Tong Xie Yao Fang relieves irritable bowel syndrome in rats via mechanisms involving regulation of 5-hydroxytryptamine and substance P

Abstract

Aim: To investigate whether the Chinese medicine Tong Xie Yao Fang (TXYF) improves dysfunction in an irritable bowel syndrome (IBS) rat model.

Methods: Thirty baby rats for IBS modeling were separated from mother rats (1 h per day) from days 8 to 21, and the rectum was expanded by angioplasty from days 8 to 12. Ten normal rats were used as normal controls. We examined the effects of TXYF on defection frequency, colonic transit function and smooth muscle contraction, and the expression of 5-hydroxytryptamine (5-HT) and substance P (SP) in colonic and hypothalamus tissues by Western blot and RT-PCT techniques in both normal rats and IBS model rats with characterized visceral hypersensitivity.

Results: Defecation frequency was 1.8 ± 1.03 in normal rats and 4.5 ± 1.58 in IBS model rats (P < 0.001). However, the defecation frequency was significantly decreased (3.0 ± 1.25 vs 4.5 ± 1.58, P < 0.05), while the time (in seconds) of colon transit function was significantly increased (256.88 ± 20.32 vs 93.36 ± 17.28, P < 0.001) in IBS + TXYF group rats than in IBS group rats. Increased colonic smooth muscle tension and contract frequency in IBS model rats were significantly decreased by administration of TXYF. Exogenous agonist stimulants increased spontaneous activity and elicited contractions of colon smooth muscle in IBS model rats, and all of these actions were significantly reduced by TXYF involving 5-HT and SP down-regulation.

Conclusion: TXYF can modulate the activity of the enteric nervous system and alter 5-HT and SP activities, which may contribute to the symptoms of IBS.

Keywords: 5-hydroxytryptamine; Irritable bowel syndrome; Substance P; Tong Xie Yao Fang; Traditional Chinese medicine.

Figure 1

Flow chart of the study design and experiment procedure. IBS: Irritable bowel syndrome; TXYF: Tong Xie Yao Fang; RT-PCR: Reverse transcription-polymerase chain reaction.

Figure 2

Tong Xie Yao Fang affects 5-hydroxytryptamine and substance P protein expression in colon (A) and hypothalamus (B) tissues in each group, as revealed by immunohistochemistry staining (× 400; inserted panel, × 1600); C: Quantitative measurements with Integral Optical Density data from A and B (n = 10 for each group). a: Normal group; b: IBS model group; c: IBS model + positive group; d: IBS model + TXYF group. IBS: Irritable bowel syndrome; TXYF: Tong Xie Yao Fang.

Figure 3

Quantitative data of 5-hydroxytryptamine (A) and substance P (B) mRNA expression via reverse transcription-polymerase chain reaction detection in rat colon of the indicated groups.

Figure 4

5-hydroxytryptamine and substance P protein expression in rat colon tissues of the indicated groups. A: Representative Western blot image; B, C: Quantitative data from collected Western blot image. IBS: Irritable bowel syndrome; TXYF: Tong Xie Yao Fang.