Chen Xin, Wang Yu-zhen, Xiu He-ming, Jiang Hui-qing
Department of Gastroenterology, Second Hospital of Hebei Medical University, Shijiazhuang 050000, China.
Zhonghua Yi Xue Za Zhi. 2006 Jul 25;86(28):1985-8.
To evaluate the effects of nerve growth factor (NGF) on the proliferation and apoptosis of hepatic stellate cells (HSCs) of fibrotic rat.
HSCs of fibrotic rat were incubated in medium with different concentrations of NGF (50 ng/ml, 100 ng/ml, and 150 ng/ml respectively) or without NGF for 24, 48 and 72 hours respectively. The proliferation rate of the HSCs was determined by MTT assay. Apoptotic HSCs were detected by TUNEL method. The apoptotic rate of HSCs was studied by flow cytometry. The conformation of HSCs was observed by microscopy.
(1) MTT method indicated that the proliferation rates of the HSCs induced with different concentrations of NGF for 24 hours was significantly reduced compared with that of the control group (0.77 +/- 0.03, all P < 0.05). The proliferation rate of the HSCs treated with NGF of the concentration of 100 ng/ml for 24 hours was the lowest (0.63 +/- 0.02, P < 0.05). (2) The proliferation rate of the HSCs treated with NGF of the concentration of 100 ng/ml, the most appropriate concentration, increased time-dependently, and the proliferation rate at the 72 th hour was the lowest in comparison with that of the control group (0.48 +/- 0.03 vs 0.89 +/- 0.01, P < 0.05). (3) After stimulation of the HSCs by NGF of the concentration of 100 ng/ml for 24 hr, TUNEL method showed a significant increased apoptotic rate of HSCs compared with the control group (10.2% +/- 1.2% vs 1.6% +/- 0.1%, P < 0.05). Meanwhile, flow cytometry showed an apoptotic rate of 6.2% +/- 0.2% among the HSCs in the experimental group, but no apoptosis in control group. (4) Microscopy did not show significant morphological change in the HSCs with the proliferation inhibited.
NGF can induce apoptosis of activated HSCs. The mechanism may be the interference with the apoptotic pathway.
评估神经生长因子（NGF）对纤维化大鼠肝星状细胞（HSCs）增殖和凋亡的影响。
将纤维化大鼠的肝星状细胞分别在含有不同浓度NGF（分别为50 ng/ml、100 ng/ml和150 ng/ml）的培养基中或不含NGF的培养基中孵育24、48和72小时。通过MTT法测定肝星状细胞的增殖率。采用TUNEL法检测凋亡的肝星状细胞。通过流式细胞术研究肝星状细胞的凋亡率。通过显微镜观察肝星状细胞的形态。
（1）MTT法表明，与对照组相比，不同浓度NGF诱导24小时的肝星状细胞增殖率显著降低（0.77±0.03，所有P<0.05）。浓度为100 ng/ml的NGF处理24小时的肝星状细胞增殖率最低（0.63±0.02，P<0.05）。（2）浓度为100 ng/ml（最适宜浓度）的NGF处理的肝星状细胞增殖率呈时间依赖性增加，与对照组相比，第72小时的增殖率最低（0.48±0.03对0.89±0.01，P<0.05）。（3）浓度为100 ng/ml的NGF刺激肝星状细胞24小时后，TUNEL法显示肝星状细胞凋亡率与对照组相比显著增加（10.2%±1.2%对1.6%±0.1%，P<0.05）。同时，流式细胞术显示实验组肝星状细胞凋亡率为6.2%±0.2%，而对照组无凋亡。（4）显微镜检查未显示增殖受抑制的肝星状细胞有明显形态变化。
NGF可诱导活化的肝星状细胞凋亡。其机制可能是干扰凋亡途径。
