摘要
目的考察巴西海木提取物对生殖系统氧化损伤的影响,初步探究其作用机制。方法采用水和不同比例(50%、70%及95%)乙醇溶液浸提巴西海木树皮,得到4种粗提物;依次用石油醚、乙酸乙酯和正丁醇萃取不同比例醇提物,得到9种有机相组分,共得到13种巴西海木提取物TCE-1~TCE-13。以小鼠睾丸支持细胞(TM4)增殖率变化为指标考察各提取物的细胞毒作用;采用H_(2)O_(2)处理TM4细胞建立氧化损伤模型,以细胞存活率及氧化指标超氧化物歧化酶(SOD)活力、丙二醛(MDA)含量和谷胱甘肽过氧化物酶(GSH-Px)活力变化为指标,邻苯二甲酸二(2-乙基己基)酯(DEHP)处理雄性小鼠建立生殖系统氧化损伤模型,造模后灌胃给予300、600、900 mg·kg^(-1)50%粗提乙酸乙酯组分(TCE-6)28 d后,以睾丸组织氧化指标变化验证TCE-6药效。最后通过实时荧光定量PCR(qRT-PCR)检测相关基因的表达情况,初步探究其作用机制。结果细胞毒性实验结果表明95%粗提物(TCE-4),50%粗提石油醚组分(TCE-5)、正丁醇组分(TCE-7),70%粗提石油醚组分(TCE-8)、正丁醇组分(TCE-10)以及95%粗提石油醚(TCE-11)、正丁醇组分(TCE-13)对TM4细胞具有杀伤作用。50%粗提物(TCE-2)、TCE-6以及70%粗提乙酸乙酯组分(TCE-9)给药组显著提高氧化损伤后TM4细胞存活率,TCE-6组SOD、GSH-Px活力显著提高、MDA含量显著下降(P<0.01)。900 mg·kg^(-1)TCE-6处理后能显著提高氧化损伤模型小鼠睾丸组织内SOD、GSH-Px活力,降低MDA含量(P<0.05)。qRT-PCR结果表明,900 mg·kg^(-1)TCE-6处理后提高了核因子E2相关因子2(Nrf2)、超氧化物歧化酶2(SOD2)及超氧化物歧化酶3(SOD3)mRNA的表达(P<0.05)。结论TCE-6可能通过上调SOD酶家族表达和调节Nrf2信号通路发挥抗生殖系统氧化损伤作用,具有开发为生殖系统损伤治疗药物的潜力。
Objective To determine the antioxidant effect of Trichilia catigua A.Juss.extract on the reproductive system and its mechanism.Methods Four crude extracts were obtained by extracting the bark of Trichilia catigua A.Juss.with water and ethanol at different proportions(50%,70%and 95%).Nine organic phase components were extracted from alcohol extracts with petroleum ether,ethyl acetate and 1-butanol at different proportions.Totally 13 Trichilia catigua A.Juss.extracts(TCE-1~TCE-13)were obtained.The cytotoxicity of the extract was evaluated by the proliferation rate of TM4 cells.The TM4 cells were treated with H_(2)O_(2) to establish oxidative damage model.The cell viability and oxidation index superoxide dismutase(SOD)activity,malondialdehyde(MDA)content and glutathione peroxidase(GSH-Px)activity were used as indicators.Male mice were treated with bis(2-ethylhexyl)phthalate(DEHP)to establish a reproductive system oxidative damage model.After the model establishment,and 28 days of intragastric administration of 300,600 and 900 mg·kg^(-1)50%crude ethyl acetate(TCE-6),the effect of TCE-6 was evaluated by the changes of testicular tissue oxidation index.Finally,the expression of related genes was detected by real-time fluorescence quantitative PCR(qRT-PCR)to explore the mechanism of protection efficacy.Results The toxicity test showed that 95%crude extract(TCE-4),50%crude petroleum ether component(TCE-5)and 1-butanol component(TCE-7),70%crude petroleum ether component(TCE-8)and 1-butanol component(TCE-10),95%crude petroleum ether component(TCE-11)and 1-butanol component(TCE-13)could kill TM4 cells.50%crude extract(TCE-2),50%crude extract ethyl acetate component(TCE-6)and 70%crude extract ethyl acetate component(TCE-9)improved the activity of TM4 cells after the oxidative damage.TCE-6(900 mg·kg^(-1))significantly increased the activities of SOD and GSH-Px in the testicular tissue of oxidative damage model mice,and decreased the content of MDA(P<0.05).qRT-PCR indicated that 900 mg·kg^(-1)TCE-6 treatment improved the expression of Nrf2,SOD2 and SOD3 mRNA(P<0.05).Conclusion TCE-6 may play an antioxidant role in the reproductive system by up-regulating the expression of SOD enzyme family and regulating Nrf2 signaling pathway,which might be a potential treatment for reproductive system damage.
作者
常馨月
米晓
董明然
王同瑾
胡美赓
沈俊宇
路娟
陈曦
CHANG Xin-yue;MI Xiao;DONG Ming-ran;WANG Tong-jin;HU Mei-geng;SHEN Jun-yu;LU Juan;CHEN Xi(Institute of Medicinal Plant Development,Peking Union Medical College,Chinese Academy of Medical Sciences,Beijing 100193;FITOTERAPIA BIOTECH LTDA,Brazil 01040-000)
出处
《中南药学》
CAS
2022年第4期751-758,共8页
Central South Pharmacy
基金
中国医学科学院医学与健康科技创新工程(No.2016-I2M-3-015)。
关键词
巴西海木
生殖系统
抗氧化活性
Trichilia catigua A.Juss.
reproductive system
antioxidant activity
