摘要
目的基于磷脂酰肌醇-3激酶(Phospoinositide 3-kinase,PI3K)/蛋白激酶B(protein kinase B,Akt)通路探讨淫羊藿素对多囊卵巢综合征颗粒细胞凋亡和自噬的影响。方法分离多囊卵巢综合征大鼠卵巢颗粒细胞,分成对照组、低、中、高剂量实验组(15,30,60μmol·L^(-1)淫羊藿素)和抑制剂组(60μmol·L^(-1)淫羊藿素和PI3K/Akt通路抑制剂LY294002处理)。以蛋白质印迹法检测PI3K/Akt通路关键蛋白、凋亡相关蛋白B细胞淋巴瘤/白血病-2(B cell lymphoma/lewkmia-2,Bcl-2)、Bcl-2相关X蛋白(Bcl-2 associated X protein,Bax)和自噬相关蛋白Beclin1、LC3Ⅱ/LC3Ⅰ表达变化,以细胞计数法-8(CCK-8)实验测定细胞增殖活性,以流式细胞术测定细胞凋亡。结果对照组、低、中、高剂量实验组和抑制剂组细胞增殖活性(OD值)分别为0.41±0.05、0.63±0.06、0.75±0.08、0.88±0.06、0.69±0.06,LC3Ⅱ/LC3Ⅰ表达量分别为1.76±0.15、1.45±0.11、1.12±0.07、1.05±0.09、1.32±0.13,细胞凋亡率分别为(42.65±6.32)%,(37.01±3.24)%,(26.47±2.11)%,(19.84±1.40)%,(35.12±0.30)%,p-PI3K表达量分别为0.35±0.05,0.48±0.05,0.68±0.07,0.89±0.09,0.41±0.06,PI3K表达量分别为0.94±0.11,0.94±0.09,0.92±0.11,0.87±0.12,0.90±0.09,以上指标,对照组与低、中、高剂量实验组比较,差异均有统计学意义(均P<0.05);低、中、高剂量实验组间比较,差异均有统计学意义(均P<0.05);高剂量实验组与抑制剂组比较,差异均有统计学意义(P<0.05)。结论淫羊藿素通过激活PI3K/Akt通路抑制多囊卵巢综合征颗粒细胞凋亡和自噬。
Objective To investigate the effects of icaritin on the apoptosis and autophagy of granulosa cells in polycystic ovary syndrome based on the phosphatidylinositol-3 kinase (PI3K)/protein kinase B(Akt) pathway.Methods The ovarian granulosa cells of rats with polycystic ovary syndrome were isolated and divided into control group,low,medium and high dose experimental groups (treated with 15,30,and 60μmol·L;icaritin),inhibitor group (treated with 60μmol·L;icaritin and PI3K/Akt pathway inhibitor LY294002).Western blot method was used to detect the PI3K/Akt pathway protein and apoptosis-related protein B-cell lymphoma/leukemia-2 (Bcl-2),Bcl-2 associated X protein (Bax) and autophagy-related protein Beclin1,LC3Ⅱ/LC3Ⅰlevels,cell counting kit-8 (CCK-8) experiment was used to determine cell proliferation activity,flow cytometry was used to determine cell apoptosis.Results The cell proliferation activity (OD value) in control group,low,medium and high dose experimental groups and inhibitor group were 0.41±0.05,0.63±0.06,0.75±0.08,0.88±0.06,0.69±0.06,the expression levels of LC3Ⅱ/LC3Ⅰwere 1.76±0.15,1.45±0.11,1.12±0.07,1.05±0.09,1.32±0.13,the apoptosis rate were (42.65±6.32)%,(37.01±3.24)%,(26.47±2.11)%,(19.84±1.40)%,(35.12±0.30)%,p-PI3K were 0.35±0.05,0.48±0.05,0.68±0.07,0.89±0.09,0.41±0.06,and the expression levels of PI3K were 0.94±0.11,0.94±0.09,0.92±0.11,0.87±0.12,0.90±0.09.For the above indicators,there were statistically significant difference between control group and low,medium and high dose experimental groups (all P<0.05);there were statistically significant difference between low,medium and high dose experimental groups (all P<0.05);there was statistically significant difference between high dose experimental group and inhibitor group (all P<0.05).Conclusion Icaritin inhibits the apoptosis and autophagy of granulosa cells in polycystic ovary syndrome by activating the PI3K/Akt pathway.
作者
邵梅
王家传
SHAO Mei;WANG Jia-chuan(The Second Department of Gynecology,Hangzhou Traditional Chinese Medicine Hospital,Hangzhou 310007,Zhejiang Province,China;Department of Pathology,Shenzhen Hospital of Traditional Chinese Medicine,Shenzhen 510083,Guangdong Province,China)
出处
《中国临床药理学杂志》
CAS
CSCD
北大核心
2021年第20期2830-2833,共4页
The Chinese Journal of Clinical Pharmacology
