Mechanistic Study On The Anti-acute Liver Injury Of Lychee Seed Flavonoids And Its Granule Preparation

Objective:Litchi chinensis Sonn.has been shown to have anti-acute liver injury and therapeutic effects on liver fibrosis,but the effects of its active ingredients are unclear and clinical application forms are lacking.In this study,we investigated the effect of Lychee seed flavonoids（TFL）extract on the anti-acute liver injury and the mechanism associated with the inhibition of NOD-like receptor family 3（NLRP3）inflammatory vesicles activation,and investigated the preparation process of Lychee seed flavonoids granules.Methods:The mice were randomly divided into blank group,model group,bifenthrin group（150 mg/kg）,MCC950 group（10 mg/kg）,TFL high（360 mg/kg）and low（180mg/kg）dose groups and administered by gavage for 10 days continuously.The acute liver injury was induced by intraperitoneal injection of CCl₄-peanut oil solution,and the liver function and morphology of each group were observed to reflect the effect of TFL against acute liver injury;in addition,the levels of reactive oxygen species（ROS）in liver tissues were measured,and the expression of NLRP3 inflammatory vesicles in liver tissues was examined by Western blot.In addition,the expression of NLRP3 inflammatory vesicles in liver tissues was examined by Western blot to investigate the mechanism of action of TFL.In order to study the formulation process of TFL granules,the formulation excipients diluent,sieving mesh and wetting agent concentration were investigated on the basis of single-factor test using forming rate,dissolution rate and resting angle as evaluation indexes;the process prescription of TFL granules was optimized by orthogonal design for the preferential selection of wet granule drying process with forming rate and moisture as evaluation indexes to form a reasonable process.The critical relative humidity of TFL granules was also determined,and the content of proanthocyanidin A2,a total flavonoid component in TFL granules,was determined by UV-visible spectrophotometry.Results:TFL significantly reduced the serum levels of alanine transaminase,aspartate aminotransferase,and total bile acids in mice with CCl₄-induced acute liver injury（P<0.05）,decreased the levels of reactive oxygen species in liver tissues（P<0.05）,and attenuated structural damage to liver tissues.High doses of total lychee kernel flavonoids significantly down-regulated NLRP3 inflammatory vesicle-associated protein TXNIP,NLRP3,pro-caspase-1,pro-IL-1β,IL-1βexpression（P<0.05）.After process research,the TFL granules were formed by taking dry paste powder and mannitol in the right amount,mixing them evenly at 1:3,making granules with 75%ethanol,laying the material at a thickness of 0.5 cm,drying at 60℃for 30 min,and then granulating.The TFL granules achieved an average molding rate of 94.21%（RSD of0.73%）,an average dissolution rate of 95.34%（RSD of 0.10%）,an average angle of repose of 23.68%（RSD of 0.38%）,an average moisture of 1.93%（RSD of 2.23%）,a critical relative humidity of 73.5%,proanthocyanidin A2 in the range of 0.0397～0.2382 mg/m L with good linearity（R2=0.9991）,mean spiked recoveries of 100.25%and mean proanthocyanidin A2 content of TFL pellets of 16.25%,respectively.Conclusion（s）:Total flavonoids from Lychee kernel can reduce CCl₄-induced acute liver injury in mice,and the mechanism may be related to the inhibition of inflammatory response involved in NLRP3 inflammatory vesicle activation.The process of forming total flavonoids from Lychee kernel pellets is feasible,and the finished product has high forming rate,good flowability and good dissolution rate.A stable UV-Vis spectrophotometric method for content determination was established,which can provide data support for the production of this preparation.