苍附导痰丸含药血清调控大鼠卵巢颗粒细胞自噬的作用

目的:研究苍附导痰丸含药血清调控大鼠卵巢颗粒细胞(GCs)自噬的作用.方法:将3月龄SD大鼠分为生理盐水组(生理盐水,灌胃)、卵巢刺激素(FSH)注射组(10.71 IU/kg,皮下注射)和苍附导痰丸高、中、低剂量灌胃组[0.5、1、2 mg/g(以生药量计),灌胃],每组6只,每天皮下注射/灌胃给药1次,连续3天;末...

Effect of Cangfu Daotan Pill Containing Serum on Autophagy of Ovarian Granulosa Cells of Rats

OBJECTIVE:To study the effect of Cangfu daotan pill(CDP)containing serum on autophagy of ovarian granulosa cells(GCs)of rat.METHODS:Three-month-old SD rat were divided into normal saline group(normal saline,ig),FSH injection group(10.71 IU/kg,ih),CDP irrigation high-dose,medium-dose and low-dse groups[0.5,1,2 mg/g(by crude drug),ig],with 6 rats in each group.They were given relevant medicine subcutaneously/intragastrically,once a day,for consecutive 3 days.After last medication,blood sample was collected from the abdominal aorta to obtain drug-containing serum.GCs of rat were divided into blank control group,model group,FSH group(positive control)and CDP high-dose,medium-dose and low-dose groups.The autophagy model was induced by giving testosterone propionate,except that the blank control group was directly added with 100μL serum of normal saline group.Then model group was given 100μL serum of normal saline group,and administration groups were given 100μL drug-containing serum of corresponding drug group.The contents of estradiol(E2)and progesterone(P)in supernatant of cells were determined by ELISA.Western blot assay was used to detect protein expression of PI3K,Akt and mTOR in cells.The mRNA expression of PI3K,Akt,mTOR,Beclin 1,LC3Ⅰ,LC3Ⅱand p62 were detected by RT-PCR.RESULTS:Compared with blank control group,the content of E2 in supernatant,relative mRNA and protein expression of PI3K,Akt and mTOR were decreased significantly in model group(P<0.01),while relative mRNA expression of Beclin 1,LC3Ⅰand LC3Ⅱwere increased significantly(P<0.01).Compared with model group,the content of E2 in supernatant were significantly increased in FSH group,CDP medium-dose and high-dose groups,while relative mRNA expression of Beclin 1,LC3Ⅰ,LC3Ⅱand p62 were decreased significantly(P<0.05 or P<0.01);relative mRNA and protein expression of PI3K,Akt and mTOR were increased significantly in administration groups(P<0.05 or P<0.01).CONCLUSIONS:CDP can inhibit autophagy of GCs by activating related protein and mRNA expression of PI3K/Akt/mTOR signaling pathway.