肝纤维化大鼠阴虚证表征的观察及一贯煎的干预作用

Features of hepatic fibrosis in rats with Yin deficiency syndrome and the intervention effects of Yiguanjian

目的检测肝纤维化大鼠的阴虚证表征,观察滋补肝肾中药一贯煎对阴虚证表征的影响。方法将48只雄性Sprague-Dawlay大鼠随机分为4组,正常对照组、模型组、阳性对照组、一贯煎组,每组12只。除正常对照组外,均建立CCl4致肝纤维化大鼠模型,在CCl4停止注射后3 d,一贯煎组每只动物给予相应浓度的一贯煎浓缩液2 m L/200 g灌胃,1次/d。阳性对照组给予秋水仙碱灌胃,0.25 mg/kg,1次/d。正常对照组和模型组按相同方法灌服等量生理盐水。4周后,检测各组大鼠血清丙氨酸氨基转移酶(ALT)、天门冬氨酸氨基转移酶(AST)等肝功能指标;通过HE染色评价肝脏炎症活动度(G)和纤维化程度(S);检测大鼠体重、舌面干湿度、舌面温度、舌微循环血流速度、饮水、大小便等阴虚证表征。结果 1一贯煎在改善肝功能、减轻炎症、抗肝纤维化方面作用较好:一贯煎组ALT[(41.66±3.72)U]较模型组[(61.00±0.00)U]降低,差异有统计学意义(P<0.05);一贯煎组AST[(146.03±46.41)U]较模型组[(218.90±12.56)U]降低,差异有统计学意义(P<0.05);一贯煎组炎症活动度[(1.00±0.00)]较模型组[(3.00±0.00)]降低,差异有统计学意义(P<0.05);一贯煎组纤维化程度[(1.00±0.00)]较模型组[(2.00±0.00)]降低,差异有统计学意义(P<0.05)。2肝纤维化大鼠具有阴虚证的表征,一贯煎能够改善阴虚证的症状。体重:模型组的体重[(380.66±37.29)g]较正常对照组[(507.00±17.69)g]减少(P<0.05);一贯煎组体重[(473.66±13.86)g]较模型组增高(P<0.05)。舌面干湿度:模型组的舌面干湿度[(0.000 36±0.000 28)g]较正常对照组[(0.007 32±0.001 26)g]减少(P<0.05);一贯煎组舌面干湿度[(0.012 83±0.005 85)g]较模型组增高(P<0.05)。舌面温度:模型组的舌温度[(34.48±1.24)℃]较正常对照组[(30.63±0.80)℃]增高(P<0.05);一贯煎组舌温度[(31.40±2.10)℃]较模型组降低(P<0.05)。舌血流速度:模型组的舌血流速度[(69.78±21.44)AU]较正常对照组[(31.43±1.58)AU]增高(P<0.05);一贯煎组舌血流速度[(40.50±3.89)AU]较模型组降低(P<0.05)。饮水:模型组饮水量[(37.52±10.46)m L]较正常对照组[(59.65±6.05)m L]显著减少(P<0.05);一贯煎组饮水量[(53.69±8.74)m L]较模型组显著增多(P<0.05)。大便:模型组大便[(1.00±0.00)分]较正常对照组[(2.00±0.00)分]分值减少(P<0.05);一贯煎组大便[(2.00±0.00)分]较模型组分值增多(P<0.05)。小便:模型组小便[(2.00±0.00)分]较正常对照组[(1.00±0.00)分]分值增高(P<0.05);一贯煎组小便[(1.33±0.57)分]较模型组分值减少(P<0.05)。结论肝纤维化大鼠具有阴虚证的特点,滋补肝肾中药一贯煎不仅能改善肝纤维化大鼠的病理,还能改善阴虚证的症状。该研究为肝纤维化大鼠阴虚证的诊断和药理机制的研究提供了新的思路。

Objective To detect the Yin deficiency symptoms of hepatic fibrosis in rats, observe the effect of traditional Chinese medicine Yiguanjian on characterization of Yin deficiency syndrome. Methods 48 male Sprague-Dawlay rats were randomly divided into 4 groups： normal group, model group, positive control group, Yiguanjian group, each group had 12 rats. Except the normal group, hepatic fibrosis model induced by CCl4 was established. After 3 days when the CCl4 injection was stopped, each rat in Yiguanjian group was given Yiguanjian（2 m L/200 g）by intragastric administration, 1 time/d. Positive control group was given Colchicine（0.25 mg/kg） by intragastric administration, 1 time/d. The normal group and model group were fed with same volume of saline daily. After 4 weeks, serum levels of aspartate aminotransferase（AST） and alanine aminotransferase（ALT） were measured by routine methods; the HE staining was used to evaluate the semi-quantitative scoring of liver inflammation（G） and liver fibrosis（S）; the features of Yin deficiency syndrome like body weight, tongue humidity,temperature of tongue, tongue microcirculation blood flow velocity, drinking, stool and urine were detected. Results1 Yiguanjian had good effect in improving liver function, reduce inflammation, anti-liver fibrosis. Compared with the model group [（61.00±0.00） U], ALT was significantly declined in Yiguanjian group [（41.66±3.72） U], with statistically significant difference（P〈0.05）. Compared with the model group [（218.90±12.56） U], AST was significantly declined in Yiguanjian group [（146.03±46.41） U], with statistically significant difference（P〈0.05）. Compared with the model group[（3.00±0.00）], the semi-quantitative scoring of liver inflammation was significantly declined in Yiguanjian group [（1.00±0.00）], with statistically significant difference（P〈0.05）. Compared with the model group [（2.00±0.00）], the semi-quantitative scoring of fibrosis was significantly declined in Yiguanjian group [（1.00±0.00）], with statistically significant difference（P〈0.05）. 2The rat of hepatic fibrosis had symptoms of Yin deficiency syndrome, Yiguanjian could improve the symptoms of Yin deficiency syndrome. Body weight： compared with the normal group [（507.00±17.69） g], the body weight was significantly declined in model group[（380.66±37.29） g, P〈0.05]. Compared with the model group, the body weight significantly increased in Yiguanjian group [（473.66±13.86） g, P〈0.05]. Tongue humidity： compared with the normal group [（0.007 32±0.001 26） g], the tongue humidity was significantly declined in model group [（0.000 36±0.000 28） g,P〈0.05]. Compared with the model group, the tongue humidity significantly increased in Yiguanjian group [（0.012 83±0.005 85） g],（P〈0.05）. Temperature of tongue： compared with the normal group [（30.63±0.80）℃], the temperature of tongue significantly increased in model group [（34.48±1.24）℃, P〈0.05]. Compared with the model group, the temperature of tongue significantly declined in Yiguanjian group [（31.40±2.10）℃, P〈0.05]. Tongue microcirculation blood flow velocity： compared with the normal group [（31.43 ±1.58）AU], the blood flow velocity significantly increased in model group [（69.78±21.44） AU, P〈0.05]. Compared with the model group, the blood flow velocity significantly declined in Yiguanjian group [（40.50±3.89） AU, P〈0.05]. Drinking： compared with the normal group [（59.65±6.05） m L], the drinking significantly declined in model group [（37.52±10.46） m L, P〈0.05]. Compared with the model group, the drinking significantly increased in Yiguanjian group [（53.69±8.74） m L, P〈0.05]. Stool compared with the normal group [（2.00±0.00）], the semi-quantitative scoring of stool significantly declined in model group [（1.00±0.00）, P〈0.05]. Compared with the model group, the semi-quantitative scoring of stool significantly increased in Yiguanjian group [（2.00±0.00）, P〈0.05]. Urine： compared with the normal group [（1.00 ±0.00）], the semi-quantitative scoring of urine significantly increased in model group [（2.00±0.00）, P〈0.05]. Compared with the model group, the semi-quantitative scoring of urine significantly declined in Yiguanjian group [（1.33±0.57）, P〈0.05]. Conclusion The rat of hepatic fibrosis has the characteristics of Yin deficiency syndrome, Yiguanjian can not only improve the liver injury in rat, but also can improve the symptoms of Yin deficiency syndrome. This study provides a new idea to study the diagnosis and research of pharmacological mechanism of Yin deficiency syndrome in rats with liver fibrosis.