Role of lactate dehydrogenase in diabetic neuropathic pain in mice: relationship with PGC-1α
王福宇 1金哲 1潘文燕 2向瀚民 1卢冠华 2贺俭 2王汉兵 2王焱林 1葛胜辉
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作者信息
1. 武汉大学中南医院麻醉科，武汉 430062
2. 佛山市第一人民医院麻醉科，佛山 528000
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摘要
目的 评价乳酸脱氢酶在小鼠糖尿病神经病理性痛(DNP)中的作用及其与过氧化物酶体增殖物激活受体γ共激活因子1α(PGC-1α)的关系。 方法 SPF级健康雄性C57BL/6J小鼠，6～8周龄，体质量18～22 g，采用腹腔注射链脲佐菌素120 mg/kg的方法制备小鼠糖尿病模型。取糖尿病造模成功小鼠24只，采用随机数字表法分为2组(n＝12)：DNP组和DNP＋草氨酸钠组(OXA组)。另取12只同月龄正常小鼠为对照组(C组)。OXA组在糖尿病造模成功后腹腔注射草氨酸钠750 mg/kg，1次/d，持续28 d；C组和DNP组腹腔注射等容量生理盐水。于链脲佐菌素注射前3 d、注射后1、2、3和4周(T0-4)时测定小鼠右侧后足机械缩足反应阈(MWT)、血糖和体质量。最后一次行为学测试完成后，麻醉小鼠后眼眶采集血样，测定血清乳酸浓度，随后处死取大脑前额叶皮质组织，测定乳酸含量，采用JC-1法检测线粒体膜电位，DHE探针检测ROS含量，采用Western blot法检测组蛋白乳酸化修饰水平及PGC-1α表达。 结果 与C组比较，DNP组和OXA组T2-4时MWT降低，血清乳酸浓度、前额叶皮质组织乳酸、ROS含量和组蛋白乳酸化修饰水平升高，线粒体膜电位降低，PGC-1α表达下调(P<0.05)；与DNP组比较，OXA组小鼠血糖与体质量差异无统计学意义(P>0.05)，T2-4时MWT升高，血清乳酸浓度、前额叶皮质组织乳酸、ROS含量和组蛋白乳酸化修饰水平降低，线粒体膜电位增加，PGC-1α表达上调(P<0.05)。 结论 乳酸脱氢酶促进小鼠DNP形成发展，其机制与促进前额叶皮质组蛋白乳酸化修饰水平增高及下调PGC-1α表达有关。 Objective To evaluate the role of lactate dehydrogenase in diabetic neuropathic pain (DNP) and the relationship with peroxisome proliferator-activated receptor gamma coactivator 1-alpha (PGC-1α) in mice. Methods SPF-grade healthy male C57BL/6J mice, aged 6-8 weeks, weighing 18-22 g, were used to establish diabetes mellitus model by intraperitoneal injection of streptozotocin (STZ) 120 mg/kg. Twenty-four mice with diabetes mellitus were divided into 2 groups (n=12 each) using a random number table method: DNP group and DNP + oxamate group (OXA group). Another 12 SPF-grade healthy male C57BL/6J mice were selected as control group (C group). In OXA group, oxamate 750 mg/kg was intraperitoneally injected once a day for 28 consecutive days. The equal volume of normal saline was given instead in C group and DNP group. The mechanical paw withdrawal threshold (MWT), blood glucose and body weight were measured at 3 days before STZ injection and at 1, 2, 3 and 4 weeks after STZ injection (T0-4). After the last behavioural test was completed, blood samples were collected from the posterior orbits of anesthetized mice for determination of serum lactate concentrations. The animals were then sacrificed and the tissues from the prefrontal cortex of the brain were taken for determination of lactate content, mitochondrial membrane potential (by the JC-1), content of reactive oxygen species (ROS) (using dihydroethidium probes), and level of histone lactylation and expression of PGC-1α (by Western blot). Results Compared with C group, the MWT was significantly decreased at T2-4, the serum lactate concentrations, contents of lactate and ROS and level of histone lactylation were increased, the mitochondrial membrane potential was decreased, and the expression of PGC-1α was down-regulated in DNP and OXA groups (P<0.05). Compared with DNP group, no significant change was found in blood glucose and body weight (P>0.05), the MWT was significantly increased at T2-4, the serum lactate concentrations, contents of lactate and ROS and level of histone lactylation were decreased, the mitochondrial membrane potential was increased, and the expression of PGC-1α was up-regulated in OXA group (P<0.05). Conclusions Lactate dehydrogenase promotes the development of DNP, and the mechanism is related to promotion of increase in histone lactfication and down-regulation of PGC-1α expression in the prefrontal cortex of mice.
Abstract
Objective To evaluate the role of lactate dehydrogenase in diabetic neuropathic pain (DNP) and the relationship with peroxisome proliferator-activated receptor gamma coactivator 1-alpha (PGC-1α) in mice. Methods SPF-grade healthy male C57BL/6J mice, aged 6-8 weeks, weighing 18-22 g, were used to establish diabetes mellitus model by intraperitoneal injection of streptozotocin (STZ) 120 mg/kg. Twenty-four mice with diabetes mellitus were divided into 2 groups (n=12 each) using a random number table method: DNP group and DNP + oxamate group (OXA group). Another 12 SPF-grade healthy male C57BL/6J mice were selected as control group (C group). In OXA group, oxamate 750 mg/kg was intraperitoneally injected once a day for 28 consecutive days. The equal volume of normal saline was given instead in C group and DNP group. The mechanical paw withdrawal threshold (MWT), blood glucose and body weight were measured at 3 days before STZ injection and at 1, 2, 3 and 4 weeks after STZ injection (T0-4). After the last behavioural test was completed, blood samples were collected from the posterior orbits of anesthetized mice for determination of serum lactate concentrations. The animals were then sacrificed and the tissues from the prefrontal cortex of the brain were taken for determination of lactate content, mitochondrial membrane potential (by the JC-1), content of reactive oxygen species (ROS) (using dihydroethidium probes), and level of histone lactylation and expression of PGC-1α (by Western blot). Results Compared with C group, the MWT was significantly decreased at T2-4, the serum lactate concentrations, contents of lactate and ROS and level of histone lactylation were increased, the mitochondrial membrane potential was decreased, and the expression of PGC-1α was down-regulated in DNP and OXA groups (P<0.05). Compared with DNP group, no significant change was found in blood glucose and body weight (P>0.05), the MWT was significantly increased at T2-4, the serum lactate concentrations, contents of lactate and ROS and level of histone lactylation were decreased, the mitochondrial membrane potential was increased, and the expression of PGC-1α was up-regulated in OXA group (P<0.05). Conclusions Lactate dehydrogenase promotes the development of DNP, and the mechanism is related to promotion of increase in histone lactfication and down-regulation of PGC-1α expression in the prefrontal cortex of mice.
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