苦参碱对瘢痕疙瘩成纤维细胞的生物学功能及相关调控蛋白的影响
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摘要
目的探讨中药苦参碱对瘢痕疙瘩成纤维细胞生物功能的影响及相关调控蛋白表达的作用,了解其对瘢痕疙瘩的相关作用机制。方法将瘢痕疙瘩行体外细胞培养后进行细胞处理。实验组加入20μmol/L和40μmol/L的苦参碱;对照组加入生理盐水。分别通过四甲基噻唑蓝(MTT)检测成纤维细胞的增殖情况;AV-PI染色检测其凋亡情况;Transwell检测其侵袭和迁移情况;Western blot检测其生物功能相关调控蛋白的表达水平。结果经苦参碱处理后,细胞增殖受到抑制,且抑制作用具有浓度、时间的依赖性;AV-PI染色发现,苦参碱能在一定程度上诱导瘢痕疙瘩成纤维细胞凋亡,且对其侵袭和迁移有抑制作用;通过Western blot实验结果发现,苦参碱对瘢痕疙瘩成纤维细胞的增殖、凋亡、自噬、侵袭和迁移等生物功能相关蛋白Cyclin D1、Bcl-2、LC3、Beclin-1、MMP2、MMP9的表达水平具有调节作用。结论苦参碱对瘢痕疙瘩成纤维细胞的生物学功能具有调节作用,可能是通过调控相关蛋白的表达来影响瘢痕疙瘩的生长。
Objective To explore the effect and mechanism of matrine on biological function and related regulatory protein expression of keloid fibroblasts. Methods The fibroblasts were extracted from keloid and cultured in vitro. Matrine of 20 μmol/L and40 μmol/L was added to fibroblasts in the experimental group, and physiological saline was added to fibroblasts in the control group. The proliferation of fibroblasts was detected by MTT assay, apoptosis was detected by AV-PI staining, invasion and migration was detected by Transwell, and biological function and related regulatory protein expression was detected by Western Blot. Results After treatment with matrine, the proliferation of fibroblasts was inhibited in a concentration-and time-dependent manner. AV-PI staining showed that matrine could induce apoptosis of keloid fibroblasts to a certain extent and inhibit their invasion and migration. The experiment results of Western Blot showed that matrine could regulate the expression of Cyclin D1, Bcl-2, LC3, Beclin-1, MMP2 and MMP9, which were related to proliferation, apoptosis, autophagy, invasion, and migration. Conclusion Matrine can regulate the biological function of keloid fibroblasts,which may affect the growth of keloids by regulating the expression of related proteins.
Objective To explore the effect and mechanism of matrine on biological function and related regulatory protein expression of keloid fibroblasts. Methods The fibroblasts were extracted from keloid and cultured in vitro. Matrine of 20 μmol/L and40 μmol/L was added to fibroblasts in the experimental group, and physiological saline was added to fibroblasts in the control group. The proliferation of fibroblasts was detected by MTT assay, apoptosis was detected by AV-PI staining, invasion and migration was detected by Transwell, and biological function and related regulatory protein expression was detected by Western Blot. Results After treatment with matrine, the proliferation of fibroblasts was inhibited in a concentration-and time-dependent manner. AV-PI staining showed that matrine could induce apoptosis of keloid fibroblasts to a certain extent and inhibit their invasion and migration. The experiment results of Western Blot showed that matrine could regulate the expression of Cyclin D1, Bcl-2, LC3, Beclin-1, MMP2 and MMP9, which were related to proliferation, apoptosis, autophagy, invasion, and migration. Conclusion Matrine can regulate the biological function of keloid fibroblasts,which may affect the growth of keloids by regulating the expression of related proteins.
引文
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[2]WANG X,LIN H,ZHANG R.The clinical efficacy and adverse effects of interferon combined with matrine in chronic hepatitis B:a systematic review and meta-analysis[J].Phytother Res,2017,31(6):849-857.
[3]XIAO X,AO M,XU F,et al.Effect of matrine against breast cancer by downregulating the vascular endothelial growth factor via the Wnt/β-catenin pathway[J].Oncol Lett,2018,15(2):1691-1697.
[4]AN G,LIANG S,SHENG C,et al.Upregulation of micro RNA-205suppresses vascular endothelial growth factor expression-mediated PI3K/Akt signaling transduction in human keloid fibroblasts[J].Exp Biol Med(Maywood),2017,242(3):275-285.
[5]GAO J,JIAO H,CAO R,et al.In vitro study of effects and mecha nism of rapamycin-induced autophagy in keloid fibrolasts[J].Zhonghua Zheng Xing Wai Ke Ze Zhi,2016,32(3):208-214.
[6]CHEN J X,SHEN H H,NIU M,et al.Anti-hepatitis B virus effect of matrine-type alkaloid and involvement of p38 mitogen-activated protein kinase and tumor necrosis factor receptor-associated factor6[J].Virus Res,2016,215:104-113.
[7]OKUNO R,ITO Y,EID N,et al.Upregulation of autophagy and glycolysis markers in keloid hypoxic-zone fibroblasts:morphological characteristics and implications[J].Histol Histopathol,2018,33(10):1075-1087.
[8]WU J,HU G,DONG Y,et al.Matrine induces Akt/mTORsignalling inhibition-mediated autophagy and apoptosis in acute myeloid leukaemia cells[J].J Cell Mol Med,2017,21(6):1171-1181.
[9]CWANG Y,LIU Y,JIANG J,et al.Antitumor effects of matrine on cancer stem like cells isolated from the human liver cancer SMMC-7721 cell line[J].Oncol Lett,2018,15(2):1777-1782.
[10]LI Q,HUANG H,HE Z,et al.Regulatory effects of antitumor agent matrine on FOXO and PI3K-AKT pathway in castration-resistant prostate cancer cells[J].Sci China Life Sci,2018,61(5):550-558.