摘要
目的分析双氢青蒿素(DHA)对加入转化生长因子-β1(TGF-β1)培养的人子宫内膜间质细胞的增殖活性以及分泌MMP-9和TIMP-1的影响,探讨DHA对子宫内膜纤维化的作用机理。方法消化子宫内膜组织并滤出其中的间质细胞。据不同的孵育方法细胞被设成4组,对照组:按照常规方法培养;DHA组:常规培养液中滴入DHA;TGF-β1组:常规培养液中滴入TGF-β1;TGF-β1+DHA组:常规培养液中滴入TGF-β1和DHA。通过吸光度值观察4组细胞的增殖结果;分析每组培养液中分泌的Ⅰ型胶原的含量;免疫细胞化学法衡量细胞里的基质金属蛋白酶-9(MMP-9)以及其抑制剂-1(TIMP-1)值的变化。结果对照组与DHA组相比无明显差异(P>0.05);TGF-β1组与对照组相比,细胞的吸光度值(0.1299±0.0023)升高,培养液中Ⅰ型胶原(0.7246±0.0036)分泌量上升,细胞内MMP-9的含量(0.0394±0.0024)稍微升高(P>0.05),TIMP-1的含量(0.7288±0.0620)明显升高(P<0.05),MMP-9/TIMP-1的值(0.0514±0.0035)减少(P<0.05)。TGF-β1+DHA组与TGF-β1组相比,细胞的吸光度值(0.1210±0.0028)下降,培养液中Ⅰ型胶原(0.5723±0.0036)分泌量降低,细胞内MMP-9的含量(0.0812±0.0031)明显升高(P<0.05),TIMP-1的含量(0.4529±0.0433)明显降低(P<0.05),MMP-9/TIMP-1的值(0.1546±0.0195)增加(P<0.05)。结论DHA能抑制TGF-β1作用的子宫内膜间质细胞增殖,并能调节TGF-β1作用的细胞内MMP-9和TIMP-1的水平来达到拮抗子宫内膜纤维化的效果。
Objective The effects of dihydroartemisinin(DHA)on the proliferation and secretion of MMP-9 and TIMP-1 of human endometrial stromal cells cultured with TGF-β1 were analyzed to explore the mechanism of DHA on endometrial fibrosis.Methods Digest endometrial tissue and filter out stromal cells.According to different incubation methods,cells were arranged for 4 groups.Control group:culture according to the conventional method;DHA group:drop DHA into the conventional culture solution;TGF-β1 group:drop TGF-β1 into the conventional culture solution;TGF-β1+DHA group:drop TGF-β1 and DHA into the conventional culture solution.The proliferation of the 4 groups of cells was observed;the content of type I collagen secreted in each group of culture medium was analyzed;the content of matrix metalloproteinase-9(MMP-9)and its antagonist-1(TIMP-1)was detected through immunoblotting.Results The control group and the DHA group was not obviously different(P>0.05);In the TGF-β1 group,compared with the control group,the cell absorbance value(0.1299±0.0023)increased,the secretion of type I collagen(0.7246±0.0036)increased,The content of MMP-9(0.0394±0.0024)increased slightly(P>0.05),the content of TIMP-1(0.7288±0.0620)increased significantly(P<0.05),and the value of MMP-9/TIMP-1(0.0514±0.0035)decreased(P<0.05);In the TGF-β1+DHA group,Compared with TGF-β1 group,the cell absorbance value(0.1210±0.0028)decreased,the secretion of type I collagen(0.5723±0.0036)decreased,The contents of MMP-9(0.0812±0.0031)and TIMP-1(0.4529±0.0433)were significantly increased(P<0.05),and the values of MMP-9/TIMP-1(0.1546±0.0195)were obviously raised(P<0.05).Conclusion DHA can inhibit the proliferation of endometrial stromal cells induced by TGF-β1,and regulate the levels of MMP-9 and TIMP-1 in the cells induced by TGF-β1 to achieve the effect of antagonizing endometrial fibrosis.
作者
栗志英
马景红
齐秀芳
单铁英
杨志红
马瑞晴
魏天慧
董付存
张素华
LI Zhi-ying;MA Jing-hon;QI Xiu-fang(Affiliated Hospital Hebei University of Engineering,Handan 056002,China)
出处
《中国实验诊断学》
2021年第8期1213-1217,共5页
Chinese Journal of Laboratory Diagnosis
基金
河北省邯郸市科学技术局立项课题(19422083011-4)。
