莲子心萃取物抑制大鼠肝纤维化的作用机制研究

Study on the effective mechanism of Plumula Nelumbinis extract against hepatic fibrosis in rats

目的以体外和动物模型探讨莲子心萃取物对于肝星状细胞活化及大鼠肝纤维化的影响。方法在细胞实验中,检测莲子心萃取物对肝星状细胞珠的影响,用荧光分析仪测定细胞内过氧化氢含量,用Western blot检测胶原蛋白沉积、核转录因子-κB(NF-κB)的活化、Iκba的磷酸化与甲型平滑肌动蛋白的表达。动物分3组:假手术组,给予0.7%的羧甲基纤维素(CMC);模型组,结扎大鼠胆管,并给予0.7%CMC;实验组,结扎大鼠胆管并给予0.5 g·kg-1莲子心萃取物,每天喂药2次。用比色分析仪检测莲子心萃取物对大鼠谷草转氨酶(AST)和谷丙转氨酶(ALT)的含量,用定量实时聚合酶反应检测肝纤维化分数、甲型平滑肌动蛋白α-SMA及胶原蛋白col1α2的基因表达的影响。结果与空白组比较,80μg·m L-1莲子心萃取物可抑制TNF-α所引发的肝星状细胞内的过氧化氢含量与甲型平滑肌动蛋白(α-SMA)蛋白质表达及细胞内核转录因子抑制蛋白α(IκBα)的磷酸化,并抑制核转录因子(NF-κB)的活化及α-SMA蛋白质的表达;80μg·m L-1莲子心萃取物可显著抑制细胞胶原蛋白生成。与模型组比较,莲子心萃取物可降低肝纤维化大鼠的AST和ALT含量,降低肝胶原蛋白含量,减少α-SMA、犬Ⅰ型胶原α2(col1α2)mRNA表达。结论莲子心萃取物能抑制肝纤维化。

Objective To study the effect of Plumula Nelumbinis extract（ PNE） on hepatic stellate cell activation and liver fibrosis in vitro and in vivo in rats model. Methods PNE impact on hepatic stellate cells was detected by cell experiments. The hydrogen peroxide content was determined in the cell by fluorescence analyzer. The ollagen deposition,nuclear factor- κB（ NF- κB） activation, Iκbα phosphorylation,α- SMA protein expression and gene transcription were detected by Western blot. Animals are divided into three groups,sham group,0. 7%carboxymethyl cellulose（ CMC） was administrated; model group,Bile duct ligation in rats,0. 7% CMC was administrated; experimental group,Bile duct ligation in rats and PNE was administrated,twice a day from3 th week. PNE impact on aspartate transaminase（ AST） and alanine transaminase（ ALT） was detected by colorimetric analyzer. The liver fibrosis scores,α- SMA and col1 α2expression was detected by quantitative real- time polymerase chain reaction. Results Compared with blank group,PNE（ 80 μg·m L- 1） can inhibit intracellular hydrogen peroxide content of hepatic stellate cells caused by TNF- α,alphasmooth muscle actin（ α- SMA） protein expression,and intracellular NF- kappa B inhibitor- α（ IκBα） phosphorylation,and inhibit NF- κB activation and α- SMA protein expression. PNE（ 80 μg·m L- 1） can significantly inhibit cell collagen production. In the experiment where Bile duct ligation（ BDL） induced liver fibrosis in rats. Compared with model group,the use of PNE can reduce AST and ALT,decrease hepatic collagen content,and reduce α- SMA,mouse collagen type I Alpha 2（ col1α2） mRNA expression. Conclusion PNE can inhibit liver fibrosis.