Functional characterization of CYP81C16 involved in the tanshinone biosynthetic pathway in Salvia miltiorrhiza / 中国天然药物

Danshen, the dried roots and rhizomes of Salvia miltiorrhiza Bunge (S. miltiorrhiza), is widely used in the treatment of cardiovascular and cerebrovascular diseases. Tanshinones, the bioactive compounds from Danshen, exhibit a wide spectrum of pharmacological properties, suggesting their potential for future therapeutic applications. Tanshinone biosynthesis is a complex process involving at least six P450 enzymes that have been identified and characterized, most of which belong to the CYP76 and CYP71 families. In this study, CYP81C16, a member of the CYP71 clan, was identified in S. miltiorrhiza. An in vitro assay revealed that it could catalyze the hydroxylation of four para-quinone-type tanshinones, namely neocryptotanshinone, deoxyneocryptotanshinone, and danshenxinkuns A and B. SmCYP81C16 emerged as a potential broad-spectrum oxidase targeting the C-18 position of para-quinone-type tanshinones with an impressive relative conversion rate exceeding 90%. Kinetic evaluations andin vivo assays underscored its highest affinity towards neocryptotanshinone among the tested substrates. The overexpression of SmCYP81C16 promoted the accumulation of (iso)tanshinone in hairy root lines. The characterization of SmCYP81C16 in this study accentuates its potential as a pivotal tool in the biotechnological production of tanshinones, either through microbial or plant metabolic engineering.

Therapeutic effect of anti-CXCL1 neutralizing antibody on acute ulcerative colitis in mice / 中南大学学报(医学版)

Objective:To evaluate the therapeutic effect of CXCL1 monoclonal antibody on dextra sulfate sodium (DSS)-induced acute ulcerative colitis (UC) in mice,and to elucidate its effect on the expressions ofTNF-α,IFN-γ,,IL-17 and IL-10 as well as neutrophil infiltration.Methods:Female BALB/c mice were randomly divided into a normal group (DSS-),a disease group (DSS+saline),an anti-CXCL1 antibody group (DSS+anti-CXCL1 Ab) and a treatment control group (DSS+IgG Ab).The DSS+saline,DSS+anti-CXCL1 Ab and DSS+anti-CXCL1 Ab groups were given 3.5％ DSS solution as drinking water to induce acute intestinal inflammation,while the normal control was given distilled water freely.The DSS+anti-CXCL1 Ab mice were intraperitoneal injected with anti-CXCL1 Ab (4 mg/kg) on the 3rd and 6th day.Same amount of rat IgG Ab was given in the DSS+IgG Ab group.The normal group and the disease group were injected with 0.9％ sodium chloride solution.The value of disease activity index (DAI) and the injury of colorectal tissue were measured.The levels of TNF-α,IFN-γ,IL-10 and IL-17 in colonic tissues of mice were detected by RT-PCR.Myeloperoxidase (MPO),a specific marker of neutrophils was measured by immunohistochemistry.Results:Compared with the normal control group,DAI score and colorectal injury score in the disease group were significantly increased,but the DAI and colorectal in the mice with acute ulcerative colitis tissue damage score were significantly reduced after anti-CXCL1 Ab intervention.Compared with the normal control group,mRNA levels of TNF-α,IFN-γ and IL-17 in the colorectal tissues were significantly elevated (P＜0.05) in the disease group while the IL-10 was decreased;these effects were attenuated by anti-CXCL1 Ab intervention (P＜0.05).Immunohistochemistry showed that the infiltration of neutrophils (MPO+) in the colon tissue was significantly increased in the disease group,while the anti-CXCL1 Ab treatment could significantly reduce the neutrophil infiltration in colon tissue (P＜0.05).Conclusion:Anti-CXCL1 Ab relieves the progression of DSS-induced acute ulcerative colitis by suppressing proinflammatory expression and neutrophil infiltration.