Department of Dermatology, The Second Affiliated Hospital of Harbin Medical University, Harbin, 150086, Heilongjiang, China.
Department of Dermatology, The First Hospital of China Medical University, Shenyang, 110001, Liaoning, China.
Arch Dermatol Res. 2024 Jun 15;316(7):401. doi: 10.1007/s00403-024-03154-2.
The adhesive properties of vitiligo melanocytes have decreased under oxidative stress., cytoskeleton proteins can control cell adhesion. Paeoniflorin (PF) was proved to resist hydrogen peroxide (HO)-induced oxidative stress in melanocytes via nuclear factorE2-related factor 2 (Nrf2)/heme oxygenase-1 (HO-1) pathway.
This study was to investigate whether PF exerts anti-oxidative effect through influencing cytoskeleton markers or potential signaling pathway.
Human Oxidative Stress Plus array was used to identify the differentially expressed genes between HO + PF group and HO only group, in PIG1 and PIG3V melanocyte cell lines respectively. Western blotting was used to verify the PCR array results and to test the protein expression levels of cytoskeleton markers including Ras homolog family member A (RhoA), Rho-associated kinase 1 (ROCK1) and antioxidative marker Nrf2. Small interfering RNA was used to knock down PDZ and LIM domain 1 (PDLIM1).
PF increased the expressions of PDLIM1, RhoA and ROCK1 in HO-induced PIG1, in contrast, decreased the expressions of PDLIM1 and ROCK1 in HO-induced PIG3V. Knockdown of PDLIM1 increased the expressions of RhoA and Nrf2 in PF-pretreated HO-induced PIG1, and ROCK1 and Nrf2 in PF-pretreated HO-induced PIG3V.
PF regulates RhoA/ROCK1 and Nrf2 pathways in PDLIM1-dependent or independent manners in HO-induced melanocytes. In PIG1, PF promotes PDLIM1 to inhibit RhoA/ROCK1 pathway or activates Nrf2/HO-1 pathway, separately. In PIG3V, PF directly downregulates ROCK1 in PDLIM1-independent manner or upregulates Nrf2 dependent of PDLIM1.
在氧化应激下，白癜风黑素细胞的黏附特性降低。细胞骨架蛋白可以控制细胞黏附。芍药苷（PF）已被证明通过核因子 E2 相关因子 2（Nrf2）/血红素加氧酶-1（HO-1）途径抵抗黑素细胞中的过氧化氢（HO）诱导的氧化应激。
本研究旨在探讨 PF 是否通过影响细胞骨架标志物或潜在信号通路发挥抗氧化作用。
分别用人氧化应激加 array 鉴定 PIG1 和 PIG3V 黑素细胞系中 HO+PF 组和 HO 组之间差异表达的基因。Western blot 用于验证 PCR 阵列结果，并检测细胞骨架标志物包括 Ras 同源家族成员 A（RhoA）、Rho 相关激酶 1（ROCK1）和抗氧化标志物 Nrf2 的蛋白表达水平。小干扰 RNA 用于敲低 PDZ 和 LIM 结构域 1（PDLIM1）。
PF 增加了 HO 诱导的 PIG1 中 PDLIM1、RhoA 和 ROCK1 的表达，相反，降低了 HO 诱导的 PIG3V 中 PDLIM1 和 ROCK1 的表达。PDLIM1 敲低增加了 PF 预处理 HO 诱导的 PIG1 中 RhoA 和 Nrf2 的表达，以及 PF 预处理 HO 诱导的 PIG3V 中 ROCK1 和 Nrf2 的表达。
PF 以 PDLIM1 依赖或不依赖的方式调节 HO 诱导的黑素细胞中 RhoA/ROCK1 和 Nrf2 通路。在 PIG1 中，PF 促进 PDLIM1 抑制 RhoA/ROCK1 通路或激活 Nrf2/HO-1 通路，分别。在 PIG3V 中，PF 以 PDLIM1 独立的方式直接下调 ROCK1 或依赖于 PDLIM1 上调 Nrf2。
