中华实验外科杂志
中華實驗外科雜誌
중화실험외과잡지
CHINESE JOURNAL OF EXPERIMENTAL SURGERY
2014年
7期
1530-1532,封4
,共4页
顾朝辉%田凤艳%李冠儒%贾占奎%孟政雷%孙科%魏金星%杨锦建%阚全程
顧朝輝%田鳳豔%李冠儒%賈佔奎%孟政雷%孫科%魏金星%楊錦建%闞全程
고조휘%전봉염%리관유%가점규%맹정뢰%손과%위금성%양금건%감전정
乙醇%睾丸损伤%睾丸特异表达基因-1
乙醇%睪汍損傷%睪汍特異錶達基因-1
을순%고환손상%고환특이표체기인-1
Ethanol%Testis injury%Testis specific expressed gene 1
目的 探讨睾丸特异表达基因-1(TSEG-1)在乙醇致小鼠睾丸损伤模型中的表达变化及意义.方法 首先建立乙醇致睾丸损伤模型,将12只雄性昆明小鼠随机分为对照组和实验组,每组6只,用生理盐水稀释无水乙醇为15％浓度.实验组小鼠每天按3 g乙醇(15％,V/V)/kg体质量,腹腔内注射14 d,对照组为等体积生理盐水.应用苏木素-伊红(HE)染色鉴定睾丸组织形态学变化,实时定量逆转录聚合酶链反应(RT-qPCR)检测TSEG-1 mRNA变化,采用免疫组织化学和Western blot检测其蛋白变化.结果 乙醇组睾丸HE染色结果提示,与对照组比较,乙醇组睾丸组织85.6％曲精小管管腔消失,Ⅰ～Ⅳ级生精细胞数目减少,78.2％精母细胞或精子细胞核皱缩.在乙醇组睾丸组织中,TSEG-1 mRNA表达水平(7.500±0.657,n=6)较对照组(0.985±0.231,n=6)显著增加,差异有统计学意义(t=22.915,P＜0.01).同时,乙醇组TSEG-1蛋白质较生理盐水组显著增加,经Quantity One软件分析灰度值后,与管家基因甘油醛-3-磷酸脱氢酶(GAPDH)比较,乙醇组TSEG-1蛋白(1.360±0.202,n=6)较生理盐水组(0.330±0.112,n=6)表达增加达4倍,差异有统计学意义(=10.69,P＜0.01).结论 乙醇致小鼠睾丸损伤模型是研究睾丸损伤的可靠模型,TSEG-1可能参与乙醇致睾丸损伤模型的分子发生过程.
目的 探討睪汍特異錶達基因-1(TSEG-1)在乙醇緻小鼠睪汍損傷模型中的錶達變化及意義.方法 首先建立乙醇緻睪汍損傷模型,將12隻雄性昆明小鼠隨機分為對照組和實驗組,每組6隻,用生理鹽水稀釋無水乙醇為15％濃度.實驗組小鼠每天按3 g乙醇(15％,V/V)/kg體質量,腹腔內註射14 d,對照組為等體積生理鹽水.應用囌木素-伊紅(HE)染色鑒定睪汍組織形態學變化,實時定量逆轉錄聚閤酶鏈反應(RT-qPCR)檢測TSEG-1 mRNA變化,採用免疫組織化學和Western blot檢測其蛋白變化.結果 乙醇組睪汍HE染色結果提示,與對照組比較,乙醇組睪汍組織85.6％麯精小管管腔消失,Ⅰ～Ⅳ級生精細胞數目減少,78.2％精母細胞或精子細胞覈皺縮.在乙醇組睪汍組織中,TSEG-1 mRNA錶達水平(7.500±0.657,n=6)較對照組(0.985±0.231,n=6)顯著增加,差異有統計學意義(t=22.915,P＜0.01).同時,乙醇組TSEG-1蛋白質較生理鹽水組顯著增加,經Quantity One軟件分析灰度值後,與管傢基因甘油醛-3-燐痠脫氫酶(GAPDH)比較,乙醇組TSEG-1蛋白(1.360±0.202,n=6)較生理鹽水組(0.330±0.112,n=6)錶達增加達4倍,差異有統計學意義(=10.69,P＜0.01).結論 乙醇緻小鼠睪汍損傷模型是研究睪汍損傷的可靠模型,TSEG-1可能參與乙醇緻睪汍損傷模型的分子髮生過程.
목적 탐토고환특이표체기인-1(TSEG-1)재을순치소서고환손상모형중적표체변화급의의.방법 수선건립을순치고환손상모형,장12지웅성곤명소서수궤분위대조조화실험조,매조6지,용생리염수희석무수을순위15％농도.실험조소서매천안3 g을순(15％,V/V)/kg체질량,복강내주사14 d,대조조위등체적생리염수.응용소목소-이홍(HE)염색감정고환조직형태학변화,실시정량역전록취합매련반응(RT-qPCR)검측TSEG-1 mRNA변화,채용면역조직화학화Western blot검측기단백변화.결과 을순조고환HE염색결과제시,여대조조비교,을순조고환조직85.6％곡정소관관강소실,Ⅰ～Ⅳ급생정세포수목감소,78.2％정모세포혹정자세포핵추축.재을순조고환조직중,TSEG-1 mRNA표체수평(7.500±0.657,n=6)교대조조(0.985±0.231,n=6)현저증가,차이유통계학의의(t=22.915,P＜0.01).동시,을순조TSEG-1단백질교생리염수조현저증가,경Quantity One연건분석회도치후,여관가기인감유철-3-린산탈경매(GAPDH)비교,을순조TSEG-1단백(1.360±0.202,n=6)교생리염수조(0.330±0.112,n=6)표체증가체4배,차이유통계학의의(=10.69,P＜0.01).결론 을순치소서고환손상모형시연구고환손상적가고모형,TSEG-1가능삼여을순치고환손상모형적분자발생과정.
Objective To explore the cellular localization and transcript levels of testis specific expressed gene 1 (TSEG-1) in ethanol-induced mouse testis injury model.Methods Twelve male Kunming mice were divided into normal saline solution group (n =6) and ethanol injection group (n =6).The morphological changes of testes were observed by hematoxylin and eosin (HE) staining.The transcriptional and protein levels of TSEG-1 were detected by real-time reverse transcriptase-polymerase chain reaction (RT-qPCR),Immunohistochemistry (IHC) and Western blotting.Results Intraperitoneal injection of 3 g ethanol (15％,V/V)/kg body weight for 14 days induced mouse testis injury.As compared with normal saline solution group,85.6％ serminous tubules disappeared,the number of Ⅰ-Ⅳ-stage spermatogenic cells was reduced,and 78.2％ spermatocytes and spermatids had nuclei shrinkage in ethanol injection group.Extensive TSEG-1 mRNA levels were observed in control testes,mainly localizing in spermatocytes and early spermatids.The transcripts of TSEG-1 (7.500 ± 0.657,n =6) were up-regulated to 8 folds in ethanol injection group as compared with those in normal saline solution control group (0.985 ±0.231,n =6),with the difference being significant (t =22.915,P ＜ 0.01).Especially,the protein expression of TSEG-1 (1.360 ±0.202,n =6) was up-regulated to 4 folds in ethanol injection group as compared with that in normal saline solution control group (0.330 ±0.112,n =6),with the difference being significant,t =10.69,P ＜ 0.01.Conclusion Ethanol-induced testis injury model was successfully constructed by intraperitoneal injection of ethanol.The novel gene TSEG-1 may play a role in the pathogenesis of ethanol-induced testis injury model,which laid the basis for further study on its function in testis diseases.
