Effect and mechanism of cryptotanshinone on early pulmonary fibrosis in LPS-induced acute respiratory distress syndrome in mice

Renzi ZHANG; Sheng LI; Daoxin WANG

doi:10.16016/j.2097-0927.202203147

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Effect and mechanism of cryptotanshinone on early pulmonary fibrosis in LPS-induced acute respiratory distress syndrome in mice

Basic Medicine | 更新时间：2026-01-09

- Effect and mechanism of cryptotanshinone on early pulmonary fibrosis in LPS-induced acute respiratory distress syndrome in mice
- Journal of Army Medical University Vol. 44, Issue 18, Pages: 1819-1825(2022)
- 作者机构：
  
  重庆 400010，重庆医科大学附属第二医院呼吸与危重症医学科
- 作者简介：
- 基金信息：
  
  the General Program of National Natural Science Foundation of China(81670071)
- DOI：10.16016/j.2097-0927.202203147
  CLC： R285.5;R363.21;R563.8
- Received：17 March 2022，
  
  Revised：2022-04-26，
  
  Published：30 September 2022
- 稿件说明：
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Renzi ZHANG, Sheng LI, Daoxin WANG. Effect and mechanism of cryptotanshinone on early pulmonary fibrosis in LPS-induced acute respiratory distress syndrome in mice[J]. Journal of Army Medical University, 2022, 44(18): 1819-1825.
DOI：

Renzi ZHANG, Sheng LI, Daoxin WANG. Effect and mechanism of cryptotanshinone on early pulmonary fibrosis in LPS-induced acute respiratory distress syndrome in mice[J]. Journal of Army Medical University, 2022, 44(18): 1819-1825. DOI： 10.16016/j.2097-0927.202203147.

摘要

目的

探讨隐丹参酮(cryptotanshinone，CTS)对脂多糖(lipopolysaccharide，LPS)诱导的急性呼吸窘迫综合征(acute respiratory distress syndrome，ARDS)小鼠的早期肺纤维化的影响及其可能的作用机制。

方法

将40只8周龄雄性C57BL/6J小鼠随机分为正常组、模型组、处理组、CTS组(

=10)。模型组采用气管滴注LPS(5 mg/kg)制备ARDS模型。处理组在ARDS建模前腹腔注射CTS(60 mg/kg) 预处理3 d，建模后再连续予以CTS 7 d。正常组和CTS组分别注射等体积的无菌磷酸盐缓冲液(phosphate-buffered saline，PBS)或CTS。末次给药24 h后处死小鼠，取支气管肺泡灌洗液(bronchoalveolar lavage fluid，BALF)及肺组织。通过HE染色、肺组织湿/干质量(wet-to-dry，W/D)比值、BALF中蛋白浓度和总细胞计数评估肺组织损伤情况；采用酶联免疫吸附试验(ELISA)检测BALF中炎症因子水平；通过Masson染色及羟脯氨酸(hydroxyproline，HYP)含量检测评估肺组织纤维化程度；利用免疫组化染色或Western blot检测VE-cadherin、CD31、波形蛋白(Vimentin)、α-平滑肌肌动蛋白(α-smooth muscle actin，α-SMA)的蛋白表达。

结果

与正常组比较，模型组的肺组织W/D、BALF中总细胞数胞计数和蛋白浓度升高，BALF中的TGF-β1、TNF-α、IL-1β、IL-6升高，IL-10降低(

0.05)，小鼠肺组织出现肺损伤和早期肺纤维化；HYP含量升高(

0.05)；Vimentin、α-SMA的表达升高，而VE-cadherin、CD31的表达隆低(

0.05)。与模型组比较，处理组的肺组织W/D、BALF中总细胞计数和蛋白浓度降低，BALF中的TGF-β1、TNF-α、IL-1β、IL-6降低，IL-10升高(

0.05)；肺损伤和早期肺纤维化程度明显减轻，肺损伤评分和肺纤维化程度评分降低(

0.05)；HYP含量显著降低(

0.05)；Vimentin、α-SMA的表达降低，但VE-cadherin、CD31的表达增强(

0.05)。

结论

CTS可能通过抑制LPS刺激后肺组织中的内皮间质转化(endothelial-to-mesenchymal transition，EndMT)过程从而减轻ARDS小鼠早期肺纤维化。

Abstract

Objective

To investigate the effects and underlying mechanism of cryptotanshinone (CTS) on early pulmonary fibrosis in mice with lipopolysaccharide (LPS)-induced acute respiratory distress syndrome (ARDS).

Methods

Forty male C57BL/6J mice (8 weeks old) were randomly divided into control group

model group

treatment group

and CTS group

with 10 mice in each group. A ARDS model was established by intratracheal instillation of LPS (5 mg/kg). The mice in the treatment group was intraperitoneally pretreated with CTS (60 mg/kg) for 3 d before modeling

followed by CTS administion for 7 consecutive days. Similarly

the mice of the control and CTS groups were injected with an equivalent volume of sterile phosphate-buffered saline (PBS) or CTS

respectively. The mice were sacrificed in 24 h after the last challenge

and bronchoalveolar lavage fluid (BALF) and lung tissues were collected. The injury of lung was observed with HE staining

lung wet-to-dry (W/D) ratio

total protein concentration and total cell number in BALF. The levels of inflammatory cytokines in BALF were determined by enzyme-linked immunosorbent assay (ELISA). The fibrosis of lung was characterized by Masson staining and content of hydroxyproline (HYP). The protein levels of VE-cadherin

CD31

Vimentin and α-smooth muscle actin(α-SMA) were detected by immunohistochemistry or Western blotting.

Results

Obviously increased lung W/D ratio and total protein concentration and total cell number in BALF

elevated contents of TNF-α

TGF-β1

IL-1β and IL-6 while reduced content of IL-10 in BALF were seen in the model group when compared with the control group (

0.05). Compared with the model group

the treatment group had significantly decreased lung W/D ratio and total protein concentration and total cell number in BALF

down-regulated levels of IL-1β

IL-6

TNF-α and TGF-β1 and up-regulated level of IL-10 in BALF (

0.05). In addition

CTS also ameliorated lung injury and early fibrosis

decreased lung injury and fibrosis score

and reduced HYP content in LPS-induced ARDS (

0.05)

up-regulated the VE-cadherin and CD31 levels

and suppressed the Vimentin and α-SMA levels (

0.05).

Conclusion

CTS alleviates LPS-induced early pulmonary fibrosis in ARDS mice

and its underlying mechanism may be related to the inhibition of endothelial-to-mesenchymal transition (EndMT).

关键词

Keywords

references

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