Intervention Mechanism of Method of Supplementing Qi, Nourishing Yin, Activating Blood Circulation and Dredging Collaterals on TGF-/Smads Signaling Pathway in Lung Fibroblasts

PANG Li-jian; LYU Xiao-dong; YUAN Quan; LIU Chuang; ZHAO Zhong-xue; ZANG Ning-zi; HUA Zhen; YANG Li

doi:10.13422/j.cnki.syfjx.2017220125

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Intervention Mechanism of Method of Supplementing Qi, Nourishing Yin, Activating Blood Circulation and Dredging Collaterals on TGF-/Smads Signaling Pathway in Lung Fibroblasts

更新时间：2024-01-04

- Intervention Mechanism of Method of Supplementing Qi, Nourishing Yin, Activating Blood Circulation and Dredging Collaterals on TGF-/Smads Signaling Pathway in Lung Fibroblasts
- Chinese Journal of Experimental Traditional Medical Formulae Vol. 23, Issue 22, Pages: 125-131(2017)
- 作者机构：
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- 基金信息：
- DOI：10.13422/j.cnki.syfjx.2017220125
  CLC： R285.5
- Published：2017
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PANG Li-jian, LYU Xiao-dong, YUAN Quan, et al. Intervention Mechanism of Method of Supplementing Qi, Nourishing Yin, Activating Blood Circulation and Dredging Collaterals on TGF-/Smads Signaling Pathway in Lung Fibroblasts[J]. Chinese journal of experimental traditional medical formulae, 2017, 23(22): 125-131.
DOI：

PANG Li-jian, LYU Xiao-dong, YUAN Quan, et al. Intervention Mechanism of Method of Supplementing Qi, Nourishing Yin, Activating Blood Circulation and Dredging Collaterals on TGF-/Smads Signaling Pathway in Lung Fibroblasts[J]. Chinese journal of experimental traditional medical formulae, 2017, 23(22): 125-131. DOI： 10.13422/j.cnki.syfjx.2017220125.

摘要

目的：探讨益气养阴活血通络法调控肺成纤维细胞转化生长因子-β1（TGF-β1）/Smads信号传导通路机制。方法：SPF级雄性Wistar大鼠30只，采用随机数字法，将30只大鼠随机分为3组，中药含药血清组，西药含药血清组，正常血清组，第4天末次给药后提取含药血清。气管内注入博来霉素造肺纤维化大鼠动物模型，采用胰蛋白酶消化液消化法培养肺成纤维细胞。细胞分为模型组，中药组，西药组，抑制剂组。免疫荧光法鉴定肺成纤维细胞，并分别于第24，48，72 h取材，采用逆转录聚合酶链式反应（RT-PCR）检测TGF-β1，Smad2，Smad4，Smad7 mRNA的表达及流式细胞仪检测细胞凋亡率。结果：第24，48，72 h检测结果显示，与模型组比较，中药组、西药组、抑制剂组细胞内TGF-β1 ，Smad2，Smad4 mRNA表达水平明显下调（P<0.01）；第24 h西药组TGF-β1 mRNA表达水平明显低于中药组（P<0.05）；第48，72 h检测结果均显示与中药组比较，西药组、抑制剂组TGF-β1 mRNA表达水平均明显下调（P<0.05，P<0.01）；第24，48 h检测结果显示与模型组比较，中药、西药、抑制剂各组细胞内Smad2 mRNA表达水平下调明显（P<0.01）；中药组、西药组、抑制剂组3者表达水平相近，无统计学意义。第72 h检测结果显示与中药组比较，西药组、抑制剂组Smad2 mRNA表达水平均明显下调（P<0.05）；第24，48，72 h检测结果均显示与中药组比较，西药组、抑制剂组Smad7 mRNA表达水平均明显下调（P<0.01）。第24，48，72 h检测结果均显示与模型组比较，中药组、西药组与抑制剂组凋亡率明显上调（P<0.01）；中药组细胞凋亡率明显高于抑制剂组（P<0.01）。结论：益气养阴活血通络法可能通过下调肺成纤维细胞TGF-β1，Smad2，Smad4含量和上调Smad7含量调控TGF-β1/Smads信号通路，从而延缓特发性肺纤维化（IPF）进展。

Abstract

Objective: To discuss the mechanism of the method of supplementing Qi

nourishing Yin

activating blood circulation and dredging collaterals in regulating transforming growth factor-β1 (TGF-β1)/Smads signaling pathway in lung fibroblasts. Method: A total of 30 SPF male rats were divided into three groups by the method of random number table

namely traditional Chinese medicine (TCM) serum group

western medicine serum group and normal serum group. Serums were extracted at the end of the fourth day after administration. Bleomycin was intratracheally injected to make animal model of pulmonary fibrosis in rats

and the lung fibroblasts were cultured by trypsin digestion method. The cells were divided into model group

TCM group

western medicine group and inhibitor group. Lung fibroblasts were identified by immunofluorescence

and materials were collected at 24

72 h. Reverse transcription-polymerase chain reaction was used to detect the mRNA expressions of TGF-β1

Smad2

Smad4

and Smad7

and flow cytometry was used to detect apoptosis rate. Result: The results of 24

72 h showed that compared with model group

the mRNA expressions of TGF-β1

Smad2

and Smad4 in the TCM group

western medicine group and inhibitor group were obviously lower (P<0.01). The mRNA expressions of TGF-β1 in 24 h western medicine group was lower than that of TCM group (P<0.05). The results of 48

72 h showed that compared with the TCM group

the mRNA expressions of TGF-β1 in the western medicine group and the inhibitor group were all obviously lower (P<0.05

P<0.01). The results of 24

48 h showed that compared with the model group

the Smad2 mRNA expression in TCM group

western medicine group and inhibitor group were obviously lower (P<0.01). The expressions of TCM group

western medicine group and inhibitor group were similar

with no statistical significance. The results of 72 h showed that compared with the TCM group

the Smad2 mRNA expressions in the western medicine group and the inhibitor group were all obviously lower (P<0.05). The results of 24

72 h showed that compared with the TCM group

the Smad7 mRNA expressions in the western medicine group and the inhibitor group were all obviously lower (P<0.01). The results of 24

72 h showed that compared with the model group

the apoptosis rates of TCM group

western medicine group and inhibitor group were obviously increased (P<0.01). The apoptosis rate of TCM group was higher than that of inhibitor group (P<0.01). Conclusion: The method of supplementing Qi

nourishing Yin

activating blood circulation and dredging collaterals may delay the progress of idiopathic pulmonary fibrosis by decreasing the content of TGF-β1

Smad2 and Smad4

increasing the content of Smad7 and regulating the TGF-β1/Smads signaling pathway.

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Related Institution

Hebei Key Laboratory of Integrated Chinese and Western Medicine for Lung Disease Research

Hebei University of Chinese Medicine

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Affiliated Hospital of Liaoning University of TCM

Collaborative Innovation Center for Chinese Medicine and Respiratory Diseases Co-Constructed by Henan & Ministry of Education of PR China，Henan Key Laboratory of Chinese Medicine for Respiratory Disease， Academy of Chinese Medical Sciences，Henan University of Chinese Medicine

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