ABSTRACT
To explore the effectiveness and safety of a Chinese medicinal decoction Wuwei Xiaodu Drink (WWXDD) in inhibiting chronic osteomyelitis via regulatory T cells signaling. The effective constitutes of WWXDD and osteomyelitis related genes were screened. Target proteins were cross-validated using the Venny database. GO function and KEGG pathway analysis were performed for target proteins, while pharmacological network was constructed. The bone properties were analyzed by HE staining and the concentrations of immune factors were measured by ELISA. The expression of CTLA-4 and Foxp3 mRNA and STAT5, p-STAT5, CTLA-4 and Foxp3 protein were detected using Real-time PCR and Western blot, respectively. FACS was used to analyze the percentages of cells. A total of 117 genes overlapped between 785 target genes of the active compounds of WWXDD and 912 osteomyelitis related genes. Inflammation-related genes, including IL-6, TNFα, IL-1β and IL-2 showed high connection degree in the drug-compound-disease-target network. GO function and KEGG pathway analysis revealed that 117 intersection genes mainly enriched in virus infection related pathways, immune related pathways and chemokine signaling pathway. Furthermore, the development of chronic osteomyelitis was suppressed in model rats after treatment with WWXDD. Meanwhile, the concentrations of IL-2 and CD4+CD25+Foxp3 Treg percentages together with the levels of p-STAT5, CTLA-4 and Foxp3 were also down-regulated. Furthermore, IL-2 and WWXDD drug-containing serum exhibited opposite effects on regulating IL-2, IL-10, TGF-β1, Foxp3, CTLA4 and STAT5. In addition, a STAT5 phosphorylation inhibitor suppressed the expression of Foxp3 and CTLA-4. WWXDD can treat chronic osteomyelitis through suppressing the main regulating factors of Tregs and interfere its immunodepression. Our results bring a new solution for chronic osteomyelitis.
Subject(s)
Animals , Rats , Forkhead Transcription Factors/metabolism , Interleukin-2/metabolism , Osteomyelitis/metabolism , STAT5 Transcription Factor/metabolism , Signal Transduction , T-Lymphocytes, RegulatoryABSTRACT
Objective:To discuss clinical effect of comprehensive Qingre Tiaoxue decoction combined with Wuwei Xiaodu drink and enema to sequelae of pelvic inflammatory disease (syndrome of dampness heat stasis) and to study the moderating effect to immune inflammatory factor. Method:One hundred and sixty patients were randomly divided into control group (80 cases) and observation group (80 cases) by random number table. The 73 patients in control group completed the therapy (4 patients were exfoliated or lost to follow-up, 3 patients were eliminate), 74 patients in observation group completed the therapy (4 patients were exfoliated or lost to follow-up, 2 patients were eliminate). Patients in control group got Fuke Qianjin capsules,2 grains/time, 3 times/day, and at the third day after menstruation, addition and subtraction therapy of Wuwei Xiaodu drink with enema for 14 days at every night, 1 dose/day, and enema continued for 2-4 h/day. Based on the treatment of enema in control group, patients in observation group added Qingre Tiaoxue decoction for 3 menstrual cycles, 1 dose/day, and stopping during menstrual cycle. Degree of pelvic pain, dysmenorrhea and the degree of pain in the lower abdomen and lumbosacral in the non menstrual period were evaluated by pain visual simulation score (VAS). Before and after treatment, scores of syndrome of dampness heat stasis, Mc Cormack scale, and Summary of the World Health Organization Quality of Life Measurement Scale (WHOQOL-BREF) scale were graded, and vaginal ultrasonography, culture of cervical secretion and routine examination of leucorrhea were detected. And levels of CD4+, CD8+, CD4+/CD8+, T lymphocyte subsets, tumor necrosis factor-α (TNF-α), interleukin-2 (IL-2) and IL-6 were detected and safety was evaluated. Result:By ANOVA of repeated measurement, after treatment, scores of VAS during dysmenorrheal and VAS during non menstrual period in two groups decreased (P<0.05), and during 3 menstrual cycles, scores of VAS during dysmenorrheal and VAS during non menstrual period in observation group were lower than those in control group (P<0.01). Scores of physical sign and syndrome of damp heat and stasis were lower than those in control group (P<0.01), score of WHOQOL-BREF was higher than that in control group (P<0.01). The depth of pelvic effusion and the volume of pelvic inflammatory mass in observation group were less than those in control group (P<0.01). Levels of CD4+, CD4+/CD8+ and IL-2 were higher than those in control group (P<0.01), and levels of CD8+, TNF-α and IL-6 were lower than those in control group (P<0.01). Comprehensive curative effect in observation group was better than that in control group (Z=2.028, P<0.05). And curative effect of traditional Chinese medicine（TCM） Syndrome in observation group was better than that in in control group (Z=2.064, P<0.05). And there were no serious adverse events and adverse reaction caused by Chinese medicine. Conclusion:Comprehensive therapy of Chinese medicine and enema to sequelae of pelvic inflammatory disease can improve the clinical symptoms and signs, improve the quality of life of patients, and regulate the cellular immune function and inflammatory factors. It has better comprehensive curative effect and TCM syndrome curative effect, and is safe for clinical use.
ABSTRACT
Objective: The effect of Wuwei Xiaodu Drink (WXD) on the metabolism of bacteria and the dose-effect relationship were investigated by MSPQC. Methods: The frequency shift-time curves of the growth and metabolism of Escherichia coli, Staphylococcus aureus, and Pseudomonas aeruginosa affected by WXD were obtained by using MSPQC. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) were gained according to FDT and the frequency shift. The dose-effect relationship was analyzed according to the curve which represented the relationship between the concentration of WXD and FDT. The parallel test was carried out with the traditional tube dilution method. Results: The results indicated that WXD inhibited the growth of E. coli, S. aureus, and P. aeruginosa, which was dose-dependent. The higher the concentration was, the stronger the antibacterial effect became. The MBC of WXD for three kinds of pathogenic bacteria was 0.9 g/mL. The MIC of WXD for S. aureus and P. aeruginosa was 0.8 g/mL, however for E. coli was 0.7 g/mL within 24 h. Conclusion: MSPQC is a sensitive, quantitative, quick method, which could provide the process information in real time for determining the dose-effect relationship of antimicrobial Chinese materia medica.