高原鼠兔组织中精子特异性乳酸脱氢酶的作用机理
The Role of the Sperm-specific Lactate Dehydrogenase in Ochotona curzoniae Tissues

中文摘要：高原鼠兔 Ochotona curzoniae具有很强的高原低氧适应能力。前期研究发现，精子特异性乳酸脱氢酶（LDH-C 4）在高原鼠兔体细胞中表达。为阐明LDH-C 4在高原鼠兔组织中的作用机理，应用RNA干扰技术沉默高原鼠兔心肌、肝脏和脑组织中的 Ldh-c基因；应用荧光定量PCR和Western Blot方法，测定 Ldh-c基因在心肌、肝脏和脑组织中的表达水平；应用生物化学方法测定沉默 Ldh-c基因后，心肌、肝脏和脑组织中LDH比活力、乳酸和ATP的含量。结果表明，腹腔注射腺病毒pMultiRNAi-Ldhc能极显著降低高原鼠兔组织中 Ldh-c基因的表达水平，在mRNA和蛋白水平，心肌中 Ldh-c基因的表达分别下降48.11%和19.27%；肝脏中 Ldh-c基因的表达分别下降70.16%和25.82%；脑中 Ldh-c基因的表达分别下降49.08%和25.36%。沉默 Ldh-c基因表达后，高原鼠兔心肌、肝脏和脑组织中LDH比活力、乳酸和ATP的含量也显著降低，分别下降25.58%、41.94%和21.23%，28.16%、15.90%和24.66%以及16.65%、12.78%和18.50%。这些结果说明，高原鼠兔在低氧环境中，其心肌、肝脏和脑组织通过LDH-C 4催化无氧糖酵解获得部分ATP，增强对低氧环境的适应能力。
英文摘要：The plateau pika ( Ochotona curzoniae) has a strong adaptability to a hypoxic plateau environment. Previous studies found that the sperm-specific lactate dehydrogenase (LDH-C 4) gene is expressed in plateau pika somatic cell. In order to shed light on the role of LDH-C 4 in plateau pika tissues, we silenced the Ldh-c gene in pika heart, liver and brain by RNA interference, and then the expression levels of Ldh-c gene in these tissues were determined by real-time PCR and Western Blot. Additionally, the activities of LDH, the contents of lactic acid and ATP were also measured by biochemical method. The results indicated that intraperitoneal injection of adenovirus pMultiRNAi-Ldhc could significantly reduce the expression of Ldh-c gene in tissues of plateau pika. Specifically, in the mRNA and protein levels, the expression levels of Ldh-c gene in heart were decreased by 48.11% and 19.27%; in liver were decreased by 70.16% and 25.82%; in brain were decreased by 49.08% and 25.36%, respectively. Meanwhile, when Ldh-c gene expression was silenced, the activities of LDH, the contents of lactic acid and ATP in each tested tissues were generally decreased by 25.58%, 41.94% and 21.23%; 28.16%, 15.90% and 24.66%; 16.65%, 12.78% and 18.50%, respectively. These results suggested that under the condition of severe hypoxia, the heart, liver and brain tissues of plateau pika can generate part of ATP uponLDH-C 4 catalyzed anaerobic glycolysis and therefore enhance the adaptation to the hypoxic environments. 2017,36(6): 624-631 收稿日期：2017-05-08 DOI：10.11984/j.issn.1000-7083.20170146 分类号：Q955 基金项目：国家自然科学基金项目（31260512，30960054，31040011）；青海省自然科学基金项目（2016-ZJ-901，2014-ZJ-714，2012-Z-905） 作者简介：王志洁(1992-),女,硕士研究生,主要从事高原动物资源保护与利用研究,E-mail:411126871@qq.com *通讯作者：魏登邦,E-mail:weidengbang@163.com 参考文献： 丁晓涛, 何秀琼, 曹玉琼, 等. 1999. 高原鼠兔寄生虫感染调查报告[J]. 四川动物, 18(1):34. 何正波, 陈斌, 冯国忠. 2009. 昆虫RNAi技术及其应用[J]. 应用昆虫学报, 46(4):525-532. 姜国良, 于晓, 徐恺, 等. 2013. 腹腔和皮下注射D-半乳糖衰老大鼠模型分析[J]. 中国老年学, 33(5):1101-1103. 姜怀春, 李宏.