苦参碱对瘢痕疙瘩成纤维细胞生长的抑制作用实验研究(1)
[摘要]目的：研究苦参碱对瘢痕疙瘩成纤维细胞增殖、凋亡和自噬的影响及其相关作用机制。方法：通过收集手术切除的瘢痕疙瘩进行体外培养成纤维细胞，将细胞分为3组，分别加入10μmol/L，20μmol/L的苦参碱以及对照组(0.9% NaCl)进行实验。MTT实验检测细胞增殖情况；流式细胞技术进行细胞周期分析；Hoechest 33258染色检测细胞凋亡情况；Western blot实验检测苦参碱对瘢痕疙瘩成纤维细胞增殖、凋亡和自噬相关调控蛋白表达的影响。结果：通过不同浓度的苦参碱处理细胞后，MTT结果发现成纤维细胞的增殖受到抑制，抑制作用具有剂量依赖性；细胞周期分析发现，苦参碱作用后细胞的G1-S期转化受到了不同程度的抑制；Hoechest 33258染色发现苦参碱可以促进凋亡细胞的增加；Western blot实验结果发现苦参碱可以抑制CDK4、CDK6、Bcl-2和LC3-I蛋白的表达，促进Bax、LC3-Ⅱ和Beclin-1的表达。结论：苦参碱对瘢痕疙瘩成纤维细胞具有抑制增殖、促进凋亡的作用，能够影响细胞增殖、凋亡和自噬相关调控蛋白的表达。
, 百拇医药
[关键词]苦参碱；成纤维细胞；增殖；凋亡；自噬；细胞周期；瘢痕疙瘩
[中图分类号]R619+.6 [文献标志码]A [文章编号]1008-6455(2018)10-0137-04
Abstract： Objective To study the effects of Matrine on the proliferation， apoptosis and autophagy of keloid fibroblasts in vitro and its related mechanisms. Methods The fibroblasts were cultured in vitro by collecting surgical excised keloids. The cells were divided into three groups and the 10μmol/L， 20μmol/L matrine and the control group (0.9% NaCl) were added to the experiment. MTT test was used to detect the proliferation of fibroblasts， flow cytometry was used for cell cycle analysis， Hoechest 33258 staining was used to detect cell apoptosis， Western blot was used to detect the effect of Matrine on the proliferation， apoptosis and autophagy related protein expression of keloid fibroblasts. Results After treating cells with different concentrations of matrine， MTT results showed that the proliferation of fibroblasts was inhibited and the inhibitory effect was dose-dependent. The cell cycle analysis showed that the G1-S phase transformation of the cells was inhibited in varying degrees after the action of matrine. Hoechest 33258 staining showed that matrine could lead to a significant increase in apoptosis. The results of Western blot showed that matrine could inhibit the expression of CDK4， CDK6， Bcl-2 and LC3-I protein， and promote the expression of Bax， LC3-Ⅱ and Beclin-1. Conclusion Matrine can inhibit the proliferation and promote apoptosis of keloid fibroblasts， and can affect the cell proliferation， apoptosis and the expression of autophagy related proteins.
, 百拇医药
Key words：Matrine； fibroblasts； proliferation； apoptosis； autophagy； cell cycle； keloid
瘢痕疙瘩(Keloid)是皮膚创伤后结缔组织过度增殖形成的良性肿块。其病理组织学特点是成纤维细胞过度增殖以及细胞外基质过量的沉积。对于瘢痕疙瘩的治疗方法有很多，药物是瘢痕疙瘩预防和治疗的一大类别，包括抗代谢药、类固醇激素、免疫抑制剂和抗肿瘤药等[1-2]。而寻找疗效确切、副作用小的药物，对于瘢痕疙瘩的预防和治疗具有重要临床意义。
苦参碱(Matrine，Mat)是从豆科植物苦参的干燥根、植株中提取得到的一种生物碱，归属于四环喹嗪啶类化合物[3]。目前已有报道[4]，苦参类生物碱具有多方面的药理活性及生物学功能。而我们常用的瘢痕软化膏的主要活性成分就是苦参类生物碱，其能够显著抑制瘢痕的增生，使增生性瘢痕软化缩小，临床效果显著。苦参碱在增生性瘢痕中的作用研究报道较多，但在瘢痕疙瘩中的作用却尚不十分清楚，而且对于苦参碱在瘢痕疙瘩中的作用机制，需要进一步研究。本次研究通过体外培养瘢痕疙瘩成纤维细胞，利用不同浓度的苦参碱对细胞进行处理，研究苦参碱对细胞增殖、凋亡、自噬等生物功能的影响。希望通过本次实验能够为临床应用苦参碱预防和治疗瘢痕疙瘩提供依据。, http://www.100md.com(徐志山 王洪一 钟黎明 回蔷 郭冰玉 陶凯)
, 百拇医药
[关键词]苦参碱；成纤维细胞；增殖；凋亡；自噬；细胞周期；瘢痕疙瘩
[中图分类号]R619+.6 [文献标志码]A [文章编号]1008-6455(2018)10-0137-04
Abstract： Objective To study the effects of Matrine on the proliferation， apoptosis and autophagy of keloid fibroblasts in vitro and its related mechanisms. Methods The fibroblasts were cultured in vitro by collecting surgical excised keloids. The cells were divided into three groups and the 10μmol/L， 20μmol/L matrine and the control group (0.9% NaCl) were added to the experiment. MTT test was used to detect the proliferation of fibroblasts， flow cytometry was used for cell cycle analysis， Hoechest 33258 staining was used to detect cell apoptosis， Western blot was used to detect the effect of Matrine on the proliferation， apoptosis and autophagy related protein expression of keloid fibroblasts. Results After treating cells with different concentrations of matrine， MTT results showed that the proliferation of fibroblasts was inhibited and the inhibitory effect was dose-dependent. The cell cycle analysis showed that the G1-S phase transformation of the cells was inhibited in varying degrees after the action of matrine. Hoechest 33258 staining showed that matrine could lead to a significant increase in apoptosis. The results of Western blot showed that matrine could inhibit the expression of CDK4， CDK6， Bcl-2 and LC3-I protein， and promote the expression of Bax， LC3-Ⅱ and Beclin-1. Conclusion Matrine can inhibit the proliferation and promote apoptosis of keloid fibroblasts， and can affect the cell proliferation， apoptosis and the expression of autophagy related proteins.
, 百拇医药
Key words：Matrine； fibroblasts； proliferation； apoptosis； autophagy； cell cycle； keloid
瘢痕疙瘩(Keloid)是皮膚创伤后结缔组织过度增殖形成的良性肿块。其病理组织学特点是成纤维细胞过度增殖以及细胞外基质过量的沉积。对于瘢痕疙瘩的治疗方法有很多，药物是瘢痕疙瘩预防和治疗的一大类别，包括抗代谢药、类固醇激素、免疫抑制剂和抗肿瘤药等[1-2]。而寻找疗效确切、副作用小的药物，对于瘢痕疙瘩的预防和治疗具有重要临床意义。
苦参碱(Matrine，Mat)是从豆科植物苦参的干燥根、植株中提取得到的一种生物碱，归属于四环喹嗪啶类化合物[3]。目前已有报道[4]，苦参类生物碱具有多方面的药理活性及生物学功能。而我们常用的瘢痕软化膏的主要活性成分就是苦参类生物碱，其能够显著抑制瘢痕的增生，使增生性瘢痕软化缩小，临床效果显著。苦参碱在增生性瘢痕中的作用研究报道较多，但在瘢痕疙瘩中的作用却尚不十分清楚，而且对于苦参碱在瘢痕疙瘩中的作用机制，需要进一步研究。本次研究通过体外培养瘢痕疙瘩成纤维细胞，利用不同浓度的苦参碱对细胞进行处理，研究苦参碱对细胞增殖、凋亡、自噬等生物功能的影响。希望通过本次实验能够为临床应用苦参碱预防和治疗瘢痕疙瘩提供依据。, http://www.100md.com(徐志山 王洪一 钟黎明 回蔷 郭冰玉 陶凯)
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