The intervention effect of nicotine on cervical fibroblast-myofibroblast differentiation in lipopolysaccharide-induced preterm birth model through activating the TGF-β1/Smad3 pathway
The intervention effect of nicotine on cervical fibroblast-myofibroblast differentiation in lipopolysaccharide-induced preterm birth model through activating the TGF-β1/Smad3 pathway
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DOI：
10.1016/j.biopha.2020.111135
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发表时间：
2021-02-01
影响因子：
7.5
通讯作者：
Yang, Jinying
中科院分区：
文献类型：
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作者：
Han, Xinjia;Cai, Chunfang;Yang, Jinying
目前，临床上治疗早产主要采用子宫收缩抑制剂，并不能从根本上降低早产（PTB）的发生率。宫颈早熟是PTB的一个重要因素。我们以前发现，尼古丁治疗的怀孕小鼠有显着的宫颈阻力拉伸和更高的胶原交联相比，对照组动物，尼古丁延长妊娠和抑制宫颈成熟。然而，尼古丁对宫颈早熟的调节作用及其在PTB中的作用仍不清楚。探讨尼古丁对PTB样模型宫颈TGF-β 1/Smad 3通路及其调控的成纤维细胞-肌成纤维细胞分化的影响。用200 μ l PBS中的15 μ g脂多糖（LPS）腹腔注射到妊娠小鼠中以诱导PTB样模型。将小鼠随机分为4组：对照组、LPS处理组、LPS +尼古丁共处理组和LPS +尼古丁+ α-BGT共处理组。监测妊娠结局。天狼星红染色法测定胶原含量。采用免疫荧光双标法和实时荧光定量PCR（qRT-PCR）检测TGF-β/Smad 3通路基因和蛋白的表达。通过双重免疫荧光染色和qRT-PCR研究肌成纤维细胞分化。透射电镜观察超微结构。LPS治疗组肺结核发生率和新生儿死亡率明显高于对照组，胶原含量明显降低，巨噬细胞TGF-β 1和成纤维细胞p-Smad 3表达减少，肺组织中TGF-β 1和p-Smad 3表达减少。α-平滑肌肌动蛋白的表达（α-SMA，活化成纤维细胞的标志物）下调，而钙调蛋白和平滑蛋白的表达（静止时成纤维细胞的标志物）被上调。尼古丁改善妊娠结局并抑制胶原蛋白降解，激活TGF-β 1/Smad 3途径并促进PTB样小鼠的宫颈成纤维细胞-肌成纤维细胞分化;这种作用可以被α-银环蛇毒素（alpha-BGT）逆转。尼古丁通过上调TGF-β/Smad 3通路和促进成纤维细胞分化为肌成纤维细胞，抑制PTB样模型中的宫颈早熟。
Currently, the clinical treatment of preterm birth, mainly using uterine contraction inhibitors, does not fundamentally reduce the incidence of premature birth (PTB). Premature cervical ripening is an important factor in PTB. We previously found that nicotine-treated pregnant murine had significant cervical resistance to stretch and higher collagen cross-links compared to the control animals, and nicotine prolonged gestation and inhibited cervical ripening. However, the regulatory effects of nicotine on premature cervical ripening and its role in PTB remain unclear. To investigate the effects of nicotine on cervical TGF-beta 1/Smad3 pathway and fibroblastmyofibroblast differentiation regulated by this pathway in PTB-like models. Intraperitoneal injection with 15 mu g lipopolysaccharide (LPS) in 200 mu l PBS into pregnant mice was used to induce the PTB-like model. Mice were randomly divided into four groups: control group, LPS-treated group, LPS + Nicotine co-treated group and LPS + Nicotine+alpha-BGT co-treated group. Pregnancy outcomes were monitored. The collagen content was assessed by Picrosirius red staining. Expressions of genes and proteins in the TGF-beta/Smad3 pathway were detected by double immunofluorescence staining and quantitative Real-time PCR (qRT-PCR). myofibroblast differentiation were investigated by double immunofluorescence staining and qRT-PCR. Ultrastructures were analyzed by conventional transmission electron microscopy. The rate of PTB and neonatal mortality at birth was significantly higher in the LPS-treated group than in the control group; collagen content also decreased remarkably; the expression of TGF-beta 1 in macrophages and p-Smad3 in fibroblasts were reduced; the expression of alpha-smooth muscle actin (alpha-SMA, markers for activated fibroblasts) was down-regulated while the expression of calponin and smoothelin (markers for fibroblasts at rest) was up-regulated. Nicotine improved pregnancy outcomes and inhibited collagen degradation, activated the TGF-beta 1/Smad3 pathway and promoted cervical fibroblast-myofibroblast differentiation in PTB-like mice; such effects could be reversed by alpha-bungarotoxin (alpha-BGT). Nicotine inhibited premature cervical ripening in PTB-like models in relation with up-regulating the TGF-beta/Smad3 pathway and promoting fibroblast to differentiate into myofibroblasts.