林丹对大鼠生精细胞活化型半胱天冬酶-3表达及血清黄体生成素、睾酮浓度的影响

Effects of lindane on cleaved caspase-3 expression in spermatogenic cells and serum luteinizing hormone and testosterone concentrations in rats

  • 摘要:
    背景 林丹是有机氯农药六六六的γ异构体,对雄性生殖系统有损害作用。
    目的 研究林丹亚慢性染毒对雄性大鼠睾丸组织活化型半胱天冬酶-3(cleaved caspase-3)表达及血清黄体生成素、睾酮浓度的影响,探讨林丹致雄性生殖系统损伤的机制。
    方法 40只雄性SD大鼠被随机分为5组,分别为低、中、高剂量(3、6、13 mg·kg-1)林丹染毒组,溶剂对照组(10 mg·kg-1玉米油),阳性对照组(100 μg·kg-1雌二醇)。每天灌胃1次,连续8周,染毒末24 h称重,禁食一晚经腹腔注射10%水合氯醛溶液麻醉后解剖取材,测定大鼠睾丸生精细胞cleaved caspase-3蛋白表达情况及血清黄体生成素、睾酮浓度。
    结果 与溶剂对照组相比:各染毒组大鼠体重差异均无统计学意义(P>0.05);13 mg·kg-1组大鼠精子密度降低(P < 0.05);6、13 mg·kg-1组大鼠精子存活率下降(P < 0.05),睾丸生精细胞cleaved caspase-3蛋白表达升高(P < 0.05);13 mg·kg-1组大鼠血清黄体生成素浓度升高(P < 0.05),睾酮浓度降低(P < 0.05)。与阳性对照组相比:各染毒组大鼠体重及精子密度差异均无统计学意义(P>0.05);3、6 mg·kg-1组大鼠精子存活率升高(P < 0.05),睾丸生精细胞cleaved caspase-3蛋白表达降低(P < 0.05),血清黄体生成素浓度降低(P < 0.05);各染毒组大鼠血清睾酮浓度升高(P < 0.05)。病理结果显示:各林丹染毒组大鼠睾丸出现不同程度的损害状态,表现为曲细精管管腔间隙增大,生精细胞排列紊乱,精子数量减少。阳性对照组亦表现为管腔增大,精子数量减少,并可在管腔内发现脱落的生精细胞。
    结论 林丹可能通过上调睾丸组织cleaved caspase-3蛋白表达来调控生精细胞凋亡,引发血清睾酮水平下降,继而导致大鼠精子生成减少,且在此实验条件下,林丹引起大鼠内分泌干扰作用较雌二醇弱。

     

    Abstract:
    Background Lindane is the gamma isomer of organochlorine pesticide hexachlorocyclohexane and can damage male reproductive system.
    Objective This experiment investigates the effects of sub-chronic exposure to lindane on cleaved caspase-3 expression in testicular tissues and serum luteinizing hormone and testosterone concentrations in male rats, aiming to explore the mechanism of lindane-induced male reproductive damage.
    Methods Forty male SD rats were randomly divided into five groups:solvent control group (10mg·kg-1 corn oil), positive control group (100 μg·kg-1 estradiol), and 3, 6, and 13mg·kg-1 lindane groups. The rats were treated with the designed dosages by gavage once a day for continuously eight weeks. After 24 h of last exposure, the rats were weighed, and after an overnight fasting, the rats were anesthetized with 10% chloral hydrate and dissected to determine the cleaved caspase-3 protein expression in spermatogenic cells and serum luteinizing hormone and testosterone concentrations.
    Results Compared with the solvent control group, there was no significant difference in body weight of rats in the exposure groups (P>0.05); the sperm density of rats in the 13 mg·kg-1 group decreased (P < 0.05); the sperm survival rate decreased and the cleaved caspase-3 protein expression level in spermatogenic cells were upregulated in the 6 mg·kg-1 and 13 mg·kg-1 groups (P < 0.05); the serum luteinizing hormone concentration increased and the serum testosterone concentration decreased in the 13 mg·kg-1 group (P < 0.05). Compared with the positive control group, there were no significant differences in body weight and sperm density in the exposure groups (P>0.05); the sperm survival rate increased, the protein expression level of cleaved caspase-3 in spermatogenic cells decreased, and the serum luteinizing hormone concentration decreased in the 3 mg·kg-1 and 6 mg·kg-1 groups (P < 0.05); the serum testosterone concentration in each exposure group increased (P < 0.05). The testicular pathological observations showed that the rats in each lindane-treated group presented varying degrees of testicular damage, including increased seminiferous tubule space, disarranged spermatogenic cells, and decreased sperm counts; the rats in the positive control group also showed enlarged seminiferous tubule space, reduced sperm counts, and spermatogenic cells in the lumen.
    Conclusion Lindane may regulate spermatogenic cell apoptosis by upregulating the cleaved caspase-3 protein expression in testicular tissues, leading to decreased serum testosterone level and spermatogenesis in rats. Under these experimental conditions, lindane may cause less endocrine disruption in rats than estradiol.

     

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