目的 探讨姜黄素介导Janus激酶2(Janus kinase 2，JAK2)/信号转导转录激活因子3(signal transducers and transcrip-tional activators 3，STAT3)信号通路对子宫内膜异位症(endometriosis，EMs)大鼠的影响。方法 随机选择10只雌性Wistar大鼠作为假手术组，剩余大鼠于第3个动情期通过子宫内膜自体移植构建EMs模型，建模成功后随机分为模型组、姜黄素低剂量组(48mg·kg-1)、姜黄素高剂量组(96mg·kg-1)及达那唑组(36 mg·kg-1)，每组10只，假手术组及模型组大鼠灌服等体积生理盐水，每日1次，连续处理15 d。评估治疗后腹腔内粘连程度;检测异位内膜体积;HE染色观察异位内膜病理形态学改变;酶联免疫吸附试验(enzyme linked immunosorbent assay，ELISA)检测大鼠腹腔灌洗液中肿瘤坏死因子-α(tumor necrosis factor-α，TNF-α)、白细胞介素-6(interleukin-6，IL-6)和血管内皮生长因子(vascular endothelial growth factor，VEGF)水平;免疫组织化学技术检测异位内膜CD31、增殖细胞核抗原(proliferating cell nuclear antigen，PCNA)阳性表达;蛋白印迹法(Western blotting)法检测子宫内膜组织中 B 细胞淋巴瘤/白血病-xl(B-cell lymphocytoma/leukemia xl，Bcl-xl)、Bcl-2 相关 X蛋白(Bcl-2 related X protein，Bax)蛋白的相对表达水平及JAK2、STAT3蛋白磷酸化情况。结果 成功构建EMs大鼠模型;模型组大鼠腹腔内粘连评分升高，异位子宫内膜体积增大，间质细胞增多，结构致密，血供丰富。与假手术组比较，模型组大鼠异位子宫内膜组织中PCNA、CD31阳性细胞比例及腹腔灌洗液中TNF-α、IL-6、VEGF水平均升高，p-JAK2/JAK2、p-STAT3/STAT3比值及Bcl-xl蛋白高表达，Bax蛋白低表达(P＜0。05);与模型组比较，姜黄素低剂量组、姜黄素高剂量组及达那唑组大鼠腹腔内粘连评分均降低，异位子宫内膜体积减小(P＜0。05)，上皮结构不完整，间质细胞稀疏、结构松散和炎性细胞浸润，供血不足，异位子宫内膜PCNA、CD31阳性细胞比例及腹腔灌洗液中TNF-α、IL-6和VEGF水平降低，且异位子宫内膜组织中p-JAK2/JAK2、p-STAT3/STAT3比值及Bcl-xl蛋白低表达，Bax蛋白高表达(P＜0。05);姜黄素低剂量组、姜黄素高剂量组及达那唑组对降低EMs大鼠异位子宫内膜体积、JAK2/STAT3信号通路相关因子表达及异位子宫内膜结构的病理损伤作用逐渐增强(P＜0。05)。结论 姜黄素可减小异位子宫内膜体积，增强对异位子宫内膜的损伤，其机制可能与抑制JAK2/STAT3信号通路激活进而降低炎性因子水平，减少细胞增殖、促进细胞凋亡并抑制血管新生有关。
Effect of curcumin on endometrial volume in endometriosis rats and its mechanism
Objective To investigate the effect of curcumin-mediated Janus kinase 2(JAK2)/signal transducers and transcriptional activators 3(STAT3)signaling pathway in endometriosis(EMs)rats.Methods Ten female Wistar rats were randomly selected as the sham operation group,and the remaining rats were randomly divided into model group,curcumin low(48 mg·kg-1),curcumin high dose group(96 mg·kg-1)and danazol group(36 mg·kg-1)after successful modeling by endometrial auto trans-plantation in the third estrous stage,with 10 rats in each group.Rats in sham operation group and model group were given equal volume of normal saline orally,once a day,for consecutive 15 days.The degree of intraperitoneal adhesion after treatment was evaluated.The volume of ectopic endometrium was detected.The pathological changes of ectopic endometrium were observed by HE staining.The levels of tumor necrosis factor-α(TNF-α),interleukin-6(IL-6)and vascular endothelial growth factor(VEGF)in abdominal lavage fluid of rats were determined by enzyme linked immunosorbent assay(ELISA).The positive ex-pression of CD31 and proliferating nuclear antigen(PCNA)in ectopic endometrium was detected by immunohistochemistry.The relative expression levels of B-cell lymphoma/leukemia-xl(Bcl-xl)and Bcl-2 associated X protein(Bax)and the phos-phorylation of JAK2 and STAT3 proteins in endometrial tissues were detected by Western blotting.Results The EMs rat model was successfully constructed.In model group,the intraperitoneal adhesion score increased,the volume of ectopic endometrium increased,the interstitial cells increased,the structure was dense,and the blood supply was rich.Compared with the sham operation group,the proportion of PCNA and CD31 positive cells in the ectopic endometrium tissues and the levels of TNF-α,IL-6 and VEGF in the peritoneal irrigation fluid in model group were increased.The p-JAK2/JAK2,p-STAT3/STAT3 ratio and the relative expression level of Bcl-xl protein were increased,while the relative expression level of Bax protein was decreased(P＜0.05).Compared with model group,the intraperitoneal adhesion score,ectopic endometrial volume decreased in curcumin low-dose,high-dose and danazol groups(P＜0.05),the epithelial structure was incomplete,the interstitial cells were sparse and loose,the inflammatory cells were infiltrated,and the blood supply was insufficient.The proportion of PCNA and CD31 positive cells in ectopic endometrium and the levels of TNF-α,IL-6 and VEGF in peritoneal perfusion fluid decreased,and the p-JAK2/JAK2,p-STAT3/STAT3 ratio and relative expression of Bcl-xl protein in ectopic endometrium tissue de-creased.The relative expression level of Bax protein was increased(P＜0.05).The effects of curcumin low-dose,high-dose and danazol group on reducing the volume of ectopic endometrium,the expression of JAK2/STAT3 signal pathway related fac-tors and the pathological damage of ectopic endometrium structure in EMs rats were gradually enhanced(P＜0.05).Conclusion Curcumin could reduce the volume of ectopic endometrium and enhance the damage to ectopic endometrium.The mechanism may be related to the inhibition of JAK2/STAT3 signaling pathway activation,thereby reducing the level of inflammatory fac-tors,reducing cell proliferation,promoting apoptosis and inhibiting angiogenesis.
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