双氢青蒿素抑制TGF-β1诱导的子宫内膜间质细胞的分泌活性

Inhibition of Dihydroartemisinin in secretion activity of endometrial stromal cells induced by TGF-β1

目的观察双氢青蒿素(DHA)对经转化生长因子-β1(TGF-β1)孵育的来源于人子宫壁的内膜组织中的间质细胞(ESCs)数目和活力的变化。方法2018年1月1日—6月1日,收集人正常的子宫内膜组织并分离出ESCs。实验设计分为4组。正常组:加入不含血清的培养液;TGF-β1组:在正常组的基础上加10 ng/mLTGF-β1;DHA组:在正常组的基础上加100 ng/mLDHA;TGF-β1+DHA组:在TGF-β1组的基础上加100 ng/mLDHA。通过噻唑蓝法检测每组细胞的增殖变化;用酶联免疫吸附(ELISA)法判定每组细胞孵育液中Ⅰ型胶原和纤黏连蛋白的水平。免疫印迹(Western blot)评估胞质内的基质金属蛋白酶-9(MMP-9)和基质金属蛋白酶抑制剂-1(TIMP-1)的含量。结果正常组和DHA组之间比较,差异无统计学意义(P>0.05);与正常组相比,TGF-β1组的ESCs数目增加,细胞分泌Ⅰ型胶原[(0.7334±0.0073)ng/L]与纤黏连蛋白[(0.6834±0.0081)ng/L]的量增多,细胞内MMP-9含量(0.471±0.030)降低,然而TIMP-1的含量(0.692±0.010)升高,差异均有统计学意义(均P<0.05);与TGF-β1组相比,TGF-β1+DHA组的ESCs数目减少,细胞分泌Ⅰ型胶原[(0.6543±0.0101)ng/L]和纤黏连蛋白[(0.5549±0.0092)ng/L]的量降低,细胞内MMP-9含量(0.593±0.026)升高,而TIMP-1含量(0.543±0.013)降低,差异均有统计学意义(均P<0.05)。结论ESCs在TGF-β1的作用下数目增多且分泌功能活跃,而DHA能抑制TGF-β1对子宫内膜间质细胞的这种作用。

Objective To observe the effect of dihydroartemisinin(DHA)on the number and activity of human endometrial stromal cells(ESCs)incubated with transforming growth factor-β1(TGF-β1).Methods Normal human endometrial tissue was collected and ESCs were isolated from January 1 to June 1 in 2018.Experiment had four groups,which were normal group:add serum free culture medium,TGF-β1 group:add 10 ng/ml TGF-β1 on the basis of normal group,DHA group:add 100 ng/ml DHA on the basis of normal group,TGF-β1+DHA group:add 100 ng/ml DHA on the basis of TGF-β1 group.The cell proliferation of each group was detected by thiazole blue method.The contents of collagen typeⅠand fibronectin in the supernatant of each group were detected by enzyme linked immunosorbent assay(ELISA).The contents of matrix metalloproteinase-9(MMP-9)and matrix metalloproteinase inhibitor-1(TIMP-1)were evaluated by Western blot.Results The difference was statistically significant between normal group and DHA group(P>0.05).Compared with normal group,the number of ESCs in TGF-β1 group increased,the amount of collagen typeⅠ[(0.7334±0.0073)ng/L]and fibronectin[(0.6834±0.0081)ng/L]secreted by the cells increased,the content of MMP-9(0.471±0.030)in the cells decreased,while TIMP-1(0.692±0.010)increased,and the differences were statistically significant(all P<0.05).Compared with TGF-β1 group,the number of ESCs in TGF-β1+DHA group decreased,the amount of collagen typeⅠ[(0.6543±0.0101)ng/L]and fibronectin[(0.5549±0.0092)ng/L]secreted by the cells decreased,the content of MMP-9(0.593±0.026)increased,while TIMP-1(0.543±0.013)decreased,and the differences were statistically significant(all P<0.05).Conclusion TGF-β1 can stimulate the proliferation and enhance the activity of ESCs,while DHA can inhibit the effect of TGF-β1 on ESCs.