Oxamate alleviates silicotic fibrosis in mice by inhibiting senescence of alveolar type II epithelial cells

LIU Wenjing; MAO Na; LI Yaqian; GAO Xuemin; WEI Zhongqiu; ZHU Ying; XU Hong; JIN Fuyu

doi:10.11836/JEOM24018

Volume 41 Issue 7

Jul. 2024

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Journal of Environmental and Occupational Medicine > 2024 > 41(7): 760-767, 779. > DOI: 10.11836/JEOM24018

LIU Wenjing, MAO Na, LI Yaqian, GAO Xuemin, WEI Zhongqiu, ZHU Ying, XU Hong, JIN Fuyu. Oxamate alleviates silicotic fibrosis in mice by inhibiting senescence of alveolar type II epithelial cells[J]. Journal of Environmental and Occupational Medicine, 2024, 41(7): 760-767, 779. DOI: 10.11836/JEOM24018

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Oxamate alleviates silicotic fibrosis in mice by inhibiting senescence of alveolar type II epithelial cells

a.
School of Public Health
b.
School of Basic Medical Sciences
c.
Health Science Center, North China University of Science and Technology, Tangshan, Hebei 063200, China

Funds: This study was funded.

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Corresponding author:
JIN Fuyu, E-mail: fuyujinjfy@163.com
Received Date: January 17, 2024
Accepted Date: May 23, 2024
Available Online: July 30, 2024

Graphical Abstract

Abstract

Abstract

Background
The senescence of alveolar type II epithelial cells is an important driving factor for the progression of silicotic fibrosis, and the regulatory effects of oxamate on the senescence of alveolar type II epithelial cells is still unclear.
Objective
To explore whether lactate dehydrogenase inhibitor oxamate can alleviate silicotic fibrosis in mice by inhibiting senescence of alveolar type II epithelial cells
Methods
This study was divided into two parts: in vivo experiments and in vitro experiments. In the first part, forty SPF C57BL/6J male mice were randomly divided into four groups with 10 in each group: control group, silicosis model group, low-dose oxamate treatment group, and high-dose oxamate treatment group. The silicotic mouse model was established by intratracheal instillation of 50 μL SiO₂ suspension (100 mg·mL⁻¹). The treatment models were prepared by intraperitoneal injection of 100 μL oxamate (225 mmol·L⁻¹ and 1125 mmol·L⁻¹). In the second part, induction of MLE-12 mouse alveolar type II epithelial cells was conducted with SiO₂. The in vitro experimental groups were ① SiO₂ induction groups: control group, 50 μg·mL⁻¹ SiO₂ group, 100 μg·mL⁻¹ SiO₂ group, and 200 μg·mL⁻¹ SiO₂ group, and ② oxamate treatment groups: control group, SiO₂ group (100 μg·mL⁻¹), low-dose oxamate (25 mmol·L⁻¹) treatment group, and high-dose oxamate (50 mmol·L⁻¹) treatment group. Pathological morphology of lung tissues was evaluated after hematoxylin-eosin (HE) staining; deposition of collagen in lung tissues was evaluated after sirius red staining; positive co-expression of prosurfactant protein C (Pro-SPC) and β-galactosidase was detected by immunofluorescence staining; positive expression of β-galactosidase in MLE-12 cells was detected by immunofluorescence staining. The protein expression levels of collagen type I (CoL I), fibronectin1 (FN1), hexokinase 2 (HK2), pyruvate kinase isozyme type M2 (PKM2), lactate dehydrogenase A (LDHA), p-ataxia telangiectasia and Rad3-related kinase (ATR), and cyclin-dependent kinase inhibitors p21, and p16 were detected by Western blotting.
Results
Compared with the control group, the protein expression levels of HK2, PKM2, LDHA, p-ATR, p21, and p16 were significantly upregulated in the silicosis model group and the SiO₂-induced MLE-12 cells (P＜0.05). The in vivo studies showed that, compared with the control group, the silicon nodule area, the collagen deposition area, the proportion of β-galactosidase positive cells, and the protein expression levels of CoL I, FN1, LDHA, p-ATR, p21, and p16 were significantly upregulated in the silicosis model group (P＜0.05). Compared with the silicosis model group, the oxamate treatment groups showed significant downregulation of the silicon nodule area, the collagen deposition area, the proportion of β-galactosidase positive cells, and the the CoL I, FN1, LDHA, p-ATR, p21, and p16 protein expression levels, and the high-dose oxamate treatment group showed a higher efficacy on these indicators than the low-dose oxamate treatment group (P＜0.05). The in vitro studies showed that, compared with the control group, the proportion of β-galactosidase positive cells and the protein expression levels of p-ATR, p21, and p16 were significantly upregulated in the SiO₂-induced group (P＜0.05). Compared with the SiO₂ group, the proportion of β-galactosidase positive cells and the LDHA, p-ATR, p21 and p16 protein expression levels were significantly downregulated in the oxamate treatment groups, and the high-dose oxamate treatment group showed a higher efficacy on these indicators than the low-dose oxamate treatment group (P＜0.05).
Conclusion
Lactate dehydrogenase inhibitor oxamate can alleviate silicotic fibrosis in mice by inhibiting the senescence of alveolar type II epithelial cells.
- oxamate,
- alveolar type II epithelial cell,
- silicosis,
- senescence,
- β-galactosidase

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References (24)

References

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