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 671000, China.
Zhongguo Zhong Yao Za Zhi. 2020 Dec;45(24):6036-6042. doi: 10.19540/j.cnki.cjcmm.20200901.405.
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-Ⅱ and P16(INK4 a). Fluorescence quantitative PCR was used to detect the changes in mRNA expressions of PI3 K, Akt, mTOR, S6 k, LC3-Ⅱ and P16(INK4 a). 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(INK4 a) increased, while the expression of autophagy signaling molecule LC3-Ⅱ decreased. 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-Ⅱ in Rg_1 group was higher than that in D-gal group, while the expression level of senescence marker P16(INK4 a) 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.
本文旨在研究人参皂苷Rg_1（Rg_1）对D - 半乳糖（D - gal）诱导的小鼠卵巢早衰模型中磷脂酰肌醇3激酶（PI3 K）、蛋白激酶B（Akt）和哺乳动物雷帕霉素靶蛋白（mTOR）的抑制作用。将54只雌性SPF级BALB/c小鼠随机分为PBS组、D - gal组和Rg_1组。D - gal组小鼠颈部和背部皮下注射D - 半乳糖（200 mg·kg⁻¹·d⁻¹），连续注射42天。PBS组小鼠颈部和背部皮下注射等量的磷酸盐缓冲液（PBS），连续注射42天。Rg_1组在第15天起除采用D - gal组的治疗方法外，腹腔注射Rg_1（20 mg·kg⁻¹·d⁻¹），持续28天，同时，D - gal组和PBS组自第15天起也腹腔注射等量的PBS，持续28天。治疗后，检测小鼠的动情周期变化，采用卵巢SA - β - Gal染色检测卵巢衰老变化。采用蛋白质免疫印迹法检测PI3 K、Akt、mTOR、S6 k、LC3 - Ⅱ和P16ⁱⁿᵏ⁴ᵃ蛋白表达变化。采用荧光定量PCR检测PI3 K、Akt、mTOR、S6 k、LC3 - Ⅱ和P16ⁱⁿᵏ⁴ᵃ mRNA表达变化。结果显示，与PBS组相比，D - gal组在第3周开始出现动情周期紊乱，D - gal组卵巢SA - β - Gal染色阳性颗粒细胞增多，衰老标志物P16ⁱⁿᵏ⁴ᵃ表达增加，而自噬信号分子LC3 - Ⅱ表达降低。Rg_1治疗后，Rg_1组卵巢SA - β - Gal染色阳性率降低，Rg_1组自噬信号分子LC3 - Ⅱ表达水平高于D - gal组，而衰老标志物P16ⁱⁿᵏ⁴ᵃ表达水平低于D - gal组。与PBS组相比，D - gal组PI3 K、Akt、mTOR和S6 k的蛋白及mRNA表达上调，Rg_1组Akt、mTOR和S6 k的蛋白表达上调，PI3 K和mTOR的mRNA表达上调。Rg_1治疗后，Rg_1组PI3 K、Akt、mTOR和S6 k的蛋白表达低于D - gal组，而Rg_1组Akt、mTOR和S6 k的mRNA表达低于D - gal组。研究结果表明，Rg_1对D - gal诱导的小鼠模型卵巢早衰具有延缓作用，PI3 K/Akt/mTOR自噬信号通路发挥重要作用。
