20(R)-人参皂苷Rg3抗角膜瘢痕机制研究
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摘要
目的:研究不同浓度的20(R)-人参皂苷Rg3(GRg3)对兔角膜成纤维细胞增殖和转化生长因子-β1(TGF-β1)表达的影响,探讨GRg3抑制角膜瘢痕增生的机制。方法:直接选用大白兔获取兔角膜成纤维细胞后进行原代培养,采用甲基噻唑基四唑(MTT)比色法观察不同浓度(10 mg·L~(-1),20 mg·L~(-1),50 mg·L~(-1),100 mg·L~(-1),150 mg·L~(-1),200 mg·L~(-1))GRg3对兔角膜成纤维细胞增殖作用的影响。应用反转录-PCR(RT-PCR)技术研究不同浓度GRg3对培养的兔角膜成纤维细胞TGF-β1m RNA表达的影响。结果:MTT法检测结果:与对照组比较,当GRg3浓度达到20 mg·L~(-1)及以上时,兔角膜成纤维细胞增殖均显著受抑制。当GRg 3浓度达到50 mg·L~(-1)及以上时,GRg3对兔角膜成纤维细胞TGF-β1 mRNA表达具有抑制作用。结论:GRg3具有抑制体外培养的兔角膜成纤维细胞增殖的作用,并随着GRg3浓度的增加,其抑制作用增强。随培养时间的延长,GRg3抑制作用增强。GRg3对兔角膜成纤维细胞TGF-β1mRNA表达具有抑制作用。
Objective To explore the effects of the different concentrations Ginsenoside Rg3 on cultured corneal fi broblasts of rabbit proliferation and the expression of TGF-β1m RNA so as to investigate the mechanism of the inhibition of corneal scar. Methods We observed the effects of the different concentrations(10 mg·L~(-1)20 mg·L~(-1),50 mg·L~(-1),100 mg·L~(-1),150 mg·L~(-1),200 mg·L~(-1))GRg3 on proliferation by tetrazotium(MTT) colorimetric assay and detected the expression of TGF-β1m RNA by RT-PCR on cultured corneal fi broblasts of rabbit. Results The OD values gained by MTT were signifi cant different among the experiment groups and control ones. When the concentrations of GRg3 is above 20 mg/L,GRg3 inhibited fi broblastic proliferation on cultured corneal fi broblasts. When the concentrations of GRg3 is above 50 mg/L, GRg3 inhibited the expression of TGF-β1m RNA on cultured corneal fi broblasts. Conclusion GRg3 signifi cantly inhibited fi broblastic proliferation and the expression of TGF-β1m RNA on cultured corneal fi broblasts of rabbit in vitro. The effect of inhibition increased with the increasing concentrations of GRg3 and cultured time.
Objective To explore the effects of the different concentrations Ginsenoside Rg3 on cultured corneal fi broblasts of rabbit proliferation and the expression of TGF-β1m RNA so as to investigate the mechanism of the inhibition of corneal scar. Methods We observed the effects of the different concentrations(10 mg·L~(-1)20 mg·L~(-1),50 mg·L~(-1),100 mg·L~(-1),150 mg·L~(-1),200 mg·L~(-1))GRg3 on proliferation by tetrazotium(MTT) colorimetric assay and detected the expression of TGF-β1m RNA by RT-PCR on cultured corneal fi broblasts of rabbit. Results The OD values gained by MTT were signifi cant different among the experiment groups and control ones. When the concentrations of GRg3 is above 20 mg/L,GRg3 inhibited fi broblastic proliferation on cultured corneal fi broblasts. When the concentrations of GRg3 is above 50 mg/L, GRg3 inhibited the expression of TGF-β1m RNA on cultured corneal fi broblasts. Conclusion GRg3 signifi cantly inhibited fi broblastic proliferation and the expression of TGF-β1m RNA on cultured corneal fi broblasts of rabbit in vitro. The effect of inhibition increased with the increasing concentrations of GRg3 and cultured time.
引文
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[13]于澎,明月,庞利民,等.人参皂苷Rg3对培养人视网膜色素上皮细胞增生的抑制作用[J].哈尔滨医科大学学报,2003,37(2):153-154.
[14]刘鹤松,赵自然,兰珊珊,等.人参皂苷Rg3对人成纤维细胞增殖和凋亡影响的实验研究[J].中国实验诊断学,2010,14(11):1697-1700.
[15]Sun X,Cheng L,Zhu W,et al.Use of ginsenoside Rg3-loaded electrospun PLGA fibrous membranes as wound cover induces healing and inhibits hypertrophic scar formation of the skin[J].Colloids Surf B Biointerfaces,2013,115(c):61-70.
[16]Cheng L,Sun X,Hu C,et al.In vivo inhibition of hypertrophic scars by implantable ginsenoside-Rg3-loaded electrospun fi brous membranes[J].Acta Biomater,2013,9(12):9461-9473.
[17]高勇,王杰军,许青,等.人参皂甙Rg3抑制肿瘤新生血管形成的研究[J].第二军医大学学报,2001,22(1):40-42.
[18]Liu WK,Xu SX,Che CT.Anti-proliferative effect of ginseng saponins onhumanprostate cancer cell line[J].Life Sciences,2000,67(11):1297-1306.
[2]Beye JA,Hart DA,Bray RC,et al.Denervation alters m RNA Levels of repair-associated genes in a rabbit medial collateral ligament injury model[J].J Orthop Res,2006,24(9):1842-1853.
[3]Nakanishi T,Nishida T,Shimo T,et al.Effects of CTGF/Hcs24,a product of a hypertrophic chondrocyte-specifi c gene,on the proliferation and differentiation of chondrocytes in culture[J].Endocrinology,2000,141(1):264-273.
[4]杨咏梅,吴欣怡,杜立群.结缔组织生长因子与转化生长因子及Smad信号通路在兔角膜创伤愈合中的作用[J].中华眼科杂志,2006,42(10):918-924.
[5]Itoh S,Itoh F,Goumans MJ,et al.Signaling of transforming growth factor-beta family members through Smad proteins[J].European Journal of Biochemistry.2000,267(24):6954-6967.
[6]张仲苗,江波,郑筱祥.人参皂苷Rg3对肿瘤放疗患者外周血淋巴细胞的体外免疫增强作用[J].中国药学杂志,2004,39(4):261-264.
[7]刘基巍,陈俊霞,于丽华,等.人参皂甙Rg3和核糖核酸酶抑制因子转基因对小鼠黑色素瘤的抑制作用研究[J].中华肿瘤杂志,2004,26(12):722-725.
[8]杨威,李博,乔世兴,等.人参皂甙Rg3抗鼠源性膀胱癌BBT739黏附、侵袭和肺转移作用的研究[J].中国老年学杂志,2005,25(5):571-573.
[9]Yue PY,Wong DY,Wu PK,et al.The angiosuppressive effects of 20(R)-ginsenoside Rg3[J].Biochem Pharmacol.2006,72(4):437-445.
[10]梁宇,崔凝,宋腾飞,等.人参皂甙Rg3抗肿瘤作用的研究进展[J].中国现代药物应用.2007,1(11):105.
[11]李平亚,郝秀华,赵春芳,等.人参皂甙-Rg3,-Rb3在抗病毒作用的研究[J].中国老年学杂志,2001,21(3):215-216.
[12]常雅萍,吴雅珍,李平亚,等.G-Rg3治疗单疱病毒性角膜炎实验观察[J].白求恩医科大学学报,2000,26(2):134-136.
[13]于澎,明月,庞利民,等.人参皂苷Rg3对培养人视网膜色素上皮细胞增生的抑制作用[J].哈尔滨医科大学学报,2003,37(2):153-154.
[14]刘鹤松,赵自然,兰珊珊,等.人参皂苷Rg3对人成纤维细胞增殖和凋亡影响的实验研究[J].中国实验诊断学,2010,14(11):1697-1700.
[15]Sun X,Cheng L,Zhu W,et al.Use of ginsenoside Rg3-loaded electrospun PLGA fibrous membranes as wound cover induces healing and inhibits hypertrophic scar formation of the skin[J].Colloids Surf B Biointerfaces,2013,115(c):61-70.
[16]Cheng L,Sun X,Hu C,et al.In vivo inhibition of hypertrophic scars by implantable ginsenoside-Rg3-loaded electrospun fi brous membranes[J].Acta Biomater,2013,9(12):9461-9473.
[17]高勇,王杰军,许青,等.人参皂甙Rg3抑制肿瘤新生血管形成的研究[J].第二军医大学学报,2001,22(1):40-42.
[18]Liu WK,Xu SX,Che CT.Anti-proliferative effect of ginseng saponins onhumanprostate cancer cell line[J].Life Sciences,2000,67(11):1297-1306.