,NE and Trappin-2 contents and mRNA expression in the inhibitor ＋ Chinese medicine group were significantly decreased（P〈0. 05 or P〈0. 01）. Conclusion YXYHP can reduce the contents of NE,Trappin-2 and the gene expression level in HaCaT cells,thus reducing inflammatory response,which may be one of the action mechanisms in treating psoriasis.