RNAseq analysis of the drug jian-yan-ling (JYL) using both in vivo and in vitro models

Xiaobo Zhang; Yunliang Zhai; Dandan Zhang; Chang Che; Yayun Zhang; Quan Li; Xue Zhang; Lingrui Zhao

doi:10.1016/j.heliyon.2023.e16143

RNAseq analysis of the drug jian-yan-ling (JYL) using both in vivo and in vitro models

Heliyon. 2023 May 19;9(5):e16143. doi: 10.1016/j.heliyon.2023.e16143. eCollection 2023 May.

Authors

Xiaobo Zhang¹, Yunliang Zhai², Dandan Zhang², Chang Che³, Yayun Zhang², Quan Li², Xue Zhang², Lingrui Zhao³

Affiliations

¹ School of Life Sciences, Fudan University, Shanghai, 200438, China.
² Lei Yun Shang Pharmaceutical Group Co.,Ltd., Suzhou, 215009, China.
³ Key Laboratory of Growth Regulation and Translational Research of Zhejiang Province, School of Life Sciences, Westlake University, Hangzhou, 310024, China.

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