Study On The Chemical Composition Content And Nephrotoxicity Of Trichosanthes Trichosanthes Before And After The Compatibility Of Trichosanthes Trichosanthes And Cao W

Purpose:The purpose of this study was to study the changes of chemical components and nephrotoxicity of Trichosanthis Fructus before and after compatibility with Aconiti Kusnezoffii Radix from the perspective of chemistry and biology.The correlation between the content of chemical components and nephrotoxicity was discussed by the content changes of primary and secondary substances and nephrotoxicity evaluation before and after the compatibility of Trichosanthis Fructus（Quangualou）,Trichosanthis Pericarpium（Gualoupi）,Trichosanthis Semen（Gualouzi）and Trichosanthis Radix（Tianhuafen）with raw and processed Aconiti Kusnezoffii Radix（Caowu）.Material and method:1 Determination of the content of primary substances（sugar,protein and fat）before and after the compatibility of Gualou and Caowu:1.1 The total sugar and total polysaccharide content of Gualou and Caowu before and after compatibility were determined by phenol-sulfuric acid colorimetry with glucose as reference substance.1.2 Coomassie brilliant blue colorimetric method was used to determine the total protein content of Gualou and Caowu before and after compatibility with bovine serum albumin as standard.1.3 The total fat content of Gualou and Caowu before and after compatibility was determined by gravimetric method.2 Determination of the content of secondary substances in Gualou before and after compatibility with Caowu:2.1 Al（NO₃）₃-Na NO₂colorimetric method was used to determine the content of total flavonoids before and after the compatibility of Gualou and Caowu with rutin as reference substance.2.2 Using vanillin-glacial acetic acid-perchloric acid colorimetry and oleanolic acid as reference substance,the content of total triterpenes before and after the compatibility of Gualou and Caowu was determined.2.3 The contents of rutin,isoquercitrin,quercetin,3,29-dibenzoyl-karounitriol and cucurbitacin B in Gualou before and after compatibility were determined by high performance liquid chromatography.3 The rats were administered by gavage,and the maximum dosage prescribed in the Pharmacopoeia was used for compatibility.The urine,blood and kidney tissues of rats were collected.Urine volume,kidney coefficient,renal pathological section,urine creatinine,serum creatinine,urea nitrogen,urine protein,urine alkaline phosphatase,blood alkaline phosphatase,N-acetyl-β-D-glucosaminidase,serum uric acid,α1-microglobulin,β2-microglobulin,urine microalbumin,neutrophil gelatinase-associated lipid carrier protein,kidney injury factor 1,serum cystatin C and glomerular filtration rate were measured and compared.4 Principal component analysis was used to calculate the comprehensive scores of total components,monomer components and nephrotoxicity,and correlation analysis was performed.Results:1 Changes in the content of primary substances in Gualou before and after compatibility with Caowu:Total sugar content:Gualoupi>Tianhuafen>Quangualou>Gualouzi;the total polysaccharide content:Tianhuafen>Gualoupi>Quangualou>Gualouzi;the total protein content:Gualouzi>Gualoupi>Quangualou>Tianhuafen;the total fat content:Quangualou>Gualouzi>Gualoupi,there was significant difference,there was no significant difference between Gualoupi and Tianhuafen.The content of total sugar increased significantly,the content of total polysaccharide had no significant difference,and the content of total protein and total fat decreased significantly.The contents of total sugar and total polysaccharide increased significantly,the content of total protein decreased significantly,and there was no significant difference in the content of total fat.The content of total sugar and total polysaccharide increased significantly,and the content of total protein decreased significantly after the compatibility of Gualou with raw and processed Caowu.The content of total fat increased significantly after the compatibility with raw Caowu,and there was no significant difference in the content of total fat after the compatibility with processed Caowu.The content of total sugar and total polysaccharide increased significantly,the content of total protein decreased significantly,and there was no significant difference in the content of total fat.2 The content changes of secondary substances in Gualou before and after compatibility with Caowu:The content of total flavonoids in Quangualou,Gualoupi,Gualouzi and Tianhuafen combined with Caowu increased significantly,and the coldness decreased.The content of total triterpenes increased in the compatibility of Quangualou and raw Caowu,and there was no significant difference between the compatibility of Quangualou and processed Caowu,.There was no significant difference in the content of total triterpenes between Gualoupi and raw Caowu,and the content decreased after compatibility with processed Caowu.There was no significant difference in the content of total triterpenes between Gualouzi and raw Caowu,but the content of total triterpenes in Gualouzi and processed Caowu decreased.The content of total triterpenes in the compatibility of Tianhuafen and raw Caowu increased,and there was no significant difference in the content of processed Caowu.The content of rutin and isoquercitrin in the compatibility of Quangualou and Caowu increased significantly.There was no significant difference before and after quercetin compatibility,and the content of cucurbitacin B decreased significantly.The contents of rutin,isoquercitrin,quercetin and cucurbitacin B in the compatibility of Gualoupi and Caowu were significantly reduced.There was no significant difference in the content of rutin between Gualouzi and Caowu,and the content of isoquercitrin was significantly increased.The contents of 3,29-dibenzoyl-karounitriol and cucurbitacin B were significantly decreased.There was no significant difference in the content of rutin and isoquercitrin between Gualouzi and Caowu.The content of triol was significantly increased and the content of cucurbitacin B was significantly decreased.The content of cucurbitacin B in the compatibility of Gualouzi and Caowu was significantly reduced.3 The changes of nephrotoxicity before and after single administration of Trichosanthis Fructus and Aconiti Kusnezoffii Radix:After administration of Quangualou,the urine volume decreased,α1-MG,β2-MG,m ALB,NGAL and KIM-1 increased,the cell structure of renal tubular wall was destroyed,the boundary was unclear,and inflammatory cells infiltrated.After administration of Gualoupi,urine volume decreased,UA,α1-MG,β2-MG,m ALB,NGAL and KIM-1increased,infiltrated inflammatory cells,proximal and distal convoluted tubules swelled,and tube wall thickened.After administration of Gualouzi,urine volume decreased,α1-MG,β2-MG,m ALB,NGAL and KIM-1 increased,renal clearance rate decreased,renal tubule boundary was unclear,and scattered inflammatory cells were observed.After administration of Tianhuafen,urinary protein,β2-MG,m ALB,NGAL and KIM-1 increased,glomerular pathological changes,glomerular pyknosis,and renal tubular swelling.After administration of raw Caowu,urine AKP,β2-MG,m ALB,NGAL and KIM-1 increased,and inflammatory cell infiltration occurred in some parts.After administration of processed Caowu,urinary creatinine,urinary AKP,β2-MG,m ALB,NGAL and KIM-1 increased,and a large number of inflammatory cells infiltrated.After administration of Quangualou-raw Caowu,urine AKP,α1-MG,β2-MG,m ALB,NGAL,KIM-1 and Cys-C increased,and there was no significant change in renal structure.Compared with Quangualou,urine AKP increased,urine volume and renal clearance rate decreased.After administration of Quangualou-processed Caowu,urine volume decreased,urine creatinine,urine AKP,α1-MG,β2-MG,m ALB,NGAL,KIM-1 and Cys-C increased,renal tubular pathological changes,unclear structure,inflammatory cell infiltration;compared with the Quangualou,urine AKP increased.After administration of Gualoupi-raw Caowu,urine volume decreased,urine AKP,α1-MG,β2-MG,m ALB,NGAL and KIM-1 increased,and a large number of infiltrated inflammatory cells were observed.Compared with Gualoupi,urine AKP increased.After administration of Gualoupi-processed Caowu,the urine volume decreased,urine AKP,α1-MG,β2-MG,m ALB,NGAL,KIM-1 and Cys-C increased,proximal and distal convoluted tubules swelled,glomerular pyknosis,inflammatory cell infiltration;compared with Gualoupi,urine AKP increased.After administration of Gualouzi-raw Caowu,urine volume decreased,urine creatinine,urine AKP,α1-MG,β2-MG,m ALB,NGAL,KIM-1 and Cys-C increased,renal tubular swelling boundary was not clear,tubular wall atrophy,and the lumen became larger.Compared with Gualouzi,urine AKP increased.After administration of Gualouzi-processed Caowu,urine volume decreased,urine creatinine,urine AKP,UA,α1-MG,β2-MG,m ALB,NGAL,KIM-1 and Cys-C increased,renal clearance rate decreased,renal interstitial and glomerular congestion,tubular swelling,inflammatory cell infiltration;compared with Gualouzi,there was no significant difference.After administration of Tianhuafen-raw Caowu,urinary protein,urinary AKP,α1-MG,β2-MG,m ALB,NGAL,KIM-1 and Cys-C increased,renal tissue showed pathological changes,and glomeruli and renal tubules showed necrotic changes.Compared with Tianhuafen,urinary AKP increased,glomeruli and renal tubular necrosis.After administration of Tianhuafen and processed Caowu,urine AKP,α1-MG,β2-MG,m ALB,NGAL,KIM-1 and Cys-C increased,inflammatory cell infiltration in glomeruli,renal tubular swelling,unclear wall structure,and congestion in some parts of renal interstitium;compared with Tianhuafen,urine AKP increased.4 The changes of nephrotoxicity after multiple administration before and after the compatibility of Gualou and Caowu:After multiple doses of Quangualou,urine volume decreased,urine protein,UA,α1-MG,β2-MG,m ALB,NGAL,KIM-1 and Cys-C increased,renal clearance rate decreased,renal tubular structure was not clear,glomerular atrophy,congestion,inflammatory cell infiltration.After multiple doses of Gualoupi,urine volume decreased,urine protein,urine AKP,UA,α1-MG,β2-MG,m ALB,NGAL,KIM-1 and Cys-C increased,and the structure of some parts of renal tubules was not clear.After multiple doses of Gualouzi,urinary protein,UA,α1-MG,β2-MG,m ALB,NGAL,KIM-1 and Cys-C increased,renal congestion,glomerular atrophy,and a small amount of infiltrating inflammatory cells.After multiple administration of Tianhuafen,urine volume decreased,urinary creatinine,urinary protein,UA,α1-MG,β2-MG,m ALB,NGAL,KIM-1 and Cys-C increased,glomerular atrophy,and renal tubular swelling structure changed.After repeated administration of raw Caowu,urinary protein,urinary AKP,UA,α1-MG,β2-MG,m ALB,NGAL and KIM-1 increased,and there was no significant change in renal tissue structure.After multiple doses of processed Caowu,urinary protein,urinary AKP,UA,α1-MG,β2-MG,m ALB,NGAL and KIM-1 increased,renal tubules were unevenly stained,and glomeruli had inflammatory cell infiltration.After repeated administration of Quangualou-raw Caowu,urinary protein,urinary AKP,α1-MG,β2-MG,m ALB,NGAL,KIM-1 and Cys-C increased,glomerular atrophy and congestion;compared with Quangualou,urine volume decreased,urine AKP increased,and renal clearance rate decreased.After multiple doses of Quangualou-processed Caowu,urine volume decreased,urine creatinine,urine protein,urine AKP,α1-MG,β2-MG,m ALB,NGAL,KIM-1 and Cys-C increased,renal tubular wall thinning structure was not clear,glomerular atrophy;compared with Quangualou,urine volume decreased,urine AKP increased,and renal clearance rate decreased.After repeated administration of Gualoupi-raw Caowu,urinary creatinine,urinary protein,urinary AKP,UA,α1-MG,β2-MG,m ALB,NGAL,KIM-1 and Cys-C increased,renal clearance rate decreased,glomerular congestion,renal tubular wall thinning,lumen enlargement;compared with Gualoupi,urine volume decreased,urine AKP increased,renal clearance rate decreased,glomerular congestion.After repeated administration of Gualoupi-processed Caowu,urine volume decreased,urinary protein,urinary AKP,α1-MG,β2-MG,m ALB,NGAL,KIM-1 and Cys-C increased,renal clearance rate decreased,and renal structure did not change significantly.Compared with Gualoupi,urine volume decreased,urine protein,urine AKP and UA increased,and renal clearance rate decreased.After repeated administration of Gualouzi-raw Caowu,urinary protein,urinary AKP,α1-MG,β2-MG,m ALB,NGAL,KIM-1 and Cys-C increased,renal tissue pathological changes,glomerular congestion,tubular structure is not clear;compared with Gualouzi,urinary AKP increased and renal tissue showed pathological changes.After repeated administration of Gualouzi-processed Caowu,urinary protein,urinary AKP,UA,α1-MG,β2-MG,m ALB,NGAL,KIM-1 and Cys-C increased,renal tubular swelling structure was not clear,and inflammatory cell infiltration was also observed.Compared with Gualouzi,urinary AKP increased,and the structure of renal tubular swelling was not clear.After repeated administration of Tianhuafen-raw Caowu,urine volume decreased,BUN,urine protein,urine AKP,UA,α1-MG,β2-MG,m ALB,NGAL,KIM-1 and Cys-C increased,renal clearance rate decreased,and renal structure did not change significantly.Compared with Tianhuafen,urine volume decreased,urine AKP and KIM-1 increased,and renal clearance rate decreased.After repeated administration of Tianhuafen-processed Caowu,urine volume decreased,BUN,urine protein,urine AKP,UA,α1-MG,β2-MG,m ALB,NGAL,KIM-1 and Cys-C increased,renal clearance rate decreased,glomerular atrophy,tubular pathological changes;compared with Tianhuafen,urine volume was decreased,urine AKP and KIM-1 were increased,renal clearance rate was decreased,and renal tubular pathological changes were observed.5 The changes of nephrotoxicity of single administration and multiple administration before and after the compatibility of Gualou and Caowu:Compared with single administration,Quangualou reduced urine volume,renal tissue congestion,and glomerular atrophy.Compared with single administration,Cys-C increased and renal tubular structure was unclear in Gualoupi.Compared with single administration,Gualouzi reduced urine volume,increased KIM-1,renal tissue congestion,and glomerular atrophy.Tianhuafen did not change significantly compared with single administration.There was no significant change in raw Caowu compared with single administration.Compared with single administration,processed Caowu showed uneven staining of renal tubules.Quangualou-raw Caowu compared with single administration,glomerular atrophy,congestion.Quangualou-processed Caowu compared with single administration,urinary protein decreased,renal tubular wall thinning structure was not clear,glomerular atrophy.Compared with single administration,Gualoupi-raw Caowu increased Cys-C,decreased renal clearance rate,glomerular congestion,thinner renal tubular wall and larger lumen.Compared with single administration,Gualoupi-processed Caowu decreased renal coefficient,increased Cys-C and decreased renal clearance rate.Compared with single administration,Gualouzi-raw Caowu increased Cys-C and pathological changes in renal tissue.Compared with single administration,Gualouzi-processed Caowu reduced urine volume,kidney coefficient and renal clearance rate.Compared with single administration,Tianhuafen-raw Caowu decreased urine volume,increased BUN,UA and KIM-1,decreased renal clearance rate,and pathological changes of renal tubules.Compared with single administration,Tianhuafen-processed Caowu decreased urine volume,increased BUN and KIM-1,decreased renal clearance rate,and pathological changes of renal tubules.6 It can be seen from the correlation heat map that total sugar and total polysaccharide were significantly positively correlated with nephrotoxicity.Conclusion:1 Quangualou,Gualoupi and Gualouzi were cold.After compatibility,the content of primary substances increased or decreased to the content of primary substances in Caowu.The proportion of Caowu in compatibility was small,and the coldness was weakened.The total sugar and total polysaccharide content of Tianhuafen were lower than that of Caowu,and the total protein content was higher than that of Caowu.The total sugar and total polysaccharide content of Tianhuafen were higher than that of Caowu,and the total protein content was lower than that of Caowu.There was no significant difference in total fat,and the heat was enhanced.2 Quangualou,Gualoupi,Gualouzi and Tianhuafen combined with Caowu increased the content of total flavonoids and decreased the coldness.The total triterpenoid content of Quangualou and raw Caowu increased,and the coldness was enhanced.There was no significant difference between Quangualou and processed Caowu,and the coldness was unchanged.There was no significant difference in the content of total triterpenoids between Gualoupi and raw Caowu,and the coldness was unchanged.After compatibility with processed Caowu,the content decreased and the coldness increased.There was no significant difference in the content of total triterpenoids between Gualouzi and raw Caowu,and the coldness was unchanged.The content of total triterpenoids in Tianhuafen and raw Caowu increased,and the coldness decreased.There was no significant difference in the content of processed Caowu,and the coldness remained unchanged.Compatibility with raw Caowu reduced the dissolution of components in Quangualou（cucurbitacin B）,Gualoupi（rutin,isoquercitrin,quercetin and cucurbitacin B）,Gualouzi（rutin,trichosanthin and cucurbitacin B）and Tianhuafen（cucurbitacin B）,but increased the dissolution of components in Quangualou（rutin,isoquercitrin）and Gualouzi（isoquercitrin）.Compatibility with processed Caowu reduced the dissolution of components in Quangualou（cucurbitacin B）,Gualoupi（rutin,isoquercitrin,quercetin and cucurbitacin B）,Gualouzi（cucurbitacin B）and Tianhuafen（cucurbitacin B）,but increased the dissolution of components in Quangualou（rutin,isoquercitrin）and Gualouzi（rutin,isoquercitrin and trichosanthin）.3 Quangualou,Gualoupi,Gualouzi,Tianhuafen,Caowu and processed Caowu all had nephrotoxicity,and the toxicity of multiple administrations was enhanced,mainly reflected in early renal injury,including renal parenchymal damage,renal tubular damage,glomerular damage,acute kidney injury,early renal injury,early renal tubular dysfunction and nephritis.Among them,Tianhuafen had the strongest nephrotoxicity and obvious pathological changes.Quangualou,Gualoupi,Gualouzi and Tianhuafen were combined with raw and processed Caowu respectively,and the toxicity was enhanced,and the damage of renal parenchyma,glomeruli and tubules was more significant.4 Total sugar and total polysaccharides were significantly positively correlated with nephrotoxicity,suggesting that total sugar and total polysaccharides were the main components of nephrotoxicity.