Mechanism of Nicotiflorin in San-Ye-Qing rhizome for anti-inflammatory effect in ulcerative colitis.

BACKGROUND: The incidence of ulcerative colitis (UC) is on the rise globally and the development of drugs targeting UC is urgent. Finding the target of action of natural products is important for drug discovery, elucidation of drug action mechanism, and disease mechanism. San-Ye-Qing (SYQ), is an ancient herbal medicine, but whether the powder of its rhizome has pharmacological effects against UC and its mechanism of action are not clear. PURPOSE: To evaluate the therapeutic effectiveness of rhizome powder of SYQ in treating UC, and conduct an isolation and characterization of the chemical constituents of the powder. Further, screen the most potent compounds among them and determine the potential mechanism for treating UC. METHODS: In vivo, the therapeutic effect of SYQ's rhizome powder on UC was assessed by mice's body weight, DAI score, colon length, tissue MPO activity, serum inflammatory markers, etc. Additionally, HPLC was used to isolate and identify the specific chemical components of SYQ's rhizome powder. Then, the most effective compounds and their therapeutic targets were analysed and screened in SYQ rhizome powder using network pharmacology, combined with CCK-8 assay, NO release assay and molecular docking assay, in conjunction with CETSA, DARTS, SPR and enzyme activity assay. Finally, the biological effects of the key compound on the targets were validated using Western blot and ELISA. RESULTS: In vivo, SYQ rhizome powder effectively restored mice's body weight, lowered DAI and pathological score, downregulated the expression of inflammatory biomarkers, and restored colon length, as well as the colonic epithelial and mucus barriers. Afterward, 9 compounds were isolated and identified from the powder of the rhizomes of SYQ by HPLC. Nicotiflorin is the primary compound in SYQ with the highest concentration. According to both CCK-8 and NO release tests, Nicotiflorin is also the most efficacious compound. Combined with network pharmacological prediction, molecular docking analysis, CETSA, DARTS, SPR and enzyme activity assay, Nicotiflorin may ultimately suppress inflammation by targeting p65 and inhibiting the NF-κB pathway, thereby attenuating the activation of NLRP3 inflammasome. To verify this conclusion, Western blot and ELISA experiments were conducted. CONCLUSIONS: Our results suggest that the extract from SYQ rhizomes has therapeutic properties for UC. Its active ingredient Nicotiflorin exerted potent anti-UC effects by binding to p65 and inhibiting the activation of NF-κB and NLRP3 inflammasomes.

Students' knowledge of and conservation attitude toward the black-necked crane (Grus nigricollis) in Guizhou, China: insights for conservation.

BACKGROUND: The persistence of threatened and protected wildlife depends not only on habitat suitability but also remarkably on local communities' acceptance. The black-necked crane (Grus nigricollis) is a flagship species for conservation on the plateau in western China. However, the human dimension has been completely ignored in the decision-making process for conservation. METHODS: In this study, a questionnaire survey aiming to assess knowledge of and conservation attitude toward this bird was carried out among 1042 students of 7th and 9th grade from 12 schools in Weining county, Guizhou province, which has a large wintering population in an urban wetland. Logistic regression was used in the generalized linear model to identify the determinants that significantly affect students' knowledge of and conservation attitude toward this species. RESULTS: Most students have positive attitudes toward conservation, which is significantly affected by awareness, knowledge of this bird and grade. However, they have somewhat limited knowledge of this bird's biology and ecosystem services (nature's contributions to people). Knowledge was found to link with observation of the bird and grade, while observation related to the experience of visiting the wetland. Social media is the most cited resource to obtain knowledge on this bird. CONCLUSION: It is suggested that local conservation experts could help introduce more information on the black-necked crane in the schools and help conduct outdoor education activities in and around the wetland. Traditional knowledge and culture could also be incorporated into the conservation awareness enhancing program. This study focuses on the human dimension for conserving the black-necked crane in China, showing significant implications in the design and application of effective measurements to improve students' perception and attitude toward its conservation. Future assessments should include other local populations, such as farmers, fishers, and urban citizens.