扶正利湿解毒法体内外逆转肝纤维化

In vivo and in vitro study on liver fibrosis reversion effect by therapy of strengthening healthy qi combined with draining dampness and relieving toxin

目的:观察扶正利湿解毒法基本方(简称基本方)对肝纤维化的逆转作用.方法:采用DMN诱导大鼠肝纤维化,造模结束后随机分为模型对照组,秋水仙碱组,基本方高剂量组,基本方中剂量组及基本方低剂量组并分组给药,同时设空白对照组.灌胃3wk后检测肝组织Hyp,ALT水平;HE,Masson染色.采用HSC-T6细胞培养,药物作用24h后光镜下观察拍照,检测药物对细胞的抑制率并在电镜下观察药物浓度为2g/L时细胞的变化.结果:基本方高、中剂量均可使大鼠肝组织Hyp,血清ALT显著下降(Hyp:213.73±58.05μg/g,245.95±67.70μg/g vs 317.37±42.06μg/g,P<0.01,P<0.05;ALT:33.56±8.16 kU/L,42.38±14.99 kU/L vs 58±10.32 kU/L,P<0.01,P<0.05);Masson染色肝纤维化分级评分显示,基本方高,中剂量均可使肝纤维化程度显著降低(2.33±0.87,2.63±0.74 vs 3.70±0.67,P<0.01,P<0.01),同时显著改善肝组织炎症、出血及纤维沉积;光镜观察及抑制率检测显示基本方呈剂量依赖性抑制HSC-T6细胞增值.电镜显示基本方主要导致HSC-T6细胞凋亡.结论:基本方显示了显著的体内外逆转肝纤维化作用.

AIM： To observe the reversing effect on liver fibrosis by the therapy of strengthening healthy qi combined with draining dampness and relieving toxin （SHQDDRT）. METHODS： Rat model of liver fibrosis was induced by dimethylnitrosamine （DMN）. All the rats were randomly divided into 6 groups, named group A （control, n = 10）, B （model, n = 10）, C （colchicine, n = 10）, D （with the basic herb complex, high dose, n =10）, E （with the basic herb complex, middle dose, n =10） and F （with the basic herb complex, low dose, n = 10）. The content of liver hydroxyproline and the serum aμnine aminotransferase （ALT） were tested. HE and Masson staining were conducted for patho-logical examination. HSC-T6 cell line was used as an in vitro model. After 24 hours of drug administration, the inhibitory effect was observed under an optical microscope. Cell prolifera- tion was detected by MTT assay. Additionally, HSC-T6 cells were observed by electron microscopy after they were treated with the basic herb complex and control drug at the concentration of 2 g/L. RESULTS： The content of liver hydroxyproline and the serum ALT in group D and E were significantly lower than those in group B （hydroxyproline： 213.73 ±58.05 μg/g, 245.95 ±67.70 μg/g vs 317.37±42.06 μg/g, P 〈 0.01, P 〈 0.05; ALT： 33.56± 8.16 kU/L, 42.38±14.99 kU/L vs 58±10.32 kU/L, P 〈 0.01, P 〈 0.05）. Masson staining showed that the fibrotic degrees were significantly decreased in group D and E in contrast with those in group B （2.33 ± 0.87, 2.63 ± 0.74 vs 3.70± 0.67, both P 〈 0.01）, and the fiber deposition, infμmmation and hemorrhage were also improved. Light endoscopy and MTT assay demonstrated that the basic herb complex inhibited the growth of HSC-T6 cells in a dose- dependent manner. Electron microscopy exhibited typical apoptosis features in HSC-T6 cells. CONCLUSION： The basic herb complex shows significant reversing effect on liver fibrosis in vitroand in vivo.