| Tiankuizi is dried roots of the genus Semiaquilegia adoxoides(DC.)Makino,which belongs to the family Ranunculaceae,Thalassiae,and subfamily,and are included in the 2015 edition of the Chinese Pharmacopoeia.They are sweet,bitter,and cold.Return to the liver and stomach.Efficacy:clearing away heat and detoxifying,reducing swelling and dissolving,used for Carbuncle and furuncle,acute mastitis,scrofula,boils,boils,snake bites.Tiankuizi is a commonly used Chinese medicine in clinical practice,but there are few studies on its chemical composition and biological activity,especially its antitumor mechanism is almost blank.Its chemical components are mainly alkaloids.At present,more than 60 chemical components have been separated from Tiankuizi,including turpentine,magnolitine,berberine and so on.Tiankuizi is one of the commonly used Chinese medicines in clinical practice,and its curative effect is definite.However,there are no reports on the antitumor mechanism of the alkaloids of Tiankuizi,and its substance basis and mechanism of action are still unclear.The tumor microenvironment refers to the internal environment in which tumor cells produce and live,including tumor cells,fibroblasts,immune and inflammatory cells,glial cells,and other cells,as well as the interstitial cells,microvessels,and biomolecules in nearby areas.The refinement can be divided into inflammatory microenvironment,vascular microcirculation microenvironment,migration microenvironment and immunosuppressive microenvironment.Among them,the inflammatory microenvironment is one of the important components,which can accelerate tumorigenesis and development,promote angiogenesis and metastasis,destroy the adaptive immune response to form immune escape,and lead to the formation of drug resistance.The inflammatory microenvironment has become an important research object in tumor treatment.According to the theory of traditional Chinese medicine,the pathogenesis of tumors is accompanied by pathological factors such as phlegm,turbidity,stasis,and heat,which have similar effects to the inflammatory cells and inflammatory factors existing in the tumor microenvironment.These pathological factors will affect the normal physiological functions of the body.Accelerate tumor development.Traditional Chinese medicine has certain curative advantages in the treatment of tumors.Under the guidance of the "cancer toxicity" theory,according to the pathological factors of tumor sputum,turbidity,stasis,and heat,traditional Chinese medicine can adjust the composition of the tumor microenvironment from various aspects to improve local Inflammatory microenvironment,and inhibit tumor growth by inhibiting angiogenesis,inducing tumor cell apoptosis,improving cell signaling pathways and conduction.Early mastitis,breast hyperplasia,and breast cancer are closely related.Studies have shown that breast hyperplasia is the pathological basis of breast cancer.Tiankuizi have significant effects in treating breast hyperplasia and mastitis.Therefore,the use of Tiankuizi to adjust breast cancer inflammatory microenvironment may be an important mechanism for its treatment of breast cancer.In order to further clarify the chemical constituents and antitumor mechanisms of sunflower seeds,we studied the chemical constituents and antitumor activities and mechanisms of Tiankuizi.Methods:In this study,the following methods were used to study the anti-tumor efficacy and mechanism of total alkaloids in Tiankuizi.1.The chemical composition of 70%ethanol extract of Tiankuizi was studied.Thirteen compounds were separated by various separation methods,and five compounds were identified by spectroscopic techniques.They are magnoflorine,semiaquilegine,thalifendine,N-methylcorydine,and tembetarine.2.The anti-tumor efficacy of total Tiankuizi alkaloids was studied.The anti-tumor activity of total Tiankuizi alkaloids in serum of KM mice were preliminarily verified using the in vivo model of S180 mouse sarcoma cells.3.To study the interference of total alkaloids from the sunflower seeds in vitro on tumor inflammatory microenvironment.THP-1 cells were used as research objects.PMA induced macrophages were used to construct a tumor inflammatory microenvironment model.TGF-?,TNF-?,and IL-6 in the cell matrix was measured to study the effect of total alkaloids in the Tiankuizi on the level of inflammatory factors in the inflammatory microenvironment of breast cancer.4.To study the effects of protein expression of inflammatory factors in tumor cells and microenvironment after treatment with total alkaloids in Tiankuizi.Western Blot method was used to detect inflammatory related pathways NF-?B and STAT3 in MDA-MB-231 cells.And SMAD-4 protein expression levels.Result:1.S180 solid tumor inhibition rate experiment results show that the tumor inhibition rate of the high-dose group of total Tiankuizi alkaloids(dose 200 mg·kg-1)reached 45.24%.The spleen index and thymus index of tumor-bearing mice in the Tiankuizi administration group had no effect compared with the model group,and the results were not significantly different(P>0.05).The result indicated that although the tumor suppression rate of the Tiankuizi high and middle dose group was lower than that of the cyclophosphamide group,it could improve the immunity of mice.The potential function of Tiankuizi alkaloids suggests that the total alkaloids of Tiankuizi have the function of regulating the immune function of mice while exerting anti-tumor effects.2.The experiment on the inflammatory microenvironment showed that the total alkaloids of Tiankuizi 60 ?g·mL-1 for 36 h can significantly reduce the expression of pro-inflammatory factors TNF-? and IL-6 in the tumor inflammatory microenvironment and promote the expression of TGF-?.It(P<0.05)indicates that total alkaloids can interfere with tumor inflammatory microenvironment by regulating the expression level of inflammatory factors.3.The effects of total alkaloids in Tiankuizi on the expression levels of NF-?B,STAT3,SMAD-4 protein indicate that Tiankuizi can reduce inflammatory proteins in tumors,and gene expression.Conclusion:This study extracted and enriched the total alkaloids of Tiankuizi,and separated the chemical constituents.Six kinds of alkaloids were identified by spectroscopy.Among them,N-methylcorydine,corytuberine and the tembetarine were isolated from Ampelopsis for the first time.Early mastitis,hyperplasia of mammary glands and breast cancer are closely related.Mastitis and hyperplasia of mammary glands are the pathological basis of breast cancer.Tiankuizi has a significant effect in the treatment of hyperplasia of mammary glands and mastitis,and no adverse reactions occur.Tiankuizi intervention to adjust the inflammatory microenvironment of breast cancer may be an important mechanism for its treatment of breast cancer.In this study,in vivo and in vitro experiments were conducted to verify that the total alkaloids of Tiankuzi had a clear inhibitory effect on tumors and had little immune effect on tumor-bearing mice.Combined with the clinical efficacy of Tiankuizi in the treatment of mastitis,we further verified the interventional regulation effect of Tiankuizi on the inflammatory microenvironment through experiments.The results of the study showed that the total alkaloids of Helianthus annuus can reduce the secretion of pro-inflammatory factors and promote the expression of tumor suppressor factors.In order to clarify the mechanism of Tiankuizi's intervention in tumor inflammatory microenvironment,we studied the expression of related proteins in tumor cells and tumor inflammatory microenvironment after administration of Tiankuizi total alkaloids.The results of the study indicate that the effective antitumor part of Tiankuizi is the total alkaloid of Tiankuizi,and its antitumor efficacy is remarkable.Its anti-tumor drug efficacy mechanism may be related to intervention in tumor inflammatory microenvironment,regulation of inflammatory factor expression and related protein expression. |
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