Research On Antioxidant Activity Of Cannabidiol And Transcriptome Analysis Under Different Environmental Conditions

Industrial hemp is an important plant with medicinal,fiber and other wider applications with a complex chemical composition.Though its major active components is cannabinoids which is reported to possessed antibacterial,anti-inflammatory and anti-tumor properties,yet chemicals such as flavonoids,organic acids,terpenes and others were isolated.Based on the optimization of different extraction methods of CBD in hemp,the exploration of antioxidant activity,regulatory network for the synthesis of CBD,as well as its physiological and biochemical effects were established.regulatory network as follows:1.Single factor optimization method and response surface test was used to optimize the extraction of CBD from solvent type,ultrasonic time,material to liquid ratio and the extraction time with the temperatures varied at 65°C,75°C,85°C,95°C,105°C and 115°C to study the decarboxylation of CBD.The results showed methanol solvent,10 min ultrasonic time,two times extraction and 1:35?g/mL?material to liquid ratio as the best condition for extracting CBD at 85°C,95°C with both temperature having the highest decarboxylation conversion of 79.7%.Using supercritical extraction optimization method,the best extraction conditions were investigated from indexes of CBD in two groups of hemp under different temperature and pressure.The results showed that:the extraction effect of cannabinoids was significantly different under different temperature and pressure,among which,a pressure and temperature of 400 bar and 45?respectively,gave the best supercritical extraction conditions.Too high or low temperature and pressure resulted in decreased extraction rate.2.The result of CBD antioxidant activities investigated against antioxidant activity,cell damage and repair showed the IC₅₀ of CBD to XOD,PPO and ABTS free radicals to be 1.14?g/mL,4.99?g/mL and 1.68?g/mL respectively.CBD to some extent inhibited XOD and PPO and its high concentration can also scavenge ABTS free radicals.CBD has good activity and less toxicity to Human esophageal cancer cell,human colon cancer cell,human breast cancer cell,human liver cancer cell,human gastric cancer cell,human normal liver cell,human gastric mucosa cell,human umbilical vein endothelial cell,which indicated great medicinal value.The results provide a basis for the study of CBD in antioxidant activity.3.Transcriptome sequencing analysis was performed on two groups of cannabis plants with large differences in CBD content cultivated by vegetative reproduction,and the GO enrichment results of the different genes showed that under different light time and different cultivation environments,2246expressed genes were up-regulated and 987 were down-regulated.According to the analysis of KEGG metabolic pathway,DEGs were significantly enriched in phenylpropionic acid biosynthesis and plant hormone signal transduction pathway.Heatmap showed an increased in the expression of CBDAS,CYP86B1 and ALDH2C4 genes.As RT-PCR was not used for expression verification,it will be very useful to investigate if the results of transcriptome were true and reliable.