Triptolide induces autophagy of ovarian granulosa cells via PI3K/AKT/m TOR pathway / 中国中药杂志

The aim of this paper was to observe the concentration,time and mechanism of autophagy induced by triptolide( TP) in ovarian granulosa cells( OGCs). CCK-8 method was used to compare the inhibitory effects of TP at different concentrations on primary cultured rat OGCs and IC50 was calculated. The effects of TP at different concentrations and time points on the expression of OGCs autophagy factor protein and the cascade of PI3 K/AKT/m TOR pathway were detected by Western blot. The effects of TP,autophagy inducer( brefeldin A) and PI3 K/m TOR inhibitor( NVP-BEZ235) on the expression of PI3 K/AKT/m TOR cascade and autophagy related factor protein were detected by Western blot. The results show that the IC50 of different concentrations of TP on OGCs of rat ovary was14. 65 μmol·L-1,and the minimum inhibitory concentration of TP was 0. 1 μmol·L-1( 100 nmol·L-1). Compared with the control group,the expression levels of beclin1 and LC3Ⅱ in each group were significantly higher than those in the control group( P<0. 05 or P<0. 01). After 12 hours of treatment with TP,brefeldin A and NVP-BEZ235,respectively,compared with the control group,TP could significantly promote the expression level of downstream autophagy effect or molecule beclin1,LC3Ⅱ and inhibit the expression level of LC3Ⅰ,p62 protein( P<0. 05 or P< 0. 01). Moreover,the expression of beclin1 and LC3Ⅱ/LC3Ⅰ in TP group was higher than that in brefeldin A group( P<0. 05 or P<0. 01),and the expression of p62 in TP group was lower than that in brefeldin A group( P<0. 05 or P<0. 01). At the same time,TP could significantly inhibit the expression of p-PI3 K,p-AKT,p-mTOR protein,and the inhibitory effect of TP was better than that of NVP-BEZ235 group. This study suggests that 100 nmol·L-1 TP could induce OGCs autophagy successfully in cultured rat ovary for 12 h; TP may induce OGCs autophagy by inhibiting PI3 k/Akt/m TOR signaling pathway.

Effect of Traditional Chinese Medicine in Regulating Apoptosis and Autophagy Through mTOR Pathway / 中国实验方剂学杂志

Mammalian target of rapamycin (mTOR) is an evolutionary conservative serine/threonine protein kinase. Its biological function is mainly to participate in cell proliferation, growth and differentiation, so as to regulate the body's metabolic process. Many domestic and foreign studies have shown that mTOR is the junction of apoptosis and autophagy signal transduction pathways. Various stimuli, such as nutrition, drugs and oxidative stress, may play a key regulatory role in cell apoptosis and autophagy through mTOR-mediated signaling pathways. At present, many studies have shown that the change in mTOR signaling pathways is closely related to the pathogenesis of many human diseases, such as cancer, metabolic disorders(obesity and type 2 diabetes), cardiovascular and neurodegenerative disease, age-related diseases and disorders of follicular. In recent years, more and more doctors have studied the regulatory effect of traditional Chinese medicine on apoptosis and autophagy, with the pathways as the starting point and cell apoptosis and autophagy as the research carriers. These studies include experimental studies on the regulation of apoptosis and autophagy by monomers of traditional Chinese medicine (TCM), Chinese patent medicine, compound prescription of TCM, acupuncture and other drugs and physiotherapy. Starting from the mTOR signaling pathways, this paper discusses the relationship between mTOR, apoptosis and autophagy, and reviews the recent progress of TCM in regulating apoptosis and autophagy through mTOR pathways, so as to provide ideas and references for further studies in this field. In the future, TCM doctors can still explore the time-effect relationship between apoptosis and autophagy based on mTOR signaling pathways under the guidance of the basic theory of Chinese medicine, in order to provide theoretical support and targets for the action mechanism of TCM on bodies.