舒肝和络醒脾方对四氯化碳诱导的肝纤维化大鼠转化生长因子β/果蝇母本抗生存因子蛋白信号通路作用的研究

Effect of Shugan Heluo Xingpi decoction on transforming growth factor-β/small mothers against decapentaplegic protein signaling pathway in hepatic fibrosis rats induced by carbon tetrachloride

目的探讨舒肝和络醒脾方(SHXD)的抗肝纤维化(HF)作用。方法将60只SPF级Wistar大鼠随机分为空白组、模型组、对照组和低、中、高剂量实验组,每组10只;除空白组外,其余组大鼠均腹腔注射40%四氯化碳构建HF大鼠模型。模型组灌胃给予与实验组等量的0.9%NaCl;对照组灌胃给予0.50 mg·kg^(-1)秋水仙碱溶液;低、中、高剂量实验组分别灌胃给予3.15,6.30和12.60 g·kg^(-1)(生药/体质量)SHXD。5组大鼠每天给药1次,连续8周。空白组不做任何处理,正常喂食喂水,作为对照观察使用。用实时荧光定量聚合酶链反应检测肝组织中转化生长因子β1(TGF-β1)和果蝇母本抗生存因子蛋白(Smad2)mRNA的表达水平,用酶联免疫吸附实验法检测血清中HF 4项含量。结果中、高剂量实验组和对照组、模型组、空白组的TGF-β1mRNA表达水平分别为1.36±0.19,1.40±0.12,1.53±0.14,1.58±0.13和1.00±0.00,Smad2 mRNA表达水平分别为1.17±0.06,1.24±0.09,1.52±0.14,1.42±0.13和1.00±0.00,层黏连蛋白的含量分别为(353.92±27.40),(309.46±22.57),(341.48±25.14),(436.71±39.51)和(205.89±27.00)ng·mL^(-1),透明质酸酶的含量分别为(117.50±9.80),(95.48±8.83),(123.24±11.02),(143.01±13.20)和(79.62±6.79)ng·mL^(-1),Ⅳ型胶原的含量分别为(60.93±6.02),(51.57±5.12),(68.91±7.35),(82.91±8.61)和(41.93±5.04)ng·mL^(-1),Ⅲ型前胶原分别为(1.85±0.33),(2.04±0.24),(2.11±0.30),(3.38±0.38)和(1.83±0.26)ng·mL^(-1)。中、高剂量实验组的上述指标与模型组比较,差异均有统计学意义(均P<0.05)。结论SHXD具有一定的抗HF作用,以高、中剂量作用效果最为显著;TGF-β/Smad通路可能为其抗HF的信号途径之一。

Objective To investigate the anti-hepatic fibrosis(HF)effect of Shugan Heluo Xingpi decoction(SHXD).Methods A total of 60 SPF Wistar rats were divided into blank,model,control groups and experimental-L,-M,-H groups with 10 rats in each group.Except for the blank group,all rats were intraperitoneally injected with 40%carbon tetrachloride to make rat model of HF.At the same time,normal saline was given to the blank group and colchicine was given to the control group at 0.5 mg·mL^(-1),experimental-L,-M,-H groups were given SHXD at 3.15,6.30 and 12.60 g·kg^(-1)respectively for 8 weeks,intragastric administration with once a day.Blank group did not give any treatment,only fed water and used as control observation.The mRNA expression of transforming growth factor-β1(TGF-β1)and small mothers against decapentaplegic 2(Smad2)were detected quantitatively by reverse transcription-polymerase chain reaction.The four HF indexes was detected by enzyme linked immunosorbent assay.Results In experimental-M,experimental-H and control,model,blank groups,the expression levels of TGF-β1 mRNA were 1.36±0.19,1.40±0.12,1.53±0.14,1.58±0.13 and 1.00±0.00;the expression levels of Smad2 mRNA were 1.17±0.06,1.24±0.09,1.52±0.14,1.42±0.13 and 1.00±0.00;laminin were(353.92±27.40),(309.46±22.57),(341.48±25.14),(436.71±39.51)and(205.89±27.00)ng·mL^(-1);hyaluronidase were(117.50±9.80),(95.48±8.83),(123.24±11.02),(143.01±13.20)and(79.62±6.79)ng·mL^(-1);collagen type Ⅳ were(60.93±6.02),(51.57±5.12),(68.91±7.35),(82.91±8.61)and(41.93±5.04)ng·mL^(-1);procollagen Ⅲ were(1.85±0.33),(2.04±0.24),(2.11±0.30),(3.38±0.38)and(1.83±0.26)ng·mL^(-1).Compared with the model group,there were significant differences in the above indexes of experimental-M and experimental-H groups(all P<0.05).Conclusion SHXD has certain effect on anti-HF,especially in experimental-M and experimental-H,and the TGF-β/Smad pathway may be a signaling pathway for its function.