出 处:《中华实验外科杂志》2020年第8期1457-1460,共4页Chinese Journal of Experimental Surgery
摘 要:目的观察GYY4137对大鼠精索静脉曲张(VC)生精功能损伤的保护作用。方法选取健康成年雄性SD大鼠32只购自湖北省疾病预防控制中心,随机分为4组:假手术组、假手术+GYY4137组、VC组及VC+GYY4137组。假手术组、假手术+GYY4137组仅分离而不结扎左肾静脉,VC组与VC+GYY4137组结扎左肾静脉制备VC模型,VC+GYY4137组、假手术+GYY4137组术后每日腹腔注射GYY4137(10 mg/kg)。采用苏木精-伊红(HE)染色观察睾丸组织病理改变;测定睾丸组织中丙二醛(MDA)、超氧化物歧化酶(SOD)含量;采用原位缺口末端标记法(TUNEL)检测细胞凋亡指数;免疫组织化学及蛋白质印迹法(Western blot)检测睾丸组织中B细胞淋巴瘤/白血病-2相关X蛋白(bax)和半胱氨酰天冬氨酸特异性蛋白酶(Caspase)-3的表达。两组间比较采用t检验。结果VC组可见各级生精细胞排列紊乱且广泛脱落,VC+GYY4137组可见生精细胞排列轻度紊乱,少数腔内见脱落的生精细胞。与假手术组[(1.86±0.23)nmol/g蛋白]和假手术+GYY4137组[(1.83±0.22)nmol/g蛋白]比较,VC组[(4.33±0.37)nmol/g蛋白]中MDA含量明显增加(t=9.820、10.059,P<0.05);VC+GYY4137组[(2.38±0.27)nmol/g蛋白]与VC组比较,MDA含量显著下降(t=-7.374,P<0.05)。VC组的SOD活性[(0.23±0.04)U/mg蛋白]与假手术组[(0.81±0.06)U/mg蛋白]和假手术+GYY4137组[(0.82±0.07)U/mg蛋白]比较明显降低(t=-9.757、-9.926,P<0.05);VC组经GYY4137处理后SOD活性增加[(0.62±0.06)U/mg蛋白](t=9.368,P<0.05)。TUNEL结果显示假手术组及假手术+GYY4137组[(3.89±0.45)%、(3.92±0.51)%]可见较少的睾丸生精细胞凋亡,VC组凋亡细胞明显增加[(17.26±0.98)%,P<0.05];与VC组比较,VC+GYY4137组生精细胞凋亡明显减少[(8.24±0.65)%,P<0.05]。免疫组织化学及Western blot结果显示Caspase-3和bax表达水平在VC组(0.41±0.04、0.59±0.06)中的表达明显高于假手术组(0.11±0.02、0.19±0.03,t=9.650、9.097,P<0.05)和假手术+GYY4137组(0.12±0.02、0.20±0.03,t=9.Objective To observe the protective effects of GYY4137 on spermatogenic function injury in varicocele(VC)rats.Methods Thirty-two adult male SD rats were randomly divided into 4 groups:sham operation group,sham operation+GYY4137 group,VC group,VC+GYY4137 group.Only abdominal cavity was opened in the sham operation group and sham operation+GYY4137 group without ligation of the left renal vein.The left renal vein was ligated in the VC group and VC+GYY4137 group to prepare the VC model.VC+GYY4137 group and sham operation+GYY4137 group received intraperitoneal injection of GYY4137[10 mg/(kg·d)].HE staining was used to observe the pathological changes of testis.Malondialdehyde(MDA)contents and superoxide dismutase(SOD)activity were determined.Cell apoptosis index was detected by dUTP nick end labeling.The expression of B cell lymphoma/leukemia-2 associated X protein(bax)and cysteinyl aspartate-specific protease(Caspase)-3 in testis tissues was detected by immunohistochemistry and Western blotting.Results The left testicular tissue showed desquamation of epithelial cells,disordered arrangement of spermatogenic cells in VC group.In VC+GYY4137 group,the arrangement of spermatogenic cells was slightly disordered,and a few of spermatogenic cells were exfoliated.As compared with sham operation group[(1.86±0.23)nmol/gprot]and sham operation+GYY4137 group[(1.83±0.22)nmol/gprot],MDA contents in VC group[(4.33±0.37)nmol/gprot]increased significantly(t=9.820,10.059,P<0.05).As compared with VC group,MDA contents in VC+GYY4137 group[(2.38±0.27)nmol/gprot]decreased significantly(t=-7.374,P<0.05).The SOD activity in VC group[(0.23±0.04)U/mgprot]was significantly lower than that in sham operation group[(0.81±0.06)U/mgprot]and sham operation+GYY4137 group[(0.82±0.07)U/mgprot,t=-9.757,-9.926,P<0.05].The SOD activity in VC group increased after GYY4137 treatment[(0.62±0.06)U/mgprot,t=9.368,P<0.05].TUNEL assay showed that there was apoptosis of less testicular spermatogenic cells in sham operation group and sham operation+GYY4137
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