Adult ; Body Temperature/drug effects* ; COVID-19/pathology* ; Drug Therapy, Combination ; Drugs, Chinese Herbal/therapeutic use* ; Ephedra sinica/chemistry* ; Female ; Fever/pathology* ; Glycyrrhiza/chemistry* ; Humans ; Indoles/administration & dosage* ; Male ; Medicine, Chinese Traditional/methods* ; Middle Aged ; Phytotherapy/methods* ; Pneumonia, Viral/pathology* ; Radiography, Thoracic ; SARS-CoV-2/drug effects*

To investigate the antipyretic effect of active components of Mahuang Decoction in febrile rats, and explore its correlation with pharmacokinetics at different time points. The feverished rat models were induced by dry yeast, and intragastrically administered with the effective components of Mahuang Decoction with different orthogonal compatibility ratios. At different time points after administration, body temperature was measured; blood was taken from orbital vena plexus, and the contents of interleukin-6(IL-6), interleukin-1β(IL-1β), and tumor necrosis factor-α(TNF-α) in rat serum were determined with the kits. Combined with the pharmacokinetic data of the seven effective components in Mahuang Decoction, PK-PD(pharmacokinetics-pharmacodynamics) data fitting was conducted by using the analysis method of non-atrioventricular model, and then the pharmacodynamic parameters were calculated to determine the optimal binding model. The results showed that the effective components of Mahuang Decoction inhibited the release of heat-causing factors IL-6, IL-1β and TNF-α, and reduced the increase of body temperature. There was a significant lag between drug effect and blood drug concentration, which was consistent with Sigmoid-E_(max) model. The model fitting value showed a good correlation with mea-sured data, which could be used to evaluate and predict the correlation between PK and PD in Mahuang Decoction, and further applied to the multiple-indicator and multiple-effect study of PK-PD in other compound traditional Chinese medicines.
Animals ; Antipyretics/therapeutic use* ; Drugs, Chinese Herbal/therapeutic use* ; Ephedra sinica/chemistry* ; Fever/drug therapy* ; Interleukin-1beta/blood* ; Interleukin-6/blood* ; Medicine, Chinese Traditional ; Rats ; Tumor Necrosis Factor-alpha/blood*

In the present paper,after the febrile rat model was prepared by injecting yeast,orthogonally compatible effective components from prescription drugs of Mahuang Decoction( Ephedra sinica total alkaloids,Cinnamomum cassia essential oil,amygdalin,Glycyrrhiza uralensis total flavonoids+G. uralensis total saponins) with nine different dosage ratios were given by gavage administration.The plasma concentrations of main active ingredients including ephedrine hydrochloride,pseudoephedrine hydrochloride,methylephedrine hydrochloride,cinnamic acid,amygdalin,liquritin and glycyrrhizin at different time points were analyzed by liquid chromatograph mass spectrometer( LC-MS). Based on the pharmacokinetic parameters of non-compartmental model,the area under curve of total quantum( AUCt) and the mean chromatographic retention time of total quantum( MRTt) were further calculated,in order to evaluate the effect of compatibility on the total statistical moment parameters. The results showed that the pharmacokinetic characteristics of main active components in febrile rats were significantly different after treatment with orthogonally compatibility of E. sinica total alkaloids,C.cassia essential oil,amygdalin,G. uralensis total flavonoids and G. uralensis total saponins. Orthogonal analysis confirmed that different compatibility components had different effects on the total statistical moment parameters. The contribution of effective components of Mahuang Decoction to AUCtwas as follows in a descending order: E. sinica total alkaloids>C. cassia essential oil>amygdalin>G. uralensis total flavonoids+G. uralensis total saponin,while the contribution to MRTtwas: E. sinica total alkaloids >G. uralensis total flavonoids+G. uralensis total saponin>amygdalin>C. cassia essential oil. The E. sinica total alkaloid had the greatest effects on both of the above parameters,and the optimal combination was A_3B_3C_2D_1 for AUCt,and A_1B_1C_1D_1 for MRTt.
Animals ; Drugs, Chinese Herbal ; pharmacokinetics ; Ephedra sinica ; chemistry ; Glycyrrhiza uralensis ; chemistry ; Oils, Volatile ; pharmacokinetics ; Phytochemicals ; pharmacokinetics ; Rats

With a great difference in therapeutic effects of Mahuang (MH, the stems of Ephedra sinica) and Mahuanggen (MHG, the roots of Ephedra sinica), chemical differences between MH and MHG should be investigated. In the present study, gas chromatography-mass spectrometry (GC-MS)-based plant metabolomics was employed to compare volatile oil profiles of MH and MHG. The antioxidant activities of volatile oils from MH and MHG were also compared. 32 differential chemical markers were identified according to the variable importance in the projection (VIP) value of orthogonal partial least squares discriminant analysis (OPLS-DA) and P value of Mann-Whitney test. Among them, chemical markers of tetramethylpyrazine (TMP) and α-terpineol were quantified. Their contents were much higher in most MH samples compared with MHG. The antioxidant assay demonstrated that MH had significantly higher free radical-scavenging activity than MHG. Although MH and MHG derived from the same medicinal plant, there was much difference in their volatile oil profiles. MH samples had significantly higher content of two reported pharmacologically important chemical markers of TMP and α-terpineol, which may account for their different antioxidant activities.
Drugs, Chinese Herbal ; chemistry ; Ephedra sinica ; chemistry ; Gas Chromatography-Mass Spectrometry ; Metabolomics ; Oils, Volatile ; chemistry ; Plant Roots ; chemistry ; Plant Stems ; chemistry

To detect the in vitro probe microdialysis recoveries based on an HPLC-DAD method for simultaneous quantification of nine active ingredients (ephedrine, pseudoephedrine, methylephedrine, amygdalin, liquiritin, cinnamyl alcohol, cinnamic acid, cinnamaldehyde and glycyrrhizic acid) in Mahuang decoction, which provides reference for in vivo pharmacokinetic study. The concentrations of nine active ingredients in dialysate were detected by HPLC-DAD, to investigate the effect of flow rates (incremental method and subtraction method) and intraday stability of the probe recoveries and medium concentrations on the recoveries. Nine active ingredients could be well separated in 52 min. At the perfusion rate of 1.0 μL x min(-1), the relative recoveries of ephedrine, pseudoephedrine, methylephedrine, amygdalin, liquiritin, cinnamyl alcohol, cinnamic acid, cinnamaldehyde and glycyrrhizic acid were (50.95 ± 0.82)%, (52.74 ± 1.13)%, (51.29 ± 0.51)%, (32.56 ± 0.84)%, (45.36 ± 0.83)%, (70.94 ± 0.99)%, (69.98 ± 2.30)%, (71.68 ± 0.63)%, and (22.14 ± 0.48)%, respectively. And the probe kept steady in 7 hours. At the same medium concentration, the probe recoveries decreased exponentially with the increase in flow rates. The recoveries of seven ingredients detected by these two methods were similar at certain flow rates, except for amygdalin and cinnamaldehyde. At the same flow rate, the relative recoveries of cinnamyl alcohol, cinnamic acid and cinnamaldehyde changed greatly (9.55%-16.2%) and the others six ingredients had less change (3.27%-5.71%) with the changes in medium concentrations. Microdialysis method could be used to detect the in vitro recoveries of nine ingredients in Mahuang decoction. Reverse dialysis method could be used for the in vivo probe recovery calibration of ephedrine, pseudoephedrine, methylephedrine, liquiritin, cinnamyl alcohol and cinnamic acid at the flow rate of 2.0 μL x min(-1).
Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Ephedra sinica ; chemistry ; Microdialysis ; methods ; Molecular Structure

OBJECTIVETo optimize the prescription dose of Mahuang decoction in a multi-target manner, in order to provide reference for the quantitative optimization of the prescription dose of the traditional Chinese medicine compound.

METHODThe number of diaphoretic spots in rats, the tracheal antispasmodic rate in guinea pigs and the writhing times by acetic acid in mice were taken as the indexes for evaluating the diaphoretic, antispasmodic and analgesic effects. According to the experimental results of the 16 orthogonal combination prescriptions, a mathematical dose-effect model was built by support vector regression (SVR) and quadratic response surface regression (RSR) respectively. The multi-target optimization was achieved by elitist non-dominated sorting genetic algorithm (NSGA-II) and entropy weight TOPSIS method.

RESULTThe optimal dose of Mahuang decoction after being optimized by SVR modeling contained 17.71 g of Ephedrae Herba, 9.57 g of Cinnamomi Ramulus, 11.75 g of Armeniacae Semen Amarum and 4.39 g of Glycyrrhizae Radix et Rhizoma Praeparata Cum Melle. The optimized result by RSR modeling contained 13.37 g of Ephedrae Herba, 11.61 g of Cinnamomi Ramulus, 11.98 g of Armeniacae Semen Amarum and 5.67 g of Glycyrrhizae Radix et Rhizoma Praeparate Cum Melle. SVR was superior to RSR in both of the forecast capacity and optimization results.

CONCLUSIONSVR-NSGA-II-TOPSIS method could be adopted for the multi-target optimization for the dose of Mahuang decoction and other traditional Chinese medicine compounds. It is proved to be the optimal prescription with the best efficacy, and could provide scientific quantitative basis for determining the dose of traditional Chinese medicine compound prescriptions and developing new traditional Chinese medicines.

Animals ; Cinnamomum ; chemistry ; Drug Compounding ; methods ; Drug Dosage Calculations ; Drug Prescriptions ; Ephedra ; chemistry ; Ephedra sinica ; chemistry ; Glycyrrhiza ; chemistry ; Guinea Pigs ; Mice ; Rats

Since the polyjuice potion ingredient is complex, we need to develop an analysis method with well separation and high stability to perform qualitative analysis. After dividing chemical components of Mahuang Decoction into fat-soluble and water-soluble constituents by gradient extraction, GC-MS was used to analyze the chemical components of the ethyl acetate extraction. The results showed that forty compounds had been identified by NIST MS search 2.0 standard mass spectrometry Library and literatures. Next, UPLC-Q-TOF-MS was applied to idendify the chemical components of the water extraction. The results showed that thirty-nine compounds had been identified by MZmine-2.9.1, Isotope Pattern, fragmentation regularity of mass spectrometry and literatures. This experiment will provide evidences for elucidation of the effective substance in Mahuang decoction and can be used as a simple, shortcut method for analysis and identification for the polyjuice potion.
Chromatography, High Pressure Liquid ; methods ; Ephedra ; chemistry ; Ephedra sinica ; chemistry ; Gas Chromatography-Mass Spectrometry ; methods

OBJECTIVETo investigate the effects of complement inhibiting component of Ephedra sinica on immunological inflammation following acute spinal cord injury (SCI) in rats.

METHODSThe complement inhibiting component of Ephedra sinica was isolated by multiple precipitation steps and thin layer chromatography, and then the activity was analyzed. Fifty healthy SD rats were selected and randomly divided into the control group and the experimental group, 25 in each group. Induction of SCI was performed following a modified Allen's weight-drop method. The complement inhibiting component from Ephedra sinica (15 mg/kg) dissolving in 5 mL normal saline was immediately administered by gastrogavage after SCI, once daily. Equal volume of normal saline was administered to rats in the control group by gastrogavage. Hematoxylin and eosin (H&E) staining and C3 immunohistochemical staining were performed in SCI tissue at 12 h, day 1, 3, 7, and 14 after SCI. C3 positive expressions and myeloperoxidase (MPO) activity were assessed. Intercellular adhesion molecule-1 (ICAM-1) mRNA expression level was evaluated by Real-time PCR technique.

RESULTSC3 positive expression, MPO activity, and ICAM-1 mRNA level were significantly weaker in the Ephedra sinica group than in the control group at all time points (12 h, day 1, day 3, day 7, and day 14 after SCI) (P < 0.01, P < 0.05).

CONCLUSIONSThere existed complement system activation following acute SCI. The complement inhibiting component of Ephedra sinica significantly reduced immunological inflammation after SCI, and played an important role in secondary SCI.

Animals ; Complement Activation ; drug effects ; immunology ; Complement Inactivating Agents ; pharmacology ; Ephedra sinica ; chemistry ; Inflammation ; immunology ; Rats ; Rats, Sprague-Dawley ; Spinal Cord Injuries ; immunology ; metabolism ; pathology

OBJECTIVETo observe the effects of Herba ephedrae, honey-fried Herba ephedrae and Maxingshigan decoction on pentobabital sodium sleep experiment in mice.

METHODSMale Kunming mice were divided into 11 groups, namely normal saline (NS) group, ephedrine group, 3 Herba ephedrae dose (high, medium, and low) groups, 3 honey-fried Herba ephedrae dose group, and 3 Maxingshigan decoction dose groups. The corresponding drugs were administered intragastrically for 6 consecutive days, and 45 min after the final administration, the mice received intraperitoneal injection of pentobabital sodium, and the latent period and continuous sleeping time were recorded.

RESULTSCompared with high- and low-dose Herba ephedrae groups, Maxinshigan decoction containing equivalent Herba ephedrae significantly increased the sleeping time of the mice (P<0.05). In comparison with NS, the decoction at medium and low doses produced no significant changes in the sleeping time, which, however, was significantly shortened in the other 8 groups (P<0.05). Compared with Herba ephedrae, Maxingshigan decoction and honey-fried Herba ephedrae at equivalent doses showed comparable effects on the sleep latency (P>0.05).

CONCLUSIONUnder the condition of this experiment and with pentobabital sodium-induced sleeping time as the index, honey-fried Herba ephedrae shows no obvious effect in reducing the excitement of the central nervous system, while Maxingshigan decoction can significantly lower the excitement level, the effect of which is inversely correlated to the dose administered.

Animals ; Central Nervous System ; drug effects ; Dose-Response Relationship, Drug ; Drugs, Chinese Herbal ; pharmacology ; Ephedra sinica ; chemistry ; Male ; Mice ; Pentobarbital ; pharmacology ; Random Allocation ; Sleep ; drug effects

OBJECTIVETo establish a method for the determination of ephedrine and pseudoephedrine in Herba Ephedrae and Maxing Shigan Tang by capillary zone electrophoresis.

METHODThe conditions of the experiment were optimized with a fused-silica capillary of 60 cm x 50 microm (50 cm effective length) in a running buffer of 50 mmol x L(-1) borax-20 mmol x L(-1) threonine (pH 9.27) and an applied voltage of 15 kV (room temperature). Samples were introduced by hydrodynamic injections (10 cm x 20 s)and determined with on-column UV monitoring at 210 nm. Phenobarbital was chosen as the internal standard.

RESULTEphedrine and pseudoephedrine are separated successfully within 8 min. The linear responses covered the ranges from 21.3 to 213 mg x L(-1) (r = 0.9996) for ephedrine and from 8.4 to 84 mg x L(-1) (r = 0.9995) for pseudoephedrine. The detection limits (S/N = 3) of ephedrine and pseudoephedrine were shown to be 1.45 and 1.48 microg x mL(-1), respectively, The quantitation limits (S/N = 10) of ephedrine and pseudoephedrine were shown to be 4.81 and 4.93 mg x L(-1), respectively. The average recoveries for ephedrine and pseudoephedrine were 97.5% and 98.6% with RSD less than 5.0%.

CONCLUSIONThe method is simple, rapid, cost-effective and precise with satisfactory results.

Drugs, Chinese Herbal ; Electrophoresis, Capillary ; methods ; Ephedra sinica ; chemistry ; Ephedrine ; analysis ; Plants, Medicinal ; chemistry ; Pseudoephedrine ; analysis ; Reproducibility of Results