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This study aims to explore the core connotation of the compatibility of Aconiti Lateralis Radix Praeparata(Fuzi)-Glycyrrhizae Radix et Rhizoma(Gancao) herb pair under physiological and pathological conditions. The biochemical indicators of serum/myocardial tissue, pathological changes of the myocardial tissue, and serum metabolic profiles of normal rats and heart failure model rats treated with Fuzi Decoction and Fuzi Gancao Decoction were determined. Network pharmacology and metabolomics were employed to establish the metabolite-target-pathway network for Glycyrrhizae Radix et Rhizoma in enhancing the efficacy and reducing the toxicity of Aconiti Lateralis Radix Praeparata, Western blotting was employed to verify the representative pathways in the network. The results showed that both decoctions lowered the levels of creatine kinase and other indicators and mitigate myocardial pathological injury in model rats. However, they caused the abnormal rises in creatine kinase and other indicators and myocardial pathological injury in normal rats. The results indicated that the compatibility reduced the toxicity in normal rats and enhanced the efficacy in model rats. The results of metabolomics showed that Fuzi Gancao Decoction recovered more metabolites in model rats and had weaker effect on interfe-ring with the metabolites in normal rats than Fuzi Decoction. The association analysis showed that the network of Glycyrrhizae Radix et Rhizoma enhancing the efficacy of Aconiti Lateralis Radix Praeparata involved 112 metabolites, 89 targets, and 15 pathways, including calcium and cAMP signaling pathways. The network of Glycyrrhizae Radix et Rhizoma reducing the cardiotoxicity of Aconiti Lateralis Radix Praeparata involved 36 metabolites, 59 targets, and 11 pathways, including adrenergic signaling and tricarboxylic acid cycle in cardiomyocytes. The experimental results of protein expression verified the reliability of the association analysis. This study demonstrated that the core connotation of the herb pair of Aconiti Lateralis Radix Praeparata-Glycyrrhizae Radix et Rhizoma changed under physio-logical and pathological states, and the compatibility results of enhancing efficacy and reducing toxicity were achieved with different metabolic pathways and biological processes.
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ObjectiveTo explore the effect and toxicity change rule of Aconiti Lateralis Radix Praeparata（ALRP） and Zingiberis Rhizoma（ZR） before and after compatibility， and to reveal the compatibility connotation of them. MethodSixty SD rats were randomly divided into blank group， blank-ALRP group， blank-ALRP-ZR group， model group， model-ALRP group and model-ALRP-ZR group， the latter three groups were injected with adriamycin via tail vein to establish the model of heart failure， and the former three groups were injected with the same amount of physiological saline via tail vein. The effects of ALRP single decoction and ALRP-ZR mixed decoction on biochemical indexes and myocardial histopathological morphology of normal rats and model rats were compared. Metabolomics analysis was performed on rat serum samples， principal component analysis（PCA） and orthogonal partial least squares-discriminant analysis（OPLS-DA） were used to screen the differential metabolites between groups， and the differential metabolic pathways were analyzed. Combined with network pharmacology technology， the metabolites and their associated targets and pathways related to enhancing anti-heart failure efficacy and reducing cardiotoxicity were screened before and after the compatibility of ALRP and ZR， the screened representative pathways were verified by Western blot. ResultCompared with the blank group， the model group showed significant increases in the contents of brain natriuretic peptide（BNP）， creatine kinase（CK）， lactate dehydrogenase（LDH） and cardiac troponin（cTn）-T（P<0.01）， the blank-ALRP group showed obvious increases in CK， LDH， and cTn-T contents（P<0.05， P<0.01）， while the normal-ALRP-ZR group showed a significant increase in CK content（P<0.01）. Compared with the blank-ALRP group， the blank-ALRP-ZR group showed a obvious decrease in LDH content（P<0.05）， and pathological sections showed that both decoctions could lead to myocardial histopathological damage in normal rats. Compared with the model group， the model-ALRP-ZR group showed obvious decreases in BNP， CK， LDH and cTn-T contents（P<0.05， P<0.01）， and the model-ALRP group showed obvious decreases in BNP， LDH and cTn-T contents（P<0.05， P<0.01）. Compared with the model-ALRP group， the model-ALRP-ZR group showed a significant decrease in CK content（P<0.01）， and both decoctions could improve the pathological morphology of myocardial tissue in the model rats. Metabolomics results showed that ALRP single decoction and ALRP-ZR mixed decoction could recover 422 and 459 metabolites in model rats， respectively. And the metabolic disruption of ALRP-ZR mixed decoction on normal rats was weaker than that of ALRP single decoction. The results of network pharmacological association analysis showed that in the aspect of ZR enhancing the anti-heart failure efficacy of ALRP， 3 metabolites such as deoxyuridylic acid were correlated to 56 metabolites， 82 targets and 13 pathways， including calcium signaling pathway， renin secretion， renin-angiotensin system， etc. In the aspect of ZR reducing the cardiotoxicity of ALRP， 3 metabolites such as tyrosol were associated with 24 metabolites， 55 targets and 14 pathways， including adrenergic signaling in cardiomyocytes and carbon metabolism and so on. Western blot results showed that the expression of angiotensin-converting enzyme（ACE）， angiotensin-converting enzyme 2（ACE2） and angiotensin Ⅱ（Ang Ⅱ） in myocardial tissues of rats from the model group was significantly elevated by comparing with the blank group（P<0.01）. Compared with the model group， the model-ALRP group and the model-ALRP-ZR group showed significantly decreased expression of ACE， ACE2 and Ang Ⅱ（P<0.01）. Compared with the model-ALRP group， the expression of ACE2 and AngⅡ was significantly decreased in the model-ALRP-ZR group. Compared with the blank group， the expression of extracellular signal regulated kinase（ERK）， protein kinase B（Akt） and cTn-I3 was significantly elevated in the blank-ALRP group and blank-ALRP-ZR group（P<0.01）. Compared with the blank-ALRP group， the blank-ALRP-ZR group showed decreased expression of ERK， Akt and cTn-I3， but there was no statistical significance. ConclusionTo a certain extent， the combination of ALRP and ZR shows synergistic relationship under pathological state， and attenuated effect of compatibility under normal physiological state， and the pharmacodynamic characteristics and compatibility relationship of ALRP and ZR are closely related to the physiological state.
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Objective:To optimize the extraction process of Shangke Huoxue Granule.Methods:Taking the factors of extraction solvent multiple, extraction time and extraction times as investigation factors, and extraction amount of ferulic acid, paeoniflorin and the ratio of extraction as comprehensive evaluation indices, one-factor experimental design and central composite design-response surface methodology were adopted to optimize the extraction process of Shangke Huoxue Granule.Results:The binomial fitting equation was Y=96.16+2.42 A+0.63 B-3.76 AB-1.57 A2-1.87 B2 ( P<0.01). The optimal extraction process parameters were confirmed to be adding 16 times of water, 64 minutes each time, twice. The deviation rates between the measured values of three verification experiments and the predicted value were 2.00%, 3.23% and 0.66%. Conclusion:The established model of central composite design-response surface methodology has high predictability and the optimized extraction process is stable and feasible.
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Objective:To establish the TLC identification method and content determination method of ferulic acid, ligustilide, hydroxysafflor yellow A and paeoniflorin in Shangke Huoxue Decoction for quality evaluation.Methods:Ferulic acid, ligustilide, hydroxysafflor yellow A and paeoniflorin were qualitatively identified by TLC method, and the content was determined by HPLC method. Waters Symmetry ShieldTM RP18 column (4.6 mm×250 mm, 5 μm) was set, the mobile phase consisted of acetonitrile-0.15% phosphoric acid water with gradient elution at a flow of 1.0 ml/min, and the column temperature was 30 ℃.The detection wavelength was 320 nm (33-50 min for ferulic acid, 55-70 min for ligustilide), 403 nm (7-31 min for hydroxysafflor yellow A) and 230 nm (7-31 min for paeoniflorin).Results:The TLC spots were clear. The linear relationships of ferulic acid, ligustilide, and hydroxysafflor yellow A were good in the range of 3.05-48.74 μg, 3.50-26.24 μg, 21.34-213.44 μg. The method was stable, repeatable with good recovery rate.Conclusion:The TLC and HPLC method for the simutanous determination of the four effective components in Shangke Huoxue Decoction were established, and the methods are suitable for the quality evaluation of Shangke Huoxue Decoction.
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Objective:To optimize the matrix formulation of Erhuang analgesic gels. Methods:Central composite design-response surface methodology was adopted to optimize the best formulation of Erhuang analgesic gels by using carbomer 940, triethanolamine and glycerine as independent variables, the appearance, stability, viscosity and in vitro release of berberine hydrochloride as comprehensive evaluation indices. Results:The fitting regressing equation was Y= 82.25 + 4.95 A+ 5.19 B + 1.41 C+ 1.51 AB + 0.904 0 AC- 0.531 9 BC- 2.92 A2-1.80 B2-0.182 1 C2. P value of the model was less than 0.000 1, and the correlation coefficient r value was 0.977. The optimal formulation of Erhuang analgesic gels consisted of 1.84% carbomer 940, 1.30 times triethanolamine of carbomer 940 and 13.99 grams of glycerine. The average comprehensive scores of three verification experiments was 88.56, and the deviations from the predicted values were 2.93%, 2.85% and 1.55%. Conclusion:The formulation process by central composite design-response surface methodology was stable and the formulation of Erhuang analgesic gels has been optimized.
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Objective:To optimize the extraction process of Huoxue-Sanyu effervescent tablets. Methods:The hydroxysafflor yellow A content, paeoniflorin and dry extract yield were used as the evaluation indexes, by using analytic hierarchy process (AHP) method, integrative graded indicators method combined with orthogonal test and back propagation (BP) artificial neural network. To optimize the technics parameters, such as the amount of water, extraction time and frequency.Results:Paeoniflorin and hydroxysafflor yellow A showed good linear relation in the range of 0.079 5-1.590 4 μg and 0.038 5-1.539 2 μg respectively, with the average recovery rates of 98.18% and 96.22%. RSD was 0.77% and 1.31% respectively. The optimum extraction technic was 9 times of water, 3 times of heating reflux extraction, and each extraction last for 1 hour. Conclusions:The optimized combined method of orthogonal experiment and BP artificial neural network is practical with high efficiency. The optimal extraction process is stable and reasonable.
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Objective:To set up a method to measure the content of baicaline, wogonoside, and paeonol in Jiedu-Zhixue Decoction extract and to measure their physico-chemical parameters. Methods:By using HPLC analytical methods to measure the content of baicaline, wogonoside, and paeonol in Jiedu-Zhixue Decoction extract. The RP18 column was adopted, gradient eluted with mobile phase of acetonitrile-1% phosphoric acid water, flow rate was 1 ml/min, detection wavelength was 274 nm, and the column temperature was 30 ℃. The method quantitatively analyze the compound extracts of baicaline, wogonoside and paeonol were used. Results:The method isrepeatable, stable with good recorey rates. The calibration curves of baicaline was in good linearity over the range of 0.191 2-1.147 2 μg. The calibration curves of wogonoside was in good linearity over the range of 0.041 5-0.207 4 μg. The calibration curves of paeonol was in good linearity over the range of 0.019 5-0.156 3 μg. There was no significant difference among the component of 3 different batches of Jiedu-Zhixue Decoction extract, and the pH value range was 5.240-5.753, relative density range was 0.846-0.889. Conclusions:The method used in this stydy to measure the effective components of Jiedu-Zhixue Decoction extract was stable and feasible, which is suitable for quality control of Jiedu-Zhixue Decoction.
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Objective@#To set up a method to measure the content of baicaline, wogonoside, and paeonol in Jiedu-Zhixue Decoction extract and to measure their physico-chemical parameters.@*Methods@#By using HPLC analytical methods to measure the content of baicaline, wogonoside, and paeonol in Jiedu-Zhixue Decoction extract. The RP18 column was adopted, gradient eluted with mobile phase of acetonitrile-1% phosphoric acid water, flow rate was 1 ml/min, detection wavelength was 274 nm, and the column temperature was 30 ℃. The method quantitatively analyze the compound extracts of baicaline, wogonoside and paeonol were used.@*Results@#The method isrepeatable, stable with good recorey rates. The calibration curves of baicaline was in good linearity over the range of 0.191 2-1.147 2 μg. The calibration curves of wogonoside was in good linearity over the range of 0.041 5-0.207 4 μg. The calibration curves of paeonol was in good linearity over the range of 0.019 5-0.156 3 μg. There was no significant difference among the component of 3 different batches of Jiedu-Zhixue Decoction extract, and the pH value range was 5.240-5.753, relative density range was 0.846-0.889.@*Conclusions@#The method used in this stydy to measure the effective components of Jiedu-Zhixue Decoction extract was stable and feasible, which is suitable for quality control of Jiedu-Zhixue Decoction.
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<p><b>OBJECTIVE</b>To observe effects and mechanism of Dinggui gel paste analgesic anti-inflammatory.</p><p><b>METHODS</b>Eighty-four male KM mice weighted from 18 to 22 g and aged 4 to 5 weeks were randomly divided into 7 groups, named blank group, model group, matrix control group, Votalin group, high dosage of Dinggui gel paste group with group, equivalent dosage of Dinggui gel paste group, Dinggui gel paste group, 12 mice in each group. Except blank and model group, the other groups were paste ointment for 7 days, and one time a day, matrix control group were pasted isodose blank matrix gel patch. Pain threshold were tested at 30, 60, 90 and 120 min after the last ad-ministration. Hot plate test were performed by injection of 5% formalin for 20 µL on right hindfoot sole after the last administration. The cumulative time of mice licking right rear foot were observed at stage of I and II, and content of IL-1, TNF-α were tested by ELISA method. Differences of weight between right and left ears were measured by ear swelling method and anti-inflammation experiment.</p><p><b>RESULTS</b>In hot plate test at 90 min, pain threshold in equivalent dosage of Dinggui gel paste group was (24.87 ± 14.67) s and (15.28 ± 8.23) s in model group; (26.33 ± 15.45) s in high dosage of Dinggui gel paste group and (15.31 ± 5.02) s in model group at 120 min in hot plate test, there were no statistical differences between two groups. Pain period at stage I, licking cumulative time in high dosage of Dinggui gel paste group was (66.70 ± 22.83) s and (101.80 ± 33.65) s in model group,and had significant differences between two groups; there were statistical differences in licking cumulative time at stage I of pain period among high dosage of Dinggui gel paste group (51.30 ± 43.60)s, equivalent dosage of Dinggui gel paste group (64.00 ± 47.27) sand model group (109.50 ± 36.78) s. Content of IL-1 in model group was (28.70 ± 8.24) ng/L and (13.33 ± 2.20) ng/L in high dosage of Dinggui gel paste group, there was obvious meaning between two groups; There were significant differences in TNF-α content among model group (93.60 ± 23.65) ng/L,high dosage of Dinggui gel paste group (63.21 ± 10.54)ng/L and equivalent dosage of Dinggui gel paste group (72.69 ± 16.26) ng/L; while there were no statistical meaning in ear swelling degree among model group (5.73 ± 0.80) mg,high dosage of Dinggui gel paste group (5.42 ± 0.68) mg and equivalent dosage of Dinggui gel paste group (4.98 ± 1.52) mg.</p><p><b>CONCLUSION</b>Dinggui gel paste could increase pain threshold, reduce licking accumulative time, and decrease ear swelling degree, and relief pain by regulating level of TNF-α and IL-1.</p>
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Animais , Humanos , Masculino , Camundongos , Analgésicos , Anti-Inflamatórios não Esteroides , Formas de Dosagem , Medicamentos de Ervas Chinesas , Interleucina-1beta , Genética , Alergia e Imunologia , Interleucina-6 , Genética , Alergia e Imunologia , Pomadas , Dor , Genética , Alergia e Imunologia , Manejo da Dor , Fator de Necrose Tumoral alfa , Genética , Alergia e ImunologiaRESUMO
Objective: To establish a method for the content determination of ginsenoside Rg1, Re, and Rb1 in Piwang Granules. Methods: HPLC-ELSD was used. The analysis was carried on Shim-pack VP-ODS (150 mm × 4.6 mm, 5 μm) column with mobile phase consisted of acetoneitrile-water, and the gradient elution (0-20 min, acetoneitrile 20%, 20-35 min, acetoneitrile 20%→30%, 35-55 min, acetoneitrile 30%, 55-56 min, acetoneitrile 30%→45%) was used. The flow rate was 0.8 mL/min. The column temperature was 25℃. The drift tube temperature was 100℃. The flow rate of gas was 3 L/min. Results: The responses of ginsenoside Rg1, Re, and Rb1 were liner in the ranges of 125.44-470.40 (r = 0.999 6), 88.96-333.60 (r = 0.999 2), and 157.12-589.20 μg/mL (r =0.999 5), respectively. The average recoveries were 100.27% (RSD = 1.713%, n = 9), 101.29% (RSD = 1.220%, n = 9), and 101.00% (RSD = 1.668%, n = 9). Conclusion: The method is simple, rapid, and with good repeatability. It can be used for the quality control of Piwang Granules.