槟榔诱导的颊黏膜成纤维细胞收缩及其信号传导：在口腔黏膜下纤维化——一种癌前病变中的潜在作用

摘要(中文译文)

咀嚼槟榔是一种口腔习惯，会增加口腔癌和口腔黏膜下纤维化（OSF）的风险，后者是一种表现为上皮萎缩和组织纤维化的癌前病变。持续的成纤维细胞收缩可能诱导组织的纤维性挛缩。在本研究中，我们发现槟榔提取物（ANE）（200-1200 µg/ml）可刺激颊黏膜成纤维细胞（OMF）填充的胶原凝胶收缩。槟榔碱而非槟榔次碱——两种槟榔生物碱——可轻度诱导OMF收缩。外源性添加羧酸酯酶（2U/ml）可阻止槟榔碱诱导的OMF收缩，但不能阻止ANE诱导的OMF收缩。OMF表达三磷酸肌醇（IP3）受体。ANE（800 µg/ml）诱导的OMF收缩分别被U73122[磷脂酶C（PLC）抑制剂]和2-氨基乙氧基二苯基硼酸酯（IP3受体拮抗剂）所抑制。乙二醇四乙酸和维拉帕米这两种钙动员调节剂也可抑制ANE诱导的OMF收缩。ANE可诱导OMF中钙/钙调蛋白激酶II和肌球蛋白轻链（MLC）磷酸化。此外，W7（一种Ca²⁺/钙调蛋白抑制剂）、HA1077（Rho激酶抑制剂）、ML-7（MLC激酶抑制剂）和细胞松弛素B（肌动蛋白丝聚合抑制剂）均可抑制ANE诱导的OMF收缩。尽管ANE升高了OMF中的活性氧（ROS）水平，但过氧化氢酶、超氧化物歧化酶和N-乙酰-L-半胱氨酸对ANE引起的OMF收缩无明显影响。这些结果表明，咀嚼槟榔可能通过ANE和槟榔生物碱诱导OMF收缩，从而影响OSF中的伤口愈合和纤维化过程。槟榔成分诱导的OMF收缩与PLC/IP3/Ca²⁺/钙调蛋白和Rho信号通路以及肌动蛋白丝聚合有关，但并非仅由ROS产生所致。

Abstract

Betel quid (BQ) chewing is an oral habit that increases the risk of oral cancer and oral submucous fibrosis (OSF), a precancerous condition showing epithelial atrophy and tissue fibrosis. Persistent fibroblast contraction may induce the fibrotic contracture of tissue. In this study, we found that areca nut extract (ANE) (200-1200 µg/ml) stimulated buccal mucosa fibroblast (OMF)-populated collagen gel contraction. Arecoline but not arecaidine-two areca alkaloids, slightly induced the OMF contraction. Exogenous addition of carboxylesterase (2U/ml) prevented the arecoline- but not ANE-induced OMF contraction. OMF expressed inositol triphosphate (IP3) receptors. ANE-induced OMF (800 µg/ml) contraction was inhibited by U73122 [phospholipase C (PLC) inhibitor] and 2-aminoethoxydiphenyl borate (IP3 receptor antagonist), respectively. Ethylene glycol tetraacetic acid and verapamil, two calcium mobilization modulators, also suppressed the ANE-induced OMF contraction. ANE induced calcium/calmodulin kinase II and myosin light chain (MLC) phosphorylation in OMF. Moreover, W7 (a Ca(2+)/calmodulin inhibitor), HA1077 (Rho kinase inhibitor), ML-7 (MLC kinase inhibitor) and cytochalasin B (actin filament polymerization inhibitor) inhibited the ANE-induced OMF contraction. Although ANE elevated reactive oxygen species (ROS) level in OMF, catalase, superoxide dismutase and N-acetyl-L-cysteine showed no obvious effect on ANE-elicited OMF contraction. These results indicate that BQ chewing may affect the wound healing and fibrotic processes in OSF via inducing OMF contraction by ANE and areca alkaloids. AN components-induced OMF contraction was related to PLC/IP3/Ca(2+)/calmodulin and Rho signaling pathway as well as actin filament polymerization, but not solely due to ROS production.

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