Abstract

Ethnopharmacological relevance

   Jian-yan-ling (JYL) is a drug used in traditional Chinese medicine
   (TCM) prescriptions for the treatment of tumors after radiotherapy and
   chemotherapy, to effectively alleviate leukocytopenia. However, the
   genetic mechanisms underlying the function of JYL remain unclear.

Aim of the study

   This study aimed to explore the RNA changes and potential biological
   processes related to the anti-aging or life-extending effects of JYL
   treatments.

Materials and methods

   In vivo treatments were performed using Canton-S Drosophila
   corresponding to three groups: control, low-concentration (low-conc.),
   and high-concentration (high-conc.) groups. The low-conc. And the
   high-conc. Groups were treated with 4 mg/mL JYL and 8 mg/mL JYL,
   respectively. Thirty Drosophila eggs were placed in each vial, and the
   third instar larvae and adults 7 and 21 days post-eclosion were
   collected for RNA sequencing, irrespective of the gender.

   In vitro treatments were conducted using humanized immune cell lines
   HL60 and Jurkat, which were divided into 3 groups: control (0 μg/mL
   JYL), low-concentration (40 μg/mL JYL), and high-concentration
   (80 μg/mL JYL). The cells were collected after 48 h of each JYL drug
   treatment. Both the Drosophila and cell samples were analyzed using RNA
   sequencing.

Results

   The in vivo experiments revealed 74 upregulated genes in the
   low-concentration group, and CG13078 was a commonly downregulated
   differential gene, which is involved in ascorbate iron reductase
   activity. Further analysis of the co-expression map identified the key
   genes: regulatory particle non-ATPase (RPN), regulatory particle
   triple-A ATPase (RPT), and tripeptidyl-peptidase II (TPP II). For the
   in vitro experiments, 19 co-differential genes were compared between
   different concentrations of the HL 60 cell line, of which three genes
   were upregulated: LOC107987457 (phostensin-like gene), HSPA1A (heat
   shock protein family A member 1 A), and H2AC19 (H2A clustered histone
   19). In the HL 60 cell line, JYL activated proteasome-related
   functions. In the Jurkat cell line, there were no common differential
   genes despite the presence of a dosage-dependent trend.

Conclusions

   The RNA-seq results showed that the traditional Chinese medicine JYL
   has longevity and anti-aging effects, indicating that further
   investigation is required.

   Keywords: Jian-yan-ling (JYL) drug, RNA sequencing, Drosophila
   melanogaster, HL60 cell line, Protein ubiquitination decomposition,
   anti-aging

1. Introduction

   The modern development of ancient Chinese medicines is embodied by
   traditional Chinese medicine (TCM) prescriptions. Historically, the use
   of Western medicines has increased, as inferred from the evidence
   provided by quantitative and standardized analyses, while the use of
   TCM prescriptions has also increased due to the accumulation of
   evidence showing their curative effects and independent folk
   collections (including traditional Chinese medical books and
   prescriptions transmitted by word of mouth) and compilations. There is
   consequently a need to improve our understanding of TCM treatments
   using current scientific techniques, such as our rapidly developing
   genetic technologies. TCM prescriptions have been successfully utilized
   for thousands of years to treat various diseases. For instance,
   artemisinin and its derivatives, an extract from Artemisia annua, are
   front-line antimalarial drugs known for their efficacy and low toxicity
   [[37]23]. Although artemisinin is the standard treatment for malaria,
   its underlying genetic mechanisms remain poorly understood. Western
   medicines are developed based on one or a few active compounds, to
   influence several specific biologically activity targets or pathways,
   so their pharmacological functions can be determined with relative
   ease. In contrast, TCM prescriptions contain multiple active compounds.
   The “multi-component, multi-target” nature of TCM prescriptions poses a
   challenge when trying to elucidate their underlying molecular
   mechanisms (see [38]Fig. 7).

Fig. 7.

   [39]Fig. 7
   [40]Open in a new tab

   Co-expression analysis and function enrichment of the HL60 cell line.

   In recent years, with the development of high-throughput technologies
   and innovative computational methods in drug discovery, the desire to
   elucidate the molecular mechanisms of TCM have been reinvigorated.
   Pharmacotranscriptomes have become a powerful tool by which to evaluate
   drug TCM drug efficacy and discover new drug targets that are based on
   several pivotal consensuses: gene expression can be accurately measured
   and could potentially be a “universal language” for disease
   characterization and prediction [[41]24]; gene expression can link
   different biological states and systems [[42]4]; different biological
   pathways can drive different disease phenotypes. Thus, TCM prescription
   research has increasingly turned to high-throughput transcriptome
   screening to understand their molecular effects and components. Many
   studies have examined the genetic targets of herbs and ingredients and
   linked them to various diseases.

   The Integrated Database of Traditional Chinese Medicine (TCMID) was
   first established in 2013 with the aim of creating a comprehensive
   database for the digitization and standardization of TCM [[43]24]. This
   has been recognized by pharmacists and Chinese medicine researchers.
   TCMID 2.0 was later updated in 2017 with data relating to 18 203 TC M
   ingredients, 15 prescriptions, 82 related targets, 1356 drugs, 842
   diseases, and various associations among the data. Considering that
   chemical changes may occur after the formula is decocted or boiled, new
   data were collected based on the composition of the formula after the
   processing steps. Meanwhile, 3895 mass spectrometry (MS) data points
   for 729 Chinese herbs were added as supplementary materials for
   ingredient identification. TCMID 2.0 will further promote the
   modernization of TCM and facilitate the exploration of their underlying
   biological processes. HERB [[44]4], another Chinese medicine database
   for high-throughput experimental references, was recently established