Uterine Metabolomics Reveals Protection of Taohong Siwu Decoction Against Abnormal Uterine Bleeding

Abstract

Incomplete abortion, a procedure for terminating pregnancy, will lead to abnormal uterine bleeding (AUB), infections, and even death. Taohong Siwu decoction (TSD) is a traditional Chinese medicine (TCM) formula, which has been developed to treat AUB for hundreds of years. However, the mechanism of the protective effect of TSD against AUB is not clear. We performed mass spectrometry (MS) of uterine samples to observe metabolic profile resulting from the treatment with TSD. An integrated gas chromatography-mass spectrometry and liquid chromatography-mass spectrometry based untargeted metabolomics approach combined with multivariate statistical analyses were used to investigate the metabolic profile of TSD against AUB. There was clear separation between pregnant and incomplete aborting rats as well as incomplete aborting and TSD administered rats. Based on random forest algorithm and receiver operator characteristic analysis, 12 biomarkers were optimized related to TSD administered. The effect of TSD on AUB are related to several pathways, such as AA metabolism, glyoxylate and dicarboxylate metabolism, alanine, aspartate, and glutamate metabolism. To our knowledge, this is the first uterine metabolomics study focusing on TSD on AUB and provide a new perspective for explaining the mechanism of TSD on AUB.

Keywords: Taohong Siwu decoction; UFLC-IT-TOF/MS; abnormal uterine bleeding; gas chromatography coupled with mass spectrometry; metabolomics.

Figure 1

Macroscopic (a–d) and Pathological (A–D) observation of uteri tissue. Samples were performed with HE as described in Methods. a, A: group P; b, B: group M; c, C: group T. d, D: group Y. Black arrows indicate dark red residue, red arrows indicate necrotic decidual cells. Duration (E) and volume (F) of uterine bleeding. The duration of uterine bleeding was recorded from the beginning of uterine bleeding after the administration of mifepristone to the cessation of bleeding completely. The unit of duration was measured in hours. Adopting the method of Alkaline Hematin Photometric to measure the volume of uterine bleeding in different groups. The unit of volume is in ml. ^#P < 0.05 vs group P. *P < 0.05 vs group M.

Figure 2

Score plots (A–C) of OPLS-DA Validation plots (D–F) obtained from 999 random permutation tests results between P (square) and M (triangle) in uterine tissues; A, D: GC/MS; B, E: UFLC-IT-TOF/MS (ESI+); C, F: UFLC-IT-TOF/MS (ESI-).

Figure 3

Score plots (A–C) of OPLS-DA Validation plots (D–F) obtained from 999 random permutation tests results between M (triangle) and T (rhombus) in uterine tissues; a, d: GC/MS; b, e: UFLC-IT-TOF/MS (ESI+); c, f: UFLC-IT-TOF/MS (ESI-).

Figure 4

Biomarkers’ heatmap related to medical abortion (A) and intervention of TSD (B) in uterine tissue. Red area: an increase of up to twofolds; blue area: a reduction of up to twofolds.

Figure 5

Analysis of metabolic pathway associated with PM (A) and MT (B) in uterine tissues based on network enrichment and network topology analysis.

Figure 6

Metabolic pathways associated with PM (A) and MT (B) in uterine tissue. Red: increased metabolites level; Green: decreased metabolites level; solid arrows: direct metabolic relationship; dashed arrows: indirect metabolic relationship; Blue box: metabolic pathway of metabolites.