Mechanisms of Lycium barbarum Treatment in Sepsis‐Induced Myocardial Injury: A Network Pharmacology and Experimental Validation Study

Figures

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  FIGURE 1. LB component‐SIMI target network construction. (A) Grouped clustering tree diagram for GEO chip data. (B) WGCNA analysis to determine SIMI's most relevant modules. (C) Venn diagram illustrating the identification of 62 candidate SIMI‐related genes. (D) Intersection of LB targets with SIMI‐related genes and reliable genes obtained from PubMed. (E) LB active ingredient‐target network.

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  FIGURE 2. PPI network construction and functional analysis of 49 common targets.

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  FIGURE 3. Core protein target screening. (A) PPI network diagram for 20 key protein targets screening. (B) GO and (C) KEGG enrichment analysis of core targets.

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  FIGURE 4. LB inhibits HL‐1 cardiomyocyte viability in the presence of LPS. (A) Cell viability of HL‐1 cells under LPS and LPS + LB treatments showing that LB restored cell viability impaired by LPS (n = 3). (B) RT‐qPCR analysis of core targets including AKT1, Bcl‐2, ESR1, JUN, and TP53, with LB reversing the expression changes of most targets induced by LPS (n = 3). (C) Western blot analysis of core targets of apoptosis (cleaved caspase‐3, Bcl‐2, and Bax) and inflammation (IL‐6, IL‐1β, and TNF‐α), demonstrating that LB attenuated LPS‐induced increases in apoptotic and inflammatory proteins (n = 3). (D) Western blot verified the effect of LB on MAPK pathway‐related proteins in HL‐1 cells (n = 3). Data were shown as mean ± SD. Statistical significance was determined by one‐way ANOVA followed by Tukey's post hoc test. *p < 0.05, **p < 0.01 and ***p < 0.001.

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  FIGURE 5. LB protected mice cardiomyocytes after CLP. (A) H&E staining and TUNEL immunohistochemistry of mouse cardiomyocytes, showing that LB alleviated myocardial damage and apoptosis (n = 6). (B) Average optical density of TUNEL immunohistochemistry in mice (n = 6). (C, D) Western blot and quantification results of inflammatory markers (IL‐6, IL‐1β) and apoptotic markers (Bax, Bcl‐2, cleaved caspase‐3) in mice, showing that LB normalized the expression of key apoptotic and inflammatory proteins altered by CLP (n = 6). Data were shown as mean ± SD. Statistical significance was determined by one‐way ANOVA followed by Tukey's post hoc test. *p < 0.05, **p < 0.01, and ***p < 0.001.

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  FIGURE 6. Combination of primary compounds and protein targets including (A) AKT1‐Q, (B) Bcl‐2‐B, (C) Bcl‐2‐Q, (D) ESRD‐G, (E) JUN‐B, (F) JUN‐Q, (G) TP53‐Q. B, β‐sitosterol; G, glycitein; Q, quercetin.

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  FIGURE 7. Immunohistochemistry staining showed alleviated SIMI by quercetin treatment, with n = 6 in each group of mice.