Ji Xiu-Jia, Zhang Xiao-Hua, Huang Can-Can, Zhang Zuo-Liang, Mao Hai-Yan, Yue Bin, Liu Bing-Yu, Wu Quan-Sheng
Gansu University of Chinese Medicine Lanzhou 730000, China.
Gansu Provincial People's Hospital Lanzhou 730030, China.
Zhongguo Zhong Yao Za Zhi. 2023 Jun;48(12):3207-3214. doi: 10.19540/j.cnki.cjcmm.20221219.402.
The present study aimed to investigate the protective role of Shaofu Zhuyu Decoction(SFZY) against endometriosis fibrosis in mice, and decipher the underlying mechanism through the phosphatase and tensin homolog deleted on chromosome ten(PTEN)/protein kinase B(Akt)/mammalian target of rapamycin(mTOR) pathway. Eighty-five BALB/c female mice were randomly assigned into a blank group, a model group, high-, medium, and low-dose SFZY(SFZY-H, SFZY-M, and SFZY-L, respectively) groups, and a gestrinone suspension(YT) group. The model of endometriosis was induced by intraperitoneal injection of uterine fragments. The mice in different groups were administrated with corresponding groups by gavage 14 days after modeling, and the blank group and model group with equal volume of distilled water by gavage. The treatment lasted for 14 days. The body weight, paw withdrawal latency caused by heat stimuli, and total weight of dissected ectopic focus were compared between different groups. The pathological changes of the ectopic tissue were observed via hematoxylin-eosin(HE) and Masson staining. Real-time PCR was employed to measure the mRNA levels of α-smooth muscle actin(α-SMA) and collagen type Ⅰ(collagen-Ⅰ) in the ectopic tissue. The protein levels of PTEN, Akt, mTOR, p-Akt, and p-mTOR in the ectopic tissue were determined by Western blot. Compared with the blank group, the modeling first decreased and then increased the body weight of mice, increased the total weight of ectopic focus, and shortened the paw withdrawal latency. Compared with the model group, SFZY and YT increased the body weight, prolonged the paw withdrawal latency, and decreased the weight of ectopic focus. Furthermore, the drug administration, especially SFZY-H and YT(P<0.01), recovered the pathological and reduced the area of collagen deposition. Compared with the blank group, the modeling up-regulated the mRNA levels of α-SMA and collagen-Ⅰ in the ectopic focus, and such up-regulation was attenuated after drug intervention, especially in the SFZY-H and YT groups(P<0.05,P<0.01). Compared with the blank group, the modeling down-regulated the protein level of PTEN and up-regulated the protein levels of Akt, mTOR, p-Akt, and p-mTOR(P<0.01, P<0.001). Drug administration, especially SFZY-H and YT, restored such changes(P<0.01). SFZY may significantly attenuate the focal fibrosis in the mouse model of endometriosis by regulating the PTEN/Akt/mTOR signaling pathway.
本研究旨在探讨少腹逐瘀汤（SFZY）对小鼠子宫内膜异位症纤维化的保护作用，并通过10号染色体缺失的磷酸酶和张力蛋白同源物（PTEN）/蛋白激酶B（Akt）/雷帕霉素哺乳动物靶蛋白（mTOR）通路阐明其潜在机制。85只BALB/c雌性小鼠随机分为空白组、模型组、高、中、低剂量SFZY组（分别为SFZY-H、SFZY-M和SFZY-L）和孕三烯酮混悬液（YT）组。通过腹腔注射子宫碎片诱导子宫内膜异位症模型。建模14天后，不同组小鼠通过灌胃给予相应药物，空白组和模型组灌胃等量蒸馏水。治疗持续14天。比较不同组之间的体重、热刺激引起的爪缩潜伏期以及解剖异位病灶的总重量。通过苏木精-伊红（HE）和Masson染色观察异位组织的病理变化。采用实时定量聚合酶链反应（Real-time PCR）检测异位组织中α-平滑肌肌动蛋白（α-SMA）和Ⅰ型胶原（collagen-Ⅰ）的mRNA水平。通过蛋白质免疫印迹法（Western blot）检测异位组织中PTEN、Akt、mTOR、p-Akt和p-mTOR的蛋白水平。与空白组相比，建模后小鼠体重先下降后上升，异位病灶总重量增加，爪缩潜伏期缩短。与模型组相比，SFZY和YT增加了体重，延长了爪缩潜伏期，并减轻了异位病灶重量。此外，给药后，尤其是SFZY-H和YT组（P<0.01），恢复了病理状态并减少了胶原沉积面积。与空白组相比，建模上调了异位病灶中α-SMA和collagen-Ⅰ的mRNA水平，药物干预后这种上调减弱，尤其是在SFZY-H和YT组（P<0.05，P<0.01）。与空白组相比，建模下调了PTEN蛋白水平，上调了Akt、mTOR、p-Akt和p-mTOR蛋白水平（P<0.01，P<0.001）。给药后，尤其是SFZY-H和YT组恢复了这些变化（P<0.01）。SFZY可能通过调节PTEN/Akt/mTOR信号通路显著减轻小鼠子宫内膜异位症模型中的病灶纤维化。
