天然牛磺酸对人肝星状细胞增殖与凋亡的影响

Effect of NTau on Proliferation and Apoptosis of HSC-LX2 Cells

目的:通过研究天然牛磺酸(Natural Taurine,NTau)抑制人肝星状细胞系(HSC-LX2)的增殖及诱导其凋亡的作用,进一步探讨NTau抗肝纤维化的相关作用机制。方法:在体外培养人肝星状细胞(HSC-LX2),将其分成空白对照组以及低剂量(3.0μg/m L)、中剂量(6.0μg/m L)、高剂量(9.0μg/m L)NTau组,分别在各培养孔内加入相应浓度的含药血清,孵育、培养2天后,采用MTT比色法测定光密度值,然后计算出各组细胞的存活率,并检测各组细胞的增殖活性;联合TUNEL法、流式细胞术计算各组细胞的凋亡数,算出凋亡比,检测各组细胞的晚期凋亡率。结果:高、中、低剂量NTau组HSC-LX2细胞光密度值(Optical Density;OD)均显著低于空白对照组(P<0.05);高、中、低剂量NTau组对HSC-LX2细胞的增殖抑制率均明显高于空白对照组(P<0.05);高、中、低剂量NTau组对HSCLX2细胞的增殖抑制率比较差异有统计学意义(P<0.05);高、中、低剂量NTau组HSC-LX2细胞G0/G1期DNA百分比均显著高于空白对照组(P<0.05);高、中、低剂量NTau组HSC-LX2细胞S期、G2/M期DNA百分比均显著低于空白对照组(P<0.05);空白对照组HSC-LX2细胞晚期凋亡率低于低、中、高剂量NTau组(P<0.05)。结论:NTau可抑制HSC-LX2的细胞增殖,并促进其凋亡,可综合干预肝纤维化的发生、发展进程,并且与剂量呈正相关性。

Objective： To study the effect of natural taurine（ NTau） on the proliferation inhibition of human hepatic stellate cell line（ HSC-LX2） and its induction of apoptosis,thus to further explore the relevant mechanism of NTau on hepatic fibrosis. Methods： HSC-LX2 were cultured in vitro and divided into the blank control group,low-dose NTau group（ 3. 0 μg/m L）,medium-dose NTau group（ 6. 0 μg/m L）,and high-dose NTau group（ 9. 0 μg/m L）. The medication groups were added the corresponding concentration of drug-containing serum to each culture well. After two-day incubation and culture,the optical density（ OD） was measured by using MTT colorimetry,then the survival rate of cells was calculated and the proliferative activity of the cells were tested in each group. Apoptotic ratio was calculated by the combination of TUNEL and flow cytometry. Results： OD of HSC-LX2 in NTau groups（ high-dose,medium-dose,low-dose） were significantly lower than that in the blank control group（ P〈0. 05）. The proliferation inhibition rate of HSC-LX2 cells in each NTau group was significantly higher than that in the blank control group（ P〈0. 05）. The percentage of DNA in G0/G1 phase of HSC-LX2 in each NTau group was significantly higher than that in the blank control group（ P〈0. 05）; however the percentage of DNA in S phase and G2/M phase of HSC-LX2 in each NTau group was significantly lower than that in the blank control group（ P〈0. 05）. The late apoptosis rate of HSC-LX2 was significantly lower in the blank control group than those in the NTau groups of low-dose,medium-dose,and high-dose（ P〈0. 05）. Conclusion： NTau can inhibit the proliferation of HSC-LX2 and promote their apoptosis,it can comprehensively interfere with the occurrence and development of liver fibrosis,which has a positive correlation with the concentration.