Traditional Chinese Medicine QPYF as Preventive Treatment for Clostridium difficile Associated Diarrhea in a Mouse Model

Abstract

Traditional Chinese medicine QPYF has a good effect for treating antibiotic-associated diarrhea in clinical practice. The aim of this study is to test its efficacy to prevent Clostridium difficile associated diarrhea (CDAD) in a mouse model. C57BL/6 mice were infected with Clostridium difficile VPI 10463 after exposure to antimicrobial mixture. QPYF was administered from 7 days prior to Clostridium difficile infection to 20 days after infection, and its effect was compared with no treatment and receiving placebo. The mice were monitored for 20 days and the percent survival, disease activity index, weight loss, colon histopathology, and the levels of toxins in the feces were measured. The expressions of TNF α, MCP-1, NF-κB p65, and phospho-NF-κB p65 in the colon were presented by immunohistochemistry. The survival rate of QPYF group (93.75%) was higher than that of model control group (65%). The mice treated with QPYF had a lower weight loss and disease activity index, compared to the mice with placebo. A significantly lower level of histopathology scores, toxins in the feces, and TNF α, MCP-1, NF-κB p65, and phospho-NF-κB p65 were detected for QPYF-treated mice. Traditional Chinese medicine QPYF showed a good preventive effect for CDAD in a mouse model.

Figure 1

Experimental scheme.

Figure 2

Kaplan-Meier survival plots for the Normal control group, QPYF group, and model control group after Clostridium difficile challenge. The survival rate of normal control group, QPYF group, and model control group was 100% (14/14), 93.75% (15/16), and 65% (13/20). The survival rate in the QPYF group was significantly higher than the model control group (the log-rank test, P = 0.013).

Figure 3

Mean relative weight (a) and disease severity score (b) were presented from days 0 to 20. Mean relative weight was the percentage of the original weight on day 0. Disease severity score was based on weight loss, the consistency of the feces, and hematochezia. On day 3 after infection, the clinical symptoms with weight loss, diarrhea, and blood in feces were the most serious. The mean relative weight of normal control group, QPYF group, and model control group were 0.99 ± 0.021, 0.94 ± 0.041, and 0.88 ± 0.045. Compared with model control group, the weight loss in QPYF group was obviously lower (P < 0.01). The mean disease severity score (DAI) was as follows: normal control group, 0.73 ± 0.033; model control mice, 4.28 ± 0.385; QPYF group, 2.53 ± 0.301. The mice in QPYF group had a lighter clinical symptoms than the model control mice (P < 0.01).

Figure 4

Colonic pathology. H&E-stained colon tissues of mice are shown (×200). (a) Normal control group (histopathological score: 0.35 ± 0.14) and (b) model control group (histopathological score: 5.31 ± 0.75) illustrate epithelial damage, neutrophil infiltration, and congestion/edema, (c) QPYF group (histopathological score: 1.99 ± 0.92); the degree of colon damage was less prominent than (b). (d) The histopathological score: the mouse treated with QPYF had a significantly lower score than model control group (^∗ P < 0.01).

Figure 5

Clostridium difficile toxin A/B levels were measured in the fecal samples on day 4 and expressed as test value (OD). The OD values of model control group and normal control group reached 0.33 ± 0.12 and 7.40 ± 0.29. Lower mean toxin level in QPYF-treated mice (4.28 ± 0.78) was detected compared to model control group (^∗ P < 0.01).

Figure 6

The immunohistochemical analysis of TNF α, MCP-1, NF-κB p65, and phospho-NF-κB p65 in the colon tissues of mice (×400). (a) The expression of TNF α, MCP-1, NF-κB p65, and phospho-NF-κB p65 in QPYF group was obviously weaker than the model control group. (b) The percent positive area was illustrated by image plus pro 6.0, and the mouse treated with QPYF had a significantly lower positive area than the model control group (^∗ P < 0.01).