Effect of Xiayuxue Tang with Different Processing Method on CCl<sub>4</sub>-induced Hepatic Fibrosis in Rats

Ding-qi ZHANG; Ying XU; Hai-lin YANG; Ping LIU; Wei LIU

doi:10.13422/j.cnki.syfjx.20200540

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Effect of Xiayuxue Tang with Different Processing Method on CCl₄-induced Hepatic Fibrosis in Rats

Classic Prescriptions | 更新时间：2021-02-09

- Effect of Xiayuxue Tang with Different Processing Method on CCl₄-induced Hepatic Fibrosis in Rats
- Chinese Journal of Experimental Traditional Medical Formulae Vol. 26, Issue 5, Pages: 18-25(2020)
- 作者机构：
  
  1.上海中医药大学　附属曙光医院　肝病研究所，上海市中医临床重点实验室，上海　 201203
  2.上海中医药大学　交叉科学研究院，上海　 201203
- 作者简介：
- 基金信息：
- DOI：10.13422/j.cnki.syfjx.20200540
  CLC： R2-0; R285; R289
- Received：08 September 2019，
  
  Published Online：25 November 2019，
  
  Published：05 March 2020
- 稿件说明：
移动端阅览
Ding-qi ZHANG, Ying XU, Hai-lin YANG, et al. Effect of Xiayuxue Tang with Different Processing Method on CCl₄-induced Hepatic Fibrosis in Rats[J]. Chinese journal of experimental traditional medical formulae, 2020, 26(5): 18-25.
DOI：

Ding-qi ZHANG, Ying XU, Hai-lin YANG, et al. Effect of Xiayuxue Tang with Different Processing Method on CCl₄-induced Hepatic Fibrosis in Rats[J]. Chinese journal of experimental traditional medical formulae, 2020, 26(5): 18-25. DOI： 10.13422/j.cnki.syfjx.20200540.

摘要

目的：

探讨蜜丸酒煎下瘀血汤与饮片酒煎下瘀血汤抗肝纤维化作用是否存在异同。

方法：

40只Wistar大鼠随机分为正常组、模型组、蜜丸酒煎下瘀血汤组、饮片酒煎下瘀血汤组、扶正化瘀方组。以50%CCl

皮下注射9周制备肝纤维化模型。第7周首日开始，正常组与模型组给予0.3%羧甲基纤维素钠灌胃，其余3组分别给予蜜丸酒煎下瘀血汤(0.58 g·kg

-1

)，饮片酒煎下瘀血汤(0.43 g·kg

-1

)，扶正化瘀方(2 g·kg

-1

)灌胃，第9周结束后收集肝组织和血清样本。分别采用苏木素-伊红(HE)染色观察各组大鼠肝组织病理学，天狼猩红染色观察纤维化程度，碱水解法测定肝组织羟脯氨酸含量情况，全自动分析法检测大鼠血清肝功能，实时荧光定量聚合酶链式反应(Real-time PCR)检测肝组织

-平滑肌肌动蛋白(

-SMA)和Ⅰ型胶原(COL-I)mRNA的表达情况，蛋白免疫印迹法(Western blot)和免疫组化测定肝组织

-SMA蛋白的表达，酶联免疫吸附测定(ELISA)测定炎症因子与氧化指标。

结果：

与正常组比较，模型组大鼠肝肝纤维化程度较严重，肝组织天狼猩红阳性染色面积比、羟脯氨酸含量，

-SMA和COL-I蛋白及mRNA表达显著增加(

0.01)，大鼠血清丙氨酸氨基转移酶(AST)，天门冬氨酸氨基转移酶(ALT)，总胆红素(TBil)，肿瘤坏死因子-

(TNF-

)，白细胞介素(IL)-1

，IL-17A及丙二醛(MDA)的水平显著增加(

0.01)，超氧化物歧化酶(SOD)及谷胱甘肽过氧化物酶(GSH-Px)的水平显著降低(

0.01)；与模型组比较，蜜丸酒煎下瘀血汤与饮片酒煎下瘀血汤可明显减轻肝纤维化程度，肝组织天狼猩红阳性染色面积比、羟脯氨酸含量，

-SMA和COL-I蛋白及mRNA表达，大鼠血清AST，ALT，TBil，TNF-

，IL-1

，IL-17A，MDA的水平明显降低(

0.05，

0.01)，SOD及GSH-Px水平明显增加(

0.05，

0.01)。其中蜜丸酒煎下瘀血汤组肝组织病理变化的改善显著优于饮片酒煎下瘀血汤，且肝组织在天狼猩红阳性染色面积比、羟脯氨酸含量、肝组织COL-I mRNA以及

-SMA mRNA和蛋白的表达降低也较饮片酒煎下瘀血汤明显(

0.05，

0.01)。血清肝功能、炎症因子及氧化相关指标2种制法下瘀血汤组间差异无统计学意义，但蜜丸酒煎下瘀血汤改善趋势更为明显。

结论：

蜜丸酒煎下瘀血汤与饮片酒煎下瘀血汤均具有较好的抗肝纤维化作用，其中以蜜丸酒煎下瘀血汤作用更为显著，提示制蜜丸一定程度上能够增加下瘀血汤抗肝纤维化作用。

Abstract

Objective:

To compare the anti-hepatic fibrosis effect of ethanol extract of honey Xiayuxue Tang (H-Xiayuxue Tang)and prepared slices of crude drug-Xiayuxue Tang (C-Xiayuxue Tang).

Method:

Totally 40 wistar rats were randomlydivided into normal group

model group

H-Xiayuxue Tang group

C-Xiayuxue Tang group

and Fuzheng Huayu group. Liver fibrosis model was induced by hypodermic injection of 50% carbon tetrachloride for 9 weeks. From the seventh week

the normal group and the model group were given 0.3% sodium carboxymethyl cellulose by gavage. The other three groups were given H-Xiayuxue Tang(0.58 g·kg

-1

)

C-Xiayuxue Tang (0.43 g·kg

-1

)

Fuzheng Huayu (2 g·kg

-1

)

respectively. At the end of the ninth week

liver tissues and serum samples were collected for follow-up studies. The pathological changes and fibrosis degree were observed by HE and Sirius red staining

hydroxyproline content in liver tissues was detected by alkaline hydrolysis

serum liver function was determined by automatic analysis

the expressions of collagen type I (COL-I) and

-smooth muscle actin (

-SMA) in liver tissues were detected by Real-time PCR. The expressions of

-SMA in liver tissues was detected by Western blot and Immunohistochemistry

and serum inflammatory factors and oxidantion indexes were determined by ELISA

respectively.

Result:

Compared with control group

the severity of liver fibrosis was observed

the area ratio of Sirius red positive staining

hydroxyproline (Hyp) content

the protein and mRNA expressions of

-SMA

COL-I in the liver tissue were significantly increased in the model group (

0.01). And the levels of aspartate aminotransferase (AST)

alanine aminotransferase (ALT)

total bilirubin (TBil)

tumor necrosis factor-

(TNF-

)

interleukin-1

(IL-1

)

interleukin-17A (IL-17A) and malondialdehyde (MDA) in the serum of the rats were markedly increased (

0.01). The levels of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) were apparently decreased (

0.01). Compared with model group

the degree of liver fibrosis was significantly improved

the area ratio of Sirius red positive staining

the content of Hyp

the protein and mRNA expression of

-SMA and COL-I

and the serum levels of AST

ALT

TBil

TNF-

IL-1

IL-17A and MDA were significantly decreased (

0.05

0.01). The serum contents of SOD and GSH-Px were markedly increased (

0.05

0.01)

respectively

by H-Xiayuxue Tang or C-Xiayuxue Tang. Among them

the improvement of liver histopathological changes was more obvious in the H-Xiayuxue Tang group than in the C-Xiayuxue Tang group. Meanwhile

the percentage of Sirius red positive staining area

the content of hydroxyproline

the expressions of COL-I and

-SMA mRNA and protein in liver tissue were significantly decreased compared with the C-Xiayuxue Tang group(

0.05

0.01). In addition

there was no significant difference in serum liver function

inflammatory factors and oxidantion related indicators between groups

but the improvement trend of H-Xiayuxue Tang was more obvious.

Conclusion:

H-Xiayuxue Tang and C-Xiayuxue Tang have a better effect in hepatic fibrosis

especially H-Xiayuxue Tang

suggesting that honey can increase the anti-hepatic fibrosis effect of Xiayuxue Tang.

关键词

Keywords

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⁰

Effect of Xiayuxue Tang with Different Processing Method on CCl4-induced Hepatic Fibrosis in Rats

DOI：10.13422/j.cnki.syfjx.20200540

摘要

Abstract

关键词

Keywords

references

Effect of Xiayuxue Tang with Different Processing Method on CCl₄-induced Hepatic Fibrosis in Rats