Exploring the effect and mechanism of action of Jinlida granules (JLD) in the treatment of diabetes-associated cognitive impairment based on network pharmacology with experimental validation

Abstract

Objectives: To explore the effect and the probable mechanisms of JLD in the treatment of type 2 diabetes mellitus (T2DM) - associated cognitive impairment (TDACI).

Methods: The effect of JLD in combating TDACI was assessed in T2DM model mice by conducting Morris water maze (MWM) behaviour testing. Active components and their putative targets, as well as TDACI-related targets, were collected from public databases. Protein-protein interactions (PPIs), Gene Ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses and molecular docking were then utilized to explore potential molecular network mechanisms. Finally, the main targets were verified in animal model experiments.

Results: MWM test showed that JLD improved aspects of behaviour in T2DM model mice. JLD improved glucose intolerance, tissue insulin sensitivity, lipid metabolism and enhanced synapse-associated protein expression in hippocampus tissue. Network pharmacology revealed 185 active components, 337 targets of JLD, and 7998 TDACI related targets were obtained . PPI network analyses revealed 39 core targets. GO and KEGG analyses suggested that JLD might improve TDACI by regulating gene expression, apoptotic processes and inflammatory responses mainly via PI3K-AKT and AGE-RAGE signaling pathways. Molecular docking revealed strong binding of the main components to core targets. JLD reduced hippocampus tissue expression of the inflammatory cytokines tumor necrosis factor-α (TNF-α) and interleukin-6 (IL6), core targets of treatment of TDACI.

Conclusions: The findings suggested that JLD has the potential to improve TDACI through multiple components, multiple targets and multiple pathways. JLD may be a promising treatment for diabetic cognitive impairment.

Keywords: Jinlida granule; Type 2 diabetes mellitus; cognitive impairment; inflammation; network pharmacology.

Figure 1.

JLD ameliorated cognitive impairment assessed by MWM testing in T2DM mice. (A) The swimming trajectory of each group tested on the sixth day. (B) Escape latency (in seconds) in the spatial probe test. (C) Number of times passing the platform. (D) Time (seconds) spent in the target quandrant. (E) Protein expression of synapse associated proteins relative to β-actin in hippocampus tissue. Data are presented as mean ± SD (n = 7 per group). *p < 0.05, **p < 0.01 and ***p < 0.001 vs. Control; #p < 0.05, ##p < 0.01 vs. T2DM.

Figure 2.

JLD improved glucose and lipid metabolism. (A) Intraperitoneal glucose tolerance test (IPGTT). (B) The area under the IPGTT curve. (C) Intraperitoneal insulin tolerance test (IPITT). (D) The area under the IPGTT curve. (E) Total cholesterol (TC). (F) Triglyceride (TG). (G) Low-density lipoprotein cholesterol (LDL-C). (H) High-density lipoprotein cholesterol (HDL-C). *p < 0.05, **p < 0.01 and ***p < 0.001 vs. Control; #p < 0.05, ##p < 0.01 and ###p < 0.001 vs. T2DM.

Figure 3.

Potential therapeutic targets and component-target-disease (C-T-D) network construction. (A) Venn diagram showing the therapeutic targets. (B) C-T-D network. Purple arrows represent herbs in JLD, blue circles are components and green ovals are the predicted therapeutic targets.

Figure 4.

Identification of core targets via PPI analysis.

Figure 5.

Gene Ontology (GO) and Kyoto Encyclopedia of genes and Genomes (KEGG) enrichment analysis. (A) Bubble chart of the biological processes from GO enrichment analysis; (B) Bubble chart of the cellular components from GO enrichment analysis; (C) Bubble chart of the molecular functions from GO enrichment analysis; (D) Bubble chart of pathways from KEGG enrichment analysis. The color and size of each bubble represent the P value and gene count, respectively. (E) Component-target-pathway (C-T-P) network. The green nodes represent the components; the purple rectangles represent the targets; the blue nodes represent the pathways.

Figure 6.

Schematic diagrams of key targets and specific components. (A) PTGS2-Quercetin; (B) PPARG-Luteolin; (C) HSP90AB1-Baicalin; (D) RELA-Quercetin; (E) CASP3-Quercetin; (F) BCL2-Quercetin; (G) AKT1-Luteolin; (H) ESR1-Quercetin; (I) TP53-Luteolin; (J) GSK3B-Quercetin; (K) TNF-Quercetin; (L) IL6-Quercetin.

Figure 7.

JLD significantly reduced the expression of inflammatory cytokines in hippocampus tissue. (A) The relative levels of TNF-α mRNA; (B) The protein expression of TNF-α; (C) The relative levels of IL6 mRNA; (D) The protein expression of IL6. *p < 0.05 and **p < 0.01 vs. Control; #p < 0.05 and ##p < 0.01 vs. T2DM.