A型肉毒毒素对增生性瘢痕JNK信号通路和成纤维细胞相关蛋白表达的影响

Effects of botulinum toxin type A on the expression of JNK signaling pathway and fibroblast-related proteins in hypertrophic scars

目的增生性瘢痕是一种真皮纤维增生性疾病。A型肉毒毒素具有预防增生性瘢痕形成的作用,但其抗皮肤纤维化的作用及其机制尚不清楚。本研究分析A型肉毒毒素(BTXA)对增生性瘢痕(HS)JNK信号通路和成纤维细胞相关蛋白表达的影响。方法本研究采用0、1、2、4U/ml BTXA对人瘢痕成纤维细胞进行干预,通过CCK8划痕法、荧光定量PCR和western印迹法检测不同浓度BTXA对HS成纤维细胞增殖、迁移、细胞凋亡和蛋白表达的变化。并对比BTXA干预前后转化生长因子-β1(TGF-β1)、α-肌动蛋白(α-SMA)、结缔组织生长因子(CTGF)、c-Jun氨基末端激酶(JNK)信号通路、磷酸化氨基末端激酶(p-JNK)等成纤维细胞相关蛋白表达的变化。结果与无BTXA干预相比,1、2、4U/ml BTXA干预后HS成纤维细胞增殖能力较低(P<0.05);与无BTXA干预相比,1、2、4U/ml BTXA干预后HS成纤维细胞凋亡率增高(P<0.05);与BTXA干预前相比,BTXA干预后TGF-β1、α-SMA、CTGF等基因水平和蛋白水平均降低(P<0.05);与JNK选择性抑制SP600125干预前相比,JNK选择性抑制SP600125干预后p-JNK表达较高(P<0.05)。结论BTXA可影响HS患者成纤维细胞增殖、迁移和凋亡,并能通过JNK信号通路诱导成纤维细胞凋亡,调控其相关蛋白表达,为临床治疗提供新靶点。

Objective To analyze the effects of botulinum toxin type A(BTXA)on the expression of JNK signaling pathway and fibroblast-related proteins in hypertrophic scars(HS).Methods A total of 24 surgical specimens of HS patients in our hospital from November 2020 to June 2021 were selected.BTXA concentrations of 0.2,0.4 and 0.8u/ml were intervened in 8 cases respectively.All patients were tested before BTXA intervention and after 0.2,0.4 and 0.8u/ml BTXA intervention.The effects of different concentrations of BTXA on the viability,proliferation cycle and apoptosis rate of HS fibroblasts were analyzed.The expression of fibroblast-related proteins(TGF-β1)and the changes of JNK signaling pathway(p-JNK)before and after BTXA intervention were compared.Results Compared with no BTXA intervention,the viability of HS fibroblasts was lower than that after 0.2,0.4 and 0.8u/ml BTXA intervention(P<0.05).Compared with no BTXA intervention,0.2,0.4,0.8 U/ml BTXA intervention could increase the number of cells in G1 phase and decrease the number of cells in S and G2 phases(P<0.05).Compared with no BTXA intervention,0.2,0.4,0.8 U/ml BTXA intervention could increase the apoptosis rate of HS fibroblasts(P<0.05).Compared with before BTXA intervention,TGF-β1 expression was lower after BTXA intervention(P<0.05).Compared with JNK selective inhibition of SP600125 before intervention,the expression of p-JNK was higher after JNK selective inhibition of SP600125(P<0.05).Conclusions BTXA can affect the fibroblast activity and cell proliferation cycle of HS patients,and can induce fibroblast apoptosis through the JNK signaling pathway,regulate the expression of related proteins,and provide a new target for clinical treatment.