Therapeutic efficacy and mechanisms of <italic style="font-style: italic">HeiBu</italic> Ointment in the treatment of chronic skin ulcers with abnormal fibrosis

YAO Wentao; PENG Bing; MA Huike; WANG Zhengchun; QU Jianhua; LI Ping; HE Xiujuan

doi:10.3969/j.issn.1006-2157.2023.12.009

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Therapeutic efficacy and mechanisms of HeiBu Ointment in the treatment of chronic skin ulcers with abnormal fibrosis

Experimental Studies | 更新时间：2024-01-03

- Therapeutic efficacy and mechanisms of HeiBu Ointment in the treatment of chronic skin ulcers with abnormal fibrosis
- Journal of Beijing University of Traditional Chinese Medicine Vol. 46, Issue 12, Pages: 1684-1693(2023)
- 作者机构：
  
  首都医科大学附属北京中医医院　北京市中医药研究所　北京　100010
- 作者简介：
  
  HE Xiujuan, Ph.D., Researcher. Beijing Institute of Chinese Medicine, Beijing Hospital of Traditional Chinese Medicine of Capital Medical University, No.23, Art Museum Back Street, Dongcheng District, Beijing 100010.E-mail: hexiujuan@bjzhongyi.com
- 基金信息：
  
  National Natural Science Foundation of China(82274524);Beijing Natural Science Foundation(7212164)
- DOI：10.3969/j.issn.1006-2157.2023.12.009
  CLC： R285.5
- Received：14 July 2023，
  
  Published Online：29 November 2023，
  
  Published：30 December 2023
- 稿件说明：
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YAO Wentao, PENG Bing, MA Huike, et al. Therapeutic efficacy and mechanisms of HeiBu Ointment in the treatment of chronic skin ulcers with abnormal fibrosis[J]. Journal of beijing university of traditional chinese medicine, 2023, 46(12): 1684-1693.
DOI：

YAO Wentao, PENG Bing, MA Huike, et al. Therapeutic efficacy and mechanisms of HeiBu Ointment in the treatment of chronic skin ulcers with abnormal fibrosis[J]. Journal of beijing university of traditional chinese medicine, 2023, 46(12): 1684-1693. DOI： 10.3969/j.issn.1006-2157.2023.12.009.

摘要

目的

研究黑布药膏对转化生长因子-β1(TGF-β1)诱导的人永生化表皮细胞(HaCaT)上皮间质转化(EMT)的影响，探讨黑布药膏治疗慢性皮肤溃疡伴有异常纤维化的作用机制。

方法

使用TGF-β1诱导人HaCa细胞EMT模型。将HaCaT细胞分为空白对照组，模型组，以及黑布药膏高、中、低剂量组。各组均加入DMEM高糖培养基(4.00 mmol/L左旋谷酰胺、4.5 g/L葡萄糖)外，模型组加入TGF-β1(0.01 mg/L)，黑布药膏高、中、低剂量组加入TGF-β1(0.01 mg/L)与不同剂量黑布药膏提取物(0.5、0.25、0.125 mg/L)。使用CCK-8法检测黑布药膏对HaCaT细胞活性的影响；划痕实验检测黑布药膏对HaCaT细胞迁移能力的影响；免疫荧光法检测EMT上皮标志物上皮钙黏素(E-cadherin)，间质标志物神经钙黏素(N-cadherin)和波形蛋白(vimentin)的表达；蛋白质印迹法检测Sma和Mad相关蛋白(Smad)和胞外信号调节激酶(ERK)信号通路相关蛋白的表达。

结果

黑布药膏提取物浓度在0.5 mg/L以下对HaCaT细胞活性无影响。与模型组相比，黑布药膏各剂量组可抑制TGF－β1刺激后的HaCaT细胞迁移，促进E-cadherin表达，抑制N-cadherin、vimentin表达，抑制TGF－β1诱导的HaCaT细胞Smad2、Smad3和ERK1/2蛋白的磷酸化(

0.05)。

结论

黑布药膏可能通过抑制Smad和ERK信号通路，抑制表皮细胞向间质细胞过度转化，从而促进创面周围异常纤维化的慢性皮肤溃疡愈合。

Abstract

Objective

We aim to investigate the impact of

HeiBu

Ointment on transforming growth factor-β1 (TGF-β1)-induced epithelial-mesenchymal transition (EMT) in human immortalized epidermal cells(HaCaT) and elucidate the mechanism underlying

HeiBu

Ointment's efficacy in treating chronic skin ulcers with abnormal fibrosis.

Methods

We employed a TGF-β1-induced EMT model using HaCaT. HaCaT cells were categorized into five groups: the blank control group

the model group

and the

HeiBu

Ointment high-

medium-

and low-dose groups. All the groups were cultured in DMEM high-glucose medium (4.00 mmol/L L-glutamine

4.5 g/L glucose). The model group was treated with TGF-β1 (0.01 mg/L)

while the

HeiBu

Ointment high-

medium-

and low-dose groups received TGF-β1 (0.01 mg/L) in combination with varying doses of

HeiBu

Ointment extract (0.5 mg/L

0.25 mg/L

and 0.125 mg/L). The CCK-8 method was used to detect the effect of

HeiBu

Ointment on HaCaT cell activity; the effect of

HeiBu

Ointment on the migration ability of HaCaT cells was determined by scratch test; the effects of

HeiBu

Ointment on the expression of the epithelial marker protein E-cadherin and the mesenchymal marker proteins N-cadherin and vimentin in HaCaT cells were observed by immuno fluorescence method; the expression of Smad and ERK signaling pathway-related proteins was detected by Western blotting.

Results

HeiBu

Ointment below 0.5 mg/L had no effect on HaCaT cell activity. Compared with the model group

HeiBu

Ointment significantly inhibited the migration of HaCaT cells after TGF-β1 stimulation (

0.05)

promoted the expression of the epithelial marker protein E-cadherin

inhibited the expression of the mesenchymal marker proteins N-cadherin and vimentin

and inhibited TGF-β1-induced phosphorylation of Smad2

Smad3

and ERK1/2 in HaCaT cells (

0.05).

Conclusion

HeiBu

Ointment may promote healing of chronic skin ulcers with abnormal fibrosis by inhibiting excessive trausition of epidermal cells to mesenchymal cells through inhibition of the Smad and ERK signaling pathways.

关键词

Keywords

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⁰

Therapeutic efficacy and mechanisms of HeiBu Ointment in the treatment of chronic skin ulcers with abnormal fibrosis

DOI：10.3969/j.issn.1006-2157.2023.12.009

摘要

Abstract

关键词

Keywords

references