原花青素对反式脂肪酸致雄性小鼠生殖毒性的保护作用

发布时间：2019-06-03 23:15

【摘要】：目的： 1.观察不同剂量反式脂肪酸（Trans Fatty Acids，TFA）对雄性小鼠的生殖毒性作用； 2.初步探讨原花青素（proanthocyanidins,PC）对反式脂肪酸致雄性小鼠生殖损伤保护效果及可能机制,为评价PC对TFA致生殖损伤的保护作用提供实验依据。方法：以TFA和PC为受试物，以健康成年昆明（KM）种雄性小鼠为实验对象。将96只小鼠随机分为8组：对照组、低、中、高剂量TFA染毒组、PC对照组、L-TFA+PC保护组、M-TFA+PC保护组、H-TFA+PC保护组；3个剂量TFA染毒组：分别按低（TFA25mg/kg）、中（TFA50mg/kg）、高（TFA100mg/kg）TFA染毒剂量给小鼠灌胃90天，隔天一次，上午灌胃TFA、下午灌胃等量的生理盐水；3个PC保护组：染毒剂量、时间同染毒组，上午灌胃TFA、下午每组灌胃同剂量的（100mg/kg）PC；对照组灌胃生理盐水、PC组灌胃（100mg/kg）PC。各组动物灌胃体积为10ml/kg．bw。实验结束后取血处死动物，取睾丸、附睾和肾脏分别称重并计算其脏器系数；常规病理学切片，HE染色观察睾丸病理形态变化；光学显微镜下观察精子质量的改变；全自动生化分析仪检测血清T、LH、FSH水平；试剂盒测定MDA、SOD、GSH-Px酶的活性。结果： 1．中、高剂量TFA染毒组小鼠末体重及净增体重明显高于对照组，经PC保护的高剂量TFA染毒组动物的净增体重低于同剂量染毒组（P＜0.05）。 2．高剂量TFA染毒组小鼠睾丸脏器系数低于对照组，经PC保护的高剂量TFA染毒组睾丸脏器系数低于同剂量染毒组（P＜0.05）；PC对其它各染毒组睾丸、附睾及肾脏脏器系数影响不大，，差异无显著性（P＞0.05）。 3．睾丸病理形态显示:对照组小鼠睾丸曲细精管完整，上皮细胞层次正常，管腔中见到大量成熟精子等；TFA染毒组睾丸组织基底膜模糊不清、核间隙增宽，部分管壁脱落缺失，腔内未见精子等；给予PC保护后，睾丸组织形态较TFA染毒组有所改善，以低、中剂量TFA染毒组改善较明显，睾丸间质有所缩小，间质细胞颗粒减少，各级生精细胞数量有所增加，曲细精管上皮细胞层次趋向正常，腔内精子较多。 4．与对照组比较，中、高剂量TFA染毒组小鼠精子数量、精子存活率、精子活力均降低，畸形率增加（P＜0.05）；经PC保护的中、高剂量TFA染毒组精子数量、精子存活率、精子活力均有不同程度的增加，畸形率减少与相同剂量染毒组比较，差别有显著性（P＜0.05）；精子畸形以尾折叠、卷尾、尾体弯曲、不规则精子、双尾精子为主。 5．高剂量TFA染毒组小鼠血清FSH、LH、T值均低于对照组（P＜0.05）；经PC保护的高剂量TFA染毒组FSH、LH、T值均较同剂量的染毒组有不同程度的增加，差别有显著性（P＜0.05）。 6．高剂量TFA染毒组小鼠睾丸LDH-X、GGT的活性均低于对照组（P＜0.05）；经PC保护高剂量TFA染毒组LDH-X、GGT活性均较同剂量的染毒组有不同程度的增加（恢复），差别有显著性（P＜0.05），其它各组对其酶的活性影响不明显（P＞0.05）。 7．与对照组比较，中、高剂量TFA染毒组小鼠睾丸MDA含量增加，SOD、GSH-Px活性降低（P＜0.05)；高剂量TFA染毒组CAT活性低于对照组(P＜0.05)；经PC保护的高剂量TFA染毒组小鼠睾丸SOD、GSH-Px、CAT活性均较同剂量的染毒组增加（P＜0.05）；PC保护的中剂量TFA染毒组睾丸GSH-Px活性高于同剂量的染毒组（P＜0.05）；其它各组酶的活性及氧化产物MDA含量影响不明显（P＞0.05）。结论： 1.不同剂量反式脂肪酸（TFA）亚慢性染毒雄性小鼠可致生殖系统损伤，高剂量TFA对生殖功能损伤严重。 2.采用100mg/kg.bw剂量原花表素（PC）可修复TFA致雄性小鼠生殖系统的损伤作用； 3.抗氧化作用是PC保护（修复）TFA所致雄性生殖损伤的重要机制之一。
[Abstract]:Purpose: 1. To observe the reproductive toxicity of trans-fatty acids (TFA) in male mice 2. The effect of procyanidins (PC) on the reproductive injury of male mice induced by trans-fatty acids and the effect of procyanidins (PC) on the reproductive injury of male mice Can be used to evaluate the protective effect of PC on the reproductive damage induced by TFA. Basis for inspection Methods: The subjects were tested with TFA and PC in healthy adult Kunming (KM) species. The mice were randomly divided into 8 groups: the control group, the low, medium, high dose TFA exposure group, the PC control group, the L-TFA + PC protection group, the M-TFA + PC protection group, the H-TFA + PC protection group, and the three dose TFA exposure groups: the mice were treated with a low (TFA25mg/ kg), medium (TFA50mg/ kg), high (TFA100mg/ kg) TFA exposure dose, respectively, 0 days, once every other day, in the morning, the rats were given the same amount of physiological saline in the morning,3 PC protection groups: the exposure dose, the time and the exposure group, the daily dose of TFA in the morning and the same dosage of (100 mg/ kg) PC in each group in the afternoon; the control group was given normal saline, and the PC group was given intragastric administration (100 m G/ kg) PC. The volume of each group of animals was 10 m. L/ kg. bw.The animals were sacrificed after the end of the experiment, and the testis, the epididymis and the kidneys were weighed and the organ coefficients of the animals were calculated; the pathological changes of the testis were observed by HE staining; the change of the quality of the sperm was observed under the optical microscope; and the full-automatic biochemical analyzer was tested. Serum T, LH, and FSH levels were measured; MDA, SOD, and GSH were determined by the kit. -Px Results:1. The body weight and net increase of the mice in the high-dose TFA-treated group were significantly higher than that in the control group. The net increase of the animals in the high-dose TFA-treated group was lower than that in the control group. 2. In the high-dose TFA-treated group, the testis-organ coefficient was lower than that of the control group (P <0.05), and the testis-organ coefficient of the high-dose TFA-treated group was lower than that of the same-dose group (P <0.05), and the PC was in the testis of other groups. There was little effect on the coefficient of the epididymis and the renal organ. There was no significant difference (P> 0.05).3. The pathological morphology of the testis showed that the testis of the control group was intact, the level of the epithelial cells was normal, and a large number of mature sperm were found in the lumen. The basal membrane of the testis of the TFA exposed group was blurred, the clearance of the nucleus was widened, There was no sperm and the like in the cavity. After administration of the PC, the form of the testis was improved with the TFA exposure group. In the low and medium-dose TFA exposure group, the number of spermatogenic cells at all levels was increased, and the interstitial cell particles decreased and the number of spermatogenic cells at all levels increased. seminiferous tubule epithelium In comparison with the control group, the number of sperm, the survival rate of the sperm and the motility of the sperm decreased and the rate of deformity increased (P <0.05). The number of sperm and the sperm of the high-dose TFA exposure group were increased by PC-protected medium and high-dose TFA. The survival rate and the motility of the sperm were increased, and the rate of deformity decreased with the same dose group. The difference was significant (P <0.05), and the sperm deformity was folded at the tail and the tail of the sperm was in the tail. The serum FSH, LH and T in the high-dose TFA-treated group were lower than that in the control group (P <0.05), and the levels of FSH, LH and T in the high-dose TFA-treated group were lower than those of the same dose group. The activity of LDH-X and GGT in the mice with high dose of TFA was lower than that in the control group (P <0.05). The difference was significant (P <0.05) and the other was significant (P <0.05). The activity of SOD and GSH-Px in the mice with high-dose TFA was lower than that in the control group (P <0.05), and the activity of SOD and GSH-Px in the high-dose TFA-treated group was lower than that of the control group (P <0.05), and the activity of the high-dose TFA in the group was lower than that of the control group (P <0.05), and the activity of the high-dose TFA in the group was lower than that of the control group (P <0.05). The activities of SOD, GSH-Px and CAT in the mice with high dose of TFA were higher than that in the same dose group (P <0.05), and the activity of GSH-Px in the rats exposed to the PC was higher than that of the same dose group (P <0.05), and the activities of other groups of enzymes and oxidation The content of MDA in the product was not significant (P> 0.05). Conclusion:1. Different doses of trans-fatty acid (TFA) subchronic exposure. In mice, the reproductive system was damaged, and the high-dose TFA had a serious damage to the reproductive function.2.100 mg/ kg.bw of the agent was used. The effect of the amount of proanthocyanidin (PC) on the reproductive system of the male mice induced by TFA;3.
【学位授予单位】：南华大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R114

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