The efficacy and mechanism of Angelica sinensis (Oliv.) Diels root aqueous extract based on RNA sequencing and 16S rDNA sequencing in alleviating polycystic ovary syndrome

Study information

Reviewed Marked as Reviewed by KateRasheed on 2025-8-11

study design

laboratory experiment

Citation

PMID PubMed identifier for scientific articles.

37639812

DOI Digital object identifier for electronic documents.

10.1016/j.phymed.2023.155013

URI Uniform resource identifier for web resources.

Authors

Gao Y, Mo S, Cao H, Zhi Y, Ma X, Huang Z, Li B, Wu J, Zhang K, Jin L

Journal

Phytomedicine : international journal of phytotherapy and phytopharmacology

Year

2023

Keywords:

16S rDNA sequencing, Angelica sinensis (oliv.) Diels root, Polycystic ovary syndrome, RNA sequencing

BACKGROUND: Polycystic ovary syndrome (PCOS) leads to persistent anovulation, hyperandrogenism, insulin resistance, and polycystic ovary, and is mainly characterized by menstrual disorders, and reproductive dysfunction. Angelica sinensis (Oliv.) Diels root has been used in many classical formulas of traditional Chinese medicine, and is commonly used to treat various gynecological diseases. PURPOSE: To investigate the protective effect of water extract of A. sinensis root (WEA) on PCOS rats, and the mechanism by RNA sequencing, and 16S rDNA sequencing. METHODS: The PCOS rat model was established by letrozole combined with high-fat diet (gavage; 2 months), and treated with WEA (gavage; 2 g/kg, 4 g/kg or 8 g/kg; 1 month). To evaluate the therapeutic effect of WEA on PCOS rats, vaginal smear, hematoxylin-eosin staining, and biochemical indicators detection were performed. The rat ovarian tissue was analyzed by RNA sequencing, and the results were verified by qRT-PCR, and Western blot. 16S rDNA sequencing was used to analyze the gut microbiota of rats. RESULTS: The results of the vaginal smear, and hematoxylin-eosin staining showed that WEA improved estrous cycle disorder, and ovarian tissue lesions. WEA (4 g/kg or 8 g/kg; 1 months) alleviated hormone disorders, insulin resistance, and dyslipidemia. RNA sequencing showed that WEA intervention significantly changed the expressions of 2756 genes, which were enriched in phosphatidylinositol3-kinase/phosphorylated protein kinase B (PI3K/AKT), peroxisome proliferator-activated receptor (PPAR), mitogen-activated protein kinase (MAPK), AMP-activated protein kinase (AMPK), and insulin signaling pathways. 16S rDNA sequencing found that WEA increased the species diversity of gut microbiota, and regulated the abundance of some microbiota (genus level: Dubosiella, Bifidobacterium, Coriobacteriaceae (UCG-002), and Treponema; species level: Bifidobacterium animalis, Lactobacillus murinus, and Lactobacillus johnsonii). CONCLUSION: WEA regulated hormone, and glycolipid metabolism disorders, thereby relieving the PCOS induced by letrozole combined with high-fat diet. The mechanism was related to the regulation of PI3K/AKT, PPAR, MAPK, AMPK, and insulin signaling pathways in ovarian tissues, and the maintenance of gut microbiota homeostasis. Clarifying the efficacy and mechanism of WEA in alleviating PCOS based on RNA sequencing and 16S rDNA sequencing will guide the more reasonable clinical use of WEA.

Experiment 1

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Reviewed Marked as Reviewed by KateRasheed on 2025-8-11

Curated date: 2025/07/16

Curator: Victoria

Revision editor(s): Victoria

Subjects

Location of subjects: China

Host species Species from which microbiome was sampled. Contact us to have more species added.: Rattus norvegicus

Body site Anatomical site where microbial samples were extracted from according to the Uber Anatomy Ontology: Feces Cow dung,Cow pat,Droppings,Dung,Excrement,Excreta,Faeces,Fecal material,Fecal matter,Fewmet,Frass,Guano,Matières fécales@fr,Merde@fr,Ordure,Partie de la merde@fr,Piece of shit,Porción de mierda@es,Portion of dung,Portion of excrement,Portion of faeces,Portion of fecal material,Portion of fecal matter,Portion of feces,Portion of guano,Portion of scat,Portionem cacas,Scat,Spoor,Spraint,Stool,Teil der fäkalien@de,Feces,feces

Condition The experimental condition / phenotype studied according to the Experimental Factor Ontology: Polycystic ovary syndrome Cystic disease of ovaries,hyperandrogenemia,Multicystic ovaries,multicystic ovaries,Ovarian Degeneration, Sclerocystic,Ovarian Syndrome, Polycystic,Ovarian Syndromes, Polycystic,Ovaries, Sclerocystic,Ovary Syndrome, Polycystic,Ovary, Sclerocystic,PCO - Polycystic ovaries,Pco1,PCOD - Polycystic ovarian disease,PCOS,Pcos,PCOS - Polycystic ovarian syndrome,PCOS1,Polycystic ovarian disease,polycystic ovarian disease,Polycystic ovarian syndrome,Polycystic ovaries,polycystic ovaries,Polycystic ovaries (disorder),polycystic ovary,polycystic ovary syndrome,polycystic ovary syndrome 1,Sclerocystic Ovarian Degeneration,Sclerocystic Ovaries,Sclerocystic Ovary,Sclerocystic Ovary Syndrome,Stein Leventhal Syndrome,Stein-Leventhal synd.,Stein-Leventhal Syndrome,Stein-Leventhal syndrome,Syndrome, Polycystic Ovary,Syndrome, Stein-Leventhal,Polycystic ovary syndrome

Group 0 name Corresponds to the control (unexposed) group for case-control studies: Control

Group 1 name Corresponds to the case (exposed) group for case-control studies: Polycystic ovary syndrome (PCOS)

Group 1 definition Diagnostic criteria applied to define the specific condition / phenotype represented in the case (exposed) group: Rats with polycystic ovary syndrome (PCOS) were induced by letrozole and a high-fat diet.

Group 0 sample size Number of subjects in the control (unexposed) group: 5

Group 1 sample size Number of subjects in the case (exposed) group: 5

Lab analysis

Sequencing type: 16S

16S variable region One or more hypervariable region(s) of the bacterial 16S gene: Not specified

Sequencing platform Manufacturer and experimental platform used for quantifying microbial abundance: Illumina

Statistical Analysis

Data transformation Data transformation applied to microbial abundance measurements prior to differential abundance testing (if any).: relative abundances

Statistical test: LEfSe

Significance threshold p-value or FDR threshold used for differential abundance testing (if any): 0.05

MHT correction Have statistical tests be corrected for multiple hypothesis testing (MHT)?: No

LDA Score above Threshold for the linear discriminant analysis (LDA) score for studies using the popular LEfSe tool: 2

Alpha Diversity

Shannon Estimator of species richness and species evenness: more weight on species richness: decreased

Chao1 Abundance-based estimator of species richness: unchanged

Simpson Estimator of species richness and species evenness: more weight on species evenness: decreased

Richness Number of species: unchanged

Signature 1

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Reviewed Marked as Reviewed by KateRasheed on 2025-8-11

Curated date: 2025/07/17

Curator: Victoria

Revision editor(s): Victoria

Source: Figure 9I

Description: The effects of WEA on the gut microbiota of PCOS rats using LEfSe.

Abundance in Group 1: increased abundance in Polycystic ovary syndrome (PCOS)

NCBI	Quality Control	Links
Erysipelotrichales
Erysipelotrichaceae
Allobaculum
Bacteroidota
Bacteroidia
Bacteroidales
Muribaculaceae
Bacteroides
Bacteroidaceae

Revision editor(s): Victoria

Signature 2

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Reviewed Marked as Reviewed by KateRasheed on 2025-8-11

Curated date: 2025/07/17

Curator: Victoria

Revision editor(s): Victoria

Source: Figure 9I

Description: The effects of WEA on the gut microbiota of PCOS rats using LEfSe.

Abundance in Group 1: decreased abundance in Polycystic ovary syndrome (PCOS)

NCBI	Quality Control	Links
Limosilactobacillus reuteri
Lactobacillus acidophilus
Lactobacillaceae
Lactobacillus
Lactobacillales

Revision editor(s): Victoria

Experiment 2

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Reviewed Marked as Reviewed by KateRasheed on 2025-8-11

Curated date: 2025/07/16

Curator: Victoria

Revision editor(s): Victoria, Ese

Differences from previous experiment shown

Subjects

Group 0 name Corresponds to the control (unexposed) group for case-control studies: Polycystic ovary syndrome

Group 1 name Corresponds to the case (exposed) group for case-control studies: Polycystic ovary syndrome + Water extract of Angelica sinensis root (PCOS+WEA)

Group 1 definition Diagnostic criteria applied to define the specific condition / phenotype represented in the case (exposed) group: Rats with polycystic ovary syndrome (PCOS) were induced by letrozole and a high-fat diet and administered intragastrically with metformin (0.3g/kg) and WEA (8g/kg) for 4 weeks as treatment.

Lab analysis

Statistical Analysis

Alpha Diversity

Shannon Estimator of species richness and species evenness: more weight on species richness: increased

Chao1 Abundance-based estimator of species richness: unchanged

Simpson Estimator of species richness and species evenness: more weight on species evenness: increased

Richness Number of species: unchanged

Signature 1

Edit History Delete

Flag for review

Your review change was submitted

Reviewed Marked as Reviewed by KateRasheed on 2025-8-11

Curated date: 2025/07/17

Curator: Victoria

Revision editor(s): Victoria

Source: Figure 9J

Description: The effects of WEA on the gut microbiota of PCOS rats using LEfSe.

Abundance in Group 1: increased abundance in Polycystic ovary syndrome + Water extract of Angelica sinensis root (PCOS+WEA)

NCBI	Quality Control	Links
Acidimicrobiia
Atopobiaceae
Bifidobacteriaceae
Bifidobacteriales
Bifidobacterium
Bifidobacterium animalis
Clostridia
Coriobacteriales
Coriobacteriia
Dubosiella
Eubacteriales
Ligilactobacillus murinus
Spirochaetaceae
Spirochaetales
Spirochaetia
Treponema
Clostridia UCG-014Clostridia UCG-014
Coriobacteriaceae UCG-002Coriobacteriaceae UCG-002