Composite Sophora Colon-Soluble Capsule Ameliorates DSS-Induced Ulcerative Colitis in Mice via Gut Microbiota-Derived Butyric Acid and NCR+ ILC3

Objective: To investigate the effects of composite Sophora colon-soluble Capsule(CSCC) on gut microbiota-mediated short-chain fatty acids(SCFAs) production and downstream group 3 innate lymphoid cells(ILC3s) of dextran sulfate sodium(DSS)-induced ulcerative colitis(UC) mice model. Methods: The main components of CSCC were analyzed by hybrid ultra-high-performance liquid chromatography ion mobility spectromety quadrupole time-of-flight mass spectrometry(UHPLC-IM-QTOF/MS). Twenty-four male BALB/c mice were randomly divided into 4 groups(n=6) by using a computer algorithm-generated random digital, including control, DSS model, mesalazine, and CSCC groups. A DSS-induced colitis mice model was established to determine the effects of CSCC by recording colonic weight, colonic length, index of colonic weight, and histological colonic score. The variations in ILC3s were assessed by immunofluorescence and flow cytometry. The results of gut microbiota and SCFAs were acquired by 16s r DNA and gas chromatography-mass spectrometry(GC-MS) analysis. The expression levels of NCR^(+)ILC3^(-), CCR6^(+)Nkp46-(Lti) ILC3^(-), and ILCreg-specific markers were detected by enzyme-linked immunosorbent assay, and real-time quantitative polymerase chain reaction and Western blot, respectively. Results: The main components of CSCC were matrine, ammothamnine, Sophora flavescens neoalcohol J, and Sophora oxytol U. After 7 days of treatment, CSCC significantly alleviated colitis by promoting the reproduction of intestinal probiotics manifested as upregulation of the abundance of Bacteroidetes species and specifically the Bacteroidales_S24-7 genus(P<0.05). Among the SCFAs, the content of butyric acid increased the most after CSCC treatment. Meanwhile, compared with the model group, Lti ILC3s and its biomarkers were significantly downregulated and NCR^(+) ILC3s were significantly elevated in the CSCC group(P<0.01). Further experiments revealed that ILC3s were differentiated from Lti ILC3s to NCR^(+)ILC3s, resulting in interleukin-22 production which regulates gut epithelial barrier function. Conclusion: CSCC may exert a therapeutic effect on UC by improving the gut microbiota, promoting metabolite butyric acid production, and managing the ratio between NCR^(+)ILC3s and Lti ILC3s.

A Multi-center,Randomized, Single-blind,Controlled Clinical Study on the Efficacy of Composite Sophora Colon-Soluble Capsules in Treating Ulcerative Colitis

Objective： To evaluate the safety and efficacy of composite sophora colon-soluble capsule （CSCC） in treating ulcerative colitis （UC） of the damp-heat accumulation syndrome pattern （DHAS） and to prepare a basis for a phase Ⅲ clinical trial.Methods： A multi-center,randomized,single-blind,and positive drug parallel-controlled design was adopted.There were 126 patients of UC-DHAS stratified and assigned equally to three groups.Patients in two CSCC treated groups,Groups T1 and T2,were treated orally with high （six capsules,thrice a day） and low （four capsules,thrice a day） doses CSCC,and patients in the control group were treated orally with Mesalazine Enteric-coated Tablets （four tablets,thrice a day）,respectively,all for eight weeks.The clinical efficacy and safety of treatments were evaluated through clinical symptom observations and colonoscopic examinations.Results： （1） Full analysis set （FAS） and per-protocol set （PPS） analyses showed the comprehensive curative effect in Groups T1,T2,and the control group,obtaining the values of 85.7%,92.9%,and 71.4% （P=0.330）,and 89.5%,92.7%,and 73.2% （P=0.552）,respectively,demonstrating no statistical significance among the three groups.（2） FAS and PPS analysis showed the efficacy on membranous lesions in Groups T1,T2,and the control group,obtaining the values of 83.3%,92.9%,and 73.8% （P=0.063）,and 86.8%,92.7%,and 75.6% （P=0.070）,respectively,showing statistical insignificance among the three groups.（3） FAS analysis showed an efficacy tendency on improving tenesmus （P=0.056）.No changes were found in improving the other symptoms,and statistical significance was not shown among the three groups （P〉0.05）.PPS analysis showed the efficacy on single item symptom in Groups T1,T2,and the control group was not statistically significant among the three groups （P=0.082）.Conclusions： The comprehensive effect of CSCC in treating UC is basically equivalent to that of Mesalazine enteric-coated tablet;however,the tendency was shown to improve symptoms.Its efficacy could not be raised by increasing the dosage used.Therefore,the recommended dosage of CSCC is four capsules,three times a day.