川芎嗪对子宫内膜异位症细胞侵袭和凋亡的影响

Effect of ligustrazine on invasion and apoptosis in endometriosis cells

目的 探讨川芎嗪调控长链非编码RNA(lncRNA)UBOX5-AS1对子宫内膜异位症细胞侵袭、凋亡和基质金属蛋白酶9(MMP-9)/基质金属蛋白酶组织抑制因子3(TIMP-3)信号通路的影响。方法 分离人子宫内膜异位症细胞,分成对照组、实验低剂量组(12μmol·L^(-1)川芎嗪处理)、实验中剂量组(24μmol·L^(-1)川芎嗪处理)、实验高剂量组(48μmol·L^(-1)川芎嗪处理)、实验高剂量+NC组(48μmol·L^(-1)川芎嗪、阴性对照载体处理)、实验高剂量+UBOX5-AS1组(48μmol·L^(-1)川芎嗪、UBOX5-AS1过表达载体处理)。以实时荧光定量逆转录聚合酶链反应(qRT-PCR)方法检测UBOX5-AS1表达,以细胞计数试剂盒-8(CCK-8)检测细胞增殖情况,以流式细胞术测定细胞凋亡情况,以蛋白质印迹法检测B淋巴细胞瘤-2相关X蛋白(Bax)、上皮钙黏素(E-cadherin)、波形蛋白(Vimentin)、MMP-9、TIMP-3蛋白表达,以Transwell小室检测细胞侵袭。结果 对照组、实验低剂量组、实验中剂量组、实验高剂量组、实验高剂量+NC组、实验高剂量+UBOX5-AS1组子宫内膜异位症细胞中UBOX5-AS1表达水平分别为1.00±0.07,0.84±0.14,0.67±0.05,0.45±0.06,0.44±0.06和0.92±0.09;细胞存活率分别为(100.00±4.38)%,(83.87±5.45)%,(68.86±2.85)%,(52.17±4.64)%,(51.15±3.54)%和(79.92±5.99)%;凋亡率分别为(2.41±0.27)%,(9.60±0.93)%,(14.89±1.15)%,(17.54±0.96)%,(17.27±1.71)%和(9.75±1.28)%;Bax蛋白表达水平分别为0.26±0.04,0.33±0.03,0.40±0.02,0.53±0.03,0.54±0.04和0.34±0.03;细胞侵袭数目分别为105.32±5.66,79.05±7.80,64.59±4.09,51.50±3.28,50.61±5.13和72.08±9.37;上述指标之间比较,实验低、中、高剂量组和对照组相比,差异均有统计学意义(均P<0.05);实验高剂量+UBOX5-AS1组与实验高剂量+NC组相比,差异均有统计学意义(均P<0.05)。结论 川芎嗪下调UBOX5-AS1抑制子宫内膜异位症细胞侵袭,诱导细胞凋亡,抑制MMP-9/TIMP-3信号激活。

Objective To investigate the effects of ligustrazine on the invasion,apoptosis and matrix metalloprotease 9 (MMP-9)/tissue inhibitors of MMP 3 (TIMP-3) of endometriosis cells through regulating long noncoding RNA (lncRNA) UBOX5-AS1.Methods Human endometriosis cells were isolated and divided into control group,experimental low-dose group (12μmol·L^(-1)ligustrazine),experimental medium-dose group (24μmol·L^(-1)ligustrazine),and experimental high-dose group (4 8μmol·L^(-1)ligustrazine),experimental high-dose+NC group (48μmol·L^(-1)ligustrazine,negative control vehicle),experimental highdose+UBOX5-AS1 group (48μmol·L^(-1)ligustrazine,UBOX5-AS1 overexpression vector processing).Realtime quantitative polymerase chain reaction (qRT-PCR) method was used to detect UBOX5-AS1 expression,cell counting kit-8(CCK-8) experiment was used to detect proliferation,flow cytometry was used to detect apoptosis,Western blot was used to detect the protein expression of B cell lymphoma-2 associated X protein (Bax),epithelial cadherin (E-cadherin),Vimentin,MMP-9,TIMP-3,Transwell chamber was used to detect cell invasion.Results The expression levels of UBOX5-AS1 in endometriosis cells in the control group,the experimental lowdose group,the experimental medium-dose group,the experimental high-dose group,the experimental high-dose+NC group,and the experimental high-dose+UBOX5-AS1 group were 1.00±0.07,0.84±0.14,0.67±0.05,0.45±0.06,0.44±0.06 and 0.92±0.09;cell survival rates were (100.00±4.38)%,(83.87±5.45)%,(68.86±2.85)%,(52.17±4.64)%,(51.15±3.54)%and (79.92±5.99)%;apoptosis rates were (2.41±0.27)%,(9.60±0.93)%,(14.89±1.15)%,(17.54±0.96)%,(17.27±1.71)%and (9.75±1.28)%;Bax protein expressions were 0.26±0.04,0.33±0.03,0.40±0.02,0.53±0.03,0.54±0.04 and 0.34±0.03;the number of cell invasion was 105.32±5.66,79.05±7.80,64.59±4.09,51.50±3.28,50.61±5.13 and 72.08±9.37;there were statistically significant differences of the above indicators between experimental low-dose group,experimental medium-dose group,experimental high-dose group and control group (all P<0.05);there were statistically significant differences between experimental high-dose+UBOX5-AS1 group and experimental highdose+NC group (all P<0.05).Conclusion Ligustrazine inhibited endometriosis cell invasion,induced cell apoptosis,and inhibited MMP-9/TIMP-3 signal activation by down-regulating UBOX5-AS1.