CAJ | 학술논문

比较环孢素A(CsA)转换为雷帕霉素(Rapa)后与CsA长期作用对移植肾大鼠睾丸功能和组织形态学的影响。方法采用标准的肾移植模型方法进行原位左肾移植,即将Fisher大鼠的供肾移植给Lewis雄性大鼠,36只大鼠分为CsA转换为Rapa组(R组,n=10)、CsA持续使用组(A组,n=10)、CsA撤离组(B组,n=8)和控制对照组(C组,n=8),另接受Lewis大鼠供肾的Lewis同系移植鼠作为同系移植组(T组,n=6)。术后前4组大鼠均给予CsA( 1.5 mg·kg-1·d-1)灌胃至第10天,术后11d至7周R、A、B3组大鼠给予CsA(10 mg·kg-1·d-1)灌胃,术后8周至24周R组改用Rapa(0.5 mg·kg-1 ·d-1)灌胃,A组仍持续使用CsA,B组则改用生理盐水(10 ml·kg-1·d-1),C组大鼠术后11d至24周、T组术后至24周均一直予以生理盐水灌胃。术后24周时处死动物,抽血测睾酮、卵泡刺激素和黄体生成素值,并计算睾酮/黄体生成素比率。记录各组大鼠右侧睾丸的质量,光镜下随机测量曲细精管直径并采用量化的组织学评分标准评估睾丸的组织学损伤;R组、A组和T组随机选取2例睾丸标本应用透射电镜行超微结构检查。结果 R组大鼠血清睾酮水平比T组、A组降低[(0.85±0.09)μg/L比(1.05±0.17) μg/L、(0.94±0.13)μg/L,均P＜0.05],其余组间比较差异均无统计学意义。卵泡雌激素、黄体生产素各组间差异无统计学意义。T组睾酮/黄体生成素比值均高于R、A、B、C4组,R组最低(0.59±0.03)。A、B、R、C组大鼠右侧睾丸质量和平均曲细精管直径比T组明显降低,R组下降最为明显。R、A、B、C组大鼠的睾丸组织学损伤评分比T组均升高。电镜检查R组睾丸组织Leydig细胞线粒体肿胀、内质网扩张和Sertoli细胞的细胞器显著减少;A组Leydig细胞内质网扩张和Sertoli细胞的细胞器减少;而T组睾丸组织的超微结构未发现明显异常。结论与单纯持续使用CsA相比,CsA转换为Rapa长期作用并未引起移植肾大鼠睾丸的功能损伤,但导致了更严重的组织学损伤。比較環孢素A(CsA)轉換為雷帕黴素(Rapa)後與CsA長期作用對移植腎大鼠睪汍功能和組織形態學的影響。方法採用標準的腎移植模型方法進行原位左腎移植,即將Fisher大鼠的供腎移植給Lewis雄性大鼠,36隻大鼠分為CsA轉換為Rapa組(R組,n=10)、CsA持續使用組(A組,n=10)、CsA撤離組(B組,n=8)和控製對照組(C組,n=8),另接受Lewis大鼠供腎的Lewis同繫移植鼠作為同繫移植組(T組,n=6)。術後前4組大鼠均給予CsA( 1.5 mg·kg-1·d-1)灌胃至第10天,術後11d至7週R、A、B3組大鼠給予CsA(10 mg·kg-1·d-1)灌胃,術後8週至24週R組改用Rapa(0.5 mg·kg-1 ·d-1)灌胃,A組仍持續使用CsA,B組則改用生理鹽水(10 ml·kg-1·d-1),C組大鼠術後11d至24週、T組術後至24週均一直予以生理鹽水灌胃。術後24週時處死動物,抽血測睪酮、卵泡刺激素和黃體生成素值,併計算睪酮/黃體生成素比率。記錄各組大鼠右側睪汍的質量,光鏡下隨機測量麯細精管直徑併採用量化的組織學評分標準評估睪汍的組織學損傷;R組、A組和T組隨機選取2例睪汍標本應用透射電鏡行超微結構檢查。結果 R組大鼠血清睪酮水平比T組、A組降低[(0.85±0.09)μg/L比(1.05±0.17) μg/L、(0.94±0.13)μg/L,均P＜0.05],其餘組間比較差異均無統計學意義。卵泡雌激素、黃體生產素各組間差異無統計學意義。T組睪酮/黃體生成素比值均高于R、A、B、C4組,R組最低(0.59±0.03)。A、B、R、C組大鼠右側睪汍質量和平均麯細精管直徑比T組明顯降低,R組下降最為明顯。R、A、B、C組大鼠的睪汍組織學損傷評分比T組均升高。電鏡檢查R組睪汍組織Leydig細胞線粒體腫脹、內質網擴張和Sertoli細胞的細胞器顯著減少;A組Leydig細胞內質網擴張和Sertoli細胞的細胞器減少;而T組睪汍組織的超微結構未髮現明顯異常。結論與單純持續使用CsA相比,CsA轉換為Rapa長期作用併未引起移植腎大鼠睪汍的功能損傷,但導緻瞭更嚴重的組織學損傷。
비교배포소A(CsA)전환위뢰파매소(Rapa)후여CsA장기작용대이식신대서고환공능화조직형태학적영향。방법 채용표준적신이식모형방법진행원위좌신이식,즉장Fisher대서적공신이식급Lewis웅성대서,36지대서분위CsA전환위Rapa조(R조,n=10)、CsA지속사용조(A조,n=10)、CsA철리조(B조,n=8)화공제대조조(C조,n=8),령접수Lewis대서공신적Lewis동계이식서작위동계이식조(T조,n=6)。술후전4조대서균급여CsA( 1.5 mg·kg-1·d-1)관위지제10천,술후11d지7주R、A、B3조대서급여CsA(10 mg·kg-1·d-1)관위,술후8주지24주R조개용Rapa(0.5 mg·kg-1 ·d-1)관위,A조잉지속사용CsA,B조칙개용생리염수(10 ml·kg-1·d-1),C조대서술후11d지24주、T조술후지24주균일직여이생리염수관위。술후24주시처사동물,추혈측고동、란포자격소화황체생성소치,병계산고동/황체생성소비솔。기록각조대서우측고환적질량,광경하수궤측량곡세정관직경병채용양화적조직학평분표준평고고환적조직학손상;R조、A조화T조수궤선취2례고환표본응용투사전경행초미결구검사。결과 R조대서혈청고동수평비T조、A조강저[(0.85±0.09)μg/L비(1.05±0.17) μg/L、(0.94±0.13)μg/L,균P＜0.05],기여조간비교차이균무통계학의의。란포자격소、황체생산소각조간차이무통계학의의。T조고동/황체생성소비치균고우R、A、B、C4조,R조최저(0.59±0.03)。A、B、R、C조대서우측고환질량화평균곡세정관직경비T조명현강저,R조하강최위명현。R、A、B、C조대서적고환조직학손상평분비T조균승고。전경검사R조고환조직Leydig세포선립체종창、내질망확장화Sertoli세포적세포기현저감소;A조Leydig세포내질망확장화Sertoli세포적세포기감소;이T조고환조직적초미결구미발현명현이상。결론 여단순지속사용CsA상비,CsA전환위Rapa장기작용병미인기이식신대서고환적공능손상,단도치료경엄중적조직학손상。
Objective To investigate the long-term effects of cyclosporine A(CsA) to rapamycin (Rapa) conversion on testicular function and morphology in rat models of kidney transplantation. Methods Left orthotopic renal transplantation in rats was performed using standard model approach, namely, using Fisher rats as donor and male Lewis rats as recipient. Thirty-six Lewis rat models were randomized into the Rapa group (group R, n=10), CsA continuous use group (group A, n=10), CsA withdrawal group (group B, n=8) and control group (group C, n=8). In addition, the Lewis rats receiving donor Lewis rat kidney were assigned into a syngeneic transplantation group (group T, n=6). Immediately after surgery, Groups R,A, B and C were subjected to gavage of CsA (1.5 mg·kg-1 ·d-1) till day 10. Thereafter, groups R, A and B were given a larger dose of CsA ( 10 mg· kg-1· d-1 ) gavage from day 11 to week 7. From weeks 8 to week 24,CsA was converted to Rapa (0.5 mg· kg-1 ·d-1 ) in group R and to normal saline (10 ml· kg-1· d-1 ) in group B,and remained unchanged in group A. Gavage of normal saline was also given to group C from day 11 to week 24, and to group T immediately after surgery till week 24. At week 24 after surgery, the rats were sacrificed and phlebotomized for determination of testosterone, follicle stimulating hormone (FSH) and luteinizing hormone (LH). Moreover, the testosterone/LH ratio was calculated. The weights of right testis were then recorded in every group. Thereafter, the seminiferous tubule diameter (STD) was randomly measured under light microscope, and a quantitative histological score criteria was used to evaluate the histological injury of testis. Every two specimens of testis were randomly selected from groups R, A and T for examination of ultrastructure using transmission electron microscopy. Results The serum level of testosterone in group R was lowered as compared with group T, group A [(0.85±0.09) μg/L vs (1.05±0.17) μg/L, (0.94+0.13) μg/L,all P＜0.05]but was not significantly different among the other groups. There were no differences in FSH and LH among the five groups. The testosterone/LH ratio was highest in T group and lowest in R group (0.59±0.03). Compared with T group, the weights of right testis and mean STD in groups R, A, B and C were significantly reduced, and were lowest in group R. Groups R, A, B and C had increased histological injury scores as compared with group T. Under electron microscopy, group R showed edematous mitochondria,dilated endoplasmic reticulum in Leydig cells and significantly reduced organelles in Sertoli cells, and group A showed edematous mitochondra in Leydig cells and reduced organelles in Sertoli cells, while group T did not show any abnormalities in ultrastructure of testis tissues. Conclusion Compared with continuous use of CsA, the long-term effect of CsA to Rapamycin conversion may not compromise the function of testis but can lead to more severe histological injury.