Based On Zebrafish Model,Safety Evaluation Of Tanshinone ?A And Its Derivatives And Exploration Of The Efficacy Of Phenanthrazole Derivatives

Objective Zebrafish has been widely used in research fields such as pharmaceutical toxicology and eco-environmental toxicology.By using the zebrafish model to evaluate the safety of the drug,we can discover the effects of the drug on the development of the heart,blood vessels,and nerves and provide a theoretical reference for the rational use of the drug and the avoidance of application risk.Salvia miltiorrhiza,as a traditional Chinese medicine,has a long application history and complex ingredients.Tanshinone IIA is its main active ingredient and has good biological activity.However,due to the phthaloquinone structure,it has certain limitations in clinical use.The Tanshinone IIA was restructured and optimized in the early stage of the laboratory and a Tanshinone IIA derivative TA20 with good screening activity was selected for experiments in this paper.Therefore,in this study,a zebrafish model was used to evaluate the effects of Tanshinone IIA and Tanshinone IIA derivative TA20 on zebrafish heart,blood vessel,and neurobehavior.The number of adverse drug reaction cases involving Chinese herbal medicines and chemicals such as anti-tumor drugs has been increasing year by year.Toxic side effects of the drugs will have adverse effects on the heart,blood vessels,nerves and prognosis.Cancer is a serious threat to our health.The anti-tumor chemotherapeutic drugs used clinically have various degrees of toxic and side effects.Some serious toxic and side effects are the direct reasons for limiting the dosage or use of drugs.The occurrence of postoperative complications is a potential hazard after cancer resection.Compounds that inhibit tumor cell proliferation and inhibit postoperative inflammation have become one of the candidates for anticancer drugs.At the same time,the antitumor drug phenanthrazole derivatives were studied for antitumor activity and postoperative inflammation complications.Methods(1)The survival rate,teratogenicity,hatchability,body length and wound healing ability of zebrafish embryos were used as indicators to observe the effects of drugs on morphological development of zebrafish.(2)The heart area,heart rate and sinus-arterial bulb(SV-BA)spacing of zebrafish embryos were used as indicators of cardiac development,and intestinal vein(SIV)growth was used as an index of vascular development to observe the effects of drugs on cardiovascular development in zebrafish.(3)The number of spontaneous tail contractions and escape behaviors after touching were observed to evaluate the effect of drugs on the neurobehavioral development of zebrafish.(4)The antitumor activity of imidazole derivatives was studied by in vitro MTT and in vivo liver cancer zebrafish models.The effect of the imidazole derivative L082 on wound inflammatory response was studied by tail-induced inflammatory response,and the mechanism of inflammatory action of the compound was further studied by lipopolysaccharide-induced ROS and NO response.Results(1)At a concentration of 40 ?M,Tanshinone IIA significantly inhibited embryo growth resulting in shortened body length,impaired wound healing and healing ability,and increased teratogenicity of embryo morphology,but it did not cause death at the highest concentration and did not cause incubation delay.In cardiovascular aspect,it reduced the heart area,reduced heart rate,shorten the distance between venous sinus and arterial glands,and inhibited the angiogenesis in the sub-intestinal vein.In neurobehavior,it inhibited the number of spontaneous tail contractions and escape behavior.In the mechanism,it inhibited the inflammatory cell aggregation response.(2)When exposed to TA20,a derivative of tanshinone IIA imidazole,at concentrations of 10 ?M and 20 ?M,the neovascularization of the subintestinal vein(SIV),of zebrafish embryos was inhibited,but the morphological development,cardiac development,neurobehavioral development and tissue repair ability of zebrafish embryos were not affected.(3)The newly synthesized imidazole derivatives L082 and L142 could inhibit the growth of a variety of tumor cells.Among them,the L082 compound inhibited the proliferation of hepatocellular carcinoma in zebrafish.It also inhibited the aggregation reaction of wound inflammatory cells and promoted the elimination of wound inflammation cells to achieve an anti-inflammatory effect.Studies have found that it can inhibit both ROS and NO production.Conclusions(1)At a concentration of 40 ?M,tanshinone ?a exhibits obvious developmental toxicity,cardiovascular toxicity,and neurobehavioral toxicity in zebrafish embryos,so we should pay more attention to the development of Tanshinone IIA products.(2)TA20,an imidazole derivative of Tanshinone IIA,was less toxic than Tanshinone IIA.It could be used as an effective low toxic agent,making further exploration of its activity more meaningful.(3)The newly synthesized and characterized imidazole derivative L082 exerted notable antitumor activity in vivo and in vitro.At the same time,at the concentration of 80 ?M,it can inhibit wound inflammation and reduce the risk of postoperative complications.