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目的 :探讨精原细胞在封闭其端粒酶RNA(mTR)后的变化情况 ,及其对细胞因子 (SCF、TGF β1 )的反应性。 方法 :以脂质体为载体将端粒酶RNA的反义寡核苷酸 (PS ASON)导入体外增殖的精原细胞后 ,加入SCF或TGF β1 ,四甲基偶氮唑盐 (MTT)比色法检测抑制前后细胞增殖情况 ,端粒重复扩增法 (TRAP)检测端粒酶活性变化。 结果 :1μmol L终浓度的PS ASON明显下调精原细胞端粒酶活性 ,抑制细胞增殖 ,而抑制结束后 ,端粒酶活性有一定程度的恢复 (P <0 .0 1)。细胞因子对抑制后的精原细胞有调控作用 ,SCF提高端粒酶活性 ,并促进细胞增殖。相反 ,TGF β1 则抑制端粒酶活性的恢复。 结论 :反义抑制精原细胞端粒酶活性能有效抑制细胞增殖 ,抑制结束后有较好的可逆性 ,并受细胞因子的调控 ,可望为男性节育提供一新型有效的方法
Objective: To investigate the changes of spermatogonia after blocking its telomerase RNA (mTR) and its reactivity to cytokines (SCF and TGF β1). METHODS: Antisense oligonucleotides of telomerase RNA (PS ASON) were transfected into spermatogonia in vitro using liposome as a carrier, and then transfected with SCF or TGF β1, MTT ratio The proliferation of cells before and after inhibition was detected by colorimetric assay. The changes of telomerase activity were detected by telomeric repeat amplification assay (TRAP). Results: PS ASON at a final concentration of 1 μmol significantly down-regulated the telomerase activity of spermatogonia and inhibited cell proliferation. However, telomerase activity recovered to a certain extent after inhibition (P <0.01). Cytokines play a regulatory role in the inhibition of spermatogonia. SCF enhances telomerase activity and promotes cell proliferation. In contrast, TGFβ1 inhibits the recovery of telomerase activity. CONCLUSIONS: Antisense inhibition of telomerase activity in spermatogonia can effectively inhibit cell proliferation, and after reversal of inhibition, it has good reversibility and is regulated by cytokines, which is expected to provide a new and effective method for male contraception