Efficacy and molecular mechanism of Qi-Tonifying preparations in rats with liver fibrosis

Xia GAO; Junxiong CHENG; Wenfu CAO

doi:10.16016/j.1000-5404.202010046

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Efficacy and molecular mechanism of Qi-Tonifying preparations in rats with liver fibrosis

Basic Medicine | 更新时间：2026-01-09

- Efficacy and molecular mechanism of Qi-Tonifying preparations in rats with liver fibrosis
- Journal of Third Military Medical University Vol. 43, Issue 8, Pages: 692-699(2021)
- 作者机构：
  
  400016 重庆，重庆医科大学中医药学院
- 作者简介：
- 基金信息：
  
  the General Project of National Natural Science Foundation of China(81573860)
- DOI：10.16016/j.1000-5404.202010046
  CLC： R285.5;R289.5;R575.2
- Received：02 October 2020，
  
  Revised：2020-12-17，
  
  Published：30 April 2021
- 稿件说明：
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Xia GAO, Junxiong CHENG, Wenfu CAO. Efficacy and molecular mechanism of Qi-Tonifying preparations in rats with liver fibrosis[J]. Journal of Third Military Medical University, 2021, 43(8): 692-699.
DOI：

Xia GAO, Junxiong CHENG, Wenfu CAO. Efficacy and molecular mechanism of Qi-Tonifying preparations in rats with liver fibrosis[J]. Journal of Third Military Medical University, 2021, 43(8): 692-699. DOI： 10.16016/j.1000-5404.202010046.

摘要

目的

探讨益气法制剂对肝纤维化大鼠的治疗作用及分子机制。

方法

7周龄Wistar大鼠，建立肝纤维化模型大鼠前，选取10只为正常对照组；建模成功后，分为模型组(

=10)、秋水仙碱组(

=9)、益气法制剂组(

=9)，后两组分别给予相应药物[秋水仙碱制剂0.45 mg/kg，益气法制剂(黄芪、白术)3.6 g/kg

]

，模型组和正常对照组大鼠给予等量生理盐水灌胃，每天1次，连续治疗12周。Western blot检测肝组织中α-平滑肌肌动蛋白(α-SMA)、磷酸化的细胞外信号调节激酶(p-ERK1/2)、磷酸化P38丝裂原活化蛋白酶(p-p38)相关指标表达。RT-qPCR检测肝组织中单核细胞趋化蛋白-1a(MCP-1a)指标的表达。免疫组化检测肝组织中基质金属蛋白酶-2(MMP-2)、基质金属蛋白酶-9(MMP-9)、金属蛋白酶组织抑制因子-1(TIMP-1)指标表达。

结果

Western blot结果显示，与正常对照组相比，模型组的α-SMA、p-ERK1/2的表达均升高(

0.05)，p-p38的表达差异无统计学意义；与模型组比较，益气法制剂组和秋水仙碱组的α-SMA、p-p38的表达均降低(

0.05)，秋水仙碱组p-ERK1/2的表达降低(

0.05)，益气法制剂组p-ERK1/2的表达差异无统计学意义，表明益气法制剂可通过抑制α-SMA、p-ERK1/2、p-p38的表达缓解肝纤维化的进程。RT-qPCR结果显示，与正常对照组相比，模型组的MCP-1a的表达上调(

0.05)；与模型组相比，益气法制剂组和秋水仙碱组的MCP-1a表达下调(

0.05)，提示益气法制剂可能通过下调MCP-1a的表达，缓解肝纤维化的进程。免疫组化结果显示，MMP-2在正常对照组明显表达，模型组中可见MMP-2的表达，与模型组相比，益气法制剂组和秋水仙碱组MMP-2的表达上调；MMP-9的表达在正常对照组可见，与正常对照组相比较，模型组上调，与模型组比较，益气法制剂组上调；TIPM-1正常对照组表达较少，与正常对照组相比，模型组的表达有所上调，与模型组相比较，益气法制剂组和秋水仙碱组均下调。

结论

益气法制剂可改善大鼠肝纤维化，可能通过抑制MCP-1来减少炎症细胞的释放，进而抑制ERK1/2、p38信号通路的激活，减少磷酸化p-ERK1/2、p-p38介导信号由胞浆向胞核传递，从而抑制TIMP-1的表达，促进MMP-2、MMP-9的释放，增加ECM降解，减少沉积，逆转肝纤维化。

Abstract

Objective

To investigate the therapeutic effect and molecular mechanism of Qi-Tonifying preparations (

Astragalus membranaceus

and

Atractylodes macrocephala

) on hepatic fibrosis in rats.

Methods

Wistar rats (7 weeks old) were divided into normal control (

=10)

model group (

=10)

colchicine group (0.45 mg/kg

=9)

and Qi-

Tonifying preparation group (3.6 g/kg

=10). Corresponding drugs or normal saline were given to the rats intragastrically

once a day

for continuous 12 weeks. Western blotting was used to detect the expression of α-smooth muscle actin (α-SMA)

phosphorylated extracellular signal regulated kinase 1/2 (p-ERK1/2) and phosphorylated p38 mitogen activated protease (p-p38) in the liver tissues. RT-qPCR was employed to detect the expression of monocyte chemoattractant protein-1a (MCP-1a) in the liver tissues. Immunohistochemical assay was adopted to detect the expression of matrix metalloproteinase-2 (MMP-2)

MMP-9 and metalloproteinase tissue inhibitor-1 (TIMP-1) in the liver tissues.

Results

The results of Western blotting showed that the expression levels of α-SMA and p-ERK1/2 were significantly higher in the model group than the normal control group (

0.05)

but no such difference was seen in that of p-p38 (

0.05). Compared with the model group

the expression levels of α-SMA and p-p38 were decreased in the Qi-Tonifying preparation group and the colchicine group (

0.05)

and that of ERK1/2 was decreased obviously in the colchicine group (

0.05)

but the level was similar in the Qi-Tonifying preparation group (

0.05). These results suggested that Qi-Tonifying preparations alleviated the progression of liver fibrosis by inhibiting the expression of α-SMA

p-ERK1/2 and p-p38. RT-qPCR results showed that the expression of MCP-1a was significantly increased in the model group than the normal control group (

0.05)

and was down-regulated in the Qi-Tonifying group and the colchicine group than the model group (

0.05). Qi-Tonifying preparations alleviated the progression of liver fibrosis by down-regulating MCP-1a. Immunohistochemical assay showed that MMP-2 was obviously expressed in the normal control group

and observ

ed in the model group. Its expression was stronger in the Qi-Tonifying group and the colchicine group than the model group. The expression of MMP-9 was mildly observed in the normal group

and was up-regulated in the model group than the normal group

but down-regulated in the Qi-Tonifying group and the colchicine group when compared with the model group.

Conclusion

Qi-Tonifying preparations can improve liver fibrosis in rats

and may reduce the release of inflammatory cells by inhibiting MCP-1

thereby suppressing the activation of ERK1/2 and p38 signaling pathways and reducing phosphorylated p-ERK1/2

P-p38 mediated signal transmission from the cytoplasm to the nucleus

and then inhibits the expression of TIMP-1

promotes the release of MMP-2 and MMP-9

increases ECM degradation

reduces deposition

and finally

reverses liver fibrosis.

关键词

Keywords

references

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