A Preliminary Study On The Mechanism And Material Basis Of Liquorice Root In Reducing Hepatorenal Toxicity Of Gardenia

Objective : In this study,the effects of different compatibility ratios between glycyrrhizae and fructus gardeniae on liver and kidney function of normal rats and icundus model rats,and the effects of different compatibility ratios between glycyrrhizae and fructus gardeniae on the dissolution rate of fructus gardeniae were determined that the compatibility between glycyrrhizae and fructus gardeniae could reduce the toxicity of fructus gardeniae.The mechanism and material basis of glycyrrhiza glycyrrhiza toxicity were clarified through the research on the compatibility of glycyrrhiza glycyrrhiza and glycyrrhiza glycyrrhiza toxicity reduction.Methods:1.The compatibility of different proportion of gardenia licorice for normal SD rats and jaundice model SD rats,gardenia,gardenia licorice 1:1,2:1 and 3:1,continuous dosing blood detection of liver and kidney function after a week: alanine aminotransferase(ALT),nmda aminotransferase(AST),total bilirubin(TBIL),urea nitrogen(BUN),creatinine(CREA)and pathological anatomy,liquorice and compatibility of gardenia attenuated.2.Ultra high performance liquid chromatography(UPLC)was used to determine the change of the dissolution rate of gardenia jasminoides and Gardenia combined with glycyrrhizae 1:1,2:1,3:1.The chromatographic column was Waters ACQUITY UPLC BEH C18(2.1×100 mm,1.7?m).The flow rate was 0.200 mL /min.The column temperature was 32.3?.The detection wavelength was 238 nm.3.Using different compatibility proportion of gardenia licorice on normal SD rats and icteric model SD rats were studied,gardenia,gardenia licorice 1:1,2:1 and 3:1,continuous dosing blood detection of liver and kidney function after a week: IL-18,IL-1? and NLRP3 in liver tissue,Caspase-1,Pro-IL-18,Pro-IL-1? protein expression,clear the mechanism of action of licorice gardenia detoxification.4.Effects of glycyrrhizin,glycyrrhetinic acid,glycyrrhizin,glycyrrhizin and isoglycyrrhizin on ALT and AST in HepG2 cell supernatant at different compatibility ratios of 1:1,1:2 and 2:1 were used to clarify the material basis of glycyrrhizin toxicity.Results:1.Compared with the blank group,ALT(P<0.01),AST(P<0.05),TBIL(P<0.05)significantly increased;Compared with the blank group,the single-zhi group showed ALT(P<0.05),AST(P<0.05),TBIL(P<0.01)all significantly increased.Compared with the single-zhi group,gardenia gardenia: licorice root(1:1)group liver index ALT(P<0.05),AST(P<0.05),TBIL(P<0.01)significantly decreased;Gardenia: glycyrrhiza glycyrrhiza(2:1)group liver index ALT(P<0.05),AST(P<0.05),TBIL(P<0.01)significantly decreased.Compared with mozhi group,M gardenia: licorice root(1:1)group liver index ALT(P<0.05),AST(P<0.01),TBIL(P<0.01)significantly decreased;Compared with mozhi group,M gardenia: licorice root group(2:1)showed ALT(P<0.05),AST(P<0.05),TBIL(P<0.01)significantly decreased.Compared with the blank group,the model group had renal index BUN(P<0.05),CREA(P<0.01)significantly increased.Compared with the blank group,the single-zhi group had kidney markers BUN(P<0.01),CREA(P<0.01)significantly increased.Compared with danzhi group,gardenia: licorice root(1:1)group,kidney index BUN(P<0.05),CREA(P<0.05)significantly reduced;Compared with the danzhi group,gardenia: licorice root(2:1)group,kidney index BUN(P<0.05)significantly reduced.Compared with mozhi group,M gardenia: kidney index BUN(P<0.01),CREA(P<0.01);M gardenia: glycyrrhiza(2:1)group kidney BUN(P<0.01),CREA(P<0.01)significantly decreased.2.The linear range is 0.0023-0.0138 maintaining g(r=0.9998).The peak area in this range shows a good linear relationship,with an average recovery rate of 102.71% and a relative standard deviation of 3.62%.The average content of geniposide was 0.1979% in the gardenia group,0.1306% in the gardenia-glycyrrhiza(1:1)group,0.1358% in the gardenia-glycyrrhiza(2:1)group,and 0.1601% in the gardenia-glycyrrhiza(3:1)group.the average content of geniposide in the gardenia-glycyrrhizae(1:1)compatibility group was0.1306%,the average content of geniposide in the gardenia-glycyrrhizae(2:1)compatibility group was 0.1358%,and the average content of geniposide in the gardenia-glycyrrhizae(3:1)compatibility group was 0.1601%.3.Compared with the blank group,model group IL-18(P<0.01),IL-1?(P<0.01)The index was significantly increased,and Zhigang IL-18(P<0.05),IL-1?(P<0.01)Significantly increased indicators.Gardenia gardenia: Glycyrrhizae(1:1)group IL-18(P<0.05),IL-1?(P<0.01)Significantly decreased indicators;Gardenia: Liquorice(2:1)group IL-1?(P<0.05)Significantly decreased indicators.M Gardenia: Radix Glycyrrhizae(1:1)group IL-18(P<0.01),IL-1?(P<0.01)Significantly decreased indicators.RT-PCR resultsshowed that compared with the blank group,the single zhigroup NLRP3(P<0.01),Caspase-1(P<0.01),the Pro-IL-18(P<0.01),Pro-IL-1?(P<0.01)mRNA expression was significantly increased.Gardenia: glycyrrhiza 1:1 groupNLRP3(P<0.01),Caspase-1(P<0.05),the Pro-IL-18(P<0.05),Pro-IL-1?(P<0.01)mRNA expression was significantly down-regulated.Gardenia: glycyrrhiza glycyrrhiza 2:1 group NLRP3(P<0.01),Caspase-1(P<0.01),Pro-IL-1?(P<0.01)mRNA expression was significantly down-regulated,gardenia: Pro-IL-1? margin of liquorice 3:1(P<0.05)mRNA expression was significantly down-regulated;Compared with mozhi group,M gardenia: glycyrrhiza 1:1 group NLRP3(P<0.01),Caspase-1(P<0.05),the Pro-IL-18(P<0.05),Pro-IL-1?(P<0.05)mRNA expression was significantly down-regulated.Gardenia M: glycyrrhiza glycyrrhiza 2:1 group NLRP3(P<0.01),Caspase-1(P<0.01)mRNA expression was significantly down-regulated.Gardenia M: glycyrrhiza 3:1 group NLRP3(P<0.05),Caspase-1(P<0.01)mRNA expression was significantly down-regulated.Western blotting results showed that compared with the blank group,NLRP3(P<0.01),Caspase-1(P<0.05),the Pro-IL-18(P<0.01),Pro-IL-1?(P<0.01)protein expression was significantly increased.Gardenia: glycyrrhiza 1:1 group NLRP3(P<0.01),the Pro-IL-18(P<0.01)and Pro-IL-1?(P<0.01)protein expression levels were significantly down-regulated.Gardenia: glycyrrhiza glycyrrhiza 2:1 group NLRP3(P<0.05),Caspase-1(P<0.01),Pro-IL-18(P<0.05),Pro-IL-1?(P<0.05)protein expression levels were significantly down-regulated.Compared with mozhi group,M gardenia:licorice 1:1 group Caspase-1(P<0.05),Pro-IL-18(P<0.01),Pro-IL-1?(P<0.01)protein expression level was significantly down-regulated.Gardenia M: glycyrrhiza glycyrrhiza2:1 group NLRP3(P<0.01),Caspase-1(P<0.01),Pro-IL-18(P<0.05),Pro-IL-1?(P<0.05)protein expression level was significantly down-regulated,and M gardenia: Caspase-1(P<0.01),Pro-IL-18(P<0.05)protein expression level was significantly down-regulated.4.Compared with the blank group,ALT(P<0.05),AST(P<0.01)the index increased significantly.Compared with the jingniping group,jingniping: glycyrrhizinate(1:1)ALT(P<0.05),AST(P<0.05)the indicators were significantly lower: ginnipin: glycyrrhizinate(1:2),ALT(P<0.01),AST(P<0.05)the index was significantly reduced;Ginnipin:glycyrrhetinic acid(1:1)ALT(P<0.01),AST(P<0.05)the index was significantly reduced;Ginnipin: glycyrrhetinic acid(1:2)ALT(P<0.01),AST(P<0.01)the index was significantly reduced;Jingniping: glycyrrhizin(1:2)ALT(P<0.05)was significantlyreduced.Jingniping: isoglycyrrhetin(1:2)ALT(P<0.01)was significantly reduced.Conclusion:1.The effect of the compatibility of glycyrrhiza gardeniae and glycyrrhiza gardeniae on the liver and kidney functions of normal rats and jaundice model rats and the analysis of the pathological section results showed that the compatibility of glycyrrhiza gardeniae and glycyrrhiza gardeniae in different proportions could reduce the hepatorenal toxicity of gardeniae in different degrees,and the compatibility ratio of 1:1 was the best.2.The mechanism of glycyrrhiza glycyrrhiza to reduce the hepatotoxicity of gardenia was mainly related to glycyrrhiza glycyrrhiza to reduce the dissolution of gardenoside;Moreover,the mechanism of glycyrrhiza glycyrrhiza in reducing hepatotoxicity of gardenia was mediated by NLRP3 pathway3.The study on the substance basis of glycyrrhiza glycyrrhiza in reducing the toxicity of gardenia gardeniae showed that the compatibility of four different components of glycyrrhiza glycyrrhiza with jingniping could reduce the toxicity of jingniping to different degrees.It was preliminarily speculated that glycyrrhiza glycyrrhetinic acid and glycyrrhetinic acid were the material basis of the hepatotoxicity of jingniping.