摘要
目的探讨正常大鼠和盆腔脏器脱垂大鼠子宫骶韧带组织中磷酸酶与张力蛋白同源基因(PTEN)/磷脂酰肌醇3-激酶(PI3K)/蛋白激酶B(Akt)/FOXO1信号通路表达差异及补中益气汤对其表达的影响。方法取健康无生育史雌性SD大鼠10只作为正常组;取20只自然分娩过3次的雌性SD大鼠制备盆腔脏器脱垂模型,造模成功后将大鼠随机分为模型组和补中益气汤组各10只。然后模型组和正常组大鼠给予生理盐水灌胃8周,补中益气汤组大鼠给予补中益气汤7 mL/(kg·d)灌胃8周。末次灌胃结束处死各组大鼠,Western blot法测定子宫骶韧带组织中PTEN、p-Akt/Akt、p-FOX1/FOX1、谷胱甘肽过氧化物酶1(GPX1)、含锰超氧化物歧化酶(Mn-SOD)、胶原蛋白Ⅰ(COL1A1)、胶原蛋白Ⅲ(COL3A1)蛋白表达情况,RT-PCR法测定子宫骶韧带组织中GPX1、Mn-SOD、COL1A1、COL3A1 mRNA表达情况。结果与正常组比较,模型组大鼠子宫骶韧带组织中PTEN、GPX1、Mn-SOD、COL1A1、COL3A1蛋白表达量及GPX1、Mn-SOD、COL1A1、COL3A1 mRNA表达量均明显降低(P均<0.05),p-Akt/Akt、p-FOX1/FOX1蛋白表达量均明显增高(P均<0.05);与模型组比较,补中益气汤组大鼠子宫骶韧带组织中PTEN、GPX1、Mn-SOD、COL1A1、COL3A1蛋白表达量及GPX1、Mn-SOD、COL1A1、COL3A1mRNA表达量均明显增高(P均<0.05),p-Akt/Akt、p-FOX1/FOX1蛋白表达量均明显降低(P均<0.05)。结论盆腔脏器脱垂大鼠子宫骶韧带组织中PTEN表达下调,胶原蛋白COL1A1、COL3A1合成减少;补中益气汤可增加子宫骶韧带组织中COL1A1、COL3A1的合成,其作用机制可能与抑制大鼠子宫骶韧带组织中PTEN/PI3K/Akt/FOXO1通路介导的氧化应激有关,即上调抗氧化酶GPX1、Mn-SOD的表达抑制氧化应激引起的细胞凋亡、衰老和胶原蛋白的破坏。
Objective It is to explore the difference of the expression of phosphatase and tensin homology deleted on chromosome ten(PTEN)/phosphatidylinositol 3 kinase(PI3K)/protein kinase B(Akt)/FOXO1 signaling pathway in uterosacral ligament between normal rats and rats with pelvic organ prolapse,and to observe the effect of Buzhong Yiqi Decoction on its expression.Methods Ten healthy female SD rats without reproductive history were selected as normal group,20 female SD rats who had been naturally delivered for 3 times were selected to establish models of pelvic organ prolapse.After the models were successfully established,the rats were randomly divided into model group and Buzhong Yiqi Decoction group with 10 rats in each group.The rats in the model group and normal group were treated with normal saline by gavage for 8 weeks,and the rats in the Buzhong Yiqi Decoction group were treated with Buzhong Yiqi Decoction 7 mL/(kg·d)by gavage for 8 weeks.At the end of the last gavage,the rats in each group were sacrificed,the protein expression of PTEN,p-Akt/Akt,p-FOX1/FOX1,glutathione peroxidase 1(GPX1),manganese containing superoxide dismutase(Mn-SOD),collagen I(COL1A1),collagen III(COL3A1)in uterosacral ligament tissue were detected by Western blot method,the mRNA expression of GPX1,Mn SOD,COL1A1,COL3A1 in uterosacral ligament tissue were detected by RT-PCR method.Results Compared with the normal group,the expression of PTEN,GPX1,Mn-SOD,COL1A1,COL3A1 protein and the expression of GPX1,Mn-SOD,COL1A1,COL3A1 mRNA in the uterosacral ligament tissue of the rats in the model group were significantly decreased(all P<0.05),while the expression of p-Akt/Akt,p-FOX1/FOX1 protein were significantly increased(all P<0.05).Compared with the model group,the expression of PTEN,GPX1,Mn-SOD,COL1A1,COL3A1 protein and the expression of GPX1,Mn-SOD,COL1A1,COL3A1 mRNA in the uterosacral ligament tissue of the rats in the Buzhong Yiqi Decoction group were significantly increased(all P<0.05),while the expression of p-Akt/Akt,p-FOX1/FOX1 protein were significantly decreased(all P<0.05).Conclusion The expression of PTEN in uterosacral ligament tissue is down regulated,and the production of collagen COL1A1 and COL3A1 is reduced in pelvic organ prolapse rats.Buzhong Yiqi Decoction can promote the production of COL1A1 and COL3A1 in uterosacral ligament tissue,its mechanism may be related to the inhibition of oxidative stress in rat uterosacral ligament tissue mediated by PTEN/PI3K/Akt/FOXO1 pathway,that is up regulation of the expression of antioxidant enzymes GPX1 and Mn-SOD which inhibit cell apoptosis,aging and collagen damage caused by oxidative stress.
作者
严晓
高立群
林艳
许岚
张忻佩
朱小丹
胡佳贞
樊伯珍
YAN Xiao;GAO Liqun;LIN Yan;XU Lan;ZHANG Xinpei;ZHU Xiaodan;HU Jiazhen;FAN Bozhen(Putuo Hospital Affiliated to Shanghai University of Traditional Chinese Medicine,Shanghai 200062,China;Shanghai University of Traditional Chinese Medicine,Shanghai 201203,China)
出处
《现代中西医结合杂志》
CAS
2022年第18期2483-2487,2501,共6页
Modern Journal of Integrated Traditional Chinese and Western Medicine
基金
上海中医药大学预算内科研项目(2020LK073)
上海市医学引导类(中、西医)科技支撑项目(18411970500)
上海市普陀区科学技术委员会卫生系统重点项目(ptkwws201906)。
