Mechanism of Mongolian mind-body interactive therapy in regulating essential hypertension through HTR2B: A metabolome- and transcriptome-based study

Abstract

Essential hypertension is a psychosomatic disease associated with emotions and behaviors. Although Mongolian mind-body interactive therapy can help patients with essential hypertension reduce their systolic blood pressure (SBP), the mechanism is unclear. We assigned patients who underwent Mongolian mind-body interactive therapy to groups that were treated with (DT) or without (NDT) antihypertensive drugs (Clinical registration no: ChiCTR2000034918). We screened differentially expressed genes (DEGs) using targeted metabolic and transcriptomic analyses of blood samples before and after intervention. Sequenced data were analyzed using quantitative polymerase chain reaction (qPCR) and validated using enzyme-linked immunosorbent assays (ELISAs). Small interfering (Si)-RNA interference on key DEGs in human umbilical vein endothelial cells (HUVECs) was experimentally verified. Omics analysis identified 187 DEGS, including human 5-hydroxytryptamine (5-HT) receptor 2B (5-HTR2B), human endothelin receptor type B (EDNRB), and the metabolite N-acetylserotonin. The qPCR and transcriptome sequencing results were consistent. Post-intervention ELISA assays revealed significantly elevated 5-HT in the NDT group after intervention (P < 0.05). Interactions between 5-HTR2B and N-acetylserotonin differed between the groups. The cellular findings showed significantly reduced G protein-coupled receptor 82 (GPR82) and phospholipid phosphatase-related protein type 4 (PLPPR4), and significantly increased S100A2 protein expression in the Si-HTR2B group, compared with the controls (P < 0.05). The biochemical results uncovered significantly decreased nitric oxide (NO) and significantly increased malondialdehyde and NO synthetase concentrations compared with the models (P < 0.05). Mongolian mind-body interactive therapy might affect SBP in patients with essential hypertension by combining 5-HT with 5-HTR2B to mediate NO relaxation.

Keywords: 5-Hydroxytryptamine; 5-Hydroxytryptamine receptor 2B; Essential hypertension; Mongolian mind–body interactive therapy; N-acetylserotonin.

Fig. 1

Study framework.

Fig. 2

Expression of targeted metabolome and transcriptome pre- and after- MMBIT intervention in EH. (A,B) Plots of PCA-targeted metabolic data in the NDT and DT groups (n = 10 each). Red and black indicate before and after intervention, respectively. (C,D) Volcano maps of DEGs in transcriptomes of both groups. (E) Trends of seven DEGs analyzed by qPCR vs. RNA-seq (n = 19). (E-G) Results of 5-HT ELISA in DT (n = 11) and NDT (n = 19) groups. ns: no difference. *P < 0.05. Down, downregulated genes; DT, drug therapy; NDT, no drug therapy; no DEGs, undifferentiated genes; up, unregulated genes.

Fig. 3

Spearman relevance network connecting DEGs and metabolites. (A) Anabolic serotonin pathway. (B) Interaction between DEGs with metabolites in the NDT group. (C) Interaction between DEGs with metabolites in DT group. Green and purple represent transcriptome DEGs and yellow and orange represent target metabolites. AAAD, aromatic L-amino acid decarboxylase; AADAC, arylacetamide deacetylase AANAT; aralkylamine-N-acetyltransferase.

Fig. 4

Effects of Si-HTR2B on MDA, NOS, and NO. (A) Protein expression of PLPPR4, GPR82, HTR2B, and S100A2 in models in vitro (B‒D) Impact of the HTR2B intervention on MDA, NOS, and NO values in the model. *P < 0.05 vs. control; ^†P < 0.05 vs. model. GPR82, G protein-coupled receptor 82; HTR2B, 5-hydroxytryptamine receptor 2B; NO, nitric oxide; NOS, nitric oxide synthase; PLPPR4, phospholipid phosphatase related 4.

Fig. 5

Intervention with MMBIT uses a mechanistic model of 5-HT to treat mild EH.