摘要
目的基于磷脂酰肌醇3激酶(PI3K)/蛋白激酶B(AKT)通路,初步探究柚皮素防治大鼠多囊卵巢综合征(PCOS)胰岛素抵抗的分子机制。方法SD大鼠颈背部每日皮下注射脱氢表雄酮建立PCOS模型,将造模成功的大鼠采用随机数字表法分为模型组、柚皮素组、PI3K抑制剂组、柚皮素+PI3K抑制剂组,每组15只;相同时间段取SD大鼠15只,于颈背部皮下注射油剂2 mL/kg,作为正常对照组。检测各组大鼠血糖、胰岛素、血脂、生殖激素水平,计算胰岛素抵抗指数;HE染色观察卵巢形态;免疫组化法检测磷酸化PI3K(p-PI3K)阳性表达;Western blot法检测PI3K/AKT通路磷酸化蛋白及通路相关蛋白胰岛素受体底物-1(IRS-1)、糖原合成酶激酶-3β(GSK-3β)、葡萄糖转运蛋白因子-4(GLUT4)、人第10号染色体上磷酸酶和张力蛋白同源缺失基因(PTEN)蛋白表达。结果与正常对照组相比,模型组大鼠卵巢组织出现卵泡囊性扩张、黄体数量及颗粒细胞层减少、闭锁卵泡增多等病理损伤症状,血脂、血糖、胰岛素水平升高,胰岛素抵抗增加,生殖激素分泌紊乱,PI3K/AKT通路磷酸化蛋白及通路相关蛋白IRS-1、GSK3β磷酸化蛋白表达、GLUT4蛋白表达降低,PTEN蛋白表达升高(P<0.05)。柚皮素可减轻卵泡囊性扩张等病理损伤,促进PI3K/AKT通路及通路相关蛋白IRS-1、GSK-3β磷酸化蛋白及GLUT4蛋白表达,改善生殖激素分泌紊乱现象,减轻胰岛素抵抗,降低血糖、血脂水平及PTEN蛋白表达(P<0.05)。PI3K抑制剂可减弱柚皮素的上述作用(P<0.05)。结论柚皮素可通过促进PI3K/AKT通路活化,降低血糖、血脂水平及胰岛素抵抗,改善PCOS大鼠生殖激素紊乱及卵巢多囊改变。
Objective To explore the molecular mechanism of naringenin in preventing and treating insulin resistance in rats with polycystic ovary syndrome(PCOS)based on the aspect of phosphatidylinositol 3 kinase(PI3K)/protein kinase B(AKT)pathway.Methods SD rats were injected with dehydroepiandrosterone subcutaneously on the back daily to establish a PCOS model.The model rats were randomly divided into the model group,the naringenin group,the PI3K inhibitor group,and the naringenin+PI3K inhibitor group,with 15 rats in each group.At the same time,15 SD rats were subcutaneously injected with oil 2 mL/kg on the neck and back,and they were used as the normal control group.The levels of blood glucose,insulin,blood lipids,and reproductive hormones were detected in each group.The morphology of ovary was observed with HE staining.The positive expression of p-PI3K was detected with immunohistochemical method.The protein expression of PI3K/AKT pathway phosphorylated protein,and pathway related protein-insulin receptor substrate-1(IRS-1),glycogen synthase-3β(GSK-3β),glucose transporter factor-4(GLUT4),phosphatase and tensin homologue deleted on chromosome 10(PTEN)were detected with Western blot assay.Results Compared with the normal control group,the ovarian tissue of the model group showed symptoms of pathological damage such as follicular cystic dilatation,decreased corpus luteum number and granulosa cell layer,and increased atretic follicles,increased levels of blood lipid and blood sugar,increased insulin resistance,and disordered secretion of reproductive hormones,decreased PI3K/AKT pathway phosphorylated proteins and pathway-related proteins IRS-1,GSK-3βphosphorylated protein and GLUT4 protein expression,and increased expression of PTEN protein(P<0.05).Naringenin could reduce pathological damage such as cystic dilatation of follicles,promote PI3K/AKT pathway phosphorylated proteins and pathway-related proteins IRS-1,GSK-3βphosphorylated protein and GLUT4 protein expression,improve the disorder of reproductive hormone secretion,reduce insulin resistance,blood glucose and blood lipid levels and PTEN protein expression(P<0.05).PI3K inhibitors could attenuate the above effects of naringenin(P<0.05).Conclusion Naringenin can promote the activation of PI3K/AKT pathway,reduce blood glucose and blood lipid levels and insulin resistance,and improve reproductive hormone disorders and ovarian polycystic changes in PCOS rats.
作者
徐晶
申丽媛
屈清华
XU Jing;SHEN Liyuan;QU Qinghua(Department of Gynecology,Chongqing Maternal and Child Health Hospital,Chongqing 400021,China;Department of Gynecological Endocrinology,Chongqing Maternal and Child Health Hospital,Chongqing 400021,China)
出处
《天津医药》
CAS
北大核心
2022年第3期270-275,共6页
Tianjin Medical Journal
基金
重庆市科卫联合医学科研项目(2018QNXM027,2019ZDXM051)。
