目的：探讨大蒜素诱导THP-1细胞凋亡中P38MAPK及Fas基因表达的变化。方法：采用四甲基偶氮唑蓝(MTT)法测定不同浓度大蒜素对THP-1细胞增殖抑制率；Annexin V- FITC/PI双染法流式细胞术(FCM)检测凋亡率的变化;免疫细胞化学法观察大蒜素处理后细胞内磷酸化P38MAPK(P-p38MAPK)表达及分布变化；Western blot检测大蒜素处理前后细胞内P-p38MAPK和Fas蛋白表达量的变化。结果：MTT显示大蒜素能抑制THP-1细胞增殖，并呈时间-剂量依赖性。其72 h中效浓度(IC50)为12.8 mg·L-1。Annexin V- FITC/PI检测凋亡率随大蒜素浓度增加而增加。免疫细胞化学法检测药物处理后P-p38MAPK表达增加，主要分布于细胞核与细胞浆，而阴性对照组仅弱表达。Western blot结果分析P-p38MAPK，Fas蛋白表达量随大蒜素浓度的增加而增加，呈剂量依赖性。其阴性组、72 h的1/2 IC50组、IC50组P-p38MAPK蛋白表达水平(相对灰度值)分别为0.259 8±0.013 2，0.361 2±0.008 3，0.505 6±0.005 5；Fas蛋白表达水平分别为0.287 4±0.008 9，0.426 8±0.007 9和0.597 1±0.010 9，差别均有显著性意义(P<0.01)。结论：大蒜素可诱导THP-1细胞凋亡，其诱导凋亡机制可能是通过激活P-p38MAPK/Fas途径实现。
Objective: The objective of this paper was to study the change of P38MAPK and Fas in the apoptosis of THP-1 cells induced by allicin. Method: The proliferation inhibition rates of THP-1 cells after various treatments were examined by MTT assay. Apoptosis rate was determined with Annexin V- FITC/PI double staining by flow cytometry. The expression and distribution change of the phosphorylation p38MAPK (P-p38MAPK) were detected by immunohistochemical staining. The changes of P-p38 MAPK and Fas proteins were detected by Western blot. Result: The proliferations of leukemia cell line THP-1 are inhibited by allicin. MTT assay showed that allicin can inhibit the proliferation of the THP-1 cell, and the inhibition was dependent on both dose and time. The IC50 of 72 hours was 12.8 mg·L-1. Apoptosis rate detected by Annexin V-FITC/PI was proportional to the concentration of the allicin. After the immunohistochemical staining test, the P-p38MAPK was located in the cell nucleus and plasma, showing deep brown, when adding allicin to THP-1 cell. Western blot test showed that the P-p38MAPK proteins expression was proportional to the concentration of Allicin and was also dose dependent. The levels of P-p38MAPK in negative control group, 1/2 IC50 of 72 hours group and IC50 of 72 hours group were 0.259 8±0.013 2, 0.361 2±0.008 3 and 0.505 6±0.005 5 respectively, and the levels of Fas proteins were 0.287 4±0.008 9, 0.426 8±0.007 9 and 0.597 1±0.010 9 respectively. The difference was statistically significant when compared with the negative control group (P<0.01). Conclusion: Allicin can significantly induce THP-1 cells apoptosis, and its mechanism may be related to the activation of P38MAPK/Fas.