假木豆提取物对四氯化碳致大鼠肝纤维化的保护作用

Protective effect of Jiamudou extract on hepatic fibrosis induced by carbon tetrachloride

目的探讨假木豆提取物抗四氯化碳(CCl_(4))所致大鼠肝纤维化的作用机制。方法将60只Sprague Dawley大鼠随机分为空白组、模型组、水飞蓟素组、低剂量假木豆组、中剂量假木豆组和高剂量假木豆组,每组10只。水飞蓟素组、低剂量假木豆组、中剂量假木豆组、高剂量假木豆组和模型组大鼠通过背部皮下注射CCl_(4)建立肝纤维化模型,空白组大鼠背部皮下注射花生油,每隔2 d注射1次,共8周。从第3周起,水飞蓟素组大鼠每日灌服水飞蓟素胶囊水溶液(50 mg·kg^(-1))1次;假木豆低、中、高剂量组大鼠每日灌服12.5、25.0、50.0 g·L^(-1)的假木豆水提液(20 mL·kg^(-1));空白组和模型组大鼠灌服等体积的蒸馏水;各组大鼠均连续给药6周。观察并记录各组大鼠饮食活动情况、体质量、毛色光亮程度、精神状况等一般情况。末次给药1 h后采集各组大鼠腹主动脉血5 mL,采用酶联免疫吸附试验检测各组大鼠血清中白细胞介素(IL)-1β、IL-6、肿瘤坏死因子-α(TNF-α)和转化生长因子-β_(1)(TGF-β_(1))水平。采血后处死大鼠,取肝脏,观察肝脏大体形态,然后取大鼠肝脏左叶,制备肝组织切片,行苏木精-伊红染色,于光学显微镜下观察大鼠肝脏病理形态学变化。结果空白组大鼠饮食正常,活动频繁,毛色洁白有光泽,精神状况正常。模型组大鼠饮食减少,活动量明显减少,毛色暗淡、不清洁,精神状况萎靡;水飞蓟素组大鼠毛色欠缺光泽但较清洁,其余一般情况同空白组;假木豆各剂量组大鼠一般情况较模型组有改善,而较空白组差。模型组、水飞蓟素组、低剂量假木豆组、中剂量假木豆组、高剂量假木豆组大鼠体质量增长量少于空白组(P<0.05),低剂量假木豆组大鼠体质量增长少于模型组(P<0.05)。模型组、水飞蓟素组、低剂量假木豆组、中剂量假木豆组、高剂量假木豆组大鼠血清中IL-1β、IL-6、TNF-α水平均高于空白组(P<0.05);水飞蓟素组与空白组大鼠血清中TGF-β_(1)水平比较差异无统计学意义(P>0.05);模型组、低剂量假木豆组、中剂量假木豆组、高剂量假木豆组大鼠血清中TGF-β_(1)水平均高于空白组(P<0.05)。水飞蓟素组、低剂量假木豆组、中剂量假木豆组、高剂量假木豆组大鼠血清中IL-1β、IL-6、TNF-α、TGF-β_(1)水平均低于模型组(P<0.05)。低剂量假木豆组大鼠血清中IL-1β、IL-6、TNF-α、TGF-β_(1)水平显著高于水飞蓟素组(P<0.05)。中剂量假木豆组与水飞蓟素组大鼠血清中IL-1β、IL-6水平比较差异无统计学意义(P>0.05),中剂量假木豆组大鼠血清中TNF-α、TGF-β_(1)水平显著高于水飞蓟素组(P<0.05);高剂量假木豆组与水飞蓟素组大鼠血清中IL-1β、IL-6、TNF-α、TGF-β_(1)水平比较差异无统计学意义(P>0.05)。低、中、高剂量假木豆组大鼠血清中IL-1β水平比较差异均无统计学意义(P>0.05)。高剂量假木豆组大鼠血清中IL-6、TGF-β_(1)水平显著低于低剂量假木豆组(P<0.05);中剂量假木豆组与低剂量假木豆组、高剂量假木豆组与中剂量假木豆组大鼠血清中IL-6、TGF-β_(1)水平比较差异均无统计学意义(P>0.05)。低、中、高剂量假木豆组大鼠血清中TNF-α水平随剂量的增加逐渐降低,两两比较差异均有统计学意义(P<0.05)。空白组大鼠肝组织结构完整,肝小叶结构正常;模型组大鼠肝组织肝纤维化明显;低剂量假木豆组大鼠肝组织病理学形态较模型组有轻微改善;水飞蓟素组和中、高剂量假木豆组大鼠肝组织病理学形态较模型组有较大的改善,肝纤维形成减轻或消失。结论假木豆提取物对大鼠肝纤维化有保护作用,其作用机制可能与降低血清中IL-1β、IL-6、TNF-α及TGF-β_(1)水平有关。

Objective To explore the mechanism of Jiamudou extract anti-hepatic fibrosis induced by carbon tetrachloride(CCl_(4))in rats.Methods Sixty Sprague Dawley rats were randomly divided into blank group,model group,silymarin group,low-dose Jiamudou group,middle-dose Jiamudou group and high-dose Jiamudou group,with 10 rats in each group.The rats in the model group,silymarin group,low-dose Jiamudou group,middle-dose Jiamudou group and high-dose Jiamudou group were injected subcutaneously with CCl_(4) every 2 days for 8 weeks to establish the liver fibrosis model;the rats in the blank group were injected subcutaneously with peanut oil every 2 days for 8 weeks.From the third week,the rats in the silymarin group were given silymarin capsule aqueous solution(50 mg·kg^(-1)),once a day;the rats in the low-dose Jiamudou group,middle-dose Jiamudou group and high-dose Jiamudou group were given 12.5,25.0,50.0 g·L^(-1) of Jiamudou water extract(20 mL·kg^(-1))by gavage,respectively;the rats in the blank group and model group were given equal volume of distilled water;all rats were continuously administered for 6 weeks.The general conditions such as diet and activity,body weight,hair color brightness and mental state of rats in each group were observed and recorded.One hour after the last administration,5 mL of abdominal aorta blood of rats in each group was collected,and the levels of serum interleukin(IL)-1β,IL-6,tumor necrosis factor-α(TNF-α)and transforming growth factor-β_(1)(TGF-β_(1))were detected by enzyme-linked immunosorbent assay.The rats were killed after blood collection,and the liver was taken to observe the general morphology.Then,the left lobe of liver was taken,and the liver tissue sections were prepared and stained with hematoxylin-eosin.The pathological changes of the liver tissues of rats were observed under the light microscope.Results The rats in the blank group had normal diet,frequent activities,white and shiny hair,and normal mental status.In the model group,the diet of rats was reduced,the activity was significantly reduced,the hair color was dark and unclean,and the mental state was depressed;in the silymarin group,the hair color of rats was less shiny but cleaner,and the other general conditions were the same as those of rats in the blank group;the general conditions of rats in the different dose of Jiamudou group were better than those in the model group but worse than those in the blank group.The growth of body mass of rats in the model group,silymarin group,low-dose Jiamudou group,middle-dose Jiamudou group and high-dose Jiamudou group was significantly less than that in the blank group(P<0.05);the growth of body mass of rats in the low-dose Jiamudou group was less than that in the model group(P<0.05).The levels of serum IL-1β,IL-6 and TNF-αof rats in the model group,silymarin group,low-dose Jiamudou group,middle-dose Jiamudou group and high-dose Jiamudou group were significantly higher than those in the blank group(P<0.05).There was no significant difference in the level of serum TGF-β_(1) of rats between silymarin group and blank group(P>0.05).The levels of serum TGF-β_(1) of rats in the model group,low-dose Jiamudou group,middle-dose Jiamudou group and high-dose Jiamudou group were significantly higher than those in the blank group(P<0.05).The levels of serum IL-1β,IL-6,TNF-αand TGF-β_(1) of rats in the silymarin group,low-dose Jiamudou group,middle-dose Jiamudou group and high-dose Jiamudou group were significantly lower than those in the model group(P<0.05).The levels of serum IL-1β,IL-6,TNF-αand TGF-β_(1) of rats in the low-dose Jiamudou group were significantly higher than those in the silymarin group(P<0.05).There was no significant difference in the levels of serum IL-1β,IL-6 of rats between the middle-dose Jiamudou group and the silymarin group(P>0.05);the levels of serum TNF-αand TGF-β_(1) of rats in the middle-dose Jiamudou group were significantly higher than those in the silymarin group(P<0.05);there was no significant difference in the levels of serum IL-1β,IL-6,TNF-αand TGF-β_(1) of rats between the high-dose Jiamudou group and the silymarin group(P>0.05).There was no significant difference in the level of serum IL-1βof rats among low-dose Jiamudou group,middle-dose Jiamudou group and high-dose Jiamudou group(P>0.05).The levels of serum IL-6,TGF-β_(1) of rats in the high-dose Jiamudou group were significantly lower than those in the low-dose Jiamudou group(P<0.05);there was no significant difference in the levels of serum IL-6,TGF-β_(1) of rats between the middle-dose Jiamudou group and the low-dose Jiamudou group,the middle-dose Jiamudou group and the high-dose Jiamudou group(P>0.05).The level of serum TNF-αof rats in the low-dose Jiamudou group,middle-dose Jiamudou group and high-dose Jiamudou group gradually decreased with the dose,and the difference was statistically significant(P<0.05).In the blank group,the liver tissue structure of rats was intact and the liver lobule structure was normal;the liver fibrosis of rats in the model group was obvious;compared with the model group,the histopathological morphology of the liver tissue of rats in the low-dose Jiamudou group was slightly improved,the histopathological morphology of liver tissue of rats in the silymarin group,middle-dose Jiamudou group and high-dose Jiamudou group were significantly improved,and the formation of liver fibers was reduced or disappeared.Conclusion Jiamudou extract has a protective effect on hepatic fibrosis of rats,and its mechanism may be related to the decrease of the levels of serum IL-1β,IL-6,TNF-αand TGF-β_(1).