Correlation Analysis Of Changes In Chemical Components Of Gardeniae Seeds With Different Processed Products And Liver And Kidney Injury

Objective:The purpose of this project is to explore the changes in chemical composition and toxicity of liver and kidney of gardenia before and after processing,to clarify whether gardenia processing has the effect of reducing liver and kidney toxicity,and to clarify the material basis of processing and reducing toxicity through the correlation analysis of chemical composition and liver and kidney toxicity.To provide a scientific basis for the reasonable use of gardenia’s different processed products and the establishment of quality standards.Methods:1.Shengzhizi uses three editions of the 2015 edition of"Chinese Pharmacopoeia",the 2010 edition of"Hunan Local Processing Standards"and the 1988 edition of"National Traditional Chinese Medicine Processing Standards"to prepare three batches of fried gardenia（CZ1-CZ3）,Jiao Gardenia（JZ1-JZ3）Gardenia charcoal（TZ1-TZ3）and ginger gardenia（JGZ1-JGZ3）medicinal materials.Use ultra-high performance liquid chromatography to compare raw gardenia（SZ1）,fried gardenia（CZ1-CZ3）,coke gardenia（JZ1-JZ3）,gardenia charcoal（TZ1-TZ3）and ginger gardenia（JGZ1-JGZ3）The content of 6 main chemical components in medicinal materials was determined,and the change rule of 6 main chemical components before and after gardenia processing was comprehensively analyzed.2.Normal SD rats were given 7.5 g·kg^-1dose of raw gardenia（SZ1）,ginger gardenia（JGZ1-JGZ3）,fried gardenia（CZ1-CZ3）,coke gardenia（JZ1-JZ3）,gardenia The sub-charcoal（TZ1-TZ3）water extract was administered continuously for 3 days,the serum ALT,AST,TBIL,CREA,BUN index levels were detected,liver and kidney indexes were calculated,and histopathological examinations were performed on the liver and kidney.At the same time,immunohistochemical detection was used to preliminarily explore the effects of different gardenia preparations on the expression of NLRP3 protein in liver tissue,and to detect the levels of IL-18 and IL-1βin serum.3.Using multivariate statistical analysis methods（clustering analysis CA,principal component analysis PCA and partial least squares analysis PLS）,the liver and kidney toxicity indexes and the contents of 6 chemical components before and after processing of gardenia were clustered and analyzed.Then the liver and kidney toxicity index+6 kinds of chemical components before and after processing of gardenia were clustered,principal component analysis and partial least squares analysis to understand the difference between 6 kinds of gardenia processed products and 6 kinds of chemical components and liver and kidney toxicity The relationship between the six chemical components and liver and kidney toxicity indicators is clear.Results:1.From the overall point of view,the total change trend of the six chemical components before and after gardenia processing is raw gardenia,ginger gardenia,fried gardenia,coke gardenia,and gardenia charcoal.The trend of the content of geniposide is raw gardenia,fried gardenia,ginger gardenia,charcoal gardenia,and gardenia charcoal.The trend of the content change of crocin I and crocinⅡis raw gardenia﹥ginger gardenia.Zi﹥Fried Gardenia﹥Jiao Gardenia﹥Garden Charcoal;Genipin Gentian Glycoside Content Variation Trends are Ginger Gardenia﹥Raw Gardenia﹥Jiao Gardenia﹥Fried Gardenia﹥Garden Charcoal;Deacetylated leaves The change trend of the content of methyl oxalate is fried gardenia,ginger gardenia,coke gardenia,raw gardenia,gardenia charcoal（content is 0）;the change trend of the content of gardenic acid is coke gardenia,gardenia charcoal,and charcoal Raw Gardenia﹥Fried Gardenia﹥Ginger Gardenia.The six chemical components before and after gardenia processing showed different trends,but the total content of the six chemical components decreased.2.Normal rats were given intragastrically the same dose(7.5 g·kg^-1)of water extracts of different gardenia preparations for 3 days.Compared with the blank group,ALT,AST and TBIL in the raw gardenia group were significantly increased（P<0.01）;compared with the Shengzhi group,the ALT and AST of the other administration groups were significantly reduced（P<0.01 or P<0.05）,and the TBIL of the Jiangzhi group,the Jiaozhi group and the carbon garden group were significant Decrease（P<0.01）.Different gardenia processed products caused different levels of liver damage to normal rats,in the order of Shengzhi group﹥Fried garden group﹥Jiangzhi group﹥Jiaozhi group﹥Carbon garden group.Liver injury is mild.Compared with the blank group,the CREA and BUN indicators of the Shengzhi group were significantly increased（P<0.01）;compared with the Shengzhi group,the CREA and BUN indicators of the Jiaozhi group and the carbon garden group were significantly reduced（P<0.01）,and the BUN index of Jiangzhi group decreased significantly（P<0.05）.Different gardenia processed products have caused different degrees of kidney damage to normal rats,in the order of Shengzhi group﹥Fried garden group﹥Jiangzhi group﹥Jiaozhi group﹥Carbon garden group,the most serious kidney damage in shengzhi group,and carbon garden group Kidney damage is the least.Compared with the blank group,the IL-18 index of Shengzhi group,Chaozhi group,Jiangzhi group,Jiaozhi group,and Tanzhi group all increased,but there was no significant difference between each group and the blank group（P﹥0.05）;compared with Shengzhi group,there was no significant difference in IL-18 index of other groups（P﹥0.05）.Compared with the blank group,the IL-1βindex of the Shengzhi group,Jiangzhi group,Jiaozhi group,and Tanzhi group were slightly increased,but there was no significant difference between the groups and the blank group（P﹥0.05）;Compared with the Shengzhi group,the IL-1βindex of the Chaozhi group was slightly lower than that of the Shengzhi group,and there was no significant difference between the other administration groups and the Shengzhi group（P﹥0.05）.Compared with the blank group,the positive rate of NLRP3 protein expression in liver tissues of different gardenia preparations was slightly increased,but there was no significant difference（P﹥0.05）;compared with the Shengzhi group,the Jiaozhi group The positive rate of NLRP3 protein expression in liver tissue of Hetanzhi group was slightly reduced,but there was no significant difference（P﹥0.05）.3.By cluster analysis method,13 different gardenia processed products were divided into 6 species（Gardenside,Genipin Gentiobiside,Gardenic Acid,Deacetylmethyl oxalate,Crocetin I and West SafflowerⅡ）The content of chemical constituents is the object of study,and it can only distinguish between raw gardenia and gardenia charcoal.Take the liver and kidney toxicity indexes（ALT,AST,TBIL,BUN,CREA）or the content of 6 chemical components+liver and kidney toxicity indexes as the research object,raw gardenia,fried gardenia,ginger gardenia,coke gardenia and gardenia char Can be clearly distinguished.Through the principal component analysis method,13 different gardenia processed products were divided into 6 species（Gardenside,Genipin Gentiobiside,Gardenic Acid,Deacetylmethyl oxalate,Crocetin I and West Safflower glycosideⅡ）The content of chemical components or liver and kidney toxicity indexes are the research objects,and only Jiao Gardenia and Gardenia charcoal can be distinguished.Taking 6 kinds of chemical components content+liver and kidney toxicity indexes as the research object,the varieties of raw gardenia,fried gardenia,ginger gardenia,coke gardenia and charcoal gardenia can be distinguished well.Through the method of partial least squares analysis,the established regression models of ALT,AST,TBIL,CREA and BUN indicators are of good quality.The contribution rate of 6 chemical components before and after processing of gardenia to liver and kidney toxicity,the order of VIP value is gardenin﹥crocin glycoside I﹥genipin gentioside glycoside﹥crocinⅡ﹥deacetylcarfolate Methyl esters﹥Gardenyl acid,and the VIP values of gardenin and crocin I are both greater than 0.8,indicating that geniposide has the largest contribution to liver and kidney toxicity,followed by crocin I.Conclusion:1.The content,total amount and composition of various chemical components before and after gardenia processing have changed greatly,which is related to the processing time and the intensity of processing conditions.With the prolongation of the processing time and the deepening of the processing intensity,the content of gardenoside,crocin I and crocinⅡdecreased to different degrees after the gardenia was processed,but the genipin gentian in Jiang Gardenia The content of diglycosides,methyl deacetylated folinate in fried gardenia,ginger gardenia and charcoal gardenia,as well as the content of gardenic acid in charcoal gardenia and gardenia charcoal are all higher than that of raw gardenia.2.A dose of 7.5 g·kg-1（equivalent to 75 g of human dose）of raw gardenia,ginger gardenia,fried gardenia,scorched gardenia,and gardenia charcoal causes varying degrees of liver and kidney damage（normal gardenia）in normal rats Zi﹥Fried Gardenia﹥Ginger Gardenia﹥Jiao Gardenia﹥Charcoal Gardenia),and the liver and kidney toxicity of gardenia reduced after varying degrees.3.Based on the NLRP3 pathway,a preliminary study of the mechanism of action of different gardenia processed products on liver toxicity in normal rats showed that the expression of NLRP3 protein in liver tissues of each group was not significantly changed by immunohistochemistry under this experimental condition.4.Different gardenia processed products were analyzed by cluster analysis and principal component analysis based on liver and kidney toxicity index+6 kinds of chemical components.5.Partial least squares analysis indicated that gardenin and crocin I had the largest contribution to liver and renal toxicity.It was preliminarily speculated that gardenin and crocin I was the material basis of liver and kidney toxicity of different processed products of gardenia.The synergistic effect of geniposide and crocin I results in liver and kidney damage in normal rats.