肝龙胶囊对CCl_(4)性肝损伤所致肝纤维化的影响

Effect of Ganlong Capsule on liver fibrosis induced by CCl_(4) liver injury

目的 探索肝龙胶囊对四氯化碳(CCl_(4))诱导的肝纤维化(HF)大鼠的影响及其可能的作用机制。方法 将48只SPF级SD大鼠随机分为正常组、模型组、对照组和低、中、高剂量实验组,每组8只,除正常组外,其余组大鼠均腹腔注射40%CCl_(4)构建HF大鼠模型。造模成功后,正常组和模型组均给予等量0.9%NaCl;对照组给予0.2 mg·kg^(-1)秋水仙碱溶液;低、中、高剂量实验组分别给予62.5,125.0和250.0 mg·kg^(-1)肝龙胶囊溶液。6组大鼠每天灌胃给药1次,连续4周。用酶联免疫吸附法检测血清中HF 4项和肝组织中炎症因子的含量,用蛋白质印迹法检测B淋巴细胞瘤-2基因(Bcl-2)、Bcl-2相关X蛋白(Bax)和半胱氨酸天冬氨酸蛋白酶3(Caspase3)的表达水平,用实时荧光定量法检测α-平滑肌肌动蛋白(α-SMA)、Ⅰ型胶原(COL-Ⅰ)和COL-Ⅲ的表达水平。结果 高剂量实验组、对照组和模型组的谷草转氨酶分别为(21.07±4.50),(20.34±3.72)和(37.88±16.30)U·L^(-1);谷丙转氨酶分别为(4.03±1.16),(7.76±7.00)和(16.43±8.39)U·L^(-1);肿瘤坏死因子-α分别为(34.93±16.44),(50.38±4.81)和(86.45±6.39)ng·L^(-1);白细胞介素-1β分别为(11.37±7.96),(15.95±4.08)和(35.12±15.31)ng·L^(-1);Bax蛋白相对表达水平分别为0.09±0.01,0.13±0.03和0.30±0.16;Bcl-2蛋白相对表达水平分别为0.11±0.01,0.09±0.01和0.08±0.01;Caspase3蛋白相对表达水平分别为0.16±0.04,0.22±0.04和0.43±0.07;α-SMA mRNA表达水平分别为2.25±1.31,2.74±1.60和8.56±4.73;COL-Ⅰ mRNA表达水平分别为1.31±0.82,5.54±2.75和7.86±4.53;COL-Ⅲ mRNA表达水平分别为1.91±1.40,3.80±2.12和5.24±3.58。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 肝龙胶囊通过控制炎症水平,抑制肝细胞凋亡,抑制胶原的合成与分泌等作用,来改善肝损伤,缓解肝纤维化。

Objective To investigate the effect of Ganlong capsule on carbon tetrachloride (CCl_(4))-induced hepatic fibrosis (HF) in rats and its possible mechanism.Methods A total of 48 SPF SD rats were randomly divided into normal,model,control groups and experimental-L,-M,-H groups,with 8 rats in each group.Except for normal group,the rats in other groups were intraperitoneally injected with 40%CCl_(4)to construct HF rat model.After successful modeling,the normal group and model group were given 0.9%Na Cl.The control group was gavaged 0.2 mg·kg^(-1)colchicine solution.Six groups were given corresponding drugs once a day for 4 weeks.The four HF indexes and inflammatory factors in the liver tissue were detected by enzyme linked immunosorbent assay.The expression levels of Blymphoblastoma-2 gene (Bcl-2),Bcl-2 associated X protein (Bax) and cysteine aspartate protease 3 (Caspase3)were detected by Western blotting.The expression levels of α-smooth muscle actin (α-SMA),typeⅠcollagen(COL-Ⅰ) and typeⅢcollagen (COL-Ⅲ) were detected by real-time fluorescence quantification.Results In experimental-H,control and model groups,glutamic oxalacetic transaminase were (21.07±4.50),(20.34±3.72)and (37.88±16.30) U·L^(-1);glutamic-pyruvic transaminase were (4.03±1.16),(7.76±7.00) and (16.43±8.39)U·L^(-1);tumor necrosis factor-α were (34.93±16.44),(50.38±4.81) and (86.45±6.39) ng·L^(-1);interleukin-1β were (11.37±7.96),(15.95±4.08) and (35.12±15.31) ng·L^(-1);the expression levels of Bax protein were 0.09±0.01,0.13±0.03 and 0.30±0.16;the expression levels of Bcl-2 protein were 0.11±0.01,0.09±0.01 and 0.08±0.01;the expression levels of Caspase3 protein were 0.16±0.04,0.22±0.04 and0.43±0.07;the expression levels of α-SMA mRNA were 2.25±1.31,2.74±1.60 and 8.56±4.73;the expression levels of COL-ⅠmRNA were 1.31±0.82,5.54±2.75 and 7.86±4.53;the expression levels of COL-ⅢmRNA were 1.91±1.40,3.80±2.12 and 5.24±3.58.The differences were statistically significant between experimental-H group and model group (all P<0.05).Conclusion Ganlong capsule can improve liver injury and alleviate HF by controlling inflammation level,inhibiting apoptosis of liver cells,inhibiting synthesis and secretion of collagen,and so on.