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Assessing Retinal Microglial Phagocytic Function In Vivo Using a Flow Cytometry-based Assay
Published on: October 18, 2016
Inhibitory effect of Houttuynia cordata Thunb on LPS-induced retinal microglial activation
Ying-Hui Zhang1, Le-Meng Ren2, Xiao-Yun Wang1
1The Second Hospital of Shandong University, Shandong University, Jinan 250033, Shandong Province, China.
Aim:
To identify the effect of Houttuynia cordata Thunb (HCT) on lipopolysaccharide (LPS)-induced microglial activation and investigate its possible molecular mechanisms.
Methods:
The primary retinal microglial cells were cultured from the retinas of newborn Sprague-Dawley rats and exposed to LPS, and/or HCT with different concentrations. The survival ability of retinal microglia cells was tested by standard MTT method. BrdU cell proliferation assay was used to evaluate the proliferation of retinal microglia. Inflammatory factors in the culture supernatants, including TNF-α, iNOS and IL-1β, were measured using ELISA. Microglia cells' migration was determined with Transwell migration assay. The total p38-MAPK and phosphorylation of p38-MAPK (p-p38-MAPK) were detected with Western blot.
Results:
Primary retinal microglia in culture exposed to LPS to induce microglia activation. Pretreatment with HCT significantly inhibited the LPS-induced cell proliferation, but not the cell viability. LPS induced inflammatory reaction in microglia and cell migration. HCT significantly reduced LPS-stimulated release of pro-inflammatory factors and decreased the number of migrating cells substantially in a concentration-dependent manner. Moreover, the protein levels of p-p38 MAPK were identified as the up regulation and co-treatment with HCT obviously inhibited the upregulation of p-p38 MAPK, but had no effect on the levels of total p38-MAPK.
Conclusion:
The data suggest that HCT inhibits LPS-induced retinal microglial activation via suppression of the p-p38-MAPK. HCT may be used for the treatment of ocular diseases characterized by over-activated microglia.
Insights
Houttuynia cordata Thunb (HCT) inhibits lipopolysaccharide (LPS)-induced microglial activation by suppressing p-p38-MAPK. This suggests HCT may treat ocular diseases involving over-activated microglia.
Area of Science:
- Neuroscience
- Pharmacology
- Immunology
Background:
- Microglial activation by lipopolysaccharide (LPS) is implicated in various ocular diseases.
- Understanding the molecular mechanisms of microglial activation is crucial for developing targeted therapies.
Purpose of the Study:
- To investigate the inhibitory effect of Houttuynia cordata Thunb (HCT) on LPS-induced microglial activation.
- To elucidate the molecular mechanisms underlying HCT's action, focusing on the p38-MAPK pathway.
Main Methods:
- Primary rat retinal microglia were cultured and treated with LPS and/or HCT.
- Cell viability (MTT), proliferation (BrdU), inflammatory factor release (ELISA), migration (Transwell), and p38-MAPK signaling (Western blot) were assessed.
Main Results:
- HCT significantly inhibited LPS-induced microglial proliferation and migration in a dose-dependent manner.
- HCT reduced the release of pro-inflammatory factors (TNF-α, iNOS, IL-1β) and suppressed the upregulation of phosphorylated p38-MAPK (p-p38-MAPK).
- HCT did not affect cell viability or total p38-MAPK levels.
Conclusions:
- HCT effectively inhibits LPS-induced retinal microglial activation.
- The mechanism involves the suppression of the p-p38-MAPK signaling pathway.
- HCT shows potential as a therapeutic agent for ocular diseases characterized by microglial overactivation.
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