Analysing pharmacodynamic interactions of traditional Chinese medicine in treating acute pancreatitis based on OPLS method.

Acute pancreatitis (AP) is a surgical abdominal disease for which the Dachengqi Decoction (DCQD) of traditional Chinese medicine (TCM) is widely used in China. This study aims to analyse the pharmacodynamic interactions and quantitative relationship of DCQD in the treatment of AP based on orthogonal partial least squares (OPLS) analysis. The experimental data show organic chemical components as candidate pharmacodynamic substances (PS) in the blood and include pharmacodynamic indicators (PIs). Taking each PI as the target and using OPLS method to construct three types of mathematical equations, including the mathematical relationship between the pharmacodynamic substances and each target pharmacodynamic indicator (PS-TPI); the mathematical relationship between the pharmacodynamic substances, the pharmacodynamics indicators and each target pharmacodynamic indicator (PS, PI-TPI); and the mathematical relationship between the pharmacodynamic indicators and each target pharmacodynamic indicator (PI-TPI). Through analysis, we find that the R2Y(cum) values and VIP values indicate that PS and PI are the follow-up factors of TPI; the coefficient value indicates that there is a quantitative relationship between the PS and the TPI; and there also is a quantitative relationship between PI and TPI. The results demonstrated that PS and other PIs are the important influencing factors of TPI, and that there are interactions and quantitative relationships among the PIs.

[Study on time-toxicity relationship and mechanism of Gardeniae Fructus extract on hepatoxicity in rats based on proteomics].

To study the time-toxicity relationship and mechanism of Gardeniae Fructus extract on the hepatoxicity in rats. Rats were randomly divided into C group(0 day), D5 group(5 days), D12 group(12 days), D19 group(19 days), and D26 group(7 days recovery after 19 days of administration). The rats in normal group received normal saline through intragastric administration, and the rats in other groups received 10 g·kg~(-1 )Gardeniae Fructus extract through intragastric administration. After the final administration, the livers were collected. Hematoxylin-eosin staining was used to observe the histopathological changes in the liver tissue. Total liver proteins were extracted for proteomic analysis, detected by the Nano-ESI liquid-mass spectrometry system and identified by Protein Disco-very software. SIEVE software was used for relative quantitative and qualitative analysis of proteins. The protein-protein interaction network was constructed based on STRING. Cytoscape software was used for cluster analysis of differential proteins. Kyoto encyclopedia of genes and genomes(KEGG) database was used to perform enrichment signal pathway analysis. Pearson correlation analysis was performed for the screened differential protein expression and liver pathology degree score. The results showed that the severity of liver injury in D5, D12 and D19 groups was significantly higher than that in group C. The degree of liver damage in D5 group was slightly higher than that in D12 and D19 groups, with no significant difference between group D26 and group C. Totally 147 key differential proteins have been screened out by proteomics and mainly formed 6 clusters, involving in drug metabolism pathways, retinol metabolism pathways, proteasomes, amino acid biosynthesis pathways, and glycolysis/gluconeogenesis pathways. The results of Pearson correlation analysis indicated that differential protein expressions had a certain temporal relationship with the change of liver pathological degree. The above results indicated that the severity of liver damage caused by Gardeniae Fructus extract did not increase with time and would recover after drug with drawal. The above pathways may be related to the mechanism of liver injury induced by Gardeniae Fructus extract.

Feature Selection Method Based on Partial Least Squares and Analysis of Traditional Chinese Medicine Data.

The partial least squares method has many advantages in multivariable linear regression, but it does not include the function of feature selection. This method cannot screen for the best feature subset (referred to in this study as the "Gold Standard") or optimize the model, although contrarily using the L1 norm can achieve the sparse representation of parameters, leading to feature selection. In this study, a feature selection method based on partial least squares is proposed. In the new method, exploiting partial least squares allows extraction of the latent variables required for performing multivariable linear regression, and this method applies the L1 regular term constraint to the sum of the absolute values of the regression coefficients. This technique is then combined with the coordinate descent method to perform multiple iterations to select a better feature subset. Analyzing traditional Chinese medicine data and University of California, Irvine (UCI), datasets with the model, the experimental results show that the feature selection method based on partial least squares exhibits preferable adaptability for traditional Chinese medicine data and UCI datasets.

A Comprehensive and Rapid Quality Evaluation Method of Traditional Chinese Medicine Decoction by Integrating UPLC-QTOF-MS and UFLC-QQQ-MS and its Application.

Decoction is one of the oldest forms of traditional Chinese medicine and it is widely used in clinical practice. However, the quality evaluation and control of traditional decoction is a challenge due to the characteristics of complicated constituents, water as solvent, and temporary preparation. ShenFu Prescription Decoction (SFPD) is a classical prescription for preventing and treating many types of cardiovascular disease. In this article, a comprehensive and rapid method for quality evaluation and control of SFPD was developed, via qualitative and quantitative analysis of the major components by integrating ultra-high-performance liquid chromatography equipped with quadrupole time-of-flight mass spectrometry and ultra-fast-performance liquid chromatography equipped with triple quadrupole mass spectrometry. Consequently, a total of 39 constituents were tentatively identified in qualitative analysis, of which 21 compounds were unambiguously confirmed by comparing with reference substances. We determined 13 important constituents within 7 min by multiple reaction monitoring. The validated method was applied for determining five different proportion SFPDs. It was found that different proportions generated great influence on the dissolution of constituents. This may be one of the mechanisms for which different proportions play different synergistic effects. Therefore, the developed method is a fast and useful approach for quality evaluation of SFPD.

[Proteomics research of intervention effect of "Qi enriching" herbs on "Qi deficiency" rats based on technology of NanoLC-LTQ-Orbitrap].

Proteomics method, based on NanoLC-LTQ-Orbitrap technology, was applied to explore the biological basis of intervention effect of "Qi enriching" herbs on "Qi deficiency" rats. The "Qi deficiency" rat model was established with caloric restriction combined with excessive swimming. Muscle proteins of vastus lateralis from the blank group, the model group and the ginseng group were detected by NanoLC-LTQ-Orbitrap system. The data were imported into Protein Discovery software to identify the proteins and all the raw datum were analyzed by SIEVE software. Compared with model group, 26 significant difference proteins were found in ginseng group, which the variation trend was consistent with the blank group. Through the biological function analysis, the found proteins could be classified into proteins involved in energy metabolism, proteins involved in glucose metabolism, electrolyte balance and material transfer related proteins, inflammation related protein and cytoskeleton protein. The above target proteins and their regulation pathways may be the biological basis which ginseng played a role of tonifying "Qi" of "Qi deficiency" symptom.

[Study on discrimination mode of cold and hot properties of traditional Chinese medicines based on biological effects].

OBJECTIVE: To observe the effect of cold or hot properties of traditional Chinese medicines (TCM) on biological effect indexes, and analyze the contribution of variables on cold or hot properties, in order to preliminarily establish the discrimination mode for the biological effects of cold or hot properties. METHOD: Rats were randomly divided into the blank control group, cold TCM groups (Coptidis Rhizoma, Scutellariae Radix, Phellodendri Cortex, Gardeniae Fructus, Sophorae Flavescentis Radix and Gentianae Radix) and hot TCM groups (Aconiti Lateralis Preparata Radix, Zingiberis Rhizoma, Alpiniae Officinarum Rhizoma, Zanthoxyli Pericarpium, Cinnamomi Cortex and Evodiae Fructus), and orally administered with 10 mL x kg(-1) of corresponding TCM water decoctions for 30 d, twice a day. Altogether 53 biological effect indexes correlated to cold or hot properties of traditional Chinese medicines were founded by searching literatures. The data warehouse were established by using data-mining software Clementine12.0. Data of the blank control group, cold TCM groups (Coptidis Rhizoma, Phellodendri Cortex, Gardeniae Fructus, Sophorae Flavescentis Radix, Gentianae Radix) and hot TCM groups (Aconiti Lateralis Preparata Radix, Zingiberis Rhizoma, Alpiniae Officinarum Rhizoma, Zanthoxyli Pericarpium, Cinnamomi Cortex) were selected into a training set. C5.0 algorithm and C&R classification and regression algorithm were adopted to define the importance of variable, create the decision trees, and test hot or cold properties of Evodiae Fructus and Scutellariae Radix. RESULT: According to C&R classification and regression algorithm, SDH activity of livers was the most important hot or cold property, with the significance closed to 30%. It was followed by triglyceride, liver Na' -K' -ATPase enzyme, muscle glycogen and platelet distribution width, with the accuracy up to 97.39% in models. C5.0 algorithm showed that liver SDH activity was the most important hot or cold property, with the significance closed to 40%. It was followed by triglyceride, GOT, muscle glycogen and liver Na(+)-K(+)-ATPase enzyme, with the accuracy up to 98.26% in models. The possibilities that Evodiae Fructus is in hot property and Scutellariae Radix is in cold property were 100. 00% and 77.78% by using both C&R classification and regression algorithm and C5.0 algorithm. CONCLUSION: The SDH activity of liver is the most important biological effect index to distinguish cold and hot properties of TCMs. The discrimination pathway or mode of cold and hot properties is closely related to energy metabolism.

Interaction of baicalin with berberine for glucose uptake in 3T3-L1 adipocytes and HepG2 hepatocytes.

ETHNOPHARMACOLOGICAL RELEVANCE: Baicalin and berberine are important coexisting constituents of the combination of Radix Scutellariae and Rhizoma Coptidis, known as scutellaria-coptis herb couple (SC), which has heat clearing and detoxifying effects. The aims of the present study were to investigate the effects of the combination of baicalin+berberine on glucose uptake in 3T3-L1 adipocytes or HepG2 cells. MATERIALS AND METHODS: Insulin-resistant adipocytes and hepatocytes models were established. Glucose consumption was assayed to evaluate the effects of berberine, baicalin, and berberine+baicalin on glucose uptake, and the interaction of baicalin with berberine for glucose uptake was evaluated in 3T3-L1 adipocytes or HepG2 cells. Moreover, the effects of baicalin on the dose-effect relationship of berberine for glucose uptake was also evaluated in 3T3-L1 adipocytes. RESULTS: The results of the present study demonstrated that berberine increased glucose consumption in 3T3-L1 adipocytes and HepG2 hepatocytes in a dose-dependent manner. In contrast, statistical analyses indicated that baicalin (in doses up to 100µmol/L) produced no obvious effect. The effect of berberine+baicalin on glucose uptake was better than that of berberine or baicalin alone, which indicated that berberine and baicalin had the trend of synergetic effect on glucose uptake. Furthermore, these results showed that the synergistic effect occurred in a specific dose range, while the antagonistic effect was present in another dose range in the presence of 10µmol/L baicalin. Interestingly, the entire dose-response curves of berberine shifted down in the presence of 100µmol/L baicalin, and baicalin antagonised the effect of berberine on glucose uptake in 3T3-L1 adipocytes. CONCLUSIONS: The results of the present study showed that berberine dose-dependently increased glucose consumption in 3T3-L1 adipocytes and HepG2 hepatocytes. Furthermore, interaction of baicalin with berberine was additive at low doses of baicalin and antagonistic at higher baicalin doses. Thus, it is possible that baicalin is a partial agonist. These results provided a basis for the study of the TCM compatibility mechanism and a new insight into the application for Gegen Qinlian Decoction (GGQLD) or SC in the clinic.

[Effect of euodiae fructus on hepatic energy metabolism-related mechanisms of reserpine-induced asthenia cold syndrome rats].

OBJECTIVE: To discuss the effect of Euodiae Fructus on hepatic energy metabolism-related mechanisms of mitochondria of hepatic tissues of asthenia cold syndrome rats. METHOD: Rats were subcutaneously injected with Reserpine to establish the model. After the oral administration with Euodiae Fructus for 12 d, the oxygen electrode method was adopted to determine the respiration efficiency. The expressions of Cox4, Atp5b, Ucp2,Pgc-1alpha, Nrf1, Tfam mRNA were assayed by using RT-PCR method. RESULT: Euodiae Fructus 4.2 g x kg(-1) could obviously increase ST3 and RCR of asthenia cold syndrome rats, and expressions of Cox4, Ucp2 Nrf1 mRNA. It could also increase expressions of Atp5b and Pgc-1alpha mRNA, but with no statistical significance. No obvious change was observed in Tfam mRNA expression. Euodiae Fructus 4.2 g x kg(-1) could significantly increase ST3 and RCR of asthenia cold syndrome rats and Pgc-1alpha mRNA and Nrf1 mRNA expressions, and significantly decrease P/O, with no obvious impact on Cox4, AtpSb, Ucp2, Tfam mRNA expressions. CONCLUSION: Euodiae Fructus can promote mitochondrial respiratory function and oxidative phosphorylation efficiency by improving Pgc-1alpha mRNA and Nrf1 mRNA expressions and regulating Cox4 and Atp5b mRNA in mitochondrial respiratory chain. It can also strengthen mitochondrial uncoupling respiration and add heat production by activating Ucp2 mRNA expression in liver.

Anti-diabetic activities of Gegen Qinlian Decoction in high-fat diet combined with streptozotocin-induced diabetic rats and in 3T3-L1 adipocytes.

Gegen Qinlian Decoction (GGQLD) is one of the well-known traditional Chinese medicines. Recently, it was reported that GGQLD had good clinical effects on type 2 diabetes mellitus. However, few studies have confirmed in detail the anti-diabetic activities of GGQLD in vivo and in vitro. In the present study, we investigated the anti-diabetic effects of GGQLD in high-fat diet combined with streptozotocin-induced diabetic rats and in 3T3-L1 adipocytes. The present results suggested GGQLD (4.95, 11.55 and 18.15 g/kg) decreased significantly fasting blood glucose, glycosylated serum protein, and glycosylated hemoglobin of diabetic rats (p<0.05), and GGQLD (4.95 and 18.15 g/kg) decreased significantly fasting serum insulin levels of diabetic rats (p<0.05); in 3T3-L1 adipocytes, Gegen Qinlian Decoction-containing serum (GGQLD-CS) (4%, 8% and 16%) enhanced glucose consumption, triglyceride (TG) content, adiponectin protein concentration and the mRNA expression of adiponectin. Adiponectin contributes to the regulation of lipid and glucose metabolism, and can play a critical role in the development of diabetes mellitus; the mechanisms of action of GGQLD might be related to augmentation of adiponectin protein concentration and up-regulation of the mRNA expression of adiponectin. However, the multi-target mechanisms of action of GGQLD need to be clarified further. The present study further validated the beneficial effects of GGQLD as an anti-diabetic agent. These findings provide a new insight into the anti-diabetic application for GGQLD in clinic and display the potential of GGQLD as a new drug candidate for the treatment of diabetes mellitus.

[Effect of cold property Chinese medicine radix scutellariae on energy metabolism of rats].

OBJECTIVE: To observe the effect on energy metabolism of rats with cold property Chinese medicine Radix Scutellariae. METHODS: The body weight gain, temperature, hydroposia content were determined before administration and every five days after administration. The activities of Na4(+)-K(+)-ATPase, Ca(2+)-ATPase and SDH, LPL, HP, the contents of NEAF, T3, T4, TSH were measured after having been administrated with water extracts of Radix Scutullaxiae at the dose of 6.0, 3.0 g/kg for 43 days. RESULTS: The body weight gains were raised and the hydroposia contents have been decreased. The activities of SDH were increased significantly while Na(+)-K(+)-ATPase, Ca(2+)-ATPase of liver had little change. The content of NEAF, the activity of LPL, HP were decreased significantly, and the contents of T3, T4, TSH and the body weight, temperature had no significant change. CONCLUSION: Radix Scutellariae can inhibit the energy metabolism of rat. The mechanism may not be related to thyroxine pathway.

[Preliminary study of metabonomcs on aqueous extract of Evodia rutaecarpa in sprague-dawley rats].

OBJECTIVE: To study the change of endogenous metabolites of SD rats administrated of aqueous extract of Evodiae rutaecarpa. METHOD: Six SD rats had been successively administrated aqueous extract of E. rutaecarpa (0.3857 g x kg(-1)) for 33 days. An agilent 1200 6410 triplequadrupole mass spectrometer was used for the analysis of endogenous metabolites in rat urine samples. These data was analyzed by the principal component analysis (PCA) and PLS-DA using the SIMCA-P 10.0 software. RESULT: The significant difference in metabolic profiles between the control group and the dosed group was well observed by PCA of the MS data. CONCLUSION: The E. rulaecarpa has changed the endogenous metabolites of SD rats. This work can provide the base for the further research on the interpretation of drug property of E. rulaecarpa.

[Study of metabonomics on pharmacological action appraisal Rhizoma coptidis in rats].

OBJECTIVE: HPLC-MS/MS-based metabonomics method was used to find the possible biomarker of Rhizoma Coptidis in rat urine. METHOD: Sprague-Dawley rats were successively administrated 7 g x kg(-1) aqueous extract of Rhizoma Coptidis for 30 days, urine were collected by metabolism cages and detected by using the HPLC-MS-MS. All dates were analyzed by the principal component analysis (PCA) through using the SIMCA-P 10.0 software. RESULT: The PCA demonstrated that the metabolome between treated group and control group had difference in rat urine sample after of 22 days administrated, for treated group 169 kinds of biomarkers were found including oxalacetic acid, malic acid, 2-ketoglutaric acid, NE, arachidonic acid, 5-HIAA and other compounds, the result was consistent with pharmacological effects of R. coptidis, such as antiinflammatory, inhibiting biosynthesis of CA biosynthesis, anticentral nerve and energy metabolism inhibition. CONCLUSION: Metabonomics may be available in pharmacological action evaluation of drugs.

[Study of hepatic energy metabolism on rats by six cold property traditional Chinese medicines].

OBJECTIVE: To investigate the common rule hepatic energy metabolism on rats by six cold property of traditional Chinese medicines (TCM). METHOD: The activities of Na+ - K+ - ATPase, Ca2+ - ATPase and SDH, the content of hepatic glycogen, and the mRNA expression of hepatic uncoupling protein 2 (UCP2) were measured after the rats and been administrated with water extracts of Radix Sophorae Flavescentis, Fructus Gardeniae, Cortex Phellodendri, Radix Scutellariae, Rhizoma Coptidis, Radix Gentianae respectively at 6.0, 7.0, 8.4, 6.0, 7.0, 4.0 g x kg(-1) doses for 30 days, twice a day. RESULT: The activities of Na+ - K+ - ATPase, Ca2+ - ATPase and SDH were depressed significantly by six cold property TCM. The decreased tendency of SDH was observed by six cold property of TCM, while statistical significance was obtained in Cortex Phellodendri, Radix Scutellariae, Rhizoma Coptidis, Radix Gentianae. The increased tendency of hepatic glycogen content was found by six cold property TCM, while statistical significance was not obtained in six groups. The decreased tendency of the mRNA expression of UCP2 was found by six cold property TCM, while statistical significance was obtained in Radix Sophorae Flavescentis, Cortex Phellodendri, and Radix Scutellariae groups. CONCLUSION: The six cold property TCM have the good effects on hepatic energy metabolism by decreasing the activity of mitochondria SDH to reduce the production of ATP by decreasing the activities of Na+ - K+ - ATPase, Ca2+ - ATPase to cut down the consumption of ATP, by decreasing the mRNA expression of hepatic UCP2 to decrease the heat production.

[Effect on factors of energy metabolism in rats skeletal muscle by cold traditional Chinese medicine].

OBJECTIVE: To investigate the effect of six cold traditional Chinese medicine on the energy metabolism factors in rats skeletal muscle. METHOD: The activity of Na(+)-K(+) -ATPase, Ca(2+) -ATPase and succinate dehydrogenase (SDH), the content of muscle glycogen, and the mRNA expression of skeletal muscle uncoupling protein 3 (UCP3) were measured after rats having been administrated with water extracts of Radix Sophorae Flavescentis, Fructus Gardeniae, Cortex Phellodendri, Radix Scutellariae, Rhizoma Coptidis, and Radix Gentianae respectively at the dose of 6.0, 7.0, 8.4, 6.0, 7.0, 4.0 g x kg(-1) for 30 days. RESULT: The activity of Na(+) -K(+) -ATPase has been depressed significantly and the content of skeletal muscle glycogen has been increased remarkably by six cold traditional Chinese medicine. The decreased tendency has been found on activity of Ca(2+) -ATPase and SDH, only the Radix scutellariaeg group decreased the Ca(2+) -ATPase activity significantly (P < 0.05), the SDH activity was decreased high significantly by Radix scutellariae, Cortex Phellodendri, Radix Gentianae and significantly by Rhizoma Coptidis. The mRNA expression of UCP3 has been decreased high significantly by all five cold traditional Chinese medicine except Cortex Phellodendri group with the decreased tendency of UCP3 mRNA expression. CONCLUSION: The cold traditional Chinese medicine has the significant effects on the skeletal muscle energy metabolism by decreasing the utlization of the glucose and the activity of mitochondria SDH to reduce the production of ATP, and depressing the activity of Na(+)-K(+) -ATPase and Ca(2+) -ATPase to cut down the consumption of ATP, by decreasing the mRNA expression of UCP3 to decrease the heat production.