摘要
目的 研究热休克蛋白 (Hsc70 /Hsp6 8)、酸性磷酸酶 (ACP)、碱性磷酸酶 (AKP)在肝再生过程中的生物学作用 ,检测这些大分子在大鼠肝大部切除后热休克处理下的动态变化。 方法 用组织化学、免疫组织化学、Western印迹、酶的原位复性电泳、体视学分析、比色分析等方法。 结果 肝切除和热休克联合处理 (PH - HS)后的0～ 144 h恢复期间 ,ACP有 3个高活性期 (12、36和 96 h) ,AKP有 2个高活性期 (12和 36 h) ,Hsc70 /Hsp6 8有 2个高表达期 (16和 48h) ;PH- HS后 ACP和 AKP活性增强与 140～ 180 k D的酶活性增加有关。与只进行热休克 (HS) (44℃ ,30 min)处理或与只进行 2 /3肝切除 (PH)后恢复期间 Hsc70 /Hsp6 8含量和磷酸酶活性动态变化的比较表明 ,PH- HS对 Hsc70 /Hsp6 8含量和磷酸酶活性的影响可持续 12 0 h;PH- HS中的 PH能略微降低 ACP和 AKP活性 ,减少 HS诱导肝细胞合成 Hsc70 /Hsp6 8的量 ;PH- HS中的 HS处理能抑制 PH诱导的 Hsc70 /Hsp6 8表达和推迟 ACP活性高峰出现的时间。 结论 ACP、AKP和 Hsc70 /Hsp6 8均在肝细胞的 HS反应和肝再生中起作用 ,其中 ,ACP可能在启动肝再生中起重要作用 ,AKP和 Hsc70 /Hsp6 8可能在
Objective We tried to probe the biological function of ACP, AKP and Hsc70/Hsp68 in liver regeneration treated by heat shock(44℃, 30min, recover 8 hours)after 2/3 hepatectomy (PH HS). Methods By histochemistry, immunohistochemistry, Western blot,non denatured electrophoresis, sterology and colorimetry, we detected the location and content of the conserved heat shock protein70/induced heat shock protein 68 and location and activity of the acid and alkaline phosphatases(ACP and AKP). Results ACP has three high activity stages(12,36 and 96h), AKP has two high activity stages(12 and 36h), and Hsc70/Hsp68 has two high expression stages (12 and 48 h) during 0\|144 h recovery time after PH HS. The enhanced ACP and AKP activity comes from 140 180 kD phosphatases after PH HS. Comparison of kinetics of Hsc70/Hsp68 content and ACP and AKP activity after PH HS with that after heat shock(HS) treatment(44℃, 30min) and 2/3 hepatectomy(PH) respectively shows that the effect of PH HS on Hsc70/Hsp68 content and phosphatase activity lasts 120 h; The PH in PH HS can reduce ACP and AKP activity and Hsc70/Hsp68 expression. The HS in PH HS can change the time of ACP and AKP enhancing activity and inhibit expression of the Hsc70/Hsp68 induced by PH. Conclusion ACP, AKP and Hsc70/Hsp68 play an important role in heat shock response of liver cells and liver regeneration, but ACP is more important in originating liver regeneration, while AKP and Hsc70/Hsp68 probably exert the effect in DNA synthesis and cell division.
出处
《解剖学报》
CAS
CSCD
北大核心
2000年第3期238-245,I009,共9页
Acta Anatomica Sinica
基金
中法先进技术项目! [国科生字 ( 1996) 13 4号 ]
关键词
肝再生
热休克
热休克蛋白
磷酸酶
大鼠
Liver regeneration
Heat shock(HS)
Conserved heat shock protein 70/induced heat shock protein 68(Hsc70/Hsp68)
ACP
AKP
Rat
