Study On The Changes In Alkaloid Content And Hepatotoxicity Of Aconi Radix Before And After The Compatibility Of Trichosanthes Nigrum And Trichosanthes Trichosanthes

Purpose:The aim of this study was to investigate the changes of chemical constituents and hepatotoxicity of trichosanthes trichosanthis.The changes of chemical composition and liver toxicity before and after the combination of raw and grass aconitum and whole trichosanthis,trichosanthis peel,trichosanthis son,trichosanthis powder were explored,and combined with the above data to analyze "aconitum trichosanthis",to provide scientific theoretical basis for18 anti.Material and method:1.Acid dye colorimetry was used to determine the content of total alkaloids in raw and processed Aconiti Kusnezoffii Radix before and after compatibility with Trichosanthis Fructus,Trichosanthis Pericarpium,Trichosanthis Fructus and Trichosanthis Radix.2.HPLC was used to determine the contents of aconitine,hypaconitine,mesaconine,benzoylmesaconine,benzoylhypaconine and benzoylaconine before and after the compatibility of raw and processed Aconiti Kusnezoffii Radix with Trichosanthis Fructus,Trichosanthis Pericarpium,Trichosanthis Fructus and Trichosanthis Radix.The blood and liver of rats were collected by intragastric administration,and the liver coefficient,liver pathological section,alanine aminotransferase(ALT),aspartate aminotransferase(AST),alkaline phosphatase(ALP),γ-glutamyltransferase(γ-GT),superoxide dismutase(SOD),malondialdehyde(MDA),alpha-fetoprotein(AFP),total bilirubin(TBil),direct bilirubin(DBil),total protein(TP),globulin(GLB)and cholinesterase(CHE)were measured and compared.4.High performance liquid chromatography was used to determine CYP3A4,CYP1A2,CYP2E1 and CYP2C9 in rats.Activity of four cytochrome P450 isoforms.5.Principal component analysis was used to analyze the correlation between total alkaloid content,monomer alkaloid content and hepatotoxicity in rats before and after the compatibility of Radix Aconiti Kusnezoffii and Fructus Trichosanthis.Results:1.Changes of total alkaloid content before and after the compatibility of Radix Aconiti Kusnezoffii and Fructus Trichosanthis: the total alkaloid content increased after the compatibility of Radix Aconiti Kusnezoffii and Fructus Trichosanthis,and the total alkaloid content decreased after the compatibility with Radix Trichosanthis;the content of total alkaloids in the compatibility of Radix Aconiti Kusnezoffii and Radix Trichosanthis decreased.2.Changes in the content of six monomer alkaloids before and after the compatibility of Radix Aconiti Kusnezoffii and Fructus Trichosanthis: the content of diester alkaloids in the compatibility of Radix Aconiti Kusnezoffii and Fructus Trichosanthis decreased,and the content of benzoylmesaconine increased;the content of benzoylmesaconine increased,and the content of other components decreased.The content of benzoylmesaconine increased,and the content of other components decreased.The content of diester alkaloids in the compatibility of Radix Aconiti Kusnezoffii and Trichosanthis Pericarpium increased;in addition to the decrease of mesaconitine content,the content of other components increased.The content of mesaconitine decreased when combined with trichosanthis.3.Changes in liver coefficient of rats before and after compatibility of Radix Aconiti Kusnezoffii and Fructus Trichosanthis: single administration,liver coefficient did not change;in multiple administrations,the organ coefficient of the compatibility of raw Aconiti Kusnezoffii Radix and Trichosanthis Pericarpium and the compatibility of processed Aconiti Kusnezoffii Radix and Trichosanthis Pericarpium decreased.4.Changes in rat liver morphology before and after the compatibility of Radix Aconiti Kusnezoffii and Fructus Trichosanthis: single administration,the liver cells of Radix Aconiti Kusnezoffii had vacuoles,the compatibility of Radix Aconiti Kusnezoffii and Fructus Trichosanthis showed congestion,the compatibility of Radix Aconiti Kusnezoffii and Fructus Trichosanthis had slight pathological changes,the compatibility of Radix Aconiti Kusnezoffii and Fructus Trichosanthis had partial liver cell necrosis,and the compatibility of Radix Aconiti Kusnezoffii and Radix Trichosanthis was blurred.The structure of liver cells of prepared Aconiti Kusnezoffii Radix changed.The compatibility of prepared Aconiti Kusnezoffii Radix and Trichosanthis Fructus showed degeneration and necrosis of liver cells.The compatibility of prepared Aconiti Kusnezoffii Radix and Trichosanthis Fructus showed apoptosis of liver cells.The compatibility of prepared Aconiti Kusnezoffii Radix and Trichosanthis Fructus showed a large number of liver cell necrosis.After many times of administration,there were necrotic hepatocytes and inflammatory cell infiltration in raw Aconitum,the structure of hepatocytes in raw Aconitum-Quangualou compatibility was not clear,inflammatory cell infiltration in raw Aconitum-Gualoupi compatibility,hepatocyte necrosis in raw Aconitum-Gualouzi compatibility,and slight changes in liver morphology in raw Aconitum-Tianhuafen compatibility.Cytoplasmic vacuoles were observed in the Aconiti Kusnezoffii Radix group.Hepatocyte necrosis was observed in the Aconiti Kusnezoffii Radix-Quangualou compatibility group.Inflammatory cell infiltration was observed in the Aconiti Kusnezoffii Radix-Gualoupi compatibility group.Hepatocyte necrosis was observed in the Aconiti Kusnezoffii Radix-Gualouzi compatibility group.Cell swelling and necrosis were observed in the Aconiti Kusnezoffii Radix-Tianhuafen compatibility group.5.Effects of Radix Aconiti Kusnezoffii and Fructus Trichosanthis on liver function in rats before and after compatibility: single administration,the AST values of Fructus Trichosanthis,Pericarpium Trichosanthis and Radix Aconiti Kusnezoffii-Tianhua Powder increased,and the AST values of Radix Aconiti Kusnezoffii,Radix Aconiti Kusnezoffii-Fructus Trichosanthis and Radix Aconiti Kusnezoffii-Tianhua Powder decreased;the ALT values of raw Aconiti Kusnezoffii Radix,processed Aconiti Kusnezoffii Radix,processed Aconiti Kusnezoffii Radix-Trichosan Fructus compatibility,trichosanthin,raw and processed Aconiti Kusnezoffii Radix-Trichosan Pollen compatibility and whole Trichosanthis Fructus decreased.ALP values did not change;the TBi L values of Fructus Trichosanthis,Radix Trichosanthis,whole Fructus Trichosanthis,raw and processed Radix Aconiti Kusnezoffii-Peel Trichosanthis,Fructus Trichosanthis,Radix Trichosanthis compatibility and whole Fructus Trichosanthis compatibility decreased.The DBi L values of Radix Aconiti Preparata,Pericarpium Trichosanthis,Radix Aconiti Preparata-Tianhuafen compatibility and Radix Aconiti Preparata-Quangualou compatibility decreased;the PA value of Tianhuafen,Shengcaowu-Tianhuafen compatibility,Quangualou and Shengcaowu-Quangualou compatibility increased.The ALB value of whole melon decreased;c HE values did not change;the MDA values of raw,processed Aconitum kusnezoffii-Trichosan Fructus compatibility,Fructus Trichosanthis and raw Aconitum kusnezoffii-Trichosan Fructus compatibility increased;the SOD values of Aconiti Kusnezoffii Radix Praeparata,Trichosanthis Pericarpium and Aconiti Kusnezoffii Radix Praeparata-trichosanthis Pericarpium decreased.After multiple administration,the AST value of Radix Aconiti Preparata,Pericarpium Trichosanthis and Radix Aconiti Preparata-Tianhua Powder increased;a LT values decreased;a LP values did not change.The TBi L values of raw Aconiti Kusnezoffii Radix,processed Aconiti Kusnezoffii Radix,Trichosanthis Pericarpium,raw Aconiti Kusnezoffii Radix-trichosanthis Pericarpium,processed Aconiti Kusnezoffii Radix-Tianhua Powder,raw Aconiti Kusnezoffii Radix-whole Trichosanthis Pericarpium and processed Aconiti Kusnezoffii Radix-whole Trichosanthis Pericarpium decreased.The DBi L values of Trichosanthis Pericarpium,raw and processed Aconiti Kusnezoffii Radix-Trichosanthis Pericarpium compatibility,raw Aconiti Kusnezoffii Radix-Tianhuafen compatibility and raw and processed Aconiti Kusnezoffii Radix-Quangualou compatibility decreased,and the DBi L value of Trichosanthis Semen increased.PA values increased;the ALB value of whole melon decreased;c HE values did not change.The MDA value of the compatibility of Radix Aconiti Kusnezoffii-Trichosanthis Pericarpium and the compatibility of Radix Aconiti Kusnezoffii-Tianhua Powder increased;the SOD values of Radix Aconiti Kusnezoffii-Trichosan Fructus,Radix Aconiti Kusnezoffii-Trichosan Fructus,Fructus Trichosanthis and Radix Trichosanthis were decreased.6.Effects of Radix Aconiti Kusnezoffii and Fructus Trichosanthis on CYP450 subtype enzymes in rats before and after compatibility: single administration,Radix Aconiti Kusnezoffii-Tianhuafen compatibility,Radix Aconiti Kusnezoffii-Quangualou compatibility had inhibitory effect on CYP3A4;the compatibility of Trichosanthis Pericarpium,Trichosanthis Fructus and processed Aconiti Kusnezoffii Radix-Trichosanthis Fructus has inhibitory effect on CYP1A2;the compatibility of Radix Aconiti Kusnezoffii-Fructus Trichosanthis,Radix Trichosanthis,Fructus Trichosanthis and Radix Aconiti KusnezoffiiFructus Trichosanthis had inhibitory effect on CYP2E1.Radix Aconiti Kusnezoffii,Fructus Trichosanthis and Radix Trichosanthis have inhibitory effects on CYP2C9.Multiple administration,the compatibility of Radix Aconiti Kusnezoffii-Tianhuafen and the compatibility of Radix Aconiti Kusnezoffii-Quangualou had inhibitory effect on CYP3A4.The compatibility of Aconiti Kusnezoffii Radix Praeparata-Trichosan Fructus and the compatibility of Aconiti Kusnezoffii Radix Praeparata-Trichosan Fructus,Trichosanthis Radix and Trichosanthis Fructus had inhibitory effect on CYP1A2.The compatibility of Radix Aconiti Kusnezoffii-Fructus Trichosanthis,Radix Trichosanthis,Fructus Trichosanthis,Radix Aconiti Kusnezoffii-Fructus Trichosanthis,Pericarpium Trichosanthis,Fructus Trichosanthis and Radix Trichosanthis inhibited CYP2E1.The compatibility of Radix Aconiti Kusnezoffii Preparata,Radix Aconiti Kusnezoffii Preparata-Fructus Trichosanthis,Radix Aconiti Kusnezoffii Preparata-Radix Trichosanthis had inhibitory effect on CYP2C9.7.Correlation between alkaloid content and hepatotoxicity in rats before and after compatibility of Aconiti Kusnezoffii Radix and Trichosanthis Fructus: total alkaloids are positively correlated with hypaconitine in monomer alkaloids,and are not related to hepatotoxicity;among the monomer alkaloids,benzoylmesaconine was negatively correlated with benzoylaconitine mesaconitine and aconitine,benzoylaconitine was positively correlated with benzoylhypaconine,mesaconitine,aconitine and hypaconitine,and mesaconitine was positively correlated with aconitine and hypaconitine.There was no correlation between monomer alkaloids and hepatotoxicity.There was also no correlation between total-monomer alkaloids and hepatotoxicity.Conclusion:1.The total alkaloid content increased after the compatibility of raw Aconiti Kusnezoffii Radix and Trichosanthis Pericarpium.2.Raw Aconiti Kusnezoffii Radix and Trichosanthis Fructus,Trichosanthis Pericarpium,Trichosanthis Fructus and Trichosanthis Fructus compatibility benzoylmesaconine content increased;the contents of new aconitine and hypaconitine increased in the compatibility of Aconiti Kusnezoffii Radix Praeparata and Fructus Trichosanthis,the content of diester alkaloids increased in the compatibility of Aconiti Kusnezoffii Radix Praeparata and Pericarpium Trichosanthis,and the contents of other alkaloids except new aconitine increased in the compatibility of Aconiti Kusnezoffii Radix Praeparata and Fructus Trichosanthis.3.A single dose of Trichosanthis Pericarpium,Fructus Trichosanthis,Radix Aconiti Kusnezoffii-Tianhua Powder and Radix Aconiti Kusnezoffii-Trichosanthes Pericarpium can cause liver damage,and the ability of the whole Trichosanthis liver to synthesize protein is weakened,leading to liver fibrosis and cirrhosis;multiple administrations,Radix Aconiti Kusnezoffii Preparata,Pericarpium Trichosanthis,Fructus Trichosanthis,Radix Aconiti Kusnezoffii Preparata-Tianhuafen and Radix Aconiti Kusnezoffii Preparata-Pericarpium Trichosanthis can lead to liver injury,and the ability of whole Trichosanthis liver to synthesize protein is weakened,leading to liver fibrosis and cirrhosis.4.After a single administration,the compatibility of Radix Aconiti Kusnezoffii-Tianhuafen and the compatibility of Radix Aconiti Kusnezoffii-Quangualou had stronger inhibitory effects on CYP3A4,CYP1A2,CYP2E1 and CYP2C9,and had serious effects on liver metabolism.After multiple administration,the compatibility of Radix Aconiti Kusnezoffii-Tianhuafen,Radix Aconiti Kusnezoffii-Quangualou,Radix Aconiti Kusnezoffii-Gualouzi and Radix Aconiti Kusnezoffii-Gualouzi had stronger inhibitory effects on CYP3A4,CYP1A2,CYP2E1 and CYP2C9,and had serious effects on liver metabolism.5.There was no correlation between total alkaloids,monomer alkaloids and hepatotoxicity.