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This study compared the effects of Curcuma longa before and after processing with vinegar on the rat model of dysmenorrhea with the syndrome of liver depression and Qi stagnation to reveal the mechanism of vinegar processing in improving the role of C. longa in soothing liver and relieving pain. The rat model of dysmenorrhea with the syndrome of liver depression and Qi stagnation was established according to the Preparation of the Animal Model of Dysmenorrhea(Draft) and the chronic unpredictable stress me-thod. The changes in the body weight, organ indexes, writhing latency, writhing score, and serum levels of six liver function indicators, sex hormones, pain factors, and blood rheological indicators were measured to evaluate the efficacy of C. longa processed with vinegar or not in treating dysmenorrhea in the rats with syndrome of liver depression and qi stagnation. Compared with the model group, the C. longa group(processed with vinegar or not) showed slow weight loss, increase in writhing latency, and decrease in writhing response(P<0.05). The inhibition rates on writhing in raw C. longa, vinegar-processed C. longa, and positive groups were 33.780%, 64.611%, and 62.466%, respectively. The significantly higher inhibition rate of the vinegar processing group indicated that vinegar-processed C. longa demonstrated more significant therapeutic effect. The vinegar-processed C. longa group showed lower levels of alanine aminotransferase(ALT), alkaline phosphatase(ALP), aspartate aminotransferase(AST), direct bilirubin(DBIL), and total bilirubin(TBIL) and higher level of albumin(ALB)(P<0.05), which indicated that vinegar processing enhanced the therapeutic effect of C. longa on liver injury. The serum levels of estradiol(E_2) and oxytocin(OT) were lower in the vinegar-processed C. longa group(P<0.05), indicating that the vinegar-processed C. longa could regulate the sex hormone levels, reduce the activity of uterine smooth muscle and contraction of uterus, and alleviate the symptoms of dysmenorrhea in rats. Moreover, the vinegar-processed C. longa group showed lower interleukin-6(IL-6) and arginine vasopressin(AVP) levels and higher beta-endorphin(β-EP) level(P<0.05), which indicated that vinegar-processed C. longa regulated the levels of pain factors to exert the pain-relieving effect. Drug intervention decreased the whole blood viscosity low-cut, medium-cut and high-cut values, plasma viscosity, whole blood reduction viscosity low-cut and high-cut values, erythrocyte cumulative pressure, and equation K value of erythrocyte sedimentation rate(P<0.05), and the vinegar-processed C. longa group outperformed other groups. This result indicated that vinegar processing enhanced the function of C. longa in improving the local blood rheology. C. longa processed with vinegar can enter the liver to relieve the da-mage to the heart, liver, kidney, and uterus, repair the liver function, and recover the sex hormone levels and immune function by regulating the levels of sex hormones and pain factors and improving the blood rheology. It activates the pain-relieving mechanism to relieve the pain, protect the liver, and fight inflammation, which is consistent with the theory that vinegar processing facilitates C. longa entering the liver to sooth liver and relieve pain.
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Plasma metabolomics based on UHPLC-Q-TOF-MS/MS technique was developed for profiling the mechanism on attenuating hepatic fibrosis of Bupleuri Radix (BR) and Paeoniae Radix Alba (PRA) before and after vinegar-processing and compatibility, and to screen potential pharmacodynamic substances by spectrum-effect correlation method in this study. Firstly, SD rats with CCl4-induced hepatic fibrosis were used as an in vivo model. The blood and tissue samples were collected for the analyses of pharmacodynamic indexes and plasma metabolomics after six weeks’ administration of BR, vinegar-processed BR (VPBR), PRA, vinegar-processed PRA (VPPRA), BR-PRA herb-pair, and VPBR-VPPRA herb-pair. The experiment was approved by the experimental animal ethics committee from Nanjing University of Chinese Medicine (No.202103A002). The results of pharmacodynamics indicated that the levels of alanine aminotransferase (ALT, P < 0.01), aspartate aminotransferase (AST, P < 0.01), and hydroxyproline (HYP, P < 0.01) were decreased significantly, while the level of glutathione peroxidase (GSH-Px, P < 0.05) was increased obviously after administration of all treatment groups. Next, UHPLC-Q-TOF-MS/MS was performed to characterize the endogenous metabolites. A total of 20 differential endogenous metabolites related to the pathogenesis of hepatic fibrosis were identified in positive and negative ion modes, mainly involving five metabolic pathways of retinol metabolism, glycerol phospholipid metabolism, glyceride metabolism, fatty acid biosynthesis, and arachidonic acid metabolism. Meanwhile, a concept named correction rate was introduced to evaluate the back-regulation effects of all treatment groups on differential metabolites, and 10 differential metabolites were corrected by all treatment groups. The correction effects of the vinegar-processed herb groups were better than those of the crude ones, and the correction effects of the herb-pair groups were better than those of the single ones. Interestingly, the best correction effect was found in the VPBR-VPPRA herb-pair group, which further verified the efficacy improvement through vinegar-processing and compatibility. Partial least square method and VIP analysis combined with spectrum-effect correlation were applied for screening pharmacodynamic markers, and 38 ingredients with higher correlation with four classical pharmacodynamic indexes (ALT, AST, HYP, and GSH-Px) were identified as pharmacodynamic markers of the anti-hepatic fibrosis effects of BR and PRA before and after vinegar-processing and compatibility. The results of the investigation could not only lay a foundation for clarifying the pharmacodynamic materials and mechanism of vinegar-processing and compatibility of BR and PRA in the treatment of hepatic fibrosis as well as provide a theoretical basis for demonstrating the scientific connotation of processing and compatibility, but also provide a reference for further drug design and development of BR and PRA in clinic.
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The present study investigated the effect of Euphorbiae Pekinensis Radix(EPR) on intestinal flora structure before and after vinegar processing and explored the detoxification mechanism of vinegar-processed EPR. In this study, the extraction efficiency of casbane diterpenes from EPR with different solvents was investigated, and the optimal solvent was selected to enrich these components. After 14 days of intragastric administration of total diterpene extract of EPR and vinegar-processed EPR, 16 S rDNA sequencing technology was used to detect the structural changes of intestinal flora. The flora related to the intestinal toxicity of EPR was screened out based on the results of intestinal pathological damage by correlation analysis. The results showed that Soxhlet extraction with chloroform as extraction solvent could enrich Casbane diterpenes in EPR. As revealed by 16 S rDNA sequencing results, EPR could significantly change the structure of intestinal flora, which could be reversed by vinegar-processing EPR. Some intestinal flora candidates might be related to detoxification of vinegar processing. The correlation analysis of intestinal flora candidates and indexes related to intestinal mucosal injury showed that compared with EPR, vinegar-processed EPR could down-regulate the abundance of some pathogenic bacteria such as Mucispirillum, Bilophila, and Ruminiclostridium, and up-regulated some probiotics such as Enterorhabdus, Ruminococcaceae_UCG-014, Barnesiella, and Candidatus. The intestinal toxicity caused by EPR may be related to the disturbance of intestinal flora, and vinegar-processed EPR can improve intestinal flora disorder by up-regulating the abundance of probiotics and down-regulating the abundance of pathogenic bacteria to remodel the intestinal mucosal barrier and reduce toxicity.
Subject(s)
Acetic Acid/chemistry , Colon , Drugs, Chinese Herbal/chemistry , Gastrointestinal Microbiome , Plant RootsABSTRACT
Objective: To screen a processing method suitable for deodorizing of Periplaneta americana (PA) through subjective and objective evaluation combined with changes in volatile components, and explore the mechanism of deodorization. Methods: Raw, vinegar-processed, wine-processed, wheat bran-processed products of PA were prepared respectively. Volunteer sensory evaluation combined with electronic nose system was used to evaluate the odor difference between raw and processed products to screen a better processed product. HS-SPME-GC-MS and ROAV were used to analyze the key odorous components of PA. The peak area normalization method combined with multivariate statistical analysis were used to analyze the difference of volatile components between raw and processed products to explore the mechanism of processing. Results|| Volunteer scores showed that the average scores of raw, vinegar-processed, wine-processed, wheat bran-processed products were 3.38, 1.25, 2.88, and 3.04, respectively. The results of the electronic nose showed that the Euclidean distance between the raw and vinegar-processed, wine-processed, wheat bran-processed products were 7.34, 3.77 and 1.60, respectively, but the direction of scatter of vinegar-processed product was opposite to that of wine-processed product and wheat bran-processed product, suggesting that the mechanism of deodorization may be different. Vinegar processing was determined as the optimum method for deodorizing by comprehensive analysis of subjective and objective evaluation data. The key odor components of PA were 3-methyl butanal, hexanal, nonyl aldehyde, heptaldehyde, decyl aldehyde, phenyl acetaldehyde, (E,E)-2,4-nonadienal, 2-pentylfuran, (+)-limonene, and myristic aldehyde. The PCA result of volatile components showed that four kinds of processed products can be clearly distinguished, and there were differences in volatile components and content. There were seven differential chemical components between raw and vinegar-processed products, including hexanal, palmitic acid, oleic acid, acetic acid, ethyl oleate, ethyl palmitate, and ethyl linoleate. The content of vinegar-processed products was lower than that of raw products, especially the key odorous component hexanal. Conclusion: The vinegar processing is a better method to improve the odor of PA, and its mechanism may be associated with reducing odorous components such as hexanal.
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Abstract This study investigated the influence of different processing methods on the oral toxicity of Sophora alopecuroides L., Fabaceae, seeds in mice and on the contents of five known toxic-effective quinolizidine alkaloids from the ethanol extracts quantified by ultra-performance liquid chromatography coupled to tandem quadrupole mass spectrometry. It provides an evidence to elucidate the possible reasons why vinegar-processing and parching methods significantly decrease the acute oral toxicity induced by S. alopecuroides and why wine-processing method increases it instead (demonstrated by measurement of LD50 and histopathological analysis). The analytical performance for the determination of the five analytes was evaluated by linearity, stability, repeatability, precision and accuracy, and recovery test. The lowest limit of quantification was determined to be 5 ng/ml for each substance and the precision and accuracy at lowest limit of quantification were below 20%. Cytisine, the most toxic alkaloid among the five alkaloids, declined 11.26, 3.98, and 2.73 folds after being vinegar-processed and fried in a ceramic or iron pan, respectively and had a very close correlation with the toxicity of S. alopecuroides seeds (r = 0.8589). Other matrine-type alkaloids with lower toxicity including matrine, sophcarpine, and sophoridine decreased after being wine-processed and fried in a ceramic pan, but increased 4.44, 7.20, and 7.23 folds when being processed by vinegar. Oxymatrine declined in all groups. It, therefore, reveals that vinegar-processing method reduces the oral toxicity of S. alopecuroides mainly due to a sharp decrease of cytisine, thus improves its clinical safety.
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To screen the toxic polar fractions of Daphne genkwa, compare the toxicity of D. genkwa on crypts epithelial cells IEC-6 before and after vinegar processing, and preliminarily investigate the mechanism of D. genkwa vinegar processing on toxicity reducing. The proliferation of IEC-6 cells was observed by MTT. The levels of superoxide dismutase (SOD), malondialdehyde (MDA), glutathione (GSH), lactate dehydrogenase (LDH), as well as the enzyme activity of Na⁺-K⁺-ATPase and Ca²⁺-Mg²⁺-ATPase were determined in IEC-6 cells to evaluate the oxidative damages degree of IEC-6 cells. The apoptosis and cell cycle were analyzed by Flow Cytometry. The results showed that the dichloromethane extraction was the toxic polar fraction of D. genkwa, and after vinegar processing, the toxicity of dichloromethane fraction was significantly reduced (<0.01). As compared with the blank control group, the dichloromethane fraction of D. genkwa can obviously decrease the levels of SOD, Na⁺-K⁺-ATPase, Ca²⁺-Mg²⁺-ATPase (<0.01) and content of GSH, but increase the level of LDH and MDA in cell supernatant (<0.01). Besides, it obviously increased the early and late apoptotic rate of IEC-6 cells, obviously decreased the proportion of G₁stage cells, increased the ratio of S stage cells and M stage cells (<0.01). After vinegar processing, as compared with D. genkwa groups of various doses, it can significantly increase the levels of SOD, Na⁺-K⁺-ATPase, Ca²⁺-Mg²⁺-ATPase (<0.01) and content of GSH, decrease the level of LDH, MDA(<0.01), significantly decrease the early and late apoptosis rate of IEC-6 cells (<0.01), increase the proportion of G₁stage cells, and decrease the ratio of S stage cells and M stage cells (<0.01). Vinegar processing can reduce the toxicity of dichloromethane fraction of D. genkwa, and its mechanism may be associated with improving the activity of antioxidant enzymes and permeability in IEC-6 cells, and decreasing the oxidative damage.
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To investigate the toxicity changes of Euphorbiae Ebracteolatae Radix (EER) before and after vinegar processing, toxic diterpenoids were concentrated with chloroform as extraction solvent from EER. Then the residue was extracted for non-chloroform extract with 95% ethanol and water after extraction with chloroform. The chloroform extraction of vinegar processed EER was prepared with the same method. The mice received the drug by oral administration. Moisture content in mice feces, duodenum and colon tissue, aquaporin AQP1, AQP3, AQP4 protein expression levels were assayed as the indexes to investigate the toxicity variation of chloroform fraction, non-chloroform fraction, as well as intestinal tract toxicity before and after vinegar processing of EER. The results showed that the chloroform fraction extracted from EER could significantly increase the moisture content in mice feces, duodenum and colon, and decrease AQP1 protein expression level, increase AQP3 and AQP4 protein expression levels in the colon. The intestinal toxicity of the chloroform extract was significantly higher than that of non-chloroform extract. The moisture content in mice feces, duodenum and colon was significantly decreased, and the AQPs protein expression tended to be normal in the colon after vinegar processing. The results showed that the chloroform fraction extracted from EER could lead to diarrhea, intestinal edema, and the intestinal toxicity action was associated with interfering AQPs protein expression and promoting intestinal fluid transport disorder in mice. Vinegar-processing could reduce intestinal toxicity of EER, so vinegar processing was considered to be the scientific processing method of EER.
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Objective: To look for anti-liver injury potential biomarkers and explore the scientific connotation of vinegar-processed Schisandra chinensis enhancing hepatoprotective effect by investigating the endogenous metabolites difference in bile before and after S. chinensis was processed with vinegar. Methods: The metabolic skills were adopted based on UPLC-Q/TOF-MS, PCA, and OPLS-DA for screening and identification of biomarkers related to liver injury. Results: Bile metabolite profile in control group, model group, raw S. chinensis (RSC) group and vinegar-processed S. chinensis (VPSC) were separated obviously, eight potential biomarkers associated with liver injury were identified, including LysoPC (20:4), PG (18:0/18:1), 12-ketodeoxycholic acid, TG (64:2), etc. Conclusion: After giving raw and vinegar-processed S. chinensis, the levels of above markers can be adjusted to the normal state, and the regulating function of VPSC is stronger than that of the raw one. VPSC might have the anti-liver injury effect on all these metabolic pathways.
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To explore the effect of vinegar-processed Curcumae Rhizoma on endogenous metabolites in bile by investigating the endogenous metabolites difference in bile before and after Curcumae Rhizoma was processed with vinegar. Alcohol extracts of crude and vinegar-processed Curcumae Rhizoma, as well as normal saline were prepared respectively, which were then given to the rats by intragastric administration for 0.5 h. Then common bile duct intubation drainage was conducted to collect 12 h bile of the rats. UPLC-TOF-MS analysis of bile samples was applied after 1∶3 acetonitrile protein precipitation; unidimensional statistics were combined with multivariate statistics and PeakView software was compared with network database to identify the potential biomarkers. Vinegar-processed Curcumae Rhizoma extracts had significant effects on metabolites spectrum in bile of the rats. With the boundaries of P<0.05, 13 metabolites with significant differences were found in bile of crude and vinegar-processed Curcumae Rhizoma groups, and 8 of them were identified when considering the network database. T-test unidimensional statistical analysis was applied between administration groups and blank group to obtain 7 metabolites with significant differences and identify them as potential biomarkers. 6 of the potential biomarkers were up-regulated in vinegar-processed group, which were related to the metabolism regulation of phospholipid metabolism, fat metabolism, bile acid metabolism, and N-acylethanolamine hydrolysis reaction balance, indicating the mechanism of vinegar-processed Curcumae Rhizoma on endogenous metabolites in bile of the rats.
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Objective:To study the influence of Rhizoma Corydalis before and after vinegar processing on the microelement content in the eye tissues of rats to verify the vinegar processing leading the herb to liver meridian. Methods: The rats were divided into the crude Corydalis group, vinegar processing group and negative group. The content of microelement Cu, Zn and Mn in rat ocular tissues was determined and compared after the administration. Results:Corydalis could significantly increase the content of Cu and Zn in fun-dus blood and that of Zn and Mn in the eyeball vitreous, and Rhizoma Corydalis after vinegar processing showed more obvious effect. Conclusion:Rhizoma Corydalis with vinegar processing can improve the distribution of microelements in eye tissues, and because liver opens at eyes, the results provide evidence for Corydalis after vinegar processing leading to liver meridian.
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Objective: To study the dissolving yield and ratio fluctuation of Sparganii Rhizoma effective composition groups, including total flavonoids, total alkaloid, and active Al3+, when processed with different vinegar processing technologies. Methods: L9(34) orthogonal test design was used with acetic acid content (A), vinegar ratio (B), soaking time (C), and frying temperature (D) as influencing factors. The differences of dissolving yield and ratio of 80% methanol extract, total flavonoids, total alkaloid, and active Al3+ of processed Sparganii Rhizoma under different vinegar processing technologies were compared. Results: Compared with crude Sparganii Rhizoma, the processed products with a little acetic acid had enhanced the dissolving yield of 80% methanol extract. Moreover, the processed Sparganii Rhizoma with vinegar had effectively enhanced the dissolving yield and ratio of total flavonoids, total alkaloid, and active Al3+. The dissolving yield and ratio of total flavonoids were greatly impacted by frying temperature. Meanwhile, the alkaloid dissolving yield and ratio were most influenced by acetic acid content. The products processed with abundant acetic acid had obviously reduced the dissolving yield of 80% methanol extract, but had increased the alkaloid and active Al3+ dissolving ratio in the extract. Conclusion: The processed Sparganii Rhizoma with vinegar could obviously affect the dissolving yield and ratio of different pharmacodynamic compositions in Sparganii Rhizoma. Suitable processed products should be adopted when Sparganii Rhizoma is used in the different treatments.