Chaixian Huashen decoction alleviates lipopolysaccharide induced acute lung injury by inhibiting TLR4/NF-κB pathway
10.3867/j.issn.1000-3002.2025.08397
- VernacularTitle:柴陷花参汤通过TLR4/NF-κB通路改善脂多糖诱导的急性肺损伤
- Author:
Guixin SU
1
;
Yulong HUANG
;
Changwei LI
;
Yu YANG
;
Yang ZHANG
;
Rui XUE
;
Shuo LI
;
Youzhi ZHANG
Author Information
1. 南京中医药大学,江苏 南京 210023;军事医学研究院,北京 100850
- Publication Type:Journal Article
- Keywords:
Chaixian Huashen decoction;
acute lung injury;
inflammation;
NF-κB signaling pathway;
TLR4 signaling pathway;
network pharmacology
- From:
Chinese Journal of Pharmacology and Toxicology
2025;39(9):651-661
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVE To investigate the mechanism through which Chaixian Huashen decoction(CXHSD)ameliorates lipopolysaccharide(LPS)-induced acute lung injury(ALI)in mice.METHODS Component analysis:the components of CXHSD extract were analyzed via ultra-high performance liquid chromatography-high resolution mass spectrometry(UPLC-Q-Exactive HFX).Network pharma-cology analysis was conducted to predict the potential active components and underlying therapeutic targets of CXHSD for ALI treatment.① Animal experiment:mice were randomly divided into the normal control group,model(LPS)group,model+dexamethasone(DEX)4 mg·kg-1 group,model+CXHSD 10 g·kg-1 group,and model+CXHSD 20 g·kg-1 group.Except for the normal control group,ALI was induced in all the mice by intratracheal instillation of LPS.Model+CXHSD groups received daily intra-gastric administration of corresponding treatments for 7 consecutive days.The model+DEX group was administered saline intragastrically for the initial 5 d,followed by DEX for the next 2 d.ALI was induced by intratracheal instillation of LPS 5 mg·kg-1 1 h after the 6th administration of CXHSD/DEX.24 h after modeling,the severity of pulmonary edema was assessed using the wet to dry weight(W/D)ratio,and hematoxylin-eosin(HE)staining was used to evaluate histopathological damage.The levels of myeloperoxidase(MPO),tumor necrosis factor-α(TNF-α),interleukin-6(IL-6),IL-1β in lung tissue homogenates and serum were measured by enzyme-linked immunosorbent assay(ELISA).The total protein concentration in bronchoalveolar lavage fluid(BALF)was measured by bicinchoninic acid(BCA)assay.Immunohistochemistry and Western blotting were used to assess the expression levels of toll-like receptor 4(TLR4),myeloid differentiation primary response 88(MyD88),zonula occludens-1(ZO-1)and occludin,as well as the phosphorylation level of nuclear factor-kappa B p65(NF-κB p65).② Cell experiment:RAW264.7 cells were divided into the cell control group,LPS 1 mg·L-1 group,LPS 1 mg·L-1+DEX 1 mg·L-1 group,and LPS 1 mg·L-1+CXHSD 50,100 and 200 mg·L-1 groups.After 24 h of culture,the nitric oxide(NO)content was measured with the nitrate reductase method,the levels of TNF-α,IL-1 β and IL-6 in the cell supernatants of each group were detected by ELISA.RESULTS Network pharmacology analysis indicated that CXHSD might alleviate ALI through the NF-κB pathway.① Com-pared with the normal control group,the W/D ratio was elevated,pathological injuries aggravated(such as alveolar wall thickening,inflammatory infiltration,and alveolar congestion),histopathological damage pronounced,MPO activity increased,and total protein concentrations in BALF raised in the model group,in which levels of TNF-α,IL-6 and IL-1 β in both lung tissue and serum became higher.Concur-rently,LPS increased the expressions of p-NF-κB p65,TLR4 and MyD88,but reduced the expressions of ZO-1 and occludin.Compared with the model group,model+CXHSD groups had their pulmonary edema and lung pathological injury ameliorated as evidenced by alleviated alveolar wall thickening,inflammatory infiltration and alveolar congestion.The levels of MPO,TNF-α,IL-1 β and IL-6 in both lung tissue and serum,and the total protein concentrations in BALF were significantly decreased in the model+CXHSD groups.Additionally,the expressions of TLR4,MyD88,and p-NF-κB p65 were significantly downregulated,while those of ZO-1 and occludin were prominently upregulated.② Compared with the cell control,the levels of TNF-α,IL-1 β,IL-6 and NO in the supernatant of RAW264.7 cells were signifi-cantly increased in the LPS group.Compared with the LPS group,in the supernatant of RAW264.7 cells treated with LPS+CXHSD at 100 mg·L-1,there was no significant difference in TNF-α levels.However,in the other groups treated with LPS+CXHSD,the levels of TNF-α,IL-1 β,IL-6,and the content of NO were significantly reduced.CONCLUSION CXHSD can alleviate LPS-induced ALI by inhibiting the TLR4/NF-κB pathway,attenuating inflammation,and preserving pulmonary barrier integrity.
