Identification Of Q-marker And Anti-ulcerative Colitis Effects Of Total Flavonoids Of Lonicera Rupicola Hook.f.et Thoms

Lonicera rupicola Hook.f.et Thoms(LRH),a natural plant of the genus Lonicera in the family Loniceraceae,is a medicinal herb commonly used in Tibetan regions for the treatment of bacon diseases,ophthalmopathy and pulmonary diseases.Till now,systematic pharmacological studies of LRH are rare.To standardize the rational use of LRH: Firstly,fingerprints of 15 batches of LRH was established by high performance liquid chromatography(HPLC)and the quality marker(Q-marker)was identified.Secondly,total flavonoids of LRH were identified by ultra performance liquid chromatograph tandem mass spectrometry(UPLC-MS/MS)and network pharmacology performed for predicting their potential targets,biological pathways and signaling pathways.Finally,Therapeutic effect of total flavonoids of LRH on dextran sulfate sodium(DSS)-induced ulcerative colitis in mice and the underlying molecular mechanisms were elucidated based on changes in body weight and colon length,disease activity index(DAI),pathological observations,cytokine expression,m RNA expression,intestinal barrier tight junction protein expression,gut microbiome and signaling pathways.And the main experimental results were as follows:1.HPLC fingerprint was established using 15 batches of LRH,and a total of 21 peaks and 7 components were identified.The Q-markers of LRH were screened for chlorogenic acid,isochlorogenic acid A and rutin based on the “five principles”,meanwhile the accurate,stable and reliable method for Q-marker contents determination was established.In order to provide a scientific basis for the development of quality standards for LRH.2.37 total flavonoid of LRH components were detected by UPLC-MS/MS.Of these,12 potential active components and 137 potential intersectional targets were predicted.Network pharmacology and molecular docking proved that total flavonoid of LRH could be used for the treatment of ulcerative colitis via “multi-component-multi-target-multi-pathway”.3.Total flavonoid of LRH revealed excellent antioxidant activity by scavenging DPPH and ·OH radicals in vitro.4.Total flavonoid of LRH was safe and non-toxic in both acute oral toxicity and accumulative toxicity.And the results of DSS-induced ulcerative colitis in mice demonstrated that total flavonoid of LRH performed the protective effects through inhibiting PI3K/AKT signaling pathway,mainly including reducing colon inflammation and oxidative stress levels,promoting colon epithelial cell proliferation,restoring colon epithelial barrier and improving gut microbiome.In summary,total flavonoid of LRH had a potential value for scientific research and development and utilization on anti-ulcerative colitis,laying the theoretical basis and data support for the further exploration of other pharmacodynamic material basis and their molecular mechanism of action.