小豆蔻明对脂多糖诱导人精子氧化损伤的保护作用  

Protective effect of cardamonin on human sperm from oxidative damage induced by lipopolysaccharide

作　　者：曾凡湘[1,2] 牛培广[1,2] 林典梁[3] 陈钢鑫 王航伟[4] 曾晓萍 吴安鹏 刘颖[1] 史道华[1,2] ZENG Fan-xiang;NIU Pei-guang;LIN Dian-liang;CHEN Gang-xin;WANG Hang-wei;ZENG Xiao-ping;WU An-peng;LIU Ying;SHI Dao-hua(Department of Pharmacy,Fujian Maternity and Child Health Hospital,Affiliated Hospital of Fujian Medical University,Fuzhou 350001,Fujian Province,China;Fujian Key Laboratory of Women and Children’s Critical Diseases Research,Fujian Maternity and Child Health Hospital,Affiliated Hospital of Fujian Medical University,Fuzhou 350001,Fujian Province,China;Reproductive Medicine Centre,Fujian Maternity and Child Health Hospital,Affiliated Hospital of Fujian Medical University,Fuzhou 350001,Fujian Province,China;Laboratory Medicine,Fujian Maternity and Child Health Hospital,Affiliated Hospital of Fujian Medical University,Fuzhou 350001,Fujian Province,China)

机构地区：[1]福建省妇幼保健院、福建医科大学附属医院药剂科,福建福州350001 [2]福建省妇幼保健院、福建医科大学附属医院福建省妇儿重大疾病研究重点实验室,福建福州350001 [3]福建省妇幼保健院、福建医科大学附属医院生殖医学中心,福建福州350001 [4]福建省妇幼保健院、福建医科大学附属医院检验科,福建福州350001

出　　处：《中国临床药理学杂志》2020年第12期1639-1642,共4页The Chinese Journal of Clinical Pharmacology

基　　金：福建省自然科学基金资助项目(2017J01239);福建省卫生和计划生育委员会青年科研课题基金资助项目(2014-1-17);福建省卫生健康青年科研课题基金资助项目(2019-2-24)。

摘　　要：目的观察小豆蔻明对脂多糖(LPS)诱导人精子氧化损伤的保护作用。方法收集健康成年男性精液,用直接上游法分离精子,以LPS 1μg·mL^-1诱导人精子氧化损伤。将精子分为5组:正常组;模型组(LPS 1μg·mL^-1);低、中、高3个剂量实验组(LPS加小豆蔻明0.5,1.0,2.0μmol·L^-1)。紫外可见分光光度法、磷钼酸比色法分别检测琥珀酸脱氢酶(SDH)和三磷酸腺苷(ATP)水平,流式细胞术检测活性氧(ROS)含量和线粒体膜电位(MMP),荧光显微镜观察顶体反应(AR)率。结果正常组、模型组和低、中、高3个剂量实验组的SDH活性分别为(8.27±0.21)×10^-6,(3.90±0.30)×10^-6,(4.56±0.28)×10^-6,(5.67±0.25)×10^-6和(7.83±0.21)×10^-6U·cell-1;这5组的ATP含量分别为(73.04±3.46)×10^-6,(54.78±2.69)×10^-6,(63.24±1.36)×10^-6,(66.46±1.91)×10^-6和(73.07±3.27)×10^-6 pmol·cell-1;这5组的ROS百分含量分别为(3.20±0.70)%,(4.50±0.21)%,(3.21±0.76)%,(3.19±0.50)%和(3.03±0.89)%;这5组的MMP分别为(35.10±4.32)%,(25.53±3.72)%,(34.21±3.92)%,(35.82±4.83)%和(36.91±4.15)%;这5组的AR率分别为(9.84±4.00)%,(42.34±4.70)%,(33.33±3.01)%,(25.42±2.59)%和(13.73±5.13)%。上述指标:模型组与正常组比较,差异均有统计学意义(P<0.05,P<0.01);低、中、高3个剂量实验组与模型组比较,差异均有统计学意义(P<0.05,P<0.01)。结论小豆蔻明能改善LPS诱导的精子氧化损伤。Objective To explore the protective effects of cardamonin on oxidative injury of human sperm induced by lipopolysaccharide(LPS).Methods Semen was separated from healthy adult males.Sperms were raised by swim-up method and then pretreated with 1μg·mL^-1 LPS.The sperms were divided into 5 groups:blank group,model group,and experimental-L,experimental-M,experimental-H groups;1μg·mL^-1 LPS was added in the model group;0.5,1.0,2.0μmol·L^-1 cardamonin and 1μg·mL^-1LPS were respectively added in experimental-L,-M,-H groups.The activity of succinate dehydrogenase(SDH)and adenosine triphosphate(ATP)were respectively detected by ultraviolet-visible spectrophotometry and molybdate colorimetry.Flow cytometry was used to analyze reactive oxygen species(ROS)and mitochondrial membrane potential(MMP).Acrosome reaction(AR)rate was observed by fluorescence microscope.Results The activity of SDH in blank group,model group,and low,medium,high dose experimental groups were(8.27±0.21)×10^-6,(3.90±0.30)×10^-6,(4.56±0.28)×10^-6,(5.67±0.25)×10^-6 and(7.83±0.21)×10^-6 U·cell-1,respectively;the ATP in the 5 groups were(73.04±3.46)×10^-6,(54.78±2.69)×10^-6,(63.24±1.36)×10^-6,(66.46±1.91)×10^-6 and(73.07±3.27)×10^-6 pmol·cell-1,respectively;the ROS in the 5 groups were(3.20±0.70)%,(4.50±0.21)%,(3.21±0.76)%,(3.19±0.50)%and(3.03±0.89)%,respectively;the MMP in the 5 groups were(35.10±4.32)%,(25.53±3.72)%,(34.21±3.92)%,(35.82±4.83)%and(36.91±4.15)%,respectively;the AR rates in the 5 groups were(9.84±4.00)%,(42.34±4.70)%,(33.33±3.01)%,(25.42±2.59)%and(13.73±5.13)%,respectively.All factors of model group were statistically significant different compared with those of blank group(P<0.05,P<0.01).The factors of low,medium,high dose experimental groups were statistically significant different compared with those of model group(P<0.05,P<0.01).Conclusion Cardamonin can reduce LPS-induced oxidative damage in human sperm.

关 键 词：小豆蔻明 脂多糖 精子 氧化损伤 活性氧 磷钼酸比色法 

分 类 号：R28[医药卫生—中药学]