丹参素治疗肝纤维化及其作用机制研究

Effect and mechanism of Tanshensu on fibrotic rats

目的 研究丹参素对大鼠免疫性肝纤维化的治疗作用及其怍用机制。 方法 用猪血清制作大鼠肝纤维化动物模型,自第8周起给予丹参素组大鼠腹腔注射丹参素300 mg·kg-1·d-1,同时给予模型对照组和正常对照组相同体积的双蒸水腹腔注射。第12周动物实验结束时,除留取一只模型组大鼠用原位循环灌流法提取肝星状细胞(HSC)外,其余大鼠全部处死提取血清检测透明质酸(HA)以及取肝脏标本切片做HE、VG染色。以MTT法观察不同浓度丹参素对HSC的增殖抑制作用,用流式细胞仪分析丹参素作用后HSC所处周期。 结果 丹参素干预组HA为(231.4±41.1)ng/ml,模型组(398.7±54.5)ng/ml,F=154.796,P<0.05。HE及VG染色示丹参素干预组肝纤维化程度明显轻于模型组;细胞试验表明50、100、200 mg/L丹参素对HSC作用后的MTT值(A值)分别为1.60×102±8.17×10-4、1.10×10-2±1.41×10-3和6.75×10-3±3.30×10-3,细胞对照组A值为7.18×10-2±1.71×10-3,F=1154.221,P<0.05。 结论丹参素对大鼠免疫性肝纤维化有明显的防治怍用。其主要机制为丹参素对HSC明显的增殖抑制作用。

Objective To investigate the effect and mechanism of Tanshensu on experimental fibrotic rats. Methods Pig s serum was used to induce liver fibrosis in Wistar rats. The rats in Tanshensu-treated group were injected peritoneally with Tanshensu solution at the dose of 300 mg·kg-1·d-1, and the rats in model-control group and normal-control group received the same volume of double distill water. At the end of the twelfth week, the hepatic stellate cells (HSCs) were isolated from the liver of one rat in model-control group using in-situ perfusion with pronase and collagenase, then density gradient centrifugation, and the other rats were killed to take the serum and liver samples. MTT colorimetric assay was used for detecting the proliferation of HSCs and flow cytometry was used for observing the cell cycles of HSCs under different concentrations of Tanshensu. The hyaluronic acid (HA) level in serum was detected and the morphological changes of liver tissue were observed. Results There was a decline of serum HA level in Tanshensu-treated group compared to that of the model-control group (231.4 ng/ml ±41.1 ng/ml vs 398.7 ng/ml ± 54.5 ng/ml, F =154.796, P < 0.05). Both HE and VG stain showed a decline of liver fibrosis degree in Tanshensu-treated group. And Tanshensu had an inhibition effect on the proliferation of HSCs at the concentrations of 50 mg/L, 100 mg/L, and 200 mg/L (1.60 × 102± 8.17 × 10-4, 1.10 × 10-2 ±1.41 × 10-3, and 6.75 × 10-3 ± 3.30 × 10-3 vs 7.18 × 10-2 ± 1.71 × 10-3, F =1154.221, P <0.01). Conclusions Tanshensu shows a therapeutic effect on liver fibrosis in rats induced by pig's serum through inhibiting the proliferation of hepatic stellate cells.