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脂质刺激的Kupffer细胞对肝星状细胞增殖的影响 |
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陆伦根 曾民德 李继强 华静 范建高 范竹萍 戴宁 邱德凯 萧树东
【摘要】 目的 探讨甘油三酯和极低密度脂蛋白(VLDL)及其刺激的Kupffer细胞对大鼠肝星状细胞(HSC)增殖的影响。方法 用链霉蛋白酶和胶原酶原位灌流，Nycodenz密度梯度离心分离大鼠HSC和Kupffer细胞，制备甘油三酯和VLDL刺激的Kupffer细胞培养的上清液(KCCM)，并以MTT法观察甘油三酯和VLDL及KCCM对HSC增殖效应。结果 甘油三酯和VLDL浓度分别在12.5 mg/L和25～100 mg/L时对HSC增殖有促进作用(P<0.05或P<0.01)，与正常和KCCM组相比，KCCM+VLDL组和KCCM+甘油三酯组对HSC增殖有促进作用(P<0.01)，KCCM+VLDL组和KCCM+甘油三酯组之间差异无显著性(P>0.05)；KCCM组高于正常组，但统计学无差异(P>0.05)。结论 甘油三酯和VLDL及其刺激的KCCM可促进HSC增殖，其与脂肪肝、肝纤维化的发生可能有关。
【关键词】 枯否细胞 脂类 肝星状细胞
Effects of Kupffer cells stimulated by triglyceride and very low-density lipoprotein on proliferation of rat hepatic stellate cells LU Lungen, ZENG Minde, LI Jiqiang, et al. Shanghai Institute of Digestive Diseases, Renji Hospital, Shanghai Second Medical University, Shanghai 200001
【Abstract】 Objective To study the effects of Kupffer cell-conditioned medium (KCCM) derived from triglyceride and very low-density lipoprotein (VLDL) treatment on proliferation of rat hepatic stellate cells (HSC). Methods HSC and Kupffer cells were isolated from liver of Wistar rats by in situ perfusion with pronase and collagenase and density gradient centrifugation with Nycodenz and then cultured. KCCM was prepared and MTT colorimetric assay was used to detect HSC proliferation. Results HSC and Kupffer cells were isolated successfully with high purity. 12.5 mg/L of triglyceride and 25～100 mg/L of VLDL promoted HSC proliferation (P<0.05 or 0.01); KCCM+triglyceride and KCCM+VLDL might promote HSC proliferation as compared with a control group (P<0.01). There was no significant difference between KCCM+triglyceride and KCCM+VLDL (P>0.05). Conclusion The technique for isolation of HSC and Kupffer cells described here is simple and reliable. Triglyceride, VLDL and KCCM stimulated by them may promote HSC proliferation and be associated with fatty liver and hepatic fibrogenesis.
【Key words】 Kupffer cells Lipids Hepatic stellate cell
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