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ObjectiveTo observe the therapeutic effect of Radix Paeoniae Rubra-Radix Aconiti Lateralis on acute-on-chronic liver failure （ACLF） rats and its effect on M1/M2 macrophage polarization. MethodMale SD rats were randomly divided into normal group， model group， positive group （lactulose， 1.8 g·kg-1） and traditional Chinese medicine （TCM） group （Radix Paeoniae Rubra-Radix Aconiti Lateralis， 5.85 g·kg-1）， six in each group. The ACLF rat model was established by subcutaneous and tail vein injection of bovine serum albumin combined with intraperitoneal injection of D-galactosamine+lipopolysaccharide. Then the modeled rats were intervened with corresponding drugs for one week. The normal group and model group were given the same dose of distilled water. The histopathological changes of liver tissue were observed by hematoxylin-eosin （HE） staining. The mRNA and protein expression levels of CD86， inducible nitric oxide synthase （iNOS）， CD206 and arginase 1 （Arg1） were detected by real-time polymerase chain reaction （Real-time PCR）， Western blot and immunohistochemistry. ResultCompared with the conditions in the normal group， pseudolobule formation in liver tissue and morphological changes and necrosis of hepatocytes were observed in ACLF rats， accompanied by a large number of inflammatory cell infiltration. Moreover， the mRNA and protein expression levels of CD86， iNOS were up-regulated（P<0.01）. Compared with the model group， the treatment groups had improved necrosis and inflammatory infiltration of hepatocytes， down-regulated mRNA and protein expression of CD86 and iNOS （P<0.01） and up-regulated mRNA and protein expression of CD206 and Arg1 （P<0.05， P<0.01）， with the up regulation in the TCM group better than that in the positive group. ConclusionACLF rats had unbalanced M1/M2 macrophage polarization， and the imbalance shifted towards M1. Radix Paeoniae Rubra-Radix Aconiti Lateralis inhibited the activation of M1 macrophages and reduced the inflammatory response of liver failure by promoting the polarization of liver macrophages towards M2.
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Objective: To investigate effects of Rhizoma Ligustici Chuanxiong and Radix Paeoniae Rubra on the relative expression of Nogo-A/NgR/RhoA/ ROCK mRNA in acute cerebral infarction rats. Methods: Healthy male SD rats were randomly divided into sham operation group, blank group, low dose, medium dose and high dose Rhizoma Ligustici Chuanxiong and Radix Paeoniae Rubra group, Ginkgo biloba group, Nimodipine group, and each group was divided into 3 days, 7 days, 14 days three time points. Real-Time quantitative polymerase chain reaction (PCR) was used to detect Nogo-A/NgR/RhoA/ROCK mRNA relative expression changes of acute cerebral infarction rats. Results: Compared with the blank group and the sham operation group, the relative expression level of Nogo-A/NgR/RhoA/ROCK mRNA was increased in the model group both at 3 days, 7 days and 14 days (P ＜ 0.05). After the treatment of Rhizoma Ligustici Chuanxiong and Radix Paeoniae Rubra, other than there was no significant difference between the low-dose group and the model group except for 7 days, the relative expression level of Nogo-A/NgR/RhoA/ROCK mRNA in Rhizoma Ligustici Chuanxiong and Radix Paeoniae Rubra groups was lower than that in the model group (P ＜ 0. 05). Conclusion: The relative expression level of Nogo-A/NgR/RhoA/ROCK mRNA in acute cerebral infarction rats can be reduced by Rhizoma Ligustici Chuanxiong and Radix Paeoniae Rubra.
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Objective To observe the effect of the three active ingredients of a Chinese traditional medicine compound named Kang Fu Ling( KFL) against PC12 cells oxidative damage induced by microwave radiation.Methods PC12 cells were differentiated into neuros induced by nerve growth factor ( NGF ) .PC12 cells were incubated for 48 hours after astragalosides,total paeony glycoside and tanshinones were added at different concentrations (1, 3, or 9 μg/ml) .The cells in the control group were cultivated with the only medium of the same volume.Then, cells were irradiated with 30 mW/cm2 microwave for 6 minutes.The morphology of PC12 cells was observed under an inverted microscope soon before and after irradiation and the cell viability was measured by methylthiazolyl tetrazolium ( MTT) colorimetry.Reactive oxygen species ( ROS ) was determined using active oxygen probe 2′, 7′-dichlorodihyarofluolescen diacetde ( DCFH-DA ) while malonyldialdehyde(MDA) was measured in the homogenate of PC12 cells through thiobarbituric acid ( TBA) reactive substance assay.Results The cell morphology of each group showed no obvious difference.6 h after irradiation, the viability of irradiation control group measured by MTT declined apparently(P<0.01)compared with the normal control group.The 3 μg/ml astragalosides treatment group increased the viability of PC12 cells after microwave exposure ( P <0.01).The contents of ROS and MDA were increased after irradiation(P<0.01).However, in the three active ingredients of Kang Fu Ling treatment groups, both ROS and MDA were much lower than in irradiation control group.Conclusion Astragalosides, total paeony glycoside and tanshinones, which are the three active ingredients of Kang Fu Ling, all have protective effect against PC12 cell injury caused by microwave radiation,possibly by scavenging free radicals and reducing oxidative stress injury.
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Objective To investigate the protective effects of Radix Paeoniae Rubra(RPR)extracts on reticulum stress and macrophage infiltration on diabetic cardiomyopathy rats and the mechanism. Methods Sprague￣Dawley rats were randomly divided into 5 groups: normol control, model control (induced by streptozotocin),RPR extracts at low dose (i.p.15 g.kg-1 ), middle dose (i.p.30 g.kg-1 ), and high dose (i.p.60 g.kg-1 ).The artery intubation was carried out in all rats for detecting cardiac function 10 weeks later.The myocardial structure was detected by masson staining.The level of myocardial tissue glucose regulatory proteins GRP78,caspase￣12 and caspase￣3 expression were measured by Western blot.The ED￣1 protein was tested by immunohistochemical staining. Results Compared with the normal controls, the model rats presented decreased LVSP, +dp/dtmax ,-dp/ dtmax , LVEF, increased expressions of GRP78, caspase￣12,caspase￣3, ED￣1 and the synthesis of collagen fiber(P<0.05 or P<0.01).RPR extract inhibited the above indexes of model control rats in a concentration￣dependent manner(P<0.05 or P<0.01). Conclusion The RPR obviously ameliorates the cardiac function and myocardial fibrosis of diabetic cardiomyopathy rats,which may be associated with suppressing the expression of GRP78,caspase￣12,caspase￣3 and macrophage infiltration.
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OBJECTIVE: To develop an HPLC method for determination of nine components in Huangqichifeng capsules. METHODS: The analysis was carried out on a Waters Symmetry Shield RP18 column (4.6 mm × 150 mm, 5 μm). The mobile phase was composed of methanol (A) and water (B) with gradient elution (0-15 min, 5%A → 20%A; 15-55 min, 20% A → 35%A; 55-100 min, 35% A → 85% A) and the flow rate was 1.0 mL · min-1. The detection wavelengths were set at 230 nm for paeoniflorin (0-25 min) and 254 nm for other components (25-100 min). The column temperature was 35°C. RESULTS: The linear ranges of paeoniflorin, prin-O-glucosylcimifugin, calycosin-7-0-β-Z)-glucopyranoside, cimifugin, 4'-0-β-Z)-glucopyranosyl-5-0-methylvisammi-nol, formononetin-7-0-β-Z)-glucopyranoside, sec-O-glucosylhamaudol, calycosin, formononetin were 1.510-15.1, 0.047-0.94, 2.192-10.96, 0.052 2-1.044, 0.0414-0.828, 1.56-7.80, 0.0422-0.844, 0.149-0.744 and 0.110-0.552 μg (r ≥ 0.9990) respectively. The average recoveries were 99.48%, 103.15%, 102.77%, 99.27%, 101.99%, 102.25%, 101.49%, 100.45% and 97. 49% (RSD < 2.5%, n=6) respectively. CONCLUSION: The method is simple, accurate, repeatable, andean be used to control the quality of Huangqichifeng capsules. Copyright 2012 by the Chinese Pharmaceutical Association.
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Objective To investigate the effect of radix paeoniae rubra (RPR) on expressions of p38MAPK, NF-κB and iNOS in ltpopolysaccharide (LPS)-induced acute lung injury (ALI) and explore its protective mechanism. Methods Forty Wistar rats with ALI were divided randomly into five groups: saline control group (Group A) , LPS group (Group B), RPR for treatment group (Group C), RPR for prevention group (Group D) and SB203580 group (Group E). The effects of RPR on protein content and the ratio of neutrophiles in bronchoalveolar lavage fluid (BALF), contents of malondidehyde (MDA) and serum NO in lung tissue were observed. Arterial blood was collected for blood gas analysis. Expressions of p38MAPK, NF-κB and iNOS in rat lung tissues were detected. Results Compared with Group A, expressions of p38MAPK, NF-κB and iNOS were significantly increased (P <0.01 orP< 0.05) , the protein content and the ratio of neutrophiles in BALF, contents of MDA and serum NO were significantly higher (P<0.01 or P<0.05) in Groups B, C, D and E. There was a significant decrease in the level of arterial bicarbonate and partial pressure of oxygen in Groups B, C, D and E (P<0.01 or P<0.05). Compared with Group B, the expressions of p38MAPK, NF-κB and iNOS were significantly lower, and the protein content and the ratio of neutrophiles in BALF, contents of MDA and serum NO were significantly decreased, while the levels of arterial bicarbonate and partial pressure of oxygen were significantly higher in Groups C, D and E (P<0.05 or P<0.01). Compared with Group B, the lunginjury of Groups C, D and E was significantly alleviated, while there was no statistical difference among Groups C, D and E in the indices of lung injury. Conclusion Protective effect of RPR on LPS-induced ALI is closely related to the inhibited expressions of p38MAPK, NF-κB and iNOS.
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Objective To observe the therapeutic effects of Radix Paeoniae Rubra decoction (RPRD) on a-cute pancreatitis (AP) in rats, and to study effects of RPRD on the expression of NF-κB, TNF-α and IL-6. Method Forty Sprague-Dawley (SD) rats were, randomly divided into five groups: normal control group (group A), AP model groups (group B, 24 h; group C,72 h) and RPRD treatment groups (group D, 24 h;group E,72 h).There were 8 rats in each group. The AP models were induced by intraperitoneally injecting with L-arginine. After the model was estabhshed, rats of group B and group C were treated with inn'a-gastric instillation of NS, and rats in group D and group E with RPRD. At 24 and 72 hours after induction of AP, rats in each group were sacri-riced and blood samples were taken for the measurement of serum amylase. The expression of NF-κB 1065 protein and TNF-α and IL-6 mRNA in the pancreas was detected by Western blotting and real-time quantitative PCR, re-spectively, and the pathological status of the pancreas was analyzed. Results The serum amylase level in group B and group D was obviously increased at 24 hours, and were significantly higher than that in group A (P < 0.05) ; the level in group D was lower thaa that in group B (P < 0.05). the level reduced at 72 hours, and there was no difference between group E and group C (P > 0.05). The pathological severity and the pathological score of pan-creatic tissue in group D and group E at 24 hours and 72 hours were more hghter than those in group B and hours C (P < 0. 05). The expression levels of NF-κB p65 protein (Western blotting) in the pancreas of group D and group E were significantly lighter than those in group B and group C at both time points (group D compared with group B, (0.07 ± 0.006) vs. (0.71 ± 0.029), P < 0.05;group D compared with B, (0.07 ± 0.006) vs. (0.71 ±0.029), P <0.05; group E compared with C, 0.18±0.014 vs. 0.65±0.026, P <0.05). The results of real-time quantitative PCR shown that the expression levels of TNF-α and IL-6 mRNA in model treatment group were significantly stronger than that in RPRD group at the two time points (P < 0.05). Conclusions RPRD is effective for the treatment of AP rats to inhibit the activity of NF-κB. Inhibition of NF-κB offers a promising strategy to treat AP by down-regulating eytokine expression.
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Objective:To observe the effect of Radix Paeoniae Rubra on intimal proliferation,activating of angiotensin Ⅱ(AgⅡ) and expression of Mitogen-activated protein kinase phosphatase-1(MKP-1) after carotid artery balloon injury in cholesterol-fed rabbits.Methods: Male rabbits were randomly divided into Radix Paeoniae Rubra(PPR) groups: high dose group(75、50、25g.kg-1.d-1;n=8),middle dose group(2g.kg-1.d-1),low dose group(1g.kg-1.d-1;n=8) and control group.Both groups received high fat forage(2% cholesterol + 5% lard).Balloon injury of carotid artery was performed.Carotid artery were harvested at the end of 10 weeks.Expression level of AgⅡwas measured by radioimmunoassay.MKP-1 expression was determined by RT-PCR.Immunohistochemical staining and morphological detection were adopted.Results: Compare with control group,expression of AgⅡ decreased obviously(P
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OBJECTIVE: To study the change of the active components in Radix Paeoniae Rubra before and after the radiation of 60Co-?-rays.METHODS: The content of paeoniflorin in Radix Paeoniae Rubra before and after the radiation of 60Co-?-rays was determined by HPLC on Spherigel ODS C18(150 mm?4.6 mm,5 ?m) chromatographic column.The mobile phase consisted of methanol-water(gradient elution) with the detection wavelength set at 260.5 nm.RESULTS: The content of paeoniflorin in Radix Paeoniae Rubra dropped by nearly 20% after the radiation of 60Co-?-rays.CONCLUSION: The effect of the radiation of 60Co?-rays on the active component should be taken into consideration in treating the Chinese drugs preparation of Radix Paeoniae Rubra.
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Objective To isolate and extract effective antiendotoxin materials from Radix Paeoniae Rubra.Methods By the biosensor technology,effective antiendotoxin materials were isolated and extracted with general separation technology for Chinese traditional medicine.By ELISA mensuration for LPS and inhibition of TNF-? release from endotoxin-stimulated cells in vitro,the antiendotoxin activity of Radix Paeoniae Rubra was elucidated.Results The materials extracted had high binding capability to LPS and inhibitory effect on TNF-? release from endotoxin-stimulated cells in vitro.Conclusion Isolated and extracted effective antiendotoxin materials targeting LPS by the biosensor technology were valuable in searching antiendotoxin agents from Radix Paeoniae Rubra.The ability of Radix Paeoniae Rubra to neutralize LPS was assayed and proved very high.
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Objective To investigate the effect of radix paeoniae rubra (RPR) on expression of heme oxygenase (HO-1) and inducible nitric oxide synthase (iNOS) in lipopolysaccharide (LPS)-induced acute lung injury (ALI) and explore its protective mechanism. Methods Forty Wistar rats were randomly and equally divided into five groups, ie, control group, LPS group, RPR treatment group, RPR prevention group and Hemin group. Arterial blood was drawn for blood gas analysis. Models of endotoxin-induced ALI were used to observe the protein content, the ratio of neutrophiles in bronchoalveolar lavage fluid (BALF), the malondialdehyde (MDA) content in lung and the activities of serum NO. Expression of HO-1 and iNOS in rat lung tissue was detected by immunohistochemistry and morphometry computer image analysis. The histological change of lung were observed under light microscope. Results Compared with control group, expression of HO-1 and iNOS was markedly increased (P
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OBJECTIVE:To investigate the protective effect of Radix Paeoniae Rubra Extracts on rats with experimental cerebral ischemia.METHODS:Middle cerebral artery occlusion(MCAO)model was prepared in rats by thread-blocking internal carotid artery,and the model rats were intragastrically administered with different doses of Radix Paeoniae Rubra Extracts with the protective effect of Radix Paeoniae Rubra Extracts monitored.Cerebral infarction tissue,brain hndex and abnovmal nevons symptoms were observed.RESULTS:The weight of cerebral infarction tissue of the MCAO rats was decreased significantly by Radix Paeoniae Rubra Extracts dose-dependently in the dose ranges of 240,120,60 and 30 mg?kg-1.Radix Paeoniae Rubra Extracts could also decrease brain index and improve abnormal nervous symptoms in MCAO rats.The preventive medication of Radix Paeoniae Rubra Extracts has significantly lowered the weight percent of cerebral infarction tissue in MCAO rats,assuming dose-dependent manner in the dose range of 240～30 mg?kg-1,decreased the brain index and significantly improved the abnormal nervous symptoms of MCAO rats.CONCLUSION:Radix Paeoniae Rubra Extracts have protective effects on cerebral ischemia in MCAO model rats.
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OBJECTIVE:To establish the quality standard for Ruheneixiao soft capsules.METHODS:Radix Paeoniae Rubra,Radix Angelicae Sinensis and Spica Prunellae were identified using TLC.The content of Paeoniflorin in Ruheneixiao Soft Capsules was determined with HPLC.HPLC analytical method was carried out using a C18 column and a mixture containing 13 volume of acetonitrile and 87 volume of water as the mobile phase.The detection wavelength was set at 230nm.RESULTS:Spots obtained from the test solutions had the same color in reference solution and medical material in the same location,and the blank solution had no interference.The linear range of Paeoniflorin was from 5.1 to 40.8mg? mL-1(r=0.997 7)and the average recovery was 98.91%(RSD=0.34%,n=5).CONCLUSIONS:The TLC method produced distinctive spots,and was highly specific for use in the determination of Ruheneixiao soft capsules.The HPLC method was simple and fast,with high accuracy and precision.The standard can be used for the quality control of the drug.
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OBJECTIVE:To apply the biosensor technology to screening of the best method to extract and separate anti-endotoxin effective materials in Radix Paeoniae rubra.METHODS:The affinities of samples extracted by three different methods binding to lipopolysaccride(LPS)were determined with biosensor technology.Extracted materials were incubated with0.25EU/ml endotoxin for determination of change of binding activity.RESULTS:Material extracted by water showed high binding capa?bility with LPS,after incubation with endotoxin,still had highly effective anti-endotoxin component;1∶40diluted water extract could neutralize78.1%endotoxin.Determination results by biosensor technology and traditional limulus reagent showed no dif?ference.CONCLUSION:Water extraction could obtain more anti-endotoxin effective materials.Compared with traditional methods,biosensor technology is a fast,highly effective,direct and accurate method.
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Objective To observe the effect of drug pairs of extracts from Rhizoma Chuanxiong(RC)and Radix Paeoniae Rubra(RPR)in different proportions on promoting blood circulation and removing blood stasis in rats and to find the optimal proportion between the two extracts.Methods The Wistar rat models of blood stasis were established by injection of adrenalin.The changes of the platelet aggregation and adhesiveness and erythrocyte aggregation were observed.Results Blood vessel contraction increased,the platelet aggregation and erythrocyte aggregation increased,erythrocyte deformation occurred and the blood viscosity increased in the model rats.The drug pairs could reduce blood viscosity,ameliorate erythrocyte ability of deformation,reduce erythrocyte aggregation and inhibit platelet aggregation.The effects were obvious especially in the groups of RC and RPR in the proportions of 0.45 g to 0.45 g,0.21 g to 0.45 g.Conclusion The drug couples from RC and RPR in different proportions could improve the blood rheology,the effect being obvious in the groups of RC and RPR in the proportions of 0.45 g to 0.45 g,0.21 g to 0.45 g.
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Objective To explore the main pharmacological actions of Penyan Qing Capsule (PQC) for chronic pelvic inflammation. Methods The analgesic action of PQC was studied by mouse hot_plate method. Experiments of mouse auricular swell ing and rat agar_induced granuloma were carried out to investigate the anti_infl ammatory action of PQC. Effects of PQC on function of mouse mononuclear phagocyt es and rat mesenteric microcirculation were also studied. Results PQC increa sed the rat pain threshold, reduced mouse auricular swelling induced by xylene a nd the weight of rat agar_induced granuloma. PQC could also improved rat mesente ric microcirculation disorder induced by noradrenline and increase the function of mouse mononuclear phagocytes. Conclusion PQC exerts good anti_inflammatory and analagisic action and can improve microcirculation. This will supply pharm acodynamic evidence for the application of PQC in clinic.
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Objective To establish a RP-HPLC method for the determination of the blood concentration of paeoniflorin which was produced by seven kinds of warming-inside drugs (WID) being used with the promoting-blood drugs (PBD) individually and to explore the mechanism of PBD and WID compound prescription. Methods The blood concentration of peaoniflorin in mouse plasma was determined by HPLC after ig seven kinds of WID being compatible with Radix Paeoniae Rubra (RPR) to mice separately. Results Fructus Piperis (FP), Cortex Cinnamoni (CC), Fructus Evodiae (FE), Fructus Foeniculi (FF), and Pericarpium Zanthoxyli (PZ) compound prescription with RPR can increase the blood concentration of paeoniflorin in different degrees (P
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Object To develop a new method of discriminating the principle components of Radix Paeoniae Rubra (RPR) from different areas, and to find the reason for forming high-quality of RPR in the place of the genuine. Methods Using Foruier transform infrared (FTIR) absorption spectrometry, the characteristic peaks of fingerprint infrared spectra of RPR samples from 18 habitats were recognized and compared. Results Frequency, intensity and shape of infrared absorption spectra were obviously different between wild and cultivated groups of RPR. The infrared absorption peaks of RPR in Duolun, a famous Chinese orthodox drug, were of distinct characteristics. Conclusion FTIR technique is first applied to rapid analysis of principle component of RPR from different areas. So an operable method in the quality control and discrimination of RPR in the place of the genuine is provided.
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Object To study the HPLC fingerprints and establish a sensitive and specific method for controlling the quality of Radix Paeoniae Rubra (RPR) Methods The HPLC method was applied for quality assessment of RPR All 18 samples collected from different places were determined A Hypersil C 18 column (200 mm?4 6 mm, 5 ?m) was used with the mobile phase being acetonitril 0 025 mol/L H 3PO 4: THF (9 5∶90 5∶1 25), flow rate being 0 8 mL/min, detecting wavelength being 254 nm, the column temperature being at room temperture Results This method had a good repeatability and reproducibility, and the ratio of peaks' area of different habitat samples were different Conclusion The method is suitable to differentiate RPR form different sources conveniently, and can be used as a quality control method for this herb
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OBJECTIVE:To isolate anti-endotoxin monomer component from Radix Paeoniae Rubra by means of biosen?sor technique.METHODS:The surface of biosensor cuvette was embedded by Lipid A;the screening target was established,tracking the silica gel column chromatogram and the binding ability of effluent component from HPLC with Lipid A with the ultraviolet scan result of the reclaimed material from biosensor as reference;anti-endotoxin monomer component was isolated;the component of monomer and the synthetic action of extrinsic lipopolysaccharides were also assayed by LAL test method.RESULTS:Components binding to Lipid A was reclaimed from cuvetee by biosensor technique,with the wavelength of UV absorption peak at194nm,215nm and275nm respectively.Anti-endotoxin monomers of higher binding activity with Lipid A isolated by HPLC method were1,2,3,4,6—O—pentagalloyl—?—D—glucose(PGG).PGG at concentration of8,4,2?g/ml respectively neutralized68.8%,43.7%and31.4%of LPS at an activity of0.1EU/ml respectively.CONCLUSION:It is fea?sible to isolate anti-endotoxin monomer component from Radix Paeoniae Rubra by means of biosensor technique,which is a fast,accurate and efficient and can be used to isolate anti-endotoxin monomer component from Radix Paeoniae Rubra on a large scale.