补肾生精调和气血法对卵巢储备功能低下大鼠氧化应激的影响  

Effect of invigorating kidney and promoting essence and reconciling qi and blood on oxidative stress in rats with diminished ovarian reserve

作　　者：黄佳梅 蔡紫璨 张花[1,2] 唐娜 万维[2] 金志春 HUANG Jiamei;CAI Zican;ZHANG Hua;TANG Na;WAN Wei;JIN Zhichun(Hubei University of Chinese Medince,Wuhan 430065,Hubei,China;Maternal and Child Healthcare Hospital of Hubei Province,Huazhong University of Science and Technology,Wuhan 430070,Hubei,China)

机构地区：[1]湖北中医药大学,湖北武汉430065 [2]华中科技大学附属湖北妇幼保健院,湖北武汉430070

出　　处：《现代中西医结合杂志》2022年第23期3224-3231,共8页Modern Journal of Integrated Traditional Chinese and Western Medicine

基　　金：湖北省卫健委中医药科研重点项目(ZY2021Z013)。

摘　　要：目的基于氧化应激途径探讨补肾生精调和气血法对卵巢储备功能低下(DOR)大鼠卵巢功能的保护作用。方法将60只成年雌性SD大鼠随机分为空白组、环磷酰胺组、戊酸雌二醇组及补肾生精调和气血方低、中、高剂量组,每组10只。除空白组外,其他组均给予环磷酰胺75 mg/kg单次腹腔注射制备DOR模型。造模后,戊酸雌二醇组给予戊酸雌二醇0.105 mg/kg灌胃,补肾生精调和气血方低、中、高剂量组分别给予补肾生精调和气血方8.075 g/kg、16.15 g/kg、32.3 g/kg灌胃,空白组和环磷酰胺组给予等量去离子水灌胃,均每日1次,连续21 d。实验过程中观察大鼠一般情况及动情周期改变;末次灌胃结束后取血,检测血清抗苗勒管激素(AMH)、雌激素(E 2)、卵泡刺激素(FSH)、黄体生成素(LH)、丙二醛(MDA)、超氧化物歧化酶(SOD)、还原型谷胱甘肽(GSH)、总抗氧化能力(T-AOC)水平,苏木素染色观察卵巢形态学改变,Western blot法、免疫组化法检测卵巢组织中血红素氧合酶1(HO-1)、醌氧化还原酶1(NQO-1)蛋白表达情况,实时荧光定量PCR法检测卵巢组织中HO-1和NQO-1 mRNA表达情况。结果与空白组比较,环磷酰胺组大鼠体重增长较慢,动情周期紊乱;血清FSH、LH、MDA水平均明显升高(P均<0.05),血清AMH、SOD、T-AOC水平均明显降低(P均<0.05);卵巢内生长卵泡数量减少,闭锁卵泡数量增多;卵巢组织中HO-1、NQO-1蛋白相对表达量均明显降低(P均<0.05),HO-1、NQO-1 mRNA相对表达量变化不明显(P均>0.05)。与环磷酰胺组比较,各药物组大鼠动情周期逐渐恢复;戊酸雌二醇组和补肾生精调和气血方中剂量组大鼠血清FSH、LH、MDA水平均明显降低(P均<0.05),血清E 2、SOD、GSH、T-AOC水平均明显升高(P均<0.05);各药物组大鼠卵巢内生长卵泡数量增多,闭锁卵泡数量减少;补肾生精调和气血方高剂量组HO-1蛋白相对表达量、补肾生精调和气血方低剂量组NQO-1蛋白相对Objective It is to investigate the protective effects of invigorating kidney and promoting essence and reconciling qi and blood therapy on ovarian function in rats with diminished ovarian reserve(DOR)based on oxidative stress pathway.Methods Sixty adult female SD rats were randomly divided into blank group,cyclophosphamide group,estradiol valerate group and the low,medium and high dose groups of decoction of invigorating kidney and promoting essence and reconciling qi and blood,10 rats in each group.All groups except for the blank group were given a single intraperitoneal injection of cyclophosphamide 75 mg/kg to prepare the DOR models.After moulding,the estradiol valerate group was given estradiol valerate 0.105 mg/kg by gavage,the low,medium and high dose groups of decoction of invigorating kidney and promoting essence and reconciling qi and blood were respectively given decoction of invigorating kidney and promoting essence and reconciling qi and blood 8.075 g/kg,16.15 g/kg and 32.3 g/kg by gavage,all once a day,continuously treated for 3 weeks.The rats were observed for general conditions and kinetic cycle changes during the experiment;their blood was collected after the last gavage to detect the levels of serum anti-mullerian hormone(AMH),estrogen(E 2),follicle-stimulating hormone(FSH),luteinizing hormone(LH),malondialdehyde(MDA),superoxide dismutase(SOD),glutathione(GSH),total antioxidant capacity(T-AOC),the morphological changes of ovaries were observed by hematoxylin staining,the expression of heme oxygenase 1(HO-1)and quinone oxidoreductase 1(NQO-1)in ovarian tissues was detected by Western blot and immunohistochemistry,and the expression of HO-1 and NQO-1 mRNA in ovarian tissues was detected by real-time fluorescence quantitative PCR.Results Compared with the blank group,the rats in the cyclophosphamide group had slower weight gain and disturbed motility cycle;the levels of serum FSH,LH and MDA were significantly increased(all P<0.05),and the levels of serum AMH,SOD and T-AOC were significantly decrease

关 键 词：卵巢储备功能低下 补肾生精调和气血法 氧化应激 血红素氧合酶1 醌氧化还原酶1 

分 类 号：R-332[医药卫生]