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Mechanistic Insights into Shenzhuo Formula for Diabetic Retinopathy: Integrating UPLC-Q-TOF-MS/MS, Network
Xiaoyu Zang1, Lili Zhang2, Jing Ma1,3
1Changchun University of Chinese Medicine, Changchun, People's Republic of China.
Purpose:
For early-stage Diabetic retinopathy (DR), various pharmacological agents and neuroprotective factors have been developed. However, these treatments often show limited efficacy, especially when initiated after retinal damage, and may cause adverse effects. Therefore, there is an urgent need to develop safer and more effective therapeutic strategies for early-stage DR. Shenzhuo Formula (SZF), a modified classical traditional Chinese medicine prescription, has shown promising clinical efficacy in early-stage DR treatment. This study aims to investigate the underlying mechanisms of SZF to expand treatment strategies for DR.
Methods:
SZF components were analyzed using Ultra Performance Liquid Chromatography-Quadrupole Time-of-Flight Mass Spectrometry/Mass Spectrometry (UPLC-Q-TOF-MS/MS). Db/db mice received three different SZF doses for 12 weeks. Physiological parameters, including water and food consumption, body weight, and urine output, were monitored. Blood samples were analyzed for fasting blood glucose and other relevant parameters. Ocular changes were assessed using fundus photography (FP), fundus fluorescein angiography (FFA), optical coherence tomography (OCT) and hematoxylin and eosin (H&E). Network pharmacology analysis (NP) identified potential SZF targets, while immunofluorescence staining evaluated SZF's mechanism in delaying DR progression. The distribution of SZF pharmacological targets in critical DR target cells was analyzed using single-cell data from the GSE245561 dataset. Molecular docking predicted SZF-target interactions.
Results:
SZF improved diabetic symptoms, increased retinal thickness, and reducedvascular leakage and microcirculation issues. The HIF-1α-VEGFA axis was suggested as a potential core target. Single-cell analysis of clinical samples suggested macrophages as a common target cell for HIF-1α and VEGFA. Molecular docking identified effective SZF components.
Conclusion:
Results indicate that SZF may impede the progression of DR by inhibiting the HIF-1α-VEGFA signaling pathway in macrophages, with quercetin and apigenin identified as significant contributors, though further experimental validation is needed to confirm these mechanistic.
Insights
Shenzhuo Formula (SZF) shows promise in treating early-stage diabetic retinopathy (DR) by targeting the HIF-1α-VEGFA pathway in macrophages. This traditional Chinese medicine may offer a safer alternative for DR management.
Area of Science:
- Ophthalmology
- Pharmacology
- Traditional Chinese Medicine
Background:
- Current treatments for early-stage diabetic retinopathy (DR) have limited efficacy and potential adverse effects.
- There is a critical need for safer and more effective therapeutic strategies for early-stage DR.
- Shenzhuo Formula (SZF), a modified traditional Chinese medicine, demonstrates potential clinical benefits in early-stage DR.
Purpose of the Study:
- To investigate the underlying mechanisms of Shenzhuo Formula (SZF) in treating early-stage diabetic retinopathy (DR).
- To identify potential therapeutic targets and active components of SZF for DR treatment.
- To expand treatment strategies for DR using traditional Chinese medicine.
Main Methods:
- SZF components were analyzed using UPLC-Q-TOF-MS/MS.
- Db/db mice were treated with SZF, and physiological and ocular parameters were assessed.
- Network pharmacology, immunofluorescence, single-cell analysis, and molecular docking were employed to elucidate SZF's mechanism of action.
Main Results:
- SZF treatment improved diabetic symptoms, increased retinal thickness, and reduced vascular leakage and microcirculation abnormalities in mice.
- The HIF-1α-VEGFA axis was identified as a potential core target pathway for SZF's efficacy.
- Macrophages were suggested as a key target cell type, with quercetin and apigenin identified as significant SZF components.
Conclusions:
- SZF may inhibit DR progression by targeting the HIF-1α-VEGFA signaling pathway in macrophages.
- Quercetin and apigenin are potential active compounds within SZF contributing to its therapeutic effects.
- Further experimental validation is required to confirm the precise mechanisms of SZF in DR treatment.
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