Three Flavonoids Or Metabolites Of Monascus Inhibit The Viability Of Glioma Cells And The Design Of Preparation Production Projects

Glioma is a common malignant intracranial tumor,which is very harmful.Chemotherapy plays a huge role in the treatment of glioma.The development and application of temozolomide(TMZ)provides a new solution for the clinical treatment of glioma,but due to its drug resistance,the subsequent treatment efficiency will be low.Therefore,we need to find new chemotherapy drugs to treat glioma.Natural substances have attracted much attention due to their low toxicity and side effects.The ethanol extract of Monascus fermentation is a mixture of various components,including Monascus pigment,Monacolin K,etc.The addition of flavonoids can reduce the content of citrinin without reducing the content of Monascus pigment and mycelium.Therefore,flavonoids,such as genistein,are added to the fermentation medium of Monascus.The ethanol extract,as natural fermentation products,have lower toxic and side effects.Taking into account the complex composition of the ethanol extract,the effect of a single component on glioma cells will be studied.Therefore,we studied the effects of flavonoids,Monacolin K(MK),one of the secondary metabolites of Monascus,and the ethanol extract of Monascus fermentation on U251 cells.The purpose is to provide a new solution for the treatment of glioma.And we designed its related preparation production engineering.The main research content of the paper are as follows:(1)Study on the inhibition of proliferation and pro-apoptosis of U251 cells treated with flavonoidsThe viability and inhibition rate of U251 cells treated with GST,GIN and DAI was tested using CCK-8 Kit;Cellular morphological treated with GST,GIN and DAI were tested with the Hoechst 33258 kit;The apoptosis rate of U251 cells treated with GST,GIN and DAI was tested with the AnnexinV/PI apoptosis kit;The expression of Bax,Caspase 3,PI3K,Akt,mTOR and other related apoptosis genes and proteins of U251 cells treated with GST,GIN and DAI were tested by RT-qPCR and Western blot method.The study found that 50,100 μM GST,GIN and DAI all had inhibitory effects on U251 cells and promoted the apoptosis of U251 cells.GST had better inhibitory effect than GIN and DAI.GST and GIN promoted the apoptosis of U251 cells by upregulating the expression of Bax and Caspase 3 proteins,while DAI promoted the apoptosis of U251 cells by inhibiting the expression of p-AKT and p-mTOR proteins.(2)Study on the inhibition of proliferation and pro-apoptosis of U251 cells treated with MK,one of the metabolites of MonascusThe detection method is the same as that of flavonoids.Studies have found that 5,10 μM MK can promote the apoptosis of U251 cells by upregulating the expression of Bax and Caspase 3 proteins.(3)Study on the inhibition of proliferation and promoting apoptosis of U251 cells treated with the ethanol extract of Monascus fermentationU251 cells were treated with the ethanol extract of Monascus fermented with no GST and the ethanol extract of Monascus fermented with 0.2 g of GST for 24 h and 48 h.It was found that these two ethanol extracts of Monascus can inhibit the proliferation and promote the apoptosis of U251 cells.And the inhibition rate of 48 h is significantly higher than that of 24 h.Among them,the ethanol extract of Monascus fermented with GST is obviously higher than the ethanol extract of Monascus fermented without GST on the inhibition rate of U251 cells.The inhibitory effects of these two ethanol extracts also showed a significant dose-dependent,and both were best at a concentration of 200 μg/mL.(4)We have carried out a preliminary design of a factory for soft capsules with metabolites of Monascus as the main ingredient.The process flow and related equipment selection of the Monascus metabolite preparations with an annual output of 8 tons have been determined.And drew the floor plan of the factory,production workshop and the flow chart of the production.