Investigation of the hemostatic mechanism of Gardeniae fructus Praeparatus based on pharmacological evaluation and network pharmacology.

Background: As documented in the Chinese Pharmacopoeia, Gardeniae fructus Praeparatus (GFP) can cool the blood during hemostasis and treat various internal hemorrhagic diseases. However, the underlying mechanisms are not yet well understood. This work was designed to decipher the possible mechanism by which GFP prevents hemorrhage. The integration of pharmacodynamics-based and bioinformatics-based methods provided evidence to support the clinical effects of GFP in treating bleeding. Methods: Using ultra-performance liquid chromatography (UPLC) analysis, we quantified the main active ingredients for a preliminary quality assessment of GFP. The pharmacology study was conducted to confirm the essential antihemorrhagic effects of GFP. A rat model of ethanol-induced gastric hemorrhage was established and was followed by intervention with GFP in low, middle, and high doses (4.5, 9, 18 g/kg). Gastric tissues were harvested for macroscopic and histological evaluation of lesions. The contents of thromboxane B2 (TXB2) and 6-keto-prostaglandin-F1α (6-keto-PGF1α) in the serum were determined. Additionally, network pharmacology was proposed to illuminate the potential mechanisms. Following the collection of GFP compositions, the compound- and hemorrhage-related targets were retrieved from public databases. The protein-protein interaction (PPI), gene ontology, pathways analysis, and molecular docking were performed for targets of GFP in gastrointestinal bleeding. Results: The study found ten main active ingredients that could be used for quality control of GFP. Importantly, the middle and high doses of GFP were found to promote the healing of gastric bleeding. The content of 6-keto-PGF1α was significantly degraded in the middle and high treated groups (P<0.05). The level of TXB2 was augmented by a middle (P<0.05) and high dose of GFP. Further, we constructed the network of candidate ingredients and hemorrhage-related targets. Pathway analysis predicted the mechanisms associated with interleukin 4 and interleukin 13 signaling and platelet activation. PPI analysis identified subnetworks with biological functions and also sifted hub targets that affected the antihemorrhagic progress. The candidate proteins had a good binding force with major components. Conclusions: GFP exhibits a promising effect in ameliorating bleeding, with the relevant molecular mechanisms possibly being related to the regulation of the immune system and platelet activation. Therefore, GFP can potentially exert a protective effect on gastrointestinal bleeding in clinic.

Determining the Stir-Frying Degree of Gardeniae Fructus Praeparatus Based on Deep Learning and Transfer Learning.

Gardeniae Fructus (GF) is one of the most widely used traditional Chinese medicines (TCMs). Its processed product, Gardeniae Fructus Praeparatus (GFP), is often used as medicine; hence, there is an urgent need to determine the stir-frying degree of GFP. In this paper, we propose a deep learning method based on transfer learning to determine the stir-frying degree of GFP. We collected images of GFP samples with different stir-frying degrees and constructed a dataset containing 9224 images. Five neural networks were trained, including VGG16, GoogLeNet, Resnet34, MobileNetV2, and MobileNetV3. While the model weights from ImageNet were used as initial parameters of the network, fine-tuning was used for four neural networks other than MobileNetV3. In the training of MobileNetV3, both feature transfer and fine-tuning were adopted. The accuracy of all five models reached more than 95.82% in the test dataset, among which MobileNetV3 performed the best with an accuracy of 98.77%. In addition, the results also showed that fine-tuning was better than feature transfer in the training of MobileNetV3. Therefore, we conclude that deep learning can effectively recognize the stir-frying degree of GFP.

A Novel Method for Quality Evaluation of Gardeniae fructus Praeparatus during Heat Processing Based on Sensory Characteristics and Chemical Compositions.

The intrinsic chemical components and sensory characteristics of Gardeniae fructus Praeparatus (GFP) directly reflect its quality and subsequently, affect its clinical curative effect. However, there is little research on the correlation between the appearance traits and chemical compositions of GFP during heat processing. In this study, the major components of five typical processed decoction pieces of GFP were determined. With the deepening of processing, the contents of geniposidic acid and 5-HMF gradually increased, while the contents of deacetyl-asperulosidic acid methyl ester, gardenoside, and two pigments declined. Moreover, the electronic eye, electronic tongue, and electronic nose were applied to quantify GFP's sensory properties. It was found that the chroma values showed a downward trend during the processing of GFP. The results of odor showed that ammonia, alkenes, hydrogen, and aromatic compounds were the material base for aroma characteristics. Complex bitterness in GF was more obvious than that in other GFP processed products. Furthermore, one mathematical model was established to evaluate the correlation between the sensory characteristics and chemical composition of GFP during five different stages. A cluster analysis and neural network analysis contributed to recognizing the processing stage of GFP. This study provided an alternative method for the exterior and interior correlation-based quality evaluation of herbs.

[Quality changes of Gardeniae Fructus Praeparatus processed by different frying methods: a color-component correlation analysis].

The chromatic values of the broken-fried and single-fried Gardeniae Fructus Praeparatus(GFP) were measured by the color analyzer to analyze the color variation rule, and the contents of 10 main components were determined by ultra-high performance liquid chromatography(UPLC). The multivariate statistical analysis, Pearson correlation analysis, and discriminant analysis were conducted to investigate the color and components of GFP samples. The experimental results revealed that L~*, a~*, b~*, and E~*ab decreased continuously during processing, and the color of samples gradually deepened. The trend and range of chromatic values during broken-frying and single-frying processes were basically identical. Gardenoside, crocin-Ⅰ(C-Ⅰ), and crocin-Ⅱ(C-Ⅱ) showed an obviously downward trend, while the contents of geniposidic acid and 5-hydroxymethylfurfural(5-HMF) increased significantly. Shanzhiside, deacetyl-asperulosidic acid methyl ester, and geniposide(G2) showed a downward trend. Scandoside methyl ester rose first and fell later. Genipin-1-O-gentiobioside(G1) went through a decrease-increase-decrease trend. The change trends of component contents during broken-frying and single-frying processes were generally consistent, but the change range was different. Among all the components, scandoside methyl ester and G1 showed obvious change. Because of different stir-frying time, the change rate of each component content in the process of broken-frying was higher than that in single-frying process. Additionally, geniposidic acid, gardenoside, scandoside methyl ester, C-Ⅰ, C-Ⅱ, and 5-HMF exhibited a higher correlation with apparent color. On the basis of above findings, the discriminant function of two frying processes was established, which could be applied to the discrimination of broken-fried and single-fried samples. This study analyzed the dynamic quality change rule of GFP during broken-frying and single-frying processes based on color-component correlation analysis, and found the two methods showed consistent change trend, yet with slight difference in the quality of samples. This study can provide data support for the processing of GFP.

Quality changes of Gardeniae Fructus Praeparatus processed by different frying methods: a color-component correlation analysis / 中国中药杂志

The chromatic values of the broken-fried and single-fried Gardeniae Fructus Praeparatus(GFP) were measured by the color analyzer to analyze the color variation rule, and the contents of 10 main components were determined by ultra-high performance liquid chromatography(UPLC). The multivariate statistical analysis, Pearson correlation analysis, and discriminant analysis were conducted to investigate the color and components of GFP samples. The experimental results revealed that L~*, a~*, b~*, and E~*ab decreased continuously during processing, and the color of samples gradually deepened. The trend and range of chromatic values during broken-frying and single-frying processes were basically identical. Gardenoside, crocin-Ⅰ(C-Ⅰ), and crocin-Ⅱ(C-Ⅱ) showed an obviously downward trend, while the contents of geniposidic acid and 5-hydroxymethylfurfural(5-HMF) increased significantly. Shanzhiside, deacetyl-asperulosidic acid methyl ester, and geniposide(G2) showed a downward trend. Scandoside methyl ester rose first and fell later. Genipin-1-O-gentiobioside(G1) went through a decrease-increase-decrease trend. The change trends of component contents during broken-frying and single-frying processes were generally consistent, but the change range was different. Among all the components, scandoside methyl ester and G1 showed obvious change. Because of different stir-frying time, the change rate of each component content in the process of broken-frying was higher than that in single-frying process. Additionally, geniposidic acid, gardenoside, scandoside methyl ester, C-Ⅰ, C-Ⅱ, and 5-HMF exhibited a higher correlation with apparent color. On the basis of above findings, the discriminant function of two frying processes was established, which could be applied to the discrimination of broken-fried and single-fried samples. This study analyzed the dynamic quality change rule of GFP during broken-frying and single-frying processes based on color-component correlation analysis, and found the two methods showed consistent change trend, yet with slight difference in the quality of samples. This study can provide data support for the processing of GFP.

[Correlation analysis of color change and Maillard reaction during processing of Gardeniae Fructus Praeparatus].

The chromaticity space parameters of the samples during the processing of Gardeniae Fructus Praeparatus(Jiaozhizi in Chinese herbal name, JZZ) were measured by the visual analyzer to analyze the color variation rule during the processing of JZZ, and the content changes of total reducing sugar, total amino acid and 5-hydroxymethylfurfural(5-HMF) related to Maillard reaction were measured. Pearson correlation analysis and linear regression analysis of the data were carried out by SPSS 24.0 software. The experimental results showed that the objective coloration of the samples in the processing of JZZ was basically consistent with the traditional subjective color judgment; the contents of total reducing sugar and total amino acids showed a decreasing trend during the processing of JZZ, and the content of 5-HMF showed an increasing trend, which was in line with Maillard reaction law. Pearson correlation analysis results showed that there was a significant correlation between the chromaticity space parameters L~*(lightness value), a~*(red green value), b~*(yellow blue value), E~*ab(total color value) and the contents of total reducing sugar, total amino acid and 5-HMF(P<0.01), among which the values of L~*, a~*, b~*, E~*ab were positively correlated with the contents of total reducing sugar and total amino acid, and negatively correlated with the contents of 5-HMF. The results of linear regression analysis also showed that these two were highly correlated. In this study, by establishing the correspondence relationship between the color change of JZZ processing and Maillard reactants, wecan not only provide a basis for the objective digital expression of subjective color of JZZ, but also provide a reference for explaining the processing mechanism of JZZ from a new perspective.

Correlation analysis of color change and Maillard reaction during processing of Gardeniae Fructus Praeparatus / 中国中药杂志

The chromaticity space parameters of the samples during the processing of Gardeniae Fructus Praeparatus(Jiaozhizi in Chinese herbal name, JZZ) were measured by the visual analyzer to analyze the color variation rule during the processing of JZZ, and the content changes of total reducing sugar, total amino acid and 5-hydroxymethylfurfural(5-HMF) related to Maillard reaction were measured. Pearson correlation analysis and linear regression analysis of the data were carried out by SPSS 24.0 software. The experimental results showed that the objective coloration of the samples in the processing of JZZ was basically consistent with the traditional subjective color judgment; the contents of total reducing sugar and total amino acids showed a decreasing trend during the processing of JZZ, and the content of 5-HMF showed an increasing trend, which was in line with Maillard reaction law. Pearson correlation analysis results showed that there was a significant correlation between the chromaticity space parameters L~*(lightness value), a~*(red green value), b~*(yellow blue value), E~*ab(total color value) and the contents of total reducing sugar, total amino acid and 5-HMF(P<0.01), among which the values of L~*, a~*, b~*, E~*ab were positively correlated with the contents of total reducing sugar and total amino acid, and negatively correlated with the contents of 5-HMF. The results of linear regression analysis also showed that these two were highly correlated. In this study, by establishing the correspondence relationship between the color change of JZZ processing and Maillard reactants, wecan not only provide a basis for the objective digital expression of subjective color of JZZ, but also provide a reference for explaining the processing mechanism of JZZ from a new perspective.

Variations of Chemical Constituents in Gardeniae Fructus Before and After Stir-frying by LCMS-IT-TOF / 中国实验方剂学杂志

Objective: To rapidly analyze chemical compositions in raw products and stir-fried products of Gardeniae Fructus,and determine the pharmacodynamic material basis of Gardeniae Fructus Praeparatus. Method: Liquid chromatography-ion trap-time-of-flight-mass spectrometry(LCMS-IT-TOF) was performed on ACQUITY UPLC HSS T3 column(2.1 mm×100 mm,1.8 μm),mobile phase was 0.005%formic acid aqueous solution(A)-acetonitrile(B) for gradient elution(0-5 min,10%-15%B;5-8 min,15%B;8-18 min,15%-95%B;18-20 min,95%B;20-22 min,95%-10%B;22-25 min,10%B),the column temperature was 30℃ and the flow rate was 0.4 mL·min-1,volume of sample injection was 2 μL;electrospray ionization(ESI) was applied for mass spectrometric analysis under positive and negative ion mode,the scanning ranges were m/z 100-1 000.The ion peaks were identified by comparison of control substances,mass spectrometry data analysis and literature references.The peak areas of some ion peaks before and after processing were compared,the changes of chemical compositions in Gardeniae Fructus after processing were investigated. Result: Based on the mass spectral data information and references,a total of 38 compounds were identified,no new compounds were produced after processing.The ion peak areas of main compounds were investigated during the stir-frying processing,the contents of geniposide,genipin-1-β-D-gentiobioside,shanzhiside,chlorogenic acid and crocin in Gardeniae Fructus Praeparatus were decreased;the contents of geniposidic acid and crocetin were increased. Conclusion: The method can rapidly and accurately identify the chemical constituents in Gardeniae Fructus Praeparatus.The changes of iridoids and crocins in Gardeniae Fructus during the stir-frying process may be related to hemostatic activity and protection of cardiovascular and cerebrovascular.