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为研究环孢酶素A(cyclosporin A,Cs A)对精原细胞增殖作用的影响,将睾丸组织小块(0.5 mm×0.5 mm×0.5 mm)分别放入终浓度为0(DMEM,溶剂对照)、0.01、0.1、1、10μg/ml的环孢酶素A溶液和卵泡雌激素(FSH,阳性对照)培养液的滤膜上面,于32℃、5%的CO2恒温箱中培养3 d。培养结束3 h之前添加溴脱氧尿苷(Bromodeoxyuridine,BrdU)进行免疫组化染色。阴性对照组直接将小断片在溴脱氧尿苷培养液中培养3 h后进行固定。测定BrdU阳性细胞数,并通过BrdU阳性细胞率考察增殖活性。结果显示,与溶剂对照组比较,0.1、1、10μm/ml环孢酶素A染毒组精原细胞的BrdU阳性细胞率均较低,差异有统计学意义(P<0.05或P<0.01);且随着环孢酶素A染毒浓度的升高,精原细胞的BrdU阳性细胞率呈下降趋势。表明环孢酶素A能抑制小鼠精原细胞的增殖活性。
In order to study the effect of cyclosporin A (CsA) on the proliferation of spermatogonial cells, small pieces of testicular tissue (0.5 mm × 0.5 mm × 0.5 mm) were placed in a final concentration of 0 (DMEM, solvent control ), 0.01, 0.1, 1, 10μg / ml cyclosporine A solution and follicular estrogen (FSH, positive control) medium for 3 days at 32 ° C in a 5% CO2 incubator. Bromodeoxyuridine (BrdU) was added for immunohistochemical staining 3 h before the end of culture. In the negative control group, small fragments were directly fixed in bromodeoxyuridine medium for 3 h. The number of BrdU positive cells was measured, and the proliferative activity was examined by the rate of BrdU positive cells. The results showed that compared with the solvent control group, the rates of BrdU positive cells in spermatogonia of 0.1, 1, 10μm / ml cyclosporin A group were lower, the difference was statistically significant (P <0.05 or P <0.01) ; And with the increase of cyclosporine A concentration, spermatogonia BrdU positive cells showed a downward trend. Cyclosporin A can inhibit the proliferation of mouse spermatogonia.