【摘要】 目的:通过检测子宫内膜异位症(EMT)痛经大鼠异位内膜PI3K/AKT信号通路相关基因蛋白表达水平,探讨敦煌疗风虚瘦弱方对EMT痛经大鼠异位内膜的影响。方法:将60只SPF级SD雌性大鼠按随机数表法分为空白组(n=10)和造模组(n=50),造模组采用自体移植法制备EMT痛经大鼠模型,将成模大鼠按照随机数表法分为模型组、孕三烯酮组(0.24g/kg)和敦煌疗风虚瘦弱方高、中、低剂量组(14、7、3.5g/kg),每组10只。疗风虚瘦弱方各剂量组每日灌胃1次,孕三烯酮组每周灌胃2次,空白组、模型组每日蒸馏水2mL灌胃1次,持续干预12d。处死前各组大鼠均给与缩宫素,观察疼痛状态,随后麻醉处死剖取大鼠异位内膜组织;采用HE染色法观察大鼠异位内膜组织病理形态学改变;采用实时荧光定量PCR法和蛋白免疫印迹法分别检测各组大鼠异位内膜组织PI3K、AKT基因、蛋白表达水平;采用ELISA法检测各组大鼠异位内膜组织COX-2、VEGF含量。结果:模型组疼痛潜伏期短,疼痛扭体次频并且剧烈;各治疗组疼痛潜伏期延长,疼痛扭体次数减少,程度明显减轻,病理提示异位内膜组织炎症细胞浸润显著减轻,血液供应、组织增生及腺体均不同程度减少;分子生物学检测示大鼠异位内膜组织,孕三烯酮组与疗风虚瘦弱方高、中剂量组PI3K、AKT基因蛋白表达及下游因子COX-2、VEGF含量均显著降低(P<0.05)。结论:敦煌疗风虚瘦弱方显著改善EMT痛经大鼠疼痛状态,其机制可能与该方有效调控PI3K/AKT信号通路进而抑制异位内膜生长有关。更多还原

【Abstract】 Objective: To investigate the effect of Dunhuang Liaofeng Xushouruo Decoction on ectopic endometrium of endometriosis(EMT) dysmenorrhea rats by detecting the protein expression level of PI3 K/AKT signaling pathway related genes.Methods: Sixty SPF female SD rats were randomly divided into blank group(n=10) and model group(n=50) according to the random number table method. Model rats were divided into model group, gestrinone group(0.24 g/kg) and Dunhuang Liaofeng Xushouruo Decoction high, medium and low groups(14, 7, 3.5 g/kg) according to the random number table method, each group 10 rats. The Liaofeng Xushouruo Decoction group was gave once a day, the gestrinone group was gave twice a week, and the blank group and model group were gave with 2 mL of distilled water once a day, and continued intervention for 12 days. Before sacrifice,rats in each group were given oxytocin to observe the state of pain, then the ectopic endometrial tissues of the rats were sacrificed by anesthesia; the pathological changes of the ectopic endometrial tissues of the rats were observed by HE staining method; realtime fluorescence quantification was used PCR and Western blotting were used to detect the expression levels of PI3 K, AKT gene and protein in ectopic endometrial tissues of rats in each group; ELISA method was used to detect the levels of COX-2 and VEGF in ectopic endometrial tissues of rats in each group. Results: The model group had a short latency period of pain, frequent and severe painful writhing; The pain latency of each treatment group was prolonged, the number of painful writhing times was reduced, and the degree was significantly reduced. Pathology suggests ectopic endometrial tissue inflammatory cell infiltration was significantly reduced, and blood supply, tissue proliferation and glands were reduced to varying degrees; Molecular biology tests showed that the rat ectopic endometrial tissue, the expression of PI3 K, AKT gene protein and the content of downstream factors COX-2 and VEGF in the gestrinone group, Dunhuang Liaofeng Xushouruo Decoction, high-, medium-groups, were significantly reduced(P<0.05). Conclusion: Dunhuang Liaofeng Xushouruo Decoction can significantly improve the pain state of EMT dysmenorrhea rats, and its mechanism may be related to its effective regulation of PI3 K/AKT signaling pathway and inhibition of ectopic endometrial growth.更多还原