卡马西平对健康及癫雄性大鼠生殖系统相关指标的影响

刘绪宏,张 辉,谭 梅,许笑天,伍春华

刘绪宏,张 辉,谭 梅,许笑天,伍春华

目的 探讨卡马西平单药以及癫与卡马西平共同作用下对雄性大鼠生殖系统相关指标的影响，为癫男性患者临床用药提供基础研究数据。方法 运用氯化锂-匹罗卡品建立Wistar雄性大鼠癫模型。选择健康清洁的Wistar雄性大鼠设造模组、非造模组及对照组，再将造模组、非造模组随机分为三个亚组，分别给予不同浓度卡马西平灌胃60 d后，所有入组大鼠取血样、睾丸、附睾、精子标本，分别观测血清性激素[(总睾酮(TT)、雌二醇(E2)、孕酮(P)、泌乳素(PRL)、黄体生成素(LH)、卵泡刺激素(FSH)]、脏器系数、精子计数、精子活力与形态等指标，分析卡马西平单药及癫与卡马西平协同作用对雄性大鼠生殖的影响。结果 造模组大鼠给予不同浓度卡马西平灌胃后，LH、FSH、E2水平均较对照组明显升高(P均<0.01)，各浓度组P水平均明显低于对照组(P均<0.05)，各浓度组TT水平明显低于对照组，PRL水平未见明显改变。非造模组大鼠给予不同浓度卡马西平药物灌胃后, LH、FSH水平均较对照组明显升高(P均<0.01)，TT水平均低于对照组(P均<0.05)，PRL水平中高浓度组明显低于对照组(P均<0.05)， P、E2未见明显改变。非造模组大鼠给予不同浓度卡马西平药物灌胃后, 各浓度组附睾重量及附睾脏器系数均较对照组明显降低(P均<0.01)。结论 卡马西平对于雄性大鼠生殖系统相关指标存在明显不良反应，且有明显药物浓度关联性，而卡马西平与癫发作共同作用的不良反应未见协同叠加效应。

卡马西平；癫；生殖；性激素

1 材料与方法

1.1.1 材料 6～8周龄(体重200～220 g)健康清洁级Wistar雄性大鼠100只(由扬州大学实验动物中心提供)，并保持饲养环境室温18～25 ℃、相对湿度50%～60%、昼夜循环照明环境中饲养，能自由摄食及饮水，每日定时清洗笼舍。

1.1.3 分组 设造模组、非造模组及对照组。将造模成功大鼠随机取24只作为造模组，再将其随机分为3组，分别为灌药高、中、低浓度组，每组8只；非造模组为相同饲养环境下的健康雄性Wistar大鼠24只，亦随机分成3组(将给予不同浓度灌药)，每组8只；对照组10只(将给予灌注盐水)。

1.1.4 给药剂量及方法 按照低剂量组30 mg/(kg· d)，中剂量组60 mg/(kg·d)，高剂量组120 mg/(kg·d)，分别给予造模组及非造模组大鼠相应浓度卡马西平连续灌药60 d，对照组给予生理盐水连续灌胃60 d，1次/d，大鼠体重每3 d称1次,以调整给药剂量。

1.2 标本取材与制备

1.2.1 血清性激素测定 所有入组大鼠给予连续灌胃60 d后，于同日早晨7：00-8:00采集外周血1次，经放射免疫法 (RIA) 测定 TT、E2、P、PRL、LH、FSH。放射免疫分析试剂盒购自北京北方生物技术研究所，FT放射免疫分析试剂盒购自比利时BIOCODE-HYCEL公司，采用科大创新股份有限公司中佳分公司GC-911γ放射免疫计数器，严格按照说明书要求进行测定。

1.2.2 性器官脏器系数测定 将所有大鼠脱臼处死，取双侧睾丸、附睾称重后计算性器官脏器系数[脏器系数=脏器重量(g)/体重(g)×100%]，以百克体重计算。

1.2.3 精子悬液制备及计数 将所有大鼠处死后分离附睾，立即取双侧附睾尾放入盛有12 ml、37 ℃的M199培养液中，沿附睾尾纵切4～5刀，放入37 ℃培养箱中扩散10 min，让精子从附睾尾中游离出，得到精子悬液(2.0×107/ml)。将制备的精子悬液用吸管取少量滴入白细胞计数板计数精子数[7]。

1.2.4 精子活动率的测定 取精子悬液1～2 滴于清洁玻片上，在(20±2)℃的室温下，20 min 内在光镜下进行观察，每只大鼠分析200个精子，并对精子的活动率进行分级。分级标准如下：(Ⅰ)不活动；(Ⅱ)原地颤动或原地转动；(Ⅲ)慢速直线或转动向前运动；(Ⅳ)快速直线向前运动。统计活动的精子总数，计算精子的活动率[活动精子率=(Ⅱ+Ⅲ+Ⅳ)/(Ⅰ+Ⅱ+Ⅲ+Ⅳ)×100%]。睾精子形态观察：用吸管吸取少量上述精子悬液，滴于载玻片上，立即用另一玻片均匀推开，干燥后用甲醇固定5 min，干后再用2%伊红水溶液染色1 h，在400倍光学显微镜下观察精子形态。

2 结 果

2.1 血清性激素 造模组高、中、低浓度卡马西平灌胃后，LH、FSH、E2均较对照组明显升高(P均<0.01)，但各浓度组间未见统计学差异。各浓度组P值均明显低于对照组(P<0.05)，造模组间比较，高浓度组降低更为显著，差异有统计学意义。各浓度组TT值明显低于对照组，差异有统计学意义(P<0.01)，而各浓度组间未见明显差异，PRL与对照组比较无统计学差异。非造模组高、中、低浓度卡马西平灌胃后，LH、FSH均较对照组显著升高(P均<0.01)，但各浓度组间未见统计学差异。TT各浓度组均低于对照组(P均<0.05)，其中高浓度组明显低于对照组(P均<0.01)，与中低浓度组存在统计学差异。PRL水平中、高浓度组明显低于对照组(P均<0.05)，其余组别无统计学差异(表1)。

2.2 性器官脏器系数 造模组大鼠给予高、中、低浓度卡马西平药物灌胃后，体重及脏器系数检测值与对照组比较无统计学意义。非造模组中各浓度组附睾重量及附睾脏器系数均较对照组降低，差异具有统计学意义(P<0.01)。余各浓度组体重、睾丸重量及睾丸脏器系数与对照组比较无统计学意义(表2)。

2.3 精子计数及精子活动率 造模组和非造模组大鼠各浓度组间精子数均未见明显异常，但精子活动率均明显低于对照组，具有显著统计学差异(P均<0.01)，其中均以高浓度组精子活动率减低最为明显(表3)。

表1 造模组和非造模组大鼠给予高、中、低浓度卡马西平药物灌胃血清性激素检测结果 ±s)

注：与对照组比较，①P<0.01；②P<0.05

表2 造模组和非造模组大鼠给予高、中、低浓度卡马西平灌胃体重及脏器系数检测结果 ±s)

注：与对照组比较，①P<0.01；②P<0.05

表3 造模组和非造模组大鼠给予高、中、低浓度卡马西平灌胃精子计数及精子活动率检测结果 ±s)

注：与对照组比较，①P<0.01；②P<0.05

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(2014-06-10收稿 2014-09-15修回)

(责任编辑 尤伟杰)

Effects of reproductive system in experimental male rats with carbamazepine

LIU Xuhong,ZHANG Hui,TAN Mei,XU Xiaotian, and WU Chunhua.

Department of Neurology，Jiangsu Provincial Corps Hospital, Chinese People’s Armed Police Forces, Yangzhou 225003, China

Objective To study the effects of CBZ and CBZ with epilepsy on reproductive system of male Wistar rats. Methods Male rat epilepsy models were established with lithium-pilocarpine and health male Wistar rats controls were used. They were given three different concentration carbamazepine infused into stomach. Blood, testis and epididymis, and sperms were sampled; serum sex hormones, viscera coefficient, sperm counts, sperm motility and morphology were observed for analyzing on effects male epileptrc rats’ reproductive system.Results In the epilepsy model rats receiving high, medium and low concentration CBZ showed that LH, FSH, E2 were significantly higher than in controls, there were but no significant differences between the three concentration groups. Progesterone (P) levels of three concentrations were obviously lower than in controls, but in low concentration group was significantly lower than in the other two concentration groups. Testosterone (T) levels in three concentration groups were significantly lower than in healthy group. No changes were found in PRL.In the health control Wistar rats receiving. high, medium and low concentration CBZ, LH and FSH were significantly higher than in controls, testosterone (T) levels in three concentration groups were significantly lower than in health group, especially the high concentration group,PRL levels in medium and low concentration group obviously lower than in controls. There was no difference in P and E2. There were no changes in weight and testicular weight and testicular viscera coefficient between male epilepsy rats and health group, but epididymis weight and epididymis viscera coefficient were obviously lower in the health rats with three concentration CBZ than in controls. No sperm count change in the three concentrations for male epilepsy rats and health group, but both of them had the lower sperm activity rate than in controls, especially in high concentration group. Conclusions Carbamazepine itself for male rats reproductive system has obvious side effects and obvious drug concentration relevance, both carbamazepine and epilepsy also present the part of the side effects of correlation concentration,but the mechanism is relatively complex, caused by many factors in joint action.

carbamazepine；epilepsy；reproduction；sex hormones

刘绪宏，本科学历，主任医师，E-mail:xuhlzy@163.com

225003扬州，武警江苏总队医院神经内科

R971.6；R742.1