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哺乳动物各种生理功能中,生殖功能最易受热应激损伤。热应激使组织产生氧化应激,破坏雄性动物精子生成,从而影响繁殖效率。为探究黄芩苷对热应激小鼠(Mus musculus)睾丸损伤的影响,探讨黄芩苷的抗氧化作用机制,本研究将小鼠分成4组,常温对照组(C组)、黄芩苷组(C+B组)、热应激组(H组)和热应激加黄芩苷组(H+B组)。C组和H组小鼠腹腔注射生理盐水,C+B组和H+B组小鼠腹腔注射黄芩苷(50 mg/kg),连续注射7 d,H组与H+B组于第8天热应激(41℃)2 h,热应激结束后立即用断颈法处死所有处理组和对照组小鼠,取睾丸及附睾检测脏器系数和制备睾丸组织切片,睾丸组织用于检测氧化损伤指标丙二醛(malondialdehyde,MDA)的含量及超氧化物歧化酶(superoxide dismutase,SOD)、谷胱甘肽过氧化物酶(glutathione peroxidase,GSH-Px)和过氧化氢酶(catalase,CAT)的活力。末端脱氧核苷酸转移酶检测法(Td T-mediated d UTP nick-end labeling,TUNEL)法检测睾丸组织细胞的凋亡。Western blot检测睾丸组织内Fas/Fas L蛋白的表达。结果显示,热应激能够显著降低睾丸和附睾脏器系数(P<0.05)并使睾丸组织结构发生病理性损伤,而黄芩苷可以显著缓解热应激引起的睾丸脏器系数降低(P<0.05)和组织损伤。与热应激组相比,热应激加黄芩苷组小鼠睾丸组织中MDA含量显著降低,SOD、CAT和GSH-Px活力则显著增加(P<0.05)。黄芩苷组与热应激加黄芩苷组相比则无显著性差异。热应组小鼠睾丸组织细胞凋亡率与对照组相比显著增加(P<0.05),而热应激黄芩苷组与热应激组相比,小鼠睾丸细胞凋亡率则显著降低(P<0.05)。此外,与对照组相比,热应激可以显著增加小鼠睾丸组织Fas(P<0.05)和/Fas L(P<0.01)蛋白表达,而黄芩苷能显著降低热应激引起的小鼠睾丸组织中Fas/Fas L蛋白表达(P<0.05)。本研究结果表明,黄芩苷能减少受热应激时小鼠睾丸组织因氧化损伤造成的凋亡,其机制可能与增加抗氧化酶活性以及抑制Fas/Fas L蛋白表达来抑制细胞凋亡有关,这为寻找缓解动物热应激方法提供理论依据。
Mammalian various physiological functions, reproductive function most vulnerable to heat stress injury. Heat stress causes tissue to oxidative stress, destroying male animal sperm production, thus affecting reproductive efficiency. In order to explore the effect of baicalin on testicular damage in heat-stressed mice (Mus musculus) and explore the anti-oxidation mechanism of baicalin, the mice were divided into 4 groups: normal control group (C), baicalin group + B group), heat stress group (H group) and heat stress plus baicalin group (H + B group). Rats in group C and group H were injected intraperitoneally with normal saline, and mice in groups C + B and H + B were intraperitoneally injected with baicalin (50 mg / kg) for 7 days. H and H + Heat shock (41 ℃) for 2 h, immediately after the end of heat stress, all treatment groups and control mice were sacrificed by cervical disintegration. Testis and epididymis were used to detect organ coefficient and testicular tissue sections were prepared. The contents of malondialdehyde (MDA) and superoxide dismutase (SOD), glutathione peroxidase (GSH-Px) and catalase (CAT) ) Vitality. TdT-mediated dUTP nick-end labeling (TUNEL) method was used to detect the apoptosis of testicular cells. Western blot was used to detect the expression of Fas / Fas L protein in testis tissue. The results showed that heat stress could significantly reduce the organ coefficient of testis and epididymis (P <0.05) and cause pathological damage to testicular tissue structure, while baicalin could significantly reduce the testis organ coefficient induced by heat stress (P <0.05 ) And tissue damage. Compared with the heat stress group, the MDA content in the heat stress plus baicalin group was significantly decreased, while the activities of SOD, CAT and GSH-Px were significantly increased (P <0.05). There was no significant difference between baicalin group and heat stress plus baicalin group. Compared with the control group, the apoptotic rate of testis in heat-induced mice increased significantly (P <0.05), while the apoptosis rate of testicular cells in heat-stressed baicalin-treated mice was significantly lower than that of heat stress group P <0.05). In addition, compared with the control group, heat stress can significantly increase the Fas (P <0.05) and / Fas (P <0.01) protein expression in the testis tissue of mice, while baicalin can significantly reduce the heat stress-induced mouse testis Fas / Fas L protein expression in the tissue (P <0.05). The results of this study show that baicalin can reduce apoptosis induced by oxidative damage in mouse testes under heat stress. The mechanism may be related to the increase of antioxidant enzyme activity and the inhibition of Fas / Fas L protein expression to inhibit apoptosis To find ways to alleviate animal heat stress provide a theoretical basis.