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目的观察内质网应激导致凋亡在子宫肌瘤及子宫平滑肌组织中表达的差异,为子宫动脉阻断术治疗子宫肌瘤提供理论依据。方法选择2011年6-12月上海市杨浦区中心医院妇科收治的子宫肌瘤患者25例,均行腹腔镜下子宫动脉阻断术。分别于子宫动脉阻断前及阻断后30 min采集子宫肌瘤及其周边平滑肌组织各2 g。在低氧状况下(1%O2)原代培养。运用RT-PCR、Western Blot法检测并比较两种组织中mRNA和内质网应激相关分子的表达情况,包括葡萄糖调节蛋白78(GRP78)、C/EBP同源蛋白(CHOP)、C-Jun氨苯末端激酶(JNK)、Bax、Bcl-2和Caspase-4,同时检测Caspase-4活性。结果肌瘤组织中内质网应激相关分子的表达显著高于平滑肌组织。在肌瘤细胞中,抗凋亡蛋白Bcl-2的表达低于平滑肌细胞。在细胞培养的起初12 h,平滑肌细胞中Caspase-4的表达和活性无明显升高。结论内质网应激导致凋亡在子宫肌瘤及平滑肌组织中的表达有差异,该系统差异表达可以为子宫动脉阻断术治疗子宫肌瘤提供理论依据。
Objective To observe the difference of endoplasmic reticulum stress-induced apoptosis in uterine leiomyoma and uterine smooth muscle, and to provide a theoretical basis for the treatment of uterine fibroids by uterine artery occlusion. Methods Twenty-five patients with uterine fibroids underwent gynecological surgery at Yangpu District Central Hospital from June to December in 2011. All patients underwent laparoscopic uterine artery occlusion. Uterine fibroids and peripheral smooth muscle tissues were collected 2 g before uterine artery occlusion and 30 min after occlusion, respectively. Primary culture was performed under hypoxic conditions (1% O2). RT-PCR and Western Blot were used to detect and compare the expression of mRNA and ER in two tissues including GRP78, CHOP, C-Jun Aminobenzenesterminal kinase (JNK), Bax, Bcl-2 and Caspase-4, simultaneously detected Caspase-4 activity. Results The expression of ER stress-related molecules in fibroids was significantly higher than that in smooth muscle. In fibroids, the expression of anti-apoptotic protein Bcl-2 is lower than that of smooth muscle cells. Caspase-4 expression and activity were not significantly increased in smooth muscle cells during the first 12 h of cell culture. Conclusion Endoplasmic reticulum stress leads to differences in the expression of apoptosis in uterine leiomyoma and smooth muscle. The differential expression of this system can provide a theoretical basis for the treatment of uterine fibroids by uterine artery occlusion.