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. 2024 Mar 26;36(2):152-165.
doi: 10.4103/tcmj.tcmj_267_23. eCollection 2024 Apr-Jun.

Protective effects of Jing-Si-herbal-tea in inflammatory cytokines-induced cell injury on normal human lung fibroblast via multiomic platform analysis

Chien-Hao Wang  1 , Jai-Sing Yang  2 , Chao-Jung Chen  3   4 , San-Hua Su  1 , Hsin-Yuan Yu  1 , Yu-Ning Juan  2 , Yu-Jen Chiu  5   6   7 , Tsung-Jung Ho  8   9
Affiliations

Protective effects of Jing-Si-herbal-tea in inflammatory cytokines-induced cell injury on normal human lung fibroblast via multiomic platform analysis

Chien-Hao Wang et al. Tzu Chi Med J. .

Abstract

Objectives: The protective effects and related mechanisms of Jing-Si herbal tea (JSHT) were investigated in cellular damage mediated by pro-inflammatory cytokines, including interleukin (IL)-1β, IL-6, and tumor necrosis factor-α, on normal human lung fibroblast by multiomic platform analysis.

Materials and methods: The in silico high-throughput target was analyzed using pharmacophore models by BIOVIA Discovery Studio 2022 with ingenuity pathway analysis software. To assess cell viability, the study utilized the MTT assay technique. In addition, the IncuCyte S3 ZOOM System was implemented for the continuous monitoring of cell confluence of JSHT-treated cytokine-injured HEL 299 cells. Cytokine concentrations were determined using a Quantibody Human Inflammation Array. Gene expression and signaling pathways were determined using next-generation sequencing.

Results: In silico high-throughput target analysis of JSHT revealed ingenuity in canonical pathways and their networks. Glucocorticoid receptor signaling is a potential signaling of JSHT. The results revealed protective effects against the inflammatory cytokines on JSHT-treated HEL 299 cells. Transcriptome and network analyses revealed that induction of helper T lymphocytes, TNFSF12, NFKB1-mediated relaxin signaling, and G-protein coupled receptor signaling play important roles in immune regulatory on JSHT-treated cytokine-injured HEL 299 cells.

Conclusion: The findings from our research indicate that JSHT holds promise as a therapeutic agent, potentially offering advantageous outcomes in treating virus infections through various mechanisms. Furthermore, the primary bioactive components in JSHT justify extended research in antiviral drug development, especially in the context of addressing coronavirus.

Keywords: Coronavirus; Cytokines; Lung injury; Multiomic; Protective effects.

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Conflict of interest statement

There are no conflicts of interest.

Figures

Figure 1
Figure 1
(a) Jing-Si herbal tea (JSHT) formula developed by Hualien Tzu Chi Hospital’s efforts in addressing coronavirus disease 2019 infection and modulating immune responses. (b) Study design and JSHT schematics. NGS: Next-generation sequencing
Figure 2
Figure 2
(a) Top 20 ingenuity canonical pathways of Jing-Si herbal tea (JSHT) by in silico high throughput target screening analysis. (b) Ingenuity pathway core analysis for potential cytokines gene targets of JSHT. JSHT: Jing-Si herbal tea, IL: Interleukin, TNF: Tumor necrosis factor
Figure 3
Figure 3
In silico pathway core analysis for potential targets of Jing-Si herbal tea in (a) lung inflammation genes; (b) lower respiratory tract disorder; (c) the viral infection genes
Figure 4
Figure 4
UPLC-Q-TOF/MS analysis data showing standard peaks of chlorogenic acid and glycyrrhetinic acid. JSHT: Jing-Si herbal tea, CA: Chlorogenic acid, GA: Glycyrrhetinic acid
Figure 5
Figure 5
Cell viability exhibited a significant concentration-dependent decrease in the absence of Jing-Si herbal tea (JSHT) extract, calceolarioside A, and chlorogenic acid treatments when exposed to tumor necrosis factor (TNF)-α, interleukin (IL)-1β, and IL-6. (a) HEL 299 cells were seeded at a density of 2.5 × 105 cells/mL/well and subsequently exposed to individual cytokines (TNF-α, IL-1β, and IL-6) at a concentration of 10 ng/mL, along with JSHT at concentrations of 250, 500, 750, and 1000 μg/mL, for a duration of 24 h. (b) The HEL 299 cells were treated with TNF-α/IL-1β/IL-6 (10 ng/mL, individual) and calceolarioside A, chlorogenic acid, glycyrrhetic acid (50–100 μM) for 24 h. The cells were treated with 0.1% DMSO as control treatment. Cell viability was measured by the MTT assay (n = 3). The obtained results were subjected to statistical analysis employing a one-way analysis of variance, followed by Tukey’s post hoc test. Significance levels were denoted as ***P < 0.001 and ###P < 0.001. (c) Real-time cellular confluence after TNF-α/IL-1β/IL-6 and JSHT treatments changed in a time-dependent manner. JSHT: Jing-Si herbal tea, IL: Interleukin, TNF: Tumor necrosis factor
Figure 6
Figure 6
The cytokines concentration of Jing-Si-herbal-tea (JSHT) by protein microarray detection analysis. Cultured HEL 299 cells at a concentration of 2.5 × 105 cells/mL/well were subjected to treatment with individual cytokines, namely tumor necrosis factor-α, interleukin (IL)-1β, and IL-6, each at a concentration of 10 ng/mL, in combination with JSHT at a concentration of 1000 μg/mL. The exposure duration for this experimental setup was 24 h. Cytokines concentration was measured by the protein microarray (n = 3). The acquired data were subjected to statistical analysis employing a one-way analysis of variance, followed by post hoc testing using Tukey’s method. ***P < 0.001 and ###P < 0.001. JSHT: Jing-Si herbal tea, IL: Interleukin, TNF: Tumor necrosis factor
Figure 7
Figure 7
Transcriptome analysis of Jing-Si herbal tea (JSHT)-treated cytokine-injured HEL 299 cells was shown. (a) Differential expression of MA and (b) volcano plots. Cytokine: Tumor necrosis factor -α/interleukin (IL)-1β/IL-6 (10 ng/mL, individual); JSHT. (c) Whole-transcriptome sequencing and comparative analysis using the ingenuity pathway analysis software on JSHT-treated cytokine-injured HEL 299 cells. Blue and red coloring indicates the activation of Z-score values. JSHT: Jing-Si herbal tea, IL: Interleukin
Figure 8
Figure 8
(a) A network depicting associations among various genes was constructed through IPA analysis. Induction of helper T lymphocytes, TNFSF12, MYD88, STIRG1, RELA, CREBBP, NFKB1-mediate relaxin signaling, and G-protein coupled receptor signaling play key roles in immune regulatory pathways. (b) The target genes and associated pro-inflammatory cytokines in the pulmonary system of JSTH-treated cytokine-injured HEL 299 cells were analyzed using IPA software to determine the transcriptional profile. (1: Cytokine (tumor necrosis factor-α/interleukin [IL]-1β/IL-6) treatment; 2: Cytokine with Jing-Si herbal tea treatment. IL: Interleukin, TNF: Tumor necrosis factor
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