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Objective:To compare the differences of chemical components between single decoction and mixed decoction with different compatibility ratio of Inulae Flos- Haematitum medicinal pair. Methods:UPLC method was used to determine the contents of 5-caffeoylquinic acid, chlorogenic acid, 4-dicaffeoylquinic acid, caffeic acid, isoquercitrin, isochlorogenic acid B, 1,5- dicaffeoyl quinic acid, isochlorogenic acid C and the fingerprints of the single decoctions and mixed decoctions of Inulae Flos- Haematitum medicinal pair in four groups of proportions. The "peak area/sample weight" value of each common peak in the fingerprints was calculated, and the SPSS 26.0 was used for independent-sample t-test analysis. Results:There are significant differences in the "peak area/weight" values of peak 1, peak 2, peak 4, peak 6 , peak 9, peak 10, peak 12, peak 13, peak 15 between mixed decoction and single decoction of Inulae Flos - Haematitum medicinal pair with different compatibility ratios ( P<0.05), with statistical significance; when the compatibility ratio of Inulae Flos- Haematitum medicinal pair was 3:1, the difference of fingerprints and index components content between single decoction and combined decoction was the largest. Except for peak 7 and peak 14, the difference of "peak area/sample weight" value of other characteristic peaks was statistically significant ( P<0.05), and the content difference of 8 index components was statistically significant ( P<0.05). Conclusion:There are differences in the chemical components of Inulae Flos - Haematitum medicinal pair for single decoction and mixed decoction.
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Objective:To establish methods for HPLC fingerprints and simultaneous determination of multi-index components before and after compatibility of Salvia miltiorrhiza and Angelica sinensis, so as to analyze the dissolution rate of the main compounds. Methods:The extracts of Salvia miltiorrhiza, Angelica sinensis and their compatibility were prepared. The separation was performed on an Eclipse XDB-C18 column (4.6 mm×250 mm,5 μm), mobile phase with 0.1% phosphoric acid aqueous solution-acetonitrile for gradient elution, flow rate at 1.0 ml/min, column temperature was maintained at 35 ℃, and the detection wavelength was set at 280 nm. The HPLC fingerprint were established before and after the compatibility of Salvia miltiorrhiza and Angelica sinensis, and the shared patterns of the fingerprint were obtained to gain chromatographic peaks. The content of 9 components Danshensu, caffeic acid, rosmarinic acid, salvianolic acid B, salvianolic acid A, tanshinone Ⅱ A, ferulic acid, chlorogenic acid and Yangchuanxiong lactone were determinated, and the changes of dissolution rate of each compound before and after the compatibility were analyzed. Results:The determination method for the multi- components with HPLC is precise and the components (waiting to be determinate) in the solution were stable within 48 hours, and the RSD values of each chromatographic peak were <5.0%. The nine components showed good linear relationships within their own ranges, and the recovery rate was in compliance with regulations. The fingerprint similarities of each sample were ?0.9. After the compatibility of Salvia miltiorrhiza and Angelica sinensis, a total of seventeen common peaks were calibrated, ten of which were from Salvia miltiorrhiza, seven from Angelica sinensis. No new components was found under this chromatographic condition. After the combination of these two material medicica decoction, the average dissolution rates of rosmarinic acid, salvianolic acids and Danshensu in Salvia miltiorrhiza were significantly lower than those of the single decoction group ( P<0.05 or P<0.01); the average dissolution rates of caffeic acid in Salvia miltiorrhiza was significantly higher than that of the single decoction group ( P<0.01); the average dissolution rates of chlorogenic acid and ferulic acid in Angelica sinensis were significantly higher than that of the single decoction group ( P<0.05 or P<0.01); the average dissolution rate of Yangchuanxiong lactone after the compatibility was not statistically different than that of single decoction group. Conclusion:The characteristic peaks of HPLC fingerprint of the compatibility of Salvia miltiorrhiza and Angelica sinensis did not increase under this chromatographic condition, which had a significant effect on the dissolution of index components.
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Objective@#To study the effects of different compatibility ratios of Centella asiatica- Rhubarb on the dissolution rates of madecassoside and asiaticoside.@*Methods@#By using the HPLC method to analyze the content of madecassoside and asiaticoside in the Centella asiatica extract solution as well as Centella asiatica - Rhubarb extract solution with 6 different kinds of compatibility ratios. The tests were carried out by Thermo C18 (250 mm×4.6 mm, 5 μm) by isocratic elution with acetonitrile and 2 mmol/L of beta-cyclodextrin solution as mobile phase at a flow rate of 1 ml/min, the column temperature was 30 ℃ and the detection wavelength was 205 nm.@*Results@#Compared with the Centella asiatica extract solution, the dissolution rate of madecassoside decreased significantly with the increase of the proportion of rhubarb. The combination of Centella asiatica and Rhubarb had little effect on the content of asiaticoside.@*Conclusions@#Considering the clinical application and efficacy of Centella asiatica, the optimal ratio of Centella asiatica-Rhubarb is 10:1 and 15:1.
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Objective@#To study the effect of different proportions of piper chinaroot and rheum palmatum on the dissolution rate of five effective components (aloe emodin, emodin acid, emodin, emodin, emodin methyl ether).@*Methods@#The high performance liquid chromatography (HPLC) method was used to analyz the contents of five effective components of rheum palmatum in the extracts of different combination of piper chinaroot and rheum palmatum. The tests were carried out by Thermo C18 (250 mm×4.6 mm, 5 μm) by gradient elution with methanol and 0.1% phosphoric acid water solution as mobile phase at a flow rate of 1ml/min, the column temperature was 30 ℃ and the detection wavelength was 254 nm.@*Results@#The linear ranges of aloe emodin, emodin acid, emodin, emodin, emodin methyl ether were 0.018 5-0.741 8, 0.017 9-0.717 8, 0.015 9-0.635 5, 0.054 2-2.167 2, 0.016 2-0.646 4 μg, respectively. The average recoveries of aloe emodin, emodin acid, emodin, emodin, emodin methyl ether were 94.35%, 95.50%, 100.61%, 96.27%, 97.39%, and the RSDs were 1.81%, 1.99%, 2.84%, 2.71%, and 1.86%, respectively. Compared with rheum palmatumby single extract, the content of aloe emodin and emodin were increased by 5 proportions (3:1, 2:1, 1:1, 1:2, 1:3), while the content of emodin and emodin methyl ether were decreased.@*Conclusions@#The optimal compatibility proportion of piper chinaroot and rheum palmatum is 1:1.
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Objective@#Comparison of effects of Zingiberis Rhizoma and Aristolochia manshuriensis on diuretic effect and acute renal injury in rats by combining two methods of co-decoction and mixed-decoction. The effects of in vitro observation on normal human renal tubular epithelial cells (HK-2) were observed.@*Methods@#The rats were randomly divided into five groups: blank group, positive control group, aristolochia manshuriensis group, combined decoction group and divided decoction group, 8 rats in each group. The water-loading rat model was established by intragastric administration of normal saline. The urine of rats was collected and the volumes of urine were measured for 24 hours after the corresponding drugs were given to each group. After 2 weeks of gavage of the corresponding drugs in each group, the serum BUN, SCr and urine UCr and PRO levels were measured by 7600P automatic biochemical analyzer, and renal histopathology were observed by HE staining. The HK-2 cells were cultured in vitro and divided into control group, Aristolochia manshuriensis group, mixed decoction group and sub-decoction group. After 24 hour intervention, the activity of cells was detected by CCK-8 method and the apoptosis was observed by Hoechst stain method.@*Results@#There was no significant difference in 24 hour urine output between the groups (P>0.05). Compared with Aristolochia manshuriensis group, the kidney coefficient (0.010 1 ± 0.005 8 vs. 0.013 3 ± 0.007 8), SCr (38.52 ± 0.58 μmol/L vs. 46.61 ± 0.72 μmol/L), BUN (8.55 ± 0.12 mmol/L vs. 10.21 ± 0.30 mmol/L), UCr (52.21 ± 0.89 μmol/L vs. 57.71 ± 0.67 μmol/L), PRO (29.89 ± 0.18 mg/L vs. 34.23 ± 6.05 mg/L) of combined decoction group significantly decreased (P<0.05). The survival rate of HK-2 cells (72.45% ± 3.70% vs. 55.92% ± 8.39%) in combined decoction group significantly increased (P<0.01), and the apoptosis rate (7.9% ± 2.6% vs. 31.6% ± 9.1%) significantly decreased (P<0.01).@*Conclusions@#The traditional co-decoction method of Aristolochia manshuriensis compatibility with Zingiberis Rhizoma can achieve a certain attenuation effect, and the mixed-decoction group can not achieve the attenuating effec.
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In the Qing dynasty Cheng Zhongling based on the Cheng's Bixie-Fenqing formula of Yang Tan, a doctor in the Southern Song dynasty, retained the original Sichuan Bixie and Acorus tatarinowii, and add Phellodendron chinense, and Semen plantaginis to form Cheng's Bixie-Fenqing formula. It had the function of get rid of dampness and heat, and is a classic prescription for stranguria, like the treatment of white turbidity. The author retrieved the related literature of Cheng's Bixie-Fenqing formula in recent years, analyzed the composition and clinical application of Cheng's Bixie-Fenqing formula, and summarized the advantages and disadvantages of the existing researches, providing reference for the further development of Cheng's Bixie-Fenqing formula.
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In the Qing dynasty Cheng Zhongling based on the Cheng's Bixie-Fenqing formula of Yang Tan, a doctor in the Southern Song dynasty, retained the original Sichuan Bixie and Acorus tatarinowii, and add Phellodendron chinense, and Semen plantaginis to form Cheng's Bixie-Fenqing formula. It had the function of get rid of dampness and heat, and is a classic prescription for stranguria, like the treatment of white turbidity. The author retrieved the related literature of Cheng's Bixie-Fenqing formula in recent years, analyzed the composition and clinical application of Cheng's Bixie-Fenqing formula, and summarized the advantages and disadvantages of the existing researches, providing reference for the further development of Cheng's Bixie-Fenqing formula.
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Objective Comparison of effects of Zingiberis Rhizoma and Aristolochia manshuriensis on diuretic effect and acute renal injury in rats by combining two methods of co-decoction and mixed-decoction. The effects of in vitro observation on normal human renal tubular epithelial cells (HK-2) were observed. Methods The rats were randomly divided into five groups: blank group, positive control group, aristolochia manshuriensis group, combined decoction group and divided decoction group, 8 rats in each group. The water-loading rat model was established by intragastric administration of normal saline. The urine of rats was collected and the volumes of urine were measured for 24 hours after the corresponding drugs were given to each group. After 2 weeks of gavage of the corresponding drugs in each group, the serum BUN, SCr and urine UCr and PRO levels were measured by 7600P automatic biochemical analyzer, and renal histopathology were observed by HE staining. The HK-2 cells were cultured in vitro and divided into control group, Aristolochia manshuriensis group, mixed decoction group and sub-decoction group. After 24 hour intervention, the activity of cells was detected by CCK-8 method and the apoptosis was observed by Hoechst stain method. Results There was no significant difference in 24 hour urine output between the groups (P>0.05). Compared with Aristolochia manshuriensis group, the kidney coefficient (0.010 1 ±0.005 8 vs. 0.013 3 ± 0.007 8), SCr (38.52 ± 0.58 μmol/L vs. 46.61 ± 0.72 μmol/L), BUN (8.55 ± 0.12 mmol/L vs. 10.21 ± 0.30 mmol/L), UCr (52.21 ± 0.89 μmol/L vs. 57.71 ± 0.67 μmol/L), PRO (29.89 ± 0.18 mg/L vs. 34.23 ± 6.05 mg/L) of combined decoction group significantly decreased (P<0.05). The survival rate of HK-2 cells (72.45% ± 3.70% vs. 55.92% ± 8.39%) in combined decoction group significantly increased (P<0.01), and the apoptosis rate (7.9% ± 2.6% vs. 31.6% ± 9.1%) significantly decreased (P<0.01). Conclusions The traditional co-decoction method of Aristolochia manshuriensis compatibility with Zingiberis Rhizoma can achieve a certain attenuation effect, and the mixed-decoction group can not achieve the attenuating effec.
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Traditional Chinese medicine(TCM)compatibility is related to the safety of clinical use of drugs and embodies the essence of interactions between drugs and organisms. This is a scientific problem thatr has attracted more attention. Currently, most of the knowledge on TCM compatibility is from predecessors′ experience. The corresponding molecular biology mechanism is poorly understood. As a powerful tool to identify a large number of proteins simultaneously,proteomics technology has the potential to reveal the protein alterations under certain conditions,and to provide more direct insights into biological processes during TCM compatibility. In this paper,we introduced the main technology of proteomics,including two dimensional gel electrophoresis,2D-DIGE,iTRAQ, QconCAT/MRM and chemical proteomics. Also,the applications of these technologies in the field of TCMM compatibility study were reviewed.
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【Objective】To observe the effect of Moschus combined with Borneol on brain water content and blood-brain barrier(BBB)permeability in rat model of cerebral focal ischemia with reperfusion.【Methods】Thirty SD rats were randomized into 6 groups: sham-operation group,model group,Moschus(1 mg?kg-1?d-1)group,Borneol(3 mg?kg-1?d-1)group,Moschus with Borneol group,nimodipine(12 mg?kg-1?d-1)group.The rat model of cerebral focal ischemia with reperfusion was established with method of putting nylon monofilament in the internal carotid artery.The brain water content was evaluated by detecting the wet weight and dry weight of brain.The detection of cerebral extra-vascular Evans blue(EB)content was used to observe the changes of BBB permeability.【Results】After cerebral ischemia for 2 hours and reperfusion for 24 hours,water content and EB content in the model group increased significantly compared with the pseudo-operation group(P
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【Objective】To compare aristolochic acid(AA)content in Caulis Aristolochiae Manshuriensis(CAM)and in the separated prescriptions of Longdan Xiegan Decoction(LXD),and to explore the effect of Chinese herbal medicine compatibility on AA content.【Methods】According to the herbs percentage compared to LXD,the herbs samples were named after: CAM group,LXD group,CAM group with heat-clearing herbs,CAM group with yin-nourishing herbs,CAM group with herbs for promoting urination,CAM group with Radix Glycyrrhizae,and LXD group with CAM removed.The AA content was determined by reverse phase HPLC.The chromatographic conditions were as follows: C18 Diamonsiltm analytical column,gradient eluted with a mixture of methyl alcohol,water and acetic acid(volume proportion as 74:24:1),and detection wavelength at 315 nm.【Results】AA content was the lowest in LXD group,and was lower in CAM group with yin-nourishing herbs than that in CAM group.【Conclusion】Subjecting to the theory of Chinese herbal medicine compatibility,it is feasible to reduce the CAM toxicity by composing a prescription with proper proportion of herbs.
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【Objective】In-vitro antibacterial activity of the combination of Herba Violae(HV)and Herba Taraxaci(HT)in different proportions was compared and analyzed for the optimization of the combination proportion of Herba Violae and Herba Taraxaci and the influencing factors.【Methods】The minimum inhibitory concentration(MIC)of the combination of HV and HT was determined by agar double dilution method and the minimum bactericidal concentration(MBC)by broth dilution method,and bactericidal curve was traced.Meanwhile,the resistance of the combination of HV and HT induced by sub-inhibitory concentration was analyzed,and the influencing factors of in-vitro antibacterial activity were investigated.【Results】The combination of HV and HT in the proportion of 1∶4 had a better antibacterial activity on four kinds of gram-positive bacteria and on four kinds of gramnegative bacteria,and exerted an bactericidal effect on Escherichia coli and Klebsiella Pneumoniae(both MBC_(50)/MIC_(50) and MBC_(90)/MIC_(90) being 2).The bactericidal curve showed that 2?Log cfu/mL bacteria strain could be killed by the combination of the proportion being 1∶4 at 8~(th) hour,the effect being superior to that of other two combinations.Inoculation dosage,serum amount and pH value had no effect on in-vitro antibacterial activity.【Conclusion】The combination of HV and HT in the proportion of 1∶4 has a better antibacterial activity.
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Objective To investigate the effect of modified Taohe Chengqi Decoction(TCD)and its separated prescriptions on tissue plasminogen activator(tPA)and plasminogen activator inhibitor-1(PAI-1)content in human umbilical vein endothelial cells(HUVEC)with diabetic damage.Methods Purified HUVEC were divided into 10 groups: normal group,model group,rosiglitazone group,high-,middle-and low-dose modified TCD groups,blood-activating prescription group,heat-clearing and purgation prescription group,Qi-strengthening and yin-nourishing group,and TCD group.The serum containing herbal medicine was prepared through seropharmacological method,and was used for co-culture of HUVEC with high glucose and high insulin in vitro.The tPA and PAI contents in the HUVEC culture of different groups were detected.Results The content of tPA was increased in blood-activating prescription group and rosiglitazone group(P0.05).The PAI-1 content in the model group was higher than that in the normal group(P0.05).Conclusion The effect of modified TCD on improving the function of endothelial cells is probably related to the regulatory effect of blood-activating,heat-clearing and purgation herbs on PAI-1.