柴胡解毒汤对人肝星状细胞LX-2增殖及凋亡的影响

目的:观察柴胡解毒汤含药血清对人肝星状细胞LX-2增殖、凋亡情况的影响,探究其抗肝纤维化作用的可能机制。方法:将Wistar大鼠分为实验组与对照组,分别以柴胡解毒汤、生理盐水灌胃5天后取血制备大鼠含药血清与正常血清。同时复苏人肝星状细胞LX-2后进行细胞培养和细胞传代。当肝星状细胞数量达到预定值时,将肝星状细胞分为对照组和药物组。观察LX-2细胞在与含药血清孵育24 h、36 h、48 h、72 h后,用CCK-8比色法测定细胞的增殖抑制率。用流式细胞术(Annexin V-FITC,PI染色法)检测细胞凋亡的概况。结果:柴胡解毒汤含药血清在给药24 h后可抑制LX-2细胞的增殖,36 h、48 h、72 h的增殖抑制率分别为0.37%、0.46%、0.44%,并可诱导LX-2细胞发生凋亡,48 h、72 h凋亡概率分别为(9.80±0.95)%、(36.40±5.09)%,差异具有统计学意义(P0.05)。结论:柴胡解毒汤具有抑制LX-2细胞增殖、诱导LX-2细胞发生凋亡的能力,这可能是柴胡解毒汤抗纤维化作用的机制之一。

Influence of Chaihu Jiedu Decoction on LX-2 Cell Proliferation and Apoptosis

ABSTRACT Objective: To observe the effects of Chaihu Jiedu decoction on human hepatic stellate LX-2 cells, and to explore the potential molecular mechanisms. Methods: The wistar rats were divided into the experimental group and the control group, respectively with Chaihu Jiedudecoction and saline lavage, then centrifugal and get the drug-containing serum and control serum. At the same time, trsuscitate cells. When we got the expected number of cells, we divided into the experimental group and the control group. The human hepatic stellate cells(LX-2) were with the drug serum for 24 h, 36 h, 48 h, 72 h. Then the cell proliferation inhibition rate was measured by CCK-8, the apoptosis were detected by flow cytometry(Annexin V-FITC, PI staining method). Results: ChaihuJiedu decoction could inhibit proliferation of LX-2 cells 24 hours after dosing. The in hibition rate in 36 h, 48 h, 72 h were 0.37 %, 0.46 %, 0.44 % respectively. It could prevent LX-2 cells into proliferation and induce the apoptosis of LX-2 cells. The apoptosis rates in 48 h, 72 h were (9.80±0.95)%, (36.40±5.09)% respectively, and there are difference of statistical significance(P<0.05). Conclusion: Chaihu Jiedu decoction can inhibit the proliferation of LX-2 cells proliferation, and induce apoptosis, so as to interfere with course of the liver fibrosis.