Effects of Jian Pi Qing Chang Hua Shi decoction on mucosal injuries in a 2,4,6-trinitrobenzene sulphonic acid-induced inflammatory bowel disease rat model

Abstract

Context: Jian Pi Qing Chang Hua Shi decoction (JPQCHSD) has been considered as an effective remedy for the treatment of inflammatory bowel disease (IBD) in Chinese traditional medicine.

Objective: We evaluated the efficacy of JPQCHSD on 2-4-6-trinitrobenzene sulphonic acid (TNBS)-induced IBD rats and the responsible mechanisms.

Materials and methods: Except the rats of the control group (50% ethanol), Sprague-Dawley rats (180 ± 20 g) induced by TNBS (150 mg/kg in 50% ethanol), received water extract of JPQCHSD daily at 0, 9.5, 19, or 38 g/kg for 12 days. The rats were sacrificed, and their colons were removed to evaluate the disease activity index. Malondialdehyde (MDA), superoxide dismutase (SOD), myeloperoxidase (MPO), immunoglobulin A (IgA), tumour necrosis factor (TNF)-α, interleukin (IL)-1β, IL-6, and nuclear factor-κB were evaluated.

Results: JPQCHSD extract significantly reduced the disease activity index of TNBS-induced colitis with a median effective dose (ED₅₀) of 26.93 g/kg. MPO and MDA were significantly reduced in the 19 and 38 g/kg groups (ED₅₀ values 37.38 and 53.2 g/kg, respectively). The ED₅₀ values for the increased SOD and IgA were 48.98 and 56.3 g/kg. ED₅₀ values for inhibition of TNF-α, IL-1β, and IL-6 were 32.66, 75.72, and 162.06 g/kg, respectively.

Discussion: JPQCHSD promoted mucosal healing in IBD rats via its anti-inflammation, immune regulation, and antioxidation properties.

Conclusions: JPQCHSD has healing function on IBD. Further clinical trials are needed to demonstrate its efficacy and tolerance to IBD.

Keywords: Superoxide dismutase; immunoglobulin A; interleukin-1β; malondialdehyde; myeloperoxidase; nuclear factor-κB; tumour necrosis factor-α.

Figure 1.

Analysis of the main components of Jian Pi Qing Chang Hua Shi decoction (JPQCHSD). Ultra-high-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) was used to identify the components of JPQCHSD and Masslynx 4.1 software was used to process and analyze the data. (A) Positive and (B) negative ion chromatograms of JPQCHSD are shown.

Figure 2.

Effect of Jian Pi Qing Chang Hua Shi decoction (JPQCHSD) on histopathological changes in 2-4-6-trinitrobenzene sulphonic acid (TNBS)-induced inflammatory bowel disease (IBD) model. (A) Representative images of haematoxylin and eosin (H&E) staining: a. control; b. model; and c-e. JPQCHSD 9.5 g/kg, JPQCHSD 19 g/kg, and JPQCHSD 38 g/kg. (B) Microscopy score was used for histological evaluation. (C) Effect of JPQCHSD on disease activity index (DAI). JPQCHSD 19 g/kg and JPQCHSD 38 g/kg, but not JPQCHSD 9.5 g/kg, significantly decreased pathological damage in IBD and reduced increased DAI and microscopy scores; ^★p < 0.05 as compared to the model group. Data are shown as the mean ± standard deviation. Control, no treatment; Model, TNBS-induced IBD rats.

Figure 3.

Effects of Jian Pi Qing Chang Hua Shi decoction (JPQCHSD) on levels of malondialdehyde (MDA) and superoxide dismutase (SOD) activity in colon in 2,4,6-trinitrobenzene sulphonic acid (TNBS)-induced inflammatory bowel disease (IBD) rats. MDA and SOD levels were measured. ^★p < 0.05 and *p < 0.05 as compared to the model group. Data are shown as the mean ± standard deviation. Control, no treatment; Model, TNBS-induced IBD rats; JPQCHSD 9.5 g/kg, JPQCHSD 19 g/kg, and JPQCHSD 38 g/kg, respectively.

Figure 4.

Detection of colonic myeloperoxidase (MPO) activity. MPO activity was significantly higher in 2,4,6-trinitrobenzene sulphonic acid (TNBS)-induced inflammatory bowel disease (IBD) rat model than in control rats. Both JPQCHSD 19 g/kg and JPQCHSD 38 g/kg groups showed low MPO activities. ^★p < 0.05 as compared to model group. Data are shown as the mean ± standard deviation. Control, no treatment; Model, TNBS-induced IBD rats.

Figure 5.

Effects of JPQCHSD on concentration of serum immunoglobulin A (IgA). Compared with the normal group, the content of IgA in the serum of the model was decreased significantly (p < 0.05). Compared with the model group, the IgA content in JPQCHSD 19 g/kg and JPQCHSD 38 g/kg groups were increased (p < 0.05), while the IgA level in the JPQCHSD 9.5 g/kg group was not significantly increased (p > 0.05). ^★p < 0.05 as compared to the model group. Data are shown as the mean ± standard deviation. Control, no treatment; Model, TNBS-induced IBD rats.

Figure 6.

Serum interleukin (IL)-1β, IL-6, and tumour necrosis factor (TNF)-α levels of the five groups. ^■p < 0.05, ^◆p < 0.05, and ^●p < 0.05 as compared to the model group. Data are shown as the mean ± standard deviation. Control, administered 0.9% saline; model, 2-4-6-trinitrobenzene sulphonic acid (TNBS)-induced inflammatory bowel disease (IBD) rats; JPQCHSD 9.5 g/kg, JPQCHSD 19 g/kg, and JPQCHSD 38 g/kg, respectively.

Figure 7.

Nuclear factor (NF)-κB mRNA and protein expression levels in colon tissue of five groups. *p < 0.05 as compared to the model group. Data are shown as the mean ± standard deviation. a. Control, no treatment; b. Model, 2,4,6-trinitrobenzene sulphonic acid (TNBS)-induced inflammatory bowel disease (IBD) rats; c–e. JPQCHSD 9.5 g/kg, JPQCHSD 19 g/kg, and JPQCHSD 38 g/kg, respectively.

Figure 8.

Jian Pi Qing Chang Hua Shi decoction (JPQCHSD) improved inflammatory damage to intestinal cells by increasing superoxide dismutase (SOD) and immunoglobulin A (IgA), suppressing malondialdehyde (MDA) and myeloperoxidase (MPO), decreasing tumour necrosis factor (TNF)-α, interleukin (IL)-1 and IL-6 levels via inhibition of nuclear factor (NF)-κB transcription in IBD rats.