摘要
研究磷脂酰肌醇3-激酶(posphoinositide 3-kinase,PI3K)、蛋白激酶B(protein kinase B,Akt)和哺乳动物雷帕霉素靶蛋白(mammalian target of rapamycin,mTOR)自噬信号通路在人参皂苷Rg_(-1)(ginsenoside Rg_(-1),Rg_(-1))延缓D-半乳糖(D-galactose,D-gal)诱导的卵巢早衰小鼠模型卵巢早衰中的作用。将SPF级雌性BALB/c小鼠54只随机分成PBS组、D-gal组、Rg_(-1)组。D-gal组给予颈背部皮下注射D-半乳糖200 mg·kg^(-1)·d^(-1),连续给药42 d;PBS组给予颈背部注射等时等量磷酸盐缓冲液(PBS),连续注射42 d;Rg_(-1)组在D-gal组处理基础上,于第15天开始给予腹腔注射Rg_(-1)20 mg·kg^(-1)·d^(-1),连续给药28 d,同时D-gal组和PBS组也在第15天开始给予腹腔注射等时等量PBS,连续给药28 d。给药后,检测小鼠动情周期改变,卵巢SA-β-Gal苷酶染色检测卵巢衰老状态改变,Western blot法检测PI3K,Akt,mTOR,S6k,LC3-Ⅱ,P16^(INK4a)蛋白表达的改变。荧光定量PCR法检测衰老调控因子PI3K,Akt,mTOR,S6k,P16^(INK4a)mRNA表达的改变。研究结果显示,与PBS组比较,D-gal组第3周开始出现动情周期紊乱,卵巢SA-β-半乳糖苷酶染色阳性率升高,衰老标记物P16^(INK4a)表达量升高,自噬信号分子LC3-Ⅱ表达量降低;Rg_(-1)作用后,Rg_(-1)组卵巢SA-β-半乳糖苷酶染色阳性率降低,衰老标记物P16^(INK4a)表达量低于D-gal组,自噬信号分子LC3-Ⅱ表达量高于D-gal组。与PBS组比较,D-gal组PI3K,Akt,mTOR,S6k蛋白和mRNA表达上调,Rg_(-1)组Akt,mTOR,S6k蛋白表达上调,PI3K,mTOR mRNA表达上调。Rg_(-1)作用后,Rg_(-1)组PI3K,Akt,mTOR,S6k蛋白表达量低于D-gal组,Akt,mTOR,S6k mRNA表达量低于D-gal组。研究结果提示,Rg_(-1)具有延缓D-gal诱导的卵巢早衰小鼠模型中卵巢早衰的作用,PI3K/Akt/mTOR自噬信号通路在其中发挥重要功能。
The aim of this paper was to study the role of phosphoinositide 3-kinase(PI3 K),protein kinase B(Akt)and mamma-lian target of rapamycin(mTOR)in the inhibition of premature ovarian failure induced by D-galactose(D-gal)in mice model by ginsenoside Rg_(-1)(Rg_(-1)).Fifty-four female SPF BALB/c mice were randomly divided into PBS group,D-gal group,and Rg_(-1) group.In the D-gal group,D-galactose(200 mg·kg^(-1)·d^(-1))was injected subcutaneously into the neck and back for 42 days.In the PBS group,an equal amount of phosphate buffered saline(PBS)was injected into the neck and back for 42 days.In addition to the therapy of D-gal group,Rg_(-1) group was given Rg_(-1)(20 mg·kg^(-1)·d^(-1))through intraperitoneal injection since the 15 th day for 28 days,at the same time,the D-gal group and the PBS group were also given an equal amount of PBS through intraperitoneal injection since the 15 th day for 28 days.After the treatment,the estrous cycle changes of the mice were detected,and the ovarian SA-β-Gal staining was used to detect the changes of ovarian aging.Western blot was used to detect the changes in protein expressions of PI3 K,Akt,mTOR,S6 k,LC3-Ⅱand P16^(INK4a).Fluorescence quantitative PCR was used to detect the changes in mRNA expressions of PI3 K,Akt,mTOR,S6 k,LC3-Ⅱand P16^(INK4a).According to the findings,compared with the PBS group,the D-gal group began to show estrous cycle disorder in the 3 rd week,the ovarian SA-β-Gal staining positive granulosa cells increased in the D-gal group,the expression of senescence marker P16^(INK4a) increased,while the expression of autophagy signaling molecule LC3-Ⅱdecreased.After treatment with Rg_(-1),the positive rate of ovarian SA-β-Gal staining in Rg_(-1) group decreased,the expression level of autophagy signaling molecule LC3-Ⅱin Rg_(-1) group was higher than that in D-gal group,while the expression level of senescence marker P16^(INK4a) was lower than that in D-gal group.Compared with the PBS group,the protein and mRNA expressions of PI3 K,Akt,mTOR and S6 k in the D-gal group were up-regulated,the protein expressions of Akt,mTOR and S6 k in the Rg_(-1) group were up-regulated,and the mRNA expressions of PI3 K and mTOR were up-regulated.After treatment with Rg_(-1),the protein expressions of PI3 K,Akt,mTOR and S6 k in the Rg_(-1) group were lower than those in the D-gal group,while the mRNA expressions of Akt,mTOR and S6 k in the Rg_(-1) group were lower than those in the D-gal group.The finding ssuggested that Rg_(-1) has the effect in delaying ovarian premature failure in D-gal-induced mouse models,and PI3 K/Akt/mTOR autophagy signaling pathways play an important role.
作者
刘小虎
赵志慧
周玥
王翠丽
韩艳军
周雯
LIU Xiao-hu;ZHAO Zhi-hui;ZHOU Yue;WANG Cui-li;HAN Yan-jun;ZHOU Wen(Department of Histology and Embryology,School of Basic Medical Sciences,Dali University,Dali 671(XM),China)
出处
《中国中药杂志》
CAS
CSCD
北大核心
2020年第24期6036-6042,共7页
China Journal of Chinese Materia Medica
基金
国家自然科学基金项目(81860038,81660731,81873103)
云南省教育厅科学研究基金项目(2020Y0541)。
