黄河包头段水源水有机提取物对雄性大鼠原代生殖细胞增殖和氧化损伤的影响

Effects of organic extracts of source water from the Baotou section of the Yellow River on proliferation and oxidative damage of primary germ cells of male rats

[目的]探讨黄河包头段水源水有机提取物对雄性大鼠原代生殖细胞的细胞增殖和氧化损伤的影响。[方法]提取黄河包头段上游水源水、下游水源水、自来水中有机物,将提取液按照低、中、高剂量(10、100、1 000 mL/mL,指1 mL有机物提取液分别来源于10、100、1 000 mL原水样)对体外培养的雄性大鼠原代生殖细胞染毒24 h,对照组给予同体积PBS液,MTT法检测细胞增殖率,透射电镜检测细胞凋亡,微板法检测乳酸脱氢酶(LDH)、总抗氧化能力(T-AOC)、超氧化物歧化酶(SOD)、丙二醛(MDA)水平。[结果]与对照组比较,自来水和黄河上游、下游水源水有机物高剂量组、中剂量组生殖细胞增殖率均降低(均P<0.05),高剂量组增殖率分别降低至(85.08±0.70)%、(90.80±0.30)%和(87.50±2.35)%。透射电镜检测显示,各高剂量染毒组细胞染色质边集、核碎裂和一些染色质块在膜散射,线粒体结构破坏,出现空泡,凋亡细胞多见。黄河下游及自来水高、中剂量组LDH活性均高于对照组(均P<0.05),自来水和黄河上游、下游水源水有机物高剂量组LDH活性分别为(2 417.48±298.50)、(1 870.37±157.95)、(2 072.90±195.11)U/L。自来水和黄河上游、下游水源水高、中剂量组T-AOC水平、SOD活性均低于对照组,MDA浓度高于对照组,差异有统计学意义(均P<0.05);在高剂量染毒条件下,自来水组SOD活性低于上游水源水[(6.52±1.77)、(8.74±0.90)U/mL,P<0.05],MDA浓度高于上游水源水[(3.14±1.18)、(2.27±0.82)nmol/L,P<0.05]。[结论]黄河上游、下游水源水有机物能够抑制雄性大鼠原代生殖细胞增殖,促进细胞凋亡,氧化损伤可能是其作用机制之一。

[Objective] To investigate the effects of organic extracts of source water from the Baotou section of the Yellow River on the proliferation and oxidative damage of primary germ cells of male rats.[Methods] Organic matters extracted from samples of the upstream and downstream of Baotou section of the Yellow River and of tap water were administered to primary germ cells in vitro of male rats at different concentrations（10, 100, and 1 000 mL/mL; 1 mL organic extracts were equivalent to 10, 100, and 1 000 mL water samples, respectively） for 24 h. The control group was given PBS at the same volume. Proliferation rate of cells was detected by MTT assay, apoptosis by transmission electron microscopy（TEM）, and lactate dehydrogenase（LDH）, total antioxidant capacity（T-AOC）, superoxide dismutase（SOD）, and malondialdehyde（MDA） by micro plate method.[Results] Compared with the control group, the proliferation rates of germ cells exposed to 100 and 1 000 mL/mL organic extracts from tap water, upstream of the Yellow River, and downstream of the Yellow River were significantly decreased（Ps 0.05）, and the proliferation rates of the 1 000 mL/mL groups were reduced to（85.08±0.70）%,（90.80±0.30）%, and（87.50±2.35）%, respectively. Under the transmission electron microscope, the 1 000 mL/mL groups showed cell chromatin clustered at the edge of nucleus, nuclear fragmentation, scattering of some chromatin blocks in membrane, broken mitochondrial structure, vacuolated cells, and cell apoptosis. The LDH activities of the 1 000 and 100 mL/mL downstream groups and the tap water groups were significantly increased compared with that of the control group（Ps 0.05）; the activities of LDH of the 1 000 mL/mL tap water, upstream, and downstream groups were（2 417.48±298.50）,（1 870.37±157.95）, and（2 072.90±195.11） U/L, respectively. The activities of T-AOC and SOD of the 1 000 and 100 mL/mL tap water, upstream, and downstream groups were significantly lower than those of the control group, and the level of MDA was higher（Ps 0.05）; the activity of SOD of the 1 000 mL/mL tap water group [（6.52±1.77）U/mL ] was significantly lower than that of the 1 000 mL/mL upstream group [（8.74±0.90） U/mL]（P 0.05）, and the level of MDA was higher [（3.14±1.18） nmol/L vs.（2.27±0.82） nmol/L]（P 0.05）.[Conclusion] The organic matters extracted from the upstream and downstream of the Baotou section of the Yellow River could inhibit proliferation and promote apoptosis of primary germ cells of male rats. Oxidative damage may be one of the potential mechanisms.