Mechanism of medicinal serum containing Tongguan Fang inhibits LPS-induced inflammation in RAW264.7 cells
10.3969/j.issn.1000-484X.2025.08.008
- VernacularTitle:通管方含药血清抑制LPS诱导的RAW264.7细胞炎症机制探讨
- Author:
Ping LI
1
;
Jia XU
;
Zichun HUANG
;
Shuting WANG
;
Jilin KUANG
Author Information
1. 湖南中医药大学第二附属医院,长沙 410005;湖南中医药大学,长沙 410208
- Publication Type:Journal Article
- Keywords:
Salpingitis infertility;
Tongguan Fang;
Apoptosis;
PI3K/Akt;
ROS
- From:
Chinese Journal of Immunology
2025;41(8):1840-1846,1852
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To investigate the effect of Tongguan Fang drug-containing serum on LPS-induced inflammation of RAW264.7 cells,so as to clarify the cellular mechanism of the Tongguan Fang in treating tubal inflammatory infertility.Methods:Lipo-polysaccharide(LPS)was used to stimulate the RAW264.7 macrophage cell line as an in vitro model,Detected levels of cellular oxida-tive stress(ROS),apoptosis and expressions of PI3K/Akt signaling pathway proteins.CCK-8 was used to detect effect of Tongguan Fang drug-containing serum on cellular inflammation.Effects of Tongguan Fang-containing serum on cell inflammation were assessed using CCK-8 assay.Cells were divided into control group,LPS-induced inflammation model group,and Tongguan Fang-containing serum-treated LPS-induced inflammation model groups of different concentrations.qPCR was performed to detect mRNA expression levels of IL-1β,IL-6 and TNF-α in different groups.Immunofluorescence and Western blot were used to evaluate phosphorylation levels of the PI3K/Akt signaling pathway and protein levels of Bcl-2,Bax and caspase-3 of apoptotic.Hoechst 33342/PI staining and flow cytometry were employed to assess the impact of Tongguan Fang-containing serum on cell apoptosis.DCFH-DA probe was used to measure cellular ROS levels affected by Tongguan Fang-containing serum.Results:①Tongguan Fang-containing serum reduced LPS-induced cellular injury in a dose-dependent manner;②Tongguan Fang-containing serum decreased production of pro-inflammatory cytokines IL-6,IL-1β and TNF-α induced by LPS;③Tongguan Fang-containing serum reduced intracellular ROS generation induced by LPS;④Pre-treatment with Tongguan Fang-containing serum activated PI3K/Akt signaling pathway;⑤Tongguan Fang-containing serum promoted cell apoptosis.Conclusion:Treatment with Tongguan Fang-containing serum can reduce cellular injury and inflam-mation,protect cells from LPS-induced cellular injury by activating the PI3K/Akt signaling pathway and promoting cell apoptosis,and further alleviate cellular inflammation levels.
