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目的:探讨丹参注射液体外对肝星状细胞HSC-T6的毒性及其作用机制。方法:利用细胞培养技术,台盼兰法检测系列丹参溶液对HSC-T6生长的影响,G im sa染色观察细胞形态,MTT法检测丹参对细胞的生长抑制率,免疫组化检测血小板衍生生长因子(PDGF-BB)表达,生物法检测转化生长因子的表达。结果:①2.5至80μg/m l系列剂量丹参作用于HSC-T6 24、48和72h后,随着丹参剂量增大,HSC细胞存活率呈明显下降。②丹参作用48 h后,10μg/m l剂量部分细胞变成缩水球状,细胞胞浆变得不透明;20μg/m l剂量,脱壁细胞增多,细胞间隙增大,细胞很多呈缩水状;40μg/m l剂量多数细胞收缩成团块状,可见成片细胞脱壁;80μg/m l剂量细胞大量脱壁,活细胞很少。③HSC-T6经10至80μg/m l丹参作用24和48h后,490nm处测得OD值随剂量增大而降低,并随时间延长OD值减少。④免疫组化分析结果显示,与对照组相比,随着丹参剂量的增大,HSC-T6 PDGF-BB表达逐渐减弱,至20μg/m l差别已很明显(P<0.05)。⑤经10至40μg/m l丹参处理后HSC-T6 TGFβ1占总TGFβ的百分比降低。结论:丹参体外能够显著抑制HST-T6细胞增殖,其抑制机制是通过拮抗细胞因子PDGF-BB和TGFβ1的表达途径达到的。
Objective: To investigate the toxicity of Salviae miltiorrhizae injection on HSC-T6 and its mechanism. METHODS: Cell culture techniques were used to examine the effects of serial Salviae miltiorrhizae solution on the growth of HSC-T6 cells by trypan blue assay, cell morphology was observed by G im sa staining, the growth inhibitory rate of Salvia miltiorrhiza was detected by MTT assay, and platelet-derived growth factor was detected by immunohistochemistry. (PDGF-BB) expression, biological detection of transforming growth factor expression. RESULTS: After 12.5 to 80 μg/ml doses of Salvia miltiorrhiza were treated with HSC-T6 for 24, 48, and 72 hours, the survival rate of HSC cells decreased significantly with increasing the dose of Salvia miltiorrhiza. 2 After 48 hours of salvia miltiorrhiza, 10μg/ml part of the cells became shrinking spheres, the cytoplasm became opaque; at the dose of 20μg/ml, the number of detachment cells increased, the intercellular space increased, and many cells were shrinking; 40μg/ml dose Most of the cells contracted into clumps, showing that the cells were detached from each other; 80 μg/ml doses of cells had a large number of detached cells and few living cells. After the HSC-T6 was treated with Salvia miltiorrhiza at 10 to 80μg/ml for 24 and 48 hours, the OD value at 490nm decreased with increasing dose, and the OD value decreased with time. 4 Immunohistochemical analysis showed that compared with the control group, the expression of HSC-T6 PDGF-BB gradually decreased as the dose of Salvia miltiorrhiza increased, and the difference to 20 μg/ml was significant (P<0.05). 5 The percentage of total TGFβ in HSC-T6 TGFβ1 decreased after treatment with 10 to 40 μg/ml Salvia miltiorrhiza. CONCLUSION: Salvia miltiorrhiza can significantly inhibit the proliferation of HST-T6 cells in vitro, and its inhibition mechanism is achieved by antagonizing the expression pathways of PDGF-BB and TGFβ1 cytokines.