Radix Kansui Stir-Fried with Vinegar Reduces Radix Kansui-Related Hepatotoxicity in Mice via Mitochondrial Pathway / 中国结合医学杂志

OBJECTIVE@#To investigate the mechanism of Radix Kansui (RK) stir-fried with vinegar (VRK) decreased hepatotoxicity in mice.@*METHODS@#According to a random number table, 40 mice were randomly divided into negative control group (0.5% carboxymethylcellulose sodium, 20 mL/kg), positive control group (0.1% mixture of carbon tetrachloride in soybean oil, 20 mL/kg), RK group (the ethyl acetate extracts of RK, 250 g crude drug/kg) and VRK group (the ethyl acetate extracts of VRK, 250 g crude drug/kg) with 10 mice per group. All mice were administered orally by gavage daily for 7 continuous days. The morphology of liver tissues was examined to assess the liver injury by a transmission electron microscope. Hepatocyte apoptosis in vivo was determined by terminal deoxynucleotidyltransferase-mediated dUTP-biotin nickend labeling (TUNEL) assay. Immunohistochemical technique was adopted to detect the expression of particular antiapoptotic and proapoptotic proteins in the mitochondrial pathways, including B-cell lymphoma (Bcl-2) and caspase-3, as well as the expression of inflammatory mediators, including nuclear factor kappa B (NF- κ B) and intercellular adhesion molecule-1 (ICAM-1).@*RESULTS@#Liver injury and hepatocyte apoptosis were observed in RK mice, and the liver injury were significantly reduced in VRK-treated mice. In immunohistochemistry study, compared with the negative control group, RK inhibited dramatically the Bcl-2 protein expression and significantly increased the expression of caspase-3, NF- κ B and ICAM-1 (all P<0.01). Compared with the RK group, VRK group induced significant increase on Bcl-2 protein expression, and decreased the caspase-3, NF- κ B and ICAM-1 protein expression (P<0.05 or P<0.01).@*CONCLUSION@#The mechanism of reduced hepatotoxicity of VRK may be associated with the reduced inflammation, regulation of antiapoptotic and proapoptotic mediators in the mitochondrial pathway.

Protective effect of different polar fractions of Carbonized Rubiae Radix et Rhizoma (cRRR) against oxidative injury / 中国中药杂志

The protective effect of different polar fractions of Carbonized Rubiae Radix et Rhizoma (cRRR) against ox-LDL-induced damage to human umbilical vein endothelial cells (HUVECs) was investigated by MTT assay, and the components were identified by using UPLC-Q-TOF-MS. According to the study, ethyl acetate extract and n-butanol extract could increase cell viability (P<0.01), while petroleum ether extract had no influence, and water extract could even inhibit the cell viability to some degree. Moreover, 32 compounds in four polar fractions were analyzed, including 31 quinones and their glycosides, and one rubiprasins C. Petroleum ether extract, ethyl acetate extract, n-butanol extract and water extract contained 23, 32, 26, 15 compounds, respectively. According to cell experiments in vitro, active fractions were ethyl acetate extract and n-butanol extract. The results could provide scientific references for further studies on effective material basic of cRRR, and lay a foundation for studies on the relationship between efficacies and materials.

Changes of chemical constituents in Rubiae Radix et Rhizoma before and after carbonized by UPLC-Q-TOF-MS method / 中国中药杂志

In order to explore the effect on chemical constituents after carbonized, the changes of chemical constituents in raw and carbonized Rubiae Radix et Rhizoma were analyzed by UPLC-Q-TOF-MS. The research also used principal component analysis(PCA) and orthogonal partial least squares discriminant analysis(OPLS-DA) for data statistics to find out the main differences on components before and after carbonized. The accurate m/z values of Q-TOF-MS and Q-TOF-MS-MS fragments were applied to identify the structures. The results showed that 6 more discrepant constituents were existed between raw and carbonized Rubiae Radix et Rhizoma. Three constituents were selected as the main discrepant components according to the peak area (276 nm) and identified, as lucidin, xanthopurpurin and 1,3,6-trihydroxy-2-methylanthraquinone. After carbonized, contents of xanthopurpurin and 1,3,6-trihydroxy-2-methylanthraquinone were observably increasing, while lucidin was obviously decreasing. They could be used as the chemical markers for the differentiation between raw and carbonized Rubiae Radix et Rhizoma. The results of this experiment played an important role in the study of processing principle of carbonized Rubiae Radix et Rhizoma. It also provided important evidences for the interpretation of effective material based on carbonized Rubiae Radix et Rhizoma.

Study on integrative technology of primary processing for Gastrodiae Rhizoma based on response surface methodology / 中草药

Objective: To optimize the integrative technology of primary processing for Gastrodiae Rhizoma by response surface methodology. Methods: The single factor experiment combined with Box-Behnken design was used to optimize the integrative technology, with five major characteristic components (gastrodin, parishin A, parishin B, parishin C, and parishin E) as indexes, in order to detect three factors (steaming time, drying time, and drying temperature), and optimize the primary processing of Gastrodiae Rhizoma. Results: Optimum percolation process was as follows: Gastrodiae Rhizoma was steamed for 30 min, and dried for 12 h at 60℃. Conclusion: This optimized integrative technology of Gastrodiae Rhizoma is reasonable and feasible, and with high accuracy. It could provide the scientific basis and innovative idea to the large-scale production of decoction pieces of Chinese Materia Medica.

Comparison between traditional processing and integration processing for Schizonepetae Herba based on chemical constituents and pharmacological effect / 中国中药杂志

The GC-MS method was adopted to determine the contents of β-myrcene, limonene, menthone, menthofuran, pulegone, β-caryophyllene, 1-octen-3-one and 3-octanone in volatile in Schizonepetae Herba processed by traditional processing and integration processing methods. The efficacies of Schizonepetae Herba with different processing methods were detected based on the inhibition of ear swelling induced by dimethylbenzene in mice. The rationality of the integration processing was expounded based on the comparison of chemical constituents and their pharmacological effects. The results showed that the contents of the eight chemical components in the products processed with the integrated processing method were higher than those processed with the other method. And both of the processing methods could reduce the degree of swelling and the content of TNF-α/IL-1β/IL-6 in mice serum. However, the anti-inflammatory efficacy of the products processed with the integration processing method was superior to that processed with the other method. Compared with the traditional processing method, the integration processing method ensures the quality of decoction pieces, with lower time and labor costs and higher efficiency.

Simultaneous determination of eight active components in Gastrodiae Rhizoma by HPLC-MS / 中草药

Objective: To establish an HPLC method for the simultaneous determination of eight active components (gastrodin, 4-hydroxybenzyl alcohol, vanillyl alcohol, vanillin, p-hydroxybenzaldehyde, parishin B, parishin C, and parishin A). Methods: The analysis was performed on an Agilent 1220 system with a Hypersil C18 column (250 mm × 4.6 mm, 5 μm). The eight components were separated in 75 min with gradient mobile phase consisting of 0.5% CH3COOH-H2O (A) and 0.5% CH3COOH-CH3OH (B): 0-10 min, 98% A; 10-60 min, 98%-60% A; 60-75 min, 60% A. The temperature of column was 30 ℃, and the injection volume was 20 μL. The multiple-reaction monitoring (MRM) scanning was employed for the quantification with switching electrospray ion source polarity in negative mode. The ion spray voltage was set at -4500 V and the turbo spray temperature was maintained at 550 ℃. Results: The eight components had the good linearity (r ≥ 0.999 2) within the linear ranges. The average recovery rate was 94.51%-102.70% with RSD < 3.50%. The contents of the eight components of Gastrodiae Rhizoma varied according to the different habits. The contents of parishins A, B, and C were high while the contents of vanillyl alcohol and vanillin were low. Conclusion: The developed HPLC-MS method is simple, sensitive, and accurate, and has the good repeatability in separation, which is available for the quality control of Gastrodiae Rhizoma.

HPLC specific chromatograms of drug pair of schizonepetae herba and saposhnikoviae radix / 中国中药杂志

In this experiment, the HPLC specific chromatogram was adopted, with Agilent Extend-C18 (4.6 mm x 250 mm, 5 microm) as the chromatographic column, and 0.5 per thousand trifluoroacetic acid and acetonitrile as the mobile phase for gradient elution, so as to establish specific chromatograms for drug pair of Schizonepetae Herba and Saposhnikoviae Radix from different producing area, identify 12 common characteristic peaks, and obtain the comparison specific chromatography of drug pair of Schizonepetae Herba and Saposhnikoviae Radix. The method is simple, accurate and highly reproducible, and thus can be used as the basis for the quality control of the drug pair.

Comparative study on effects of Rubiae Radix et Rhizoma and carbonized Rubiae Radix et Rhizoma on acute blood stasis rat model / 中国中药杂志

<p><b>OBJECTIVE</b>To observe the effects of Rubiae Radix et Rhizoma (RRR) and carbonized Rubiae Radix et Rhizoma (CRRR) on the acute blood stasis rat model, and reveal their differences in efficacy.</p><p><b>METHOD</b>The acute blood stasis model was induced by subcutaneously injecting adrenaline hydrochloride and soaking in ice water. Yunnan Baiyao was used as the positive control drug, and administered for consecutively seven days. This model was adopted to observe the effect of high, middle and low dose RRR and CRRR groups on hemorheology, thrombin activity, and blood platelet system.</p><p><b>RESULT</b>RRR could significantly reduce the wholeblood viscosity and plasma viscosity of blood stasis rats under different shear rates, and showed certain two-way regulating function in hemostasis. It also showed certain effect on ADP-induced platelet aggregation rate, but which was lower than CRRR. CRRR achieved the main hemostatic mechanism by stimulating intrinsic and extrinsic blood coagulation and fibrinogen, and could significantly enhance the platelet aggregation rate of rats in the acute blood stasis model (P <0. 01).</p><p><b>CONCLUSION</b>RRR had the effect of removing blood stasis and hemostasis, while CRRR mainly has the hemostatic effect. This further demonstrates the traditional processing theory of "promoting blood circulation with crude herbs and stopping bleeding with processed herbs".</p>

Protective effect of different solvent extracts from platycladi cacumen carbonisatum on LPS-induced human umbilical vein endothelial cells damage / 中国中药杂志

<p><b>OBJECTIVE</b>To study the protective effect of different solvent extracts from Platycladi Cacumen Carbonisatum (PCC) on LPS-induced human umbilical vein endothelial cell damage, and discuss the effective extracts from PCC for protecting vascular endothelial cells and their possible active substances.</p><p><b>METHOD</b>HUVECs were cultured in vitro; And LPS was adopted to establish the human umbilical vein endothelial cell damage model. MTT colorimetric method was used to determine cell activity; Xanthine oxidase method was adopted to detect the activity of superoxide dismutases (SOD) in the cell culture fluid; The TBA method was adopted to determine the content of malondialdehyde (MDA); The nitrate reductase method was used to detect the content of nitric oxide (NO); And UPLC/Q-TOF-MS was used to analyze the difference in flavonoids components among different solvent extracts from PCC.</p><p><b>RESULT</b>Compared with the model group, N-butanol extract (100 mg x L(-1)) and ethylacetate extract (100, 50 mg x L(-1)) could significantly enhance the cell activity (P < 0.05), significantly reduce MDA and NO content, and increase SOD activity (P < 0.05). Among the four solvent extracts, the content of total flavonids were the highest in ethyl acetate extract, the lowest in water extract and equivalent in N-butanol and petroleum benzene extract. In terms of the contents of quercitrin and myricitrin, N-butanol extract were second only to ethyl acetate extract.</p><p><b>CONCLUSION</b>Ethylacetate extract from PCC has a notable antagonistic effect in the damage induced by LPS to HUVECs, and thus is the most effective extract from PCC in protecting vascular endothelial cells. Quercitrin, myricitrin or multiple flavonoids that it contains may be their active substances for protecting vascular endothelial cells. Its mechanism may be related to the decrease in the production of NO and the inhibition of lipid peroxidation in cells.</p>

Effects and mechanisms of platycladi cacumen carbonisatum on rats with blood-heat and hemorrhage syndrome / 中国中药杂志

<p><b>OBJECTIVE</b>To discuss the effect and mechanism of Platycladi Cacumen Carbonisatum (PCC) on rats with blood heat and hemorrhage syndromes.</p><p><b>METHOD</b>Rats were fed with 15 g x kg(-1) water decoctions of Zingiberis Rhizoma and 5% alcohol for 15 days to establish the blood-heat and hemorrhage syndrome model. Yunnan Baiyao was taken as the positive control drug, and PCC decoctions (5.0, 10.0 g x kg(-1)) were given simultaneously, in order to detect changes in general physical signs of rats, such as body weight, daily diet, volume of daily drinking and urine and stool, and rectal temperature. Automatic hematology analyzers was used to determine white blood cell (WBC), red blood cell (RBC), hemoglobin (HGB), and hematocrit (HCT), blood time by docking (BT). Blood rheometers was used to detect whole blood and plasma viscosities, thrombin time (TT), activated partial thromboplastin time (APTT), prothrombin time (PT) and fibrinogen content (FIB). Indexes related to thyroid functions, such as triiodothyronine (T3), tetraiodothyronine (T4), reverse triiodothyronine (rT3) and thyroid stimulating hormone (TSH) were measured by radio-immunoassay, and changes in lung tissues were observed by hematoxylin-eosin (HE) stain.</p><p><b>RESULT</b>After modeling, rats witnessed slow-down in weight growth rate, significant increase in daily diet, volume of daily drinking, urine and temperature, significant decrease in stools and their water content (P < 0.05, P < 0.01), rise in plasma T4 level, notable growth in T3 and rT3 concentrations (P < 0.05), decline in TSH concentration. Additionally, their WBC, RBC, HGB and HCT remarkably increased (P < 0.05, P < 0.01), with significant increase in high, middle and low whole blood viscosities and plasma viscosity (P < 0.01); their BT, TT, APTT were notably prolonged (P < 0.01), with significant increase in FIB content (P < 0.01). After oral administration of Yunnan Baiyao or PCC, rats of all groups showed significant improvement in blood heat syndromes (P < 0.05, P < 0.01), and their blood coagulation indexes including BT, TT, APTT, FIB, thyroid function indexes including T4, T3, rT3, TSH, WBC, RBC, HGB, HCT, whole blood viscosity and plasma viscosity were getting normal (P < 0.05, P < 0.01).</p><p><b>CONCLUSION</b>PCC can ameliorate blood heat symptoms and pathologic hemorrhage among rats with blood heat and hemorrhage syndromes by inhibiting thyroid functions and correcting hemorheological and coagulation disorders.</p>

Simultaneous determination of four quinone compounds in carbonized Rubiae Radix et Rhizoma by UPLC / 中国中药杂志

<p><b>OBJECTIVE</b>To establish an UPLC method for simultaneous determination of purpuroxanthine, purpurin, 1,3,6-trihydroxy-2-methylanthraquinone, rubimaillin in carbonized Rubiae Radix et Rhizoma.</p><p><b>METHOD</b>The components were separated on acquity BEHC18 (2.1 mm x 50 mm, 1.7 microm) using methanol and 0.3% formic acid solution as the mobile phase; The flow rate was 0.2 mL x min(-1) and the volume of injection was 2 microL; the column temperature was maintained at 30 degrees C and the detective wavelength was set at 276 nm.</p><p><b>RESULT</b>There were good liner relationships between the peak area and concentration at ranges of 0.68-34.44 mg x L(-1) (r = 0.9999), 0.66-33.2 mg x L(-1) (r = 0.9997), 0.68-34.08 mg x L(-1) (r = 0.9999), 1.07-53.52 mg x L(-1) (r = 0.9999) for purpuroxanthine, purpurin, 1,3,6-trihydroxy-2-methylanthraquinone, rubimaillin, respectively; the average recovery rates of purpuroxanthine, purpurin, 1,3,6-trihydroxy-2-methylanthraquinone, rubimaillin were 96.95%, 95.75%, 102.5%, 96.15%, respectively, with RSD less than 3%.</p><p><b>CONCLUSION</b>The established method was rapid and simple with good accuracy and reproducibility for the determination of carbonized Rubiae Radix et Rhizoma, the method was suitable for the quality control of carbonized Rubiae Radix et Rhizoma.</p>

Determination of five flavonoids in Cacumen Platycladi Carbonisatum by UPLC / 中草药

Objective: To establish a UPLC method for simultaneous determination of five flavonoids (myricetin, quercitrin, quercetin, kaempferol, and amentoflavone) in Cacumen Platycladi Carbonisatum. Methods: The analysis was performed on a Waters Acquity UPLC system with a Waters BEH C18 column (50 mmx2.1 mm, 1.7 mm). The five flavonoids were separated with gradient mobile phase consisting of 0.05% aqueous formic acid and methanol. The temperature of column was 30°C, and the injection volume was 2 μl/min, detection wavelength was 360 nm. Results: The five flavonoids including myricetin, quercitrin, quercetin, kaempferol, and amentoflavone had good linearity (r≥0.999 5) within the linear ranges. The average recovery rate was 96.85% - 99.01% with RSD ≤4.00%. Conclusion: The developed UPLC method is simple, sensitive and accurate and has the good repeatability in separation, which is available for the quality control of Cacumen Platycladi Carbonisatum.