黄芩苷通过增强抗氧化和抑制细胞凋亡减轻哮喘小鼠雄性生殖损伤

Male Reproductive Injury is Blocked by Baicalin by Modulating Oxidative Stress and Apoptosis in Asthmatic Mice

生殖健康是人口与健康领域的重要议题。作为全球最常见的呼吸道疾病哮喘会影响男性生殖功能,但相关机制鲜有报道。本文研究了黄酮类化合物黄芩苷(baicalin,BA)对哮喘小鼠睾丸损伤的干预作用及相关机制。选择雄性BALB/c小鼠随机分为对照组(CK组)、卵清蛋白(ovalbumin,OVA)致敏的哮喘组(OVA组)和黄芩苷干预哮喘组(OVA+BA组)。结果发现,3组小鼠体重无明显差异。OVA组小鼠睾丸系数和精子数量显著降低(P<0.05),精子畸形率显著增加(P<0.05);黄芩苷干预组小鼠睾丸系数显著增加(P<0.05),精子畸形率显著降低(P<0.05)。HE染色观察到OVA组小鼠睾丸组织生精小管结构损伤,精子发生异常,生精细胞减少,Johnson得分显著降低;BA干预组生精小管直径及生精上皮细胞高度显著增加,生精小管基膜结构较完整,Johnson得分显著提高(P<0.05);试剂盒法检测氧化还原指标发现,OVA组睾丸组织过氧化氢(H_(2)O_(2))和丙二醛(MDA)含量显著增加(P<0.05),总超氧化物歧化酶(T-SOD)活性和谷胱甘肽(GSH)含量显著降低(P<0.05);OVA+BA组睾丸组织H_(2)O_(2)和MDA含量显著降低(P<0.05),T-SOD活性显著增加(P<0.05);实时荧光定量RT-PCR检测发现,OVA组睾丸组织中促凋亡基因p53、Casp-3转录上调,抗凋亡基因Bcl2转录显著下调,胱天蛋白酶3(caspase-3)活性显著增加(P<0.05);OVA+BA组p53和Casp-3转录下调,Bcl2转录上调,胱天蛋白酶3活性显著降低(P<0.05)。结果表明,哮喘小鼠睾丸组织发生了氧化应激和结构损伤,细胞凋亡途径被激活,BA干预可有效减轻哮喘小鼠睾丸组织的氧化胁迫,抑制凋亡通路,保护睾丸组织的功能和结构。结果提示,黄芩苷能缓解哮喘小鼠的生殖毒性,该效应与机体抗氧化能力提高、细胞凋亡途径抑制有关。

Reproductive health is an important issue in the field of population and health.As the most common respiratory disease in the world,asthma is linked with male reproductive dysfunction,but the related study is rarely reported.In this study,we analyzed the interventional effect of baicalin(BA) on male reproductive injury in asthmatic mice and its mechanism.Male BALB/c mice were randomly divided into three groups,namely as control group(CK group),OVA-induced asthma model group(OVA group) and baicalin intervention asthma group(OVA+BA group).The results showed that no significant difference was found in body weight among the three groups.In the OVA group,the testicular coefficient and sperm count decreased significantly,with sperm malformation rate increased significantly(P < 0.05).In the baicalin intervention group,the testicular coefficient was increased significantly and sperm malformation rate decreased significantly(P < 0.05);Hematoxylin-eosin(HE) staining showed that damaged basement membrane of seminiferous tubules,decreased the number of spermatogenic cells and reduced Johnson score were observed in OVA group.The seminiferous tubule diameter and seminiferous epithelium height were significantly increased(P < 0.05),the basement membrane of seminiferous tubules was relatively complete and Johnson score gains in the OVA+BA group.Compared with control group,the contents of hydrogen peroxide(H_(2)O_(2)) and malondialdehyde(MDA) were significantly increased(P < 0.05),and the total superoxide dismutase(T-SOD) activity and glutathione(GSH) content were decreased(P < 0.05) in the testis of mice in OVA group.Compared with the OVA group,the contents of H_(2)O_(2) and MDA in OVA+BA group were significantly reduced(P < 0.05),and the T-SOD activity was significantly increased(P < 0.05).These results indicated that the testicular cells of asthmatic mice encounter oxidative stress and oxidative damage,and baicalin intervention can inhibit oxidative damage to testicular tissue in asthmatic mice.Analysis of real-time quantitative PCR showed that the transcription of pro-apoptotic genes p53 and caspase-3 were up-regulated and the activity of caspase-3 enzyme was increased significantly(P < 0.05) in the testes of asthmatic mice,while the transcription of apoptosis-suppressing gene Bcl2 was down-regulated(P < 0.05).In the OVA+BA group,the transcription of p53 and caspase-3 were down-regulated and the activity of caspase-3 enzyme was significantly reduced(P < 0.05),accompanied by up-regulated Bcl2 expression(P < 0.05),indicating that baicalin can inhibit the apoptotic pathway in testicular cells activated by OVA.Baicalin is a drug and food homologous substance that can alleviate the reproductive toxicity of asthmatic mice via reducing oxidative stress and inhibiting the expression of apoptotic factors.