The Immune Change of the Lung and Bowel in an Ulcerative Colitis Rat Model and the Protective Effect of Sodium Houttuyfonate Combined With Matrine

Abstract

Objective: To explore the immune change of lung injury of Ulcerative colitis (UC) by observing the changes of inherent immunity and adaptive immunity of the lung and bowel in UC rat models after the treatment of Sodium Houttuyfonate combined with Matrine.

Method: UC rat models were established with the mucous membrane of colon allergize combined with TNBS-alcohol enteroclysis for 1 week and 5 weeks. 1-week experimental rats were divided into normal group and model group, 5/each group. 5-weeks experimental rats were divided into normal group, model group, Sodium Houttuyfonate (2.9mg/ml) combined with Matrine (1.47mg/ml), and positive control sulfasalazine (10mg/ml), 5/each group. All rats were administered by gavage for 5 weeks. The histopathological and fibrotic changes in the lung and bowel were observed, and the expressions of Tumor Necrosis Factor (TNF)- α, interleukin (IL)-8 in the lung, bowel, and serum were detected by radio-immunity and immunohistochemistry, and the mRNA expressions of Toll-like receptor (TLR)-4, nuclear factor kappa (NF-κB), Macrophage migration inhibitory factor (MIF), Mucosal addressing cell adhesion molecule-1 (MadCAM1) and Pulmonary surfactant protein-A (SP-A) in the lung and bowel were detected by Real time-PCR.

Result: Compared with the normal group, the model rats had significant histopathological and fibrotic changes both in the lung and bowel, and all treatment groups were improved. After treatment, TLR4, IL-8, MIF, and TNF-α in the lung decreased (P<0.05); NF-KB, IL-8, and MIF in the bowel increased (P<0.05); MadCAM1 both in lung and bowel decreased (P<0.05); SP-A decreased in bowel and increased in the lung (P<0.05).

Conclusion: The cause of lung injury in this model was found to be related to inherent immunity and adaptive immunity, while the cause of bowel injury in this model was found to be mainly related to adaptive immunity. Sodium Houttuyfonate combined with Matrine could improve bowel and lung injury.

Keywords: adaptive immunity; inherent immunity; lung injury; sodium houttuyfonate combined with matrine; ulcerative colitis.

Figure 1

Pathological and fibrosis changes of lung and bowel tissues in rats with ulcerative colitis. UC models were established with the mucous membrane of colon allergize combined with TNBS-alcohol enteroclysis. (A) Intestinal and (B) lung tissues with HE staining; (C) Intestinal and (D) lung tissues with Ma Song dyeing.

Figure 2

Expression of TLR4 mRNA, NF-kB mRNA, and MIF mRNA in the lung and bowel. UC models were established with the mucous membrane of colon allergize combined with TNBS-alcohol enteroclysis. TLR4 mRNA in Lung (A) and Bowel (B); NF-kB mRNA in Lung (C) and Bowel (D). Compared with Normal group, **P < 0.01; ***P < 0.001. Compared with model group, ^#P < 0.05.

Figure 3

Expression of TNF-a, IL-8 in the lung, bowel, and serum. UC models were established with the mucous membrane of colon allergize combined with TNBS-alcohol enteroclysis. (A) Expression of TNF-a (Radioimmunoassay); (B) Expression of IL-8 (Radioimmunoassay); (C) Expression of TNF-a (Immunohistochemistry); (D) Expression of IL-8 (Immunohistochemistry). Compared with Normal group, *P < 0.05; **P < 0.01; ***P < 0.001. Compared with model group, #P < 0.05, ##P < 0.01 , ^###P < 0.001.

Figure 4

Expression of MIF mRNA in the lung and bowel. UC models were established with the mucous membrane of colon allergize combine with TNBS-alcohol enteroclysis. MIF mRNA in the lung (A) and bowel (B). Compared with Normal group, *P < 0.05; **P < 0.01; ***P < 0.001. Compared with model group, ^#P < 0.05, ^###P < 0.001

Figure 5

The Expression of MadCAM1 mRNA in the lung and bowel, and the Expression of MadCAM1 protein in the serum. UC models were established with the mucous membrane of colon allergize combined with TNBS-alcohol enteroclysis. MadCam-1 mRNA expression in lung (A) and bowel (B) (PCR); (C) MadCam-1 protein expression in surem (Elisa). Compared with Normal group, *P < 0.05; **P < 0.01; ***P < 0.001. Compared with model group, ^#P < 0.05, ^##P < 0.01, ^###P < 0.001.

Figure 6

Changes of SP-A in the lung and the bowel. UC models were established with the mucous membrane of colon allergize combine with TNBS-alcohol enteroclysis. (A) SP-A mRNA expression in the bowel (PCR); (B) SP-A protein expression in the bowel (Western Blotting); (C) SP-A relative protein level in the bowel; (D) SP-A mRNA expression in the lung (PCR); (E) SP-A protein expression in the lung (Western Blotting); (F) SP-A relative protein level in lung. Compared with Normal group, *P < 0.05; **P < 0.01; ***P < 0.001. Compared with model group, ^#P < 0.05, ^##P < 0.01, ^###P < 0.001.

Figure 7

Evaluation of toxicity of Sodium Houttuyfonate combined with Matrine in rats. Ulcerative colitis models were established with mucous membranes of colons sensitized with TNBS-alcohol enteroclysis. Treatments were administered with SASP, Sodium Houttuyfonate combined with Matrine. (A) Body weight changes. (B) Hepatorenal function changes. ALT, AST, BUN, and Cr were evaluated. (C) HE staining of liver and kidney. (D) Fecal characteristics. TNBS, immune- 2,4,6-trinitrobenzene sulfonic acid; SASP, sulfasalazine; ALT, alanine aminotransferase; AST, aspartate aminotransferase; BUN, blood urea nitrogen; Cr, Creatinine. and Compared with the model group, ^#P < 0.05.