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纸质出版日期:2017
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刘雪萍, 韦啟球, 李振明, 等. 排钱草对食蟹猴肝纤维化的影响[J]. 中国实验方剂学杂志, 2017,23(24):133-140.
LIU Xue-ping, WEI Qi-qiu, LI Zhen-ming, et al. Effect of Phyllodii Pulchelli Radix on Hepatic Fibrosis of Cynomolgus Monkeys[J]. Chinese journal of experimental traditional medical formulae, 2017, 23(24): 133-140.
刘雪萍, 韦啟球, 李振明, 等. 排钱草对食蟹猴肝纤维化的影响[J]. 中国实验方剂学杂志, 2017,23(24):133-140. DOI: 10.13422/j.cnki.syfjx.2017240133.
LIU Xue-ping, WEI Qi-qiu, LI Zhen-ming, et al. Effect of Phyllodii Pulchelli Radix on Hepatic Fibrosis of Cynomolgus Monkeys[J]. Chinese journal of experimental traditional medical formulae, 2017, 23(24): 133-140. DOI: 10.13422/j.cnki.syfjx.2017240133.
目的:探讨排钱草对四氯化碳(CCl4)诱导的食蟹猴肝纤维化的防治作用及其机制。方法:雄性食蟹猴30例,随机分为正常组、模型组、排钱草高、低剂量组(4.0,2.4 g·kg-1)和秋水仙碱组(0.08 mg·kg-1),每组6例。除正常组外,其余各组灌胃40% CCl4橄榄油混合液1.6 mL·kg-1,每周2次,建立CCl4肝纤维化模型。第四周开始,给药组分别灌胃给药12周。每4周称1次体重,采血检测肝功能,酶联免疫吸附法(ELISA)检测血清中透明质酸(HA),层粘连蛋白(LN),基质金属蛋白酶-1(MMP-1)和金属蛋白酶抑制因子-1(TIMP-1)含量。第8,16周进行肝脏B超检查,在B超下进行肝脏穿刺活检,活检组织进行苏木素-伊红(HE)染色和Van-Gieson胶原纤维染色病理检查,采用蛋白免疫印迹法(Western blot)检测肝脏平滑肌肌动蛋白-α(α-SMA)和转化生长因子-β1(TGF-β1)表达水平。结果:造模8周,与正常组和造模前比较,模型组、秋水仙碱组、排钱草高剂量组和排钱草低剂量组血清丙氨酸氨基转移酶(ALT),天门冬氨酸氨基转移酶(AST),总胆红素(TBIL),碱性磷酸酶(ALP),HA和LN水平显著升高,总蛋白(TP)和白蛋白(ALB)水平显著下降(P<0.05,P<0.01);造模12周,与正常组比较,模型组、秋水仙碱组、排钱草高剂量组和排钱草低剂量血清TIMP-1水平显著升高(P<0.05);造模16周,B超显示,模型组肝脏光点增粗,回声增强,HE染色显示肝细胞坏死、成纤维细胞增生,Van-Gieson染色显示胶原纤维含量增加,肝脏α-SMA和TGF-β1表达水平显著升高(P<0.05)。给药8周后,排钱草高剂量组和秋水仙碱组均显著降低血清ALT,AST,TBIL,ALP和HA水平,升高TP水平(P<0.05,P<0.01);给药12周后,排钱草高剂量组和秋水仙碱组能显著降低血清LN和TIMP-1水平显著(P<0.05),升高血清ALB水平(P<0.05);B超和病理结果均显示排钱草高剂量组和秋水仙碱组肝细胞坏死减少、肝脏纤维化程度降低;Western blot显示排钱草高剂量组和秋水仙碱组肝脏α-SMA和TGF-β1表达水平显著下降(P<0.05)。结论:排钱草(4.0 g·kg-1)对CCl4诱导的食蟹猴肝纤维化有较好的治疗作用,能明显改善肝功能,降低纤维化程度,治疗作用与其降低肝细胞损伤和抑制纤维化有关。
Objective: To explore the effect of Phyllodii Pulchelli Radix on carbon tetrachloride CCl4-induced hepatic fibrosis of cynomolgus monkeys and the underling mechanism. Method: The 30 male cynomolgus monkeys were randomly divided into 5 groups
namely normal group
model group
colchicine group (0.08 mg·kg-1)
Phyllodii Pulchelli Radix high and low dose groups (4.0
2.4 g·kg-1)
with 6 monkeys in each group. Except the monkeys in normal group
CCl4-induced hepatic fibrosis models were established by intragastrical administration of 40% carbon tetrachloride (CCl4) olive oil mixture at dose of 1.6 mL·kg-1 in other animals
2 times a week. From week 4
intragastrical administration was given for 12 weeks in various treatment groups. They were weighed every 4 weeks
and blood was taken to detect liver function. The levels of hyaluronic acid (HA)
laminin(LN)
metal matrix proteinase-1(MMP-1) and tissue inhibitor of metalloproteinase-1(TIMP-1) in serum were detected by using ELISA. At week 8 and week 16
livers of all animals were detected by B ultrasound
then aspiration biopsy was conducted under B ultrasound. Hematoxylin-eosin (HE) staining and Van-Gienson staining were conducted on the aspiration biopsy samples
and transforming growth factor-β1 (TGF-β1) and smooth muscle actin-α (α-SMA) in liver were also detected in the samples by Western blot. Result: After administration of CCl4 for 8 weeks
the levels of alanine amino transferase (ALT)
aspartate amino transferase (AST)
total bilirubin (TBIL)
alkaline phosphatase(ALP)
HA and LN in serum were increased in model group
colchicines group
and Phyllodii Pulchelli Radix high and low dose groups as compared with control group and before modeling before
while total protein (TP) and albumin (ALB) in serum were decreased significantly (P<0.05
P<0.01). After modeling for 12 weeks
TIMP-1 was increased significantly in model group and all treatment groups as compared with the normal group (P<0.05). After modeling for 16 weeks
the strong echo was enhanced in liver of model group in detecting by B ultrosound
and HE staining showed hepatocyte necrosis and fibroblast proliferation; Van-Gieson staining showed that the content of collagen fiber was increased
and the expression levels of TGF-β1 and α-SMA in liver were increased significantly in model group (P<0.05). After administration of colchicines or Phyllodii Pulchelli Radix for 8 weeks
the levels of AST
ALT
TBIL
ALP and HA in serum were decreased significantly in colchicines group and high dose Phyllodii Pulchelli Radix group as compared with model group
while the level of TP was increased differently (P<0.05
P<0.01). After administration of colchicines or Phyllodii Pulchelli Radix for 12 weeks
the levels of LN and TIMP-1 were decreased significantly in colchicines group and high dose Phyllodii Pulchelli Radix group (P<0.05)
while the level of ALB was increased (P<0.05). The results from B ultrasound and pathological examination showed that the fibrosis in liver was relieved in colchicines group and high dose Phyllodii Pulchelli Radix group
and the levels of TGF-β1 and α-SMA in liver were also decreased (P<0.05). Conclusion: Administration of Phyllodii Pulchelli Radix at dose of 4.0 g·kg-1 showed significant efficacy for liver fibrosis of cynomolgus monkeys
could obviously improve the liver function and reduce the degree of fibrosis. The underlying mechanism may be correlated with reduction of liver cell damage and prevention of collagen fiber formation.
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