【摘要】：目的： 1.觀察不同劑量反式脂肪酸（Trans Fatty Acids，TFA）對(duì)雄性小鼠的生殖毒性作用； 2.初步探討原花青素（proanthocyanidins,PC）對(duì)反式脂肪酸致雄性小鼠生殖損傷保護(hù)效果及可能機(jī)制,為評(píng)價(jià)PC對(duì)TFA致生殖損傷的保護(hù)作用提供實(shí)驗(yàn)依據(jù)。 方法： 以TFA和PC為受試物，以健康成年昆明（KM）種雄性小鼠為實(shí)驗(yàn)對(duì)象。將96只小鼠隨機(jī)分為8組：對(duì)照組、低、中、高劑量TFA染毒組、PC對(duì)照組、L-TFA+PC保護(hù)組、M-TFA+PC保護(hù)組、H-TFA+PC保護(hù)組；3個(gè)劑量TFA染毒組：分別按低（TFA25mg/kg）、中（TFA50mg/kg）、高（TFA100mg/kg）TFA染毒劑量給小鼠灌胃90天，隔天一次，上午灌胃TFA、下午灌胃等量的生理鹽水；3個(gè)PC保護(hù)組：染毒劑量、時(shí)間同染毒組，上午灌胃TFA、下午每組灌胃同劑量的（100mg/kg）PC；對(duì)照組灌胃生理鹽水、PC組灌胃（100mg/kg）PC。各組動(dòng)物灌胃體積為10ml/kg．bw。實(shí)驗(yàn)結(jié)束后取血處死動(dòng)物，取睪丸、附睪和腎臟分別稱重并計(jì)算其臟器系數(shù)；常規(guī)病理學(xué)切片，HE染色觀察睪丸病理形態(tài)變化；光學(xué)顯微鏡下觀察精子質(zhì)量的改變；全自動(dòng)生化分析儀檢測(cè)血清T、LH、FSH水平；試劑盒測(cè)定MDA、SOD、GSH-Px酶的活性。 結(jié)果： 1．中、高劑量TFA染毒組小鼠末體重及凈增體重明顯高于對(duì)照組，經(jīng)PC保護(hù)的高劑量TFA染毒組動(dòng)物的凈增體重低于同劑量染毒組（P＜0.05）。 2．高劑量TFA染毒組小鼠睪丸臟器系數(shù)低于對(duì)照組，經(jīng)PC保護(hù)的高劑量TFA染毒組睪丸臟器系數(shù)低于同劑量染毒組（P＜0.05）；PC對(duì)其它各染毒組睪丸、附睪及腎臟臟器系數(shù)影響不大，，差異無(wú)顯著性（P＞0.05）。 3．睪丸病理形態(tài)顯示:對(duì)照組小鼠睪丸曲細(xì)精管完整，上皮細(xì)胞層次正常，管腔中見(jiàn)到大量成熟精子等；TFA染毒組睪丸組織基底膜模糊不清、核間隙增寬，部分管壁脫落缺失，腔內(nèi)未見(jiàn)精子等；給予PC保護(hù)后，睪丸組織形態(tài)較TFA染毒組有所改善，以低、中劑量TFA染毒組改善較明顯，睪丸間質(zhì)有所縮小，間質(zhì)細(xì)胞顆粒減少，各級(jí)生精細(xì)胞數(shù)量有所增加，曲細(xì)精管上皮細(xì)胞層次趨向正常，腔內(nèi)精子較多。 4．與對(duì)照組比較，中、高劑量TFA染毒組小鼠精子數(shù)量、精子存活率、精子活力均降低，畸形率增加（P＜0.05）；經(jīng)PC保護(hù)的中、高劑量TFA染毒組精子數(shù)量、精子存活率、精子活力均有不同程度的增加，畸形率減少與相同劑量染毒組比較，差別有顯著性（P＜0.05）；精子畸形以尾折疊、卷尾、尾體彎曲、不規(guī)則精子、雙尾精子為主。 5．高劑量TFA染毒組小鼠血清FSH、LH、T值均低于對(duì)照組（P＜0.05）；經(jīng)PC保護(hù)的高劑量TFA染毒組FSH、LH、T值均較同劑量的染毒組有不同程度的增加，差別有顯著性（P＜0.05）。 6．高劑量TFA染毒組小鼠睪丸LDH-X、GGT的活性均低于對(duì)照組（P＜0.05）；經(jīng)PC保護(hù)高劑量TFA染毒組LDH-X、GGT活性均較同劑量的染毒組有不同程度的增加（恢復(fù)），差別有顯著性（P＜0.05），其它各組對(duì)其酶的活性影響不明顯（P＞0.05）。 7．與對(duì)照組比較，中、高劑量TFA染毒組小鼠睪丸MDA含量增加，SOD、GSH-Px活性降低（P＜0.05)；高劑量TFA染毒組CAT活性低于對(duì)照組(P＜0.05)；經(jīng)PC保護(hù)的高劑量TFA染毒組小鼠睪丸SOD、GSH-Px、CAT活性均較同劑量的染毒組增加（P＜0.05）；PC保護(hù)的中劑量TFA染毒組睪丸GSH-Px活性高于同劑量的染毒組（P＜0.05）；其它各組酶的活性及氧化產(chǎn)物MDA含量影響不明顯（P＞0.05）。 結(jié)論： 1.不同劑量反式脂肪酸（TFA）亞慢性染毒雄性小鼠可致生殖系統(tǒng)損傷，高劑量TFA對(duì)生殖功能損傷嚴(yán)重。 2.采用100mg/kg.bw劑量原花表素（PC）可修復(fù)TFA致雄性小鼠生殖系統(tǒng)的損傷作用； 3.抗氧化作用是PC保護(hù)（修復(fù)）TFA所致雄性生殖損傷的重要機(jī)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.
【學(xué)位授予單位】：南華大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2012
【分類號(hào)】：R114

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