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Aim To investigate the mechanism of eata- pol (CAT) inhibiting differentiation and glyeolysis of Thl7 eel Is through miR-143-3p.Method The peripheral hloorl CD4 ∗ T eells of HA patients were obtained to deteet the expression of miR-143-3p and the mRNA levels of key glycolytic enzymes, ineluding glucose transporter 1 ( Glutl ) , hexokinase 2 ( HK2 ) , pyruvate kinase 2 (PKM2) , laetate dehydrogenase A ( LDHA).The differentiation of Thl7 eells was induced in vitro, and the ShRNA/lentivirus was applied to achieve the overexpression or knockdown of miR- 143-3 p.Un-transfected eells were divided into control group and CAT group (20, 40, 80 mg • L 1 ) , and transfected eells were divided into four groups: negative control group, miR-143-3p inhibitor group, miR- 143-3p mimies group, miR-143-3p inhibitor + CAT group.The percentage of Thl7 eells was deteeted by flow cytometry, and the level of IL-17A was detected by EL1SA.Quantitative real-time PCR was used to detect the mRNA expression of miR-143-3p and key glycolytic enzymes, and the levels of pyruvate and lactate were also detected.Results The mRNA expression of miR-143-3p in RA peripheral blood CD4 ∗ T cells was negatively correlated with disease severity ( DAS28 ) , transcription factor ROR-yt, and the key glycolytic enzymes Glutl/HK2/LDHA.Compared with negative control group, the down-expression of miR-143-3p markedly elevated the mRNA expression of ROR-yt, Glutl, HK2, LDHA, and the levels of IL-17A, pyruvate, lactate.Catalpol groups significantly up-regula- ted the expression of miR-143-3p, decreased the mRNA expression of HK2/LDHA and the levels of pvru- vate/lactate, and inhibited Thl7 cells differentiation.Compared with miR - 1 4 3 - 3 p inhibitor group , catapol could significantly inhibit the abnormal up-regulated of HK2/LDHA mRNA relative expression, pyruvate/lactate levels and the abnormal differentiation of Thl7 eells.Conclusion MiR-143-3p inhibits the differentiation and glycolysis of Thl7 cells.Catalpol could sup-press the glycolysis and differentiation of Thl7 eells by regulating mill-143-3p.
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Objective To analyze the treatment and medication rules of formulae containing prepared rehmannia root in Pharmaceutical Standard of Ministry of Public Health of the People's Republic of China-Traditional Chinese Patent Medicine (referred to as Traditional Chinese Patent Medicine). Methods Through Traditional Chinese Medicine Inheritance Support Platform (V2.5), prescriptions containing prepared rehmannia root in traditional Chinese patent medicine were selected and collected to build the database. Frequency counts,association rules and other data mining methods were used to analyze the frequency of indication syndromes and medicine combination of prescription containing prepared rehmannia root, and the compatibility rule of high frequency drug pairs and the rescription rule of core syndromes were analyzed. Results There were a total of 467 prescriptions containing pepared rehmannia root, involving 28 combinations of commonly used Chinese herbals (support degree 30%). The compatibility source of high-frequency drug pairs of prepared rehmannia root and Angelicae Sinensis, prepared rehmannia root and Astragalus embranaceus were both of Shiquan-Dabu decoction in Taiping Huimin Heji Ju Fang (support degree 40%). There were a total of 81 indication syndromes, and among them, there were 16 kinds of high-frequency indication syndromes (the frequency greater than or equal to 10). The core drug combination of high-frequency indication syndromes of "deficiency of both qi and blood" (137 times) and "deficiency of kidney yang" (63 times) were the subtract of Shiquan-Dabu decoction and Yougui pill in Jingyue Quanshu respectively. Conclusions Prepared rehmannia root is mainly compatible with herbs of nourishing blood, supplementing qi and invigorating yang in the traditional Chinese patent medicine containing prepared rehmannia root. The traditional Chinese patent medicine containing prepared rehmannia root were based mainly on classical prescriptions. This study can provide reference for clinical application and new drug development of pepared rehmannia root.
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OBJECTIVE: To observe the change of EPO in brain of aging rat induced by D-galactose (D-gal) and the EPO-based antiaging of the water extract of Radix Rehmanniae Preparata (WERRP). METHODS: D-gal-treated groups were received subcutaneous injection of D-gal at dose of 50, 150 and 250 mg·kg-1 daily for 8 weeks to imitate an aging model that was induced by oxidative stress. After the detection of EPO in hippocampus, the 150 mg
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This study was aimed to analyze the content variations of chemical constituents produced in fresh Radix Rehmanniae (Xian-Di-Huang) after processing into Radix Rehmanniae Recens (Sheng-Di-Huang) and Radix Rehmanniae Preparata (Shu-Di-Huang). HPLC was used in the study of preparing Xian-Di-Huang into Sheng-Di-Huang, and the processing into Shu-Di-Huang. The contents of two newly produced chemical components, which were 2,3-dihydro-3,5-dihydroxy-6-methyl-4H-pyran-4-one (DDMP) and 5-hydryoxymethyl-furfural (5-HMF). The contents of both chemical components were determined in Sheng-Di-Huang and Shu-Di-Huang bought from the market. The Zorbax SB-C18 column (250 mm í 4.6 mm, 5 μm) was used. The mobile phase was methanol-wa-ter (5:95). The flow rate was 1.0 mL·min-1. The detection wavelength was set at 280 nm. The results showed that no DDMP or 5-HMF was detected in Xian-Di-Huang. However, after processing, DDMP and 5-HMF can be de-tected in both Sheng-Di-Huang and Shu-Di-Huang. The content in Shu-Di-Huang was higher than that in Sheng-Di-Huang. Both contents in Shu-Di-Huang were gradually increased along with processing time. The con-tent reached to the highest level after processing for 24 h and 32 h, respectively. And then, the content decreased. Both DDMP and 5-HMF were detected from three batches of Sheng-Di-Huang and ten batches of Shu-Di-Huang bought from the market. The content of 5-HMF was higher in Shu-Di-Huang than in Sheng-Di-Huang. There was no obvious difference in the content of DDMP between Sheng-Di-Huang and Shu-Di-Huang. It was concluded that DDMP and 5-HMF were produced in the processing Sheng-Di-Huang and Shu-Di-Huang. The contents were gradually increased along with the prolonging of processing time. There was obvious difference in the content of 5-HMF in Shu-Di-Huang and Sheng-Di-Huang.
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OBJECTIVE: To investigate the active substances with immunoenhancement effect in Prepared Radix Rehmanniae and its mechanism of action. METHODS: Balb/c mice lymphocytes were isolated from thymus and spleens with conventional methods. MTT assay was carried out for proliferation detection, and then ELISA assay was used to measure the levels of Th1 cytokines IL-2, IFN-γ and Th2 cytokines IL-4, IL-5 of lymphocytes from thymus and spleens of the mice. RESULTS: The water extract (0.049, 0.49 and 4. 9 mg · mL-1) and crude polysaccharide extract (0.032, 0.32 and 3.2 mg · mL-1) of Prepared Radix Rehmanniae resulted in strong promotion of thymus and spleen lymphocytes proliferation and obvious increase of IL-2, IFN-γ, IL-4 and IL-5 levels with or without Con A stimulation. These effects were dose-related. CONCLUSION: Prepared Radix Rehmanniae can significantly enhance immune responses, and the active substance might be crude polysaccharide. The mechanism of their immunoenhancement effects is partly due to enhancement of gene expression of Th1/Th2 cytokines of T cells.
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Objective To observe the therapeutic effect of Radix Astragali,Radix Rehmanniae combined with glucocorticoid(GC) in treating patients with systemic lupus erythematosus(SLE) and its influence on some experimental indexes.Methods 72 cases of SLE were randomly recruited into the integrated traditional Chinese and western medicine treated group (treated group) and the western medicine treated group (control group).The control group was treated with prednisone and cyclophosphamide,while the treated group was treated with Radix Astragali and Radix Rehmanniae on the basis of the control group.Both groups had been treated for 6 months as a therapy course.The use of prednisone dosage were recorded and its influence on indexes of clinical and laboratory value were observed.Results 27 SLE patients in the treated group and 34 in the control group were completely observed.The dosages of prednisone in both groups were decreased gradually after the state of disease had been relieved.The predrisone dosage in the treated group was lower than that in the control group in the therapy course(15.41 ±7.65)mg vs (17.25±8.83)mg,but without significant difference (t=0.85,P＞0.05).The total decreased prednisone dosage in the treated group (50.12±12.53 mg) was slightly higher than that in control group(48.96± 13.71) mg (t=0.34,P＞0.05).The ratio of cases who had to add prednisone for aggravating disease in treated group(15.92％) was less than that in the control group (38.24％),the difference was significant (x2=3.24,P＜0.05).The incidence rate of cushing syndrome,infection,diseases of digestive,cardiovascular anomalies in the treated group was slightly less than the control group (x2=0.42,0.98,0.01,0.03respectivly,P＞0.05),but the number of insomnia and hot flush were significantly larger in the control group than the treatment group (x2=4.24,5.93,P＜0.05).No difference about the SLEDAI,ESR,CRP,Blood Complement3,Blood imune globinG was found between the two groups (t=2.16,1.04,1.18,0.26,1.61 respectively,P＞0.05),and 24 hours urinary protein count decreased significantly in the treated group (1.06 ± 0.12) g/L,compared with control group (1.42 ± 0.54) g/L,(t =3.78,P ＜ 0.05 ).Conclusion Radix Astragal,Radix Rehmanniae combined with conventional treatment of western medicine could withdraw corticosteroid smoothly,relieve symptoms and alleviate some side-effects of western medicine.
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Aim To investigate the effect of catalpol from Radix rehmanniae on the proliferation,differentiation and matrix mineralization of MC3T3-E1 cells in vitro.Methods Catalpol from Radix rehmanniae of different concentration preparations were extracted with ethanol(catalpol ethanol-extract),respectively.A mouse osteoblast cell line MC3T3-E1 was used to determine the potency of catalpol.MTT were applied to determine proliferation of the cell treated by catalpol at different concentrations.Differentiating effects of the catalpol with different concentrations in the cell were evaluated through the examinations of alkaline phosphatase(ALP)activities and bone gla protein(BGP)levels.Von kossa staining method was used to demonstrate the effects of the catalpol on calcification of the cells.Results Catalpol at concentration from 1×10~(-7) to 1×10~(-9) mol·L~(-1) for 24 hours and 48 hours effective promoted the proliferation of osteoblasts of MC3T3-E1 cells line.Catalpol at concentration from 1×10~(-5) to 1×10~(-6) mol·L~(-1) for 48 hours and 72 hours effective stimulated the activity of ALP of osteoblasts of MC3T3-E1 cells line.Catalpol at concentration from 1×10~(-5) to 1×10~(-6) mol·L~(-1) for 8 days and 12 days effective increased the synthesis and secretion of bone gla protein(BGP) of osteoblasts of MC3T3-E1 cells line.Catalpol at concentration from 1×10~(-5) to 1×10~(-6) mol·L~(-1) for 19 days effective increased the mineralized bone nodular structure number of osteoblasts of MC3T3-E1 cells line.Conclusion Catalpol could promote proliferation and differentiation of MC3T3-E1 cells in vitro.Catalpol may be one of effective monomer of Radix rehmanniae on treatment of osteoporosis.
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Purpose To investigate the anti-inflammatory effect of Radix Rehmanniae Praeparata(RRP)on the footpad inflammation induced by complete Freund's adjuvant(CFA)in rats.Methods CFA 100 μL were injected subcutaneously to the Wistar rats at the pad of right hindfoots.19 days later,the rats were daily siven RRP water extract(0.625,1.250,2.500 g/kg·bw)or dexamethasone(0.5mg/ks·bw)intragastrically.The changes of body weight and foot volume were measured.The indexes of organ and blood were determined at the 29th day,the foot pad was removed,and routine paraffin section was performed.Results The model rats kept foot swelling and lymphocyte infiltrating,and the platelet number decreased.The other indexes were statistically insignificant when compared to the controls.RRP did not display any anti-inflammatory effect on the swollon foots,but thoracic gland and spleen indexes were rescued,and platelet number and creatinine content were increased by RRP administration in a dose-dependent manner.The anti-inflammation of dexamethasone was conspicuous,but the side effects were also significant.Conclusion RRP may be plays an adjunctive action in herbal recipes to treat rheumatoid arthritis.
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Objective To investigate the effect of catalpol from Radix Rehmanniae on A?25-35-induced apoptosis of PC12 cells.Methods PC12 cells were routinely cultivated and treated by A?25-35(final concentration,20 ?mol/L) 24 hours after the addition of catalpol or saline.Forty-eight hours later,cells were examined for viability and apoptosis by MTT method and TUNEL method,respectively,while Bax and Bcl-2 mRNA expression were analyzed by semi-quantitive RT-PCR. Results Catalpol could significantly elevate the viability at 1?10-5 mol/L and 1?10-4 mol/L(P
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Objective:To approach the determination method of 5-hydroxymethyl-furfural in Radix Rehmanniae,and to investigate influence of steam processing on catalpol and 5-hydroxymethyl-furfural. Methods:Catalpol and 5-hydroxymethyl-furfural from Radix Rehmanniae at different processing time was determined by high performance liquid chromatography. Results:In the steam processing,content of 5-hydroxymethyl-furfural increased with the increase of time in the beginning,when steamed to 52h,the levels began to decline; content of catalpol reduced with increase of the processing time. Conclusion:Content of catalpol and 5-hydroxymethyl-furfural changed signifi cantly in the steam processing.
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Objective: To probe the effects of gross polysaccharide of Radix Rehmanniae Preparata(GPRRP) on the morphology changes of immune organ of blood deficiency model mice.Methods: Using the blood deficiency mice models induced by releasing blood and injection of Cyclophosphamide as the experimental objects,the model animals were divided into 5 groups,that was the large,middle and small dose of GPRRP groups,Danggui Buxue Oral Liquid group,model group,at the same time,giving a blank control group.It was given successively relating drugs to the animals of several groups for 10 days.At the end of the experiment,the hemogram was measured and the thymus and spleen were taken,and making pathological section,then these tissues morphology changes were observed under microscope.Results: GPRRP can obviously resist the atrophy of thymus and spleen induced by the model establishing,can obviously increase the depth of thymus cortex and the number of cortex cells,can enlarge the size of spleen nodules and obviously increase the number of lymphocyte.Conclusion: GPRRP can resist the atrophy of thymus and spleen in blood deficiency model mice.
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Objective To optimize the technique of isolating and extracting oligosaccharide from Radix Rehmanniae.Methods The optimum conditions for the water extracting technique of Radix Rehmanniae were studied by orthogonal test design.Then the water extract was isolated by macroporous resins and active charcoal successively,meanwhile the pattern of macroporous resins,the volume of the eluents,and the decolorizing methods of active charcoal were studied.And then the oligosaccharide was obtained after being purified by Sephadex gel column chromatography.The content of stachyose was determined by HPLC.Results The optimal preparation process was as follows: 12 fold water,three times,0.5 h for each time.Then the water extract was isolated and purified by D-101 macroporous resins,0.1% active charcoal and Sephadex G15 column chromatography were successively used to obtain the oligosaccharide.The analysis of oligosaccharide indicated its main constituents are stachyose,rafinose,and manninotriose,and the content of stachyose is more than 60%.Conclusion This preparation technique of extracting oligosaccharide from Radix Rehmanniae is reasonable and feasible.And the yield of the oligosaccharide from Radix Rehmanniae achieves to 26%.
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Object To investigate the saccharide changes of Radix Rehmanniae when being processed. Methods The fresh roots were torrefied at 65 ℃ and sampled at 0,1, and 6 d. The samples and 4 h steamed slices of those dried roots were extracted with hot water respectively. The extracts were subjected to HPLC analysis, Sugar pak 1 column at 80 ℃,with water as the mobile phase at 0 7 mL/min and detected by RID. Results There are three principal components in the fresh roots, i.e. stachyose (11%-15%), sucrose (0 30%-0 92%), and catalpol (0 27%-0 88%). In the processing of the fresh roots, the HPLC chromatorams of the extracts differed to each other remarkbly. On the chromatogram of roots torrefied for 1 d, one distinct monosaccharide peak displayed at 10 2 min, which is likely to be galactose, and it become prominent in the dried roots together with raffinose. In steamed ones, the peaks of fructose and glucose became outstanding. Conclusion According to the HPLC chromatograms, the degalacto sylation of stachyose may take place during torrifying, while defructosylation during steaming. One of the principal purposes of the processing may be for stachyose degradation for its flatulence causing character.
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Objective To set up the standard for the quality control of Chongcaojing Oral Liquid.Methods Radix Astragal-i,Radix Ginseng,Radix Angelicae Si nensis,Radix Rehmanniae were ident ified quantitatively by thin -layer chromatog-raphy(TLC)and astragalosideⅣcontent in Radix Astragali was determined by TLC scanning quantitiatvely.Results The TLC spots were highly specific,clea r and concentrated without the interference of negative controls.Linearity was ob-tained in the range of 1.03～6.15?g(r =0.9994).The average recovery was 99.2%with RSD =1.8%.Conclu-sion This method is sensitive,simple,sp ecific and accurate for the quality c ontrol of Chongcaojing Oral Liquid.
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Objective To improve the quality standard for quality control of Baji Bushen Pills.Methods Radix Rehmanniae Praeparata and Radix Angelicae Sinensis in this medicine were identified by TLC,the 2,3,5,4’-Tetrahydroxystilbene-2-O-?-D-glucoside content in this medicine was determined by HPLC.The acetonitril-water(18:82)solution was used as the mobile phase,the column was C18,the flow rate was 1.0 mL/min,and the detective wavelength was at 320 nm.Results Radix Rehmanniae Praeparata and Radix Angelicae Sinensis could be identified by TLC.2,3,5,4’-Tetrahydroxystilbene-2-O-?-D-glucoside showed a good linear relationship at the range of 11.28 ?g/mL～282.00 ?g/mL(r=0.999 9).The average recovery was 96.1 %with RSD of 0.8 %(n=6).Conclusion The method is simple,reliable,accurate and can be applied to the quality control of the preparation.
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Objective To establish a TLC identification method for Erhuang Capsule and to provide basis for the establishment of its quality standard.Methods TLC method was adopted.Results Nine kinds of medicinal materials such as Radix Rehmanniae,Radix Polygoni Multiflori,Rhizoma Gastrodiae had discriminating characteristics and distinctive spots.Conclusion TLC is a simple,reliable and specific method with good reproducibility,and can be used for the quality control of Erhuang Capsule.
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AIM:To improve the quality standard of Fuke Yangkun Pill. METHODS: Fructus Viticis was(identified) by micrology,Radix Angelica sinensis,Rhizoma Chuanxiong,Rhizoma Cyperi,Radix Aucklandiae,Rhizoma Corydalis,Radix Rehmanniae,Radix Rehmanniae preparata,Radix Paeoniae alba,Radix et Rhizoma Glycyrrhizae,Fructus Amomi were identified by TLC,baicalin content was determined by HPLC. RESULTS: The TLC spots were fairly clear and distinguishable,the linear range of baicalin was within 3.28-131.2 ?g/mL.The average recovery was 100.4% with RSD 1.0%(n=6). CONCLUSION: The method is simple,and specific,and can be used for quality control of Fuke Yangkun Pill.
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AIM:To investigate the effect of Rehmannia glutinosa Libosch on collagen Ⅰ and Ⅲ of in pulmonary interstitial fibroblast in rat. METHODS: The components of Radix Rehmanniae were extracted and isolated to several parts.Primary pulmonary fibroblasts were separated from lung interstitial tissue of rat.After being reproduced for several generations,fibroblasts were cultured in the extracts of Radix Rehmanniae and Shengdi Solution for injection in several concentrations respectively for 72 hours,and then contents of Col Ⅰ and Col Ⅲ of these fibroblast were determined by RT-PCR. RESULTS: The part containing mainly oligosaccharide from preparation that decocted with water and deposited with alcohol could inhibit Col Ⅰ with a few exceptions concentration while it could inhibit Col Ⅲ at high concentration.The part of deposit inhibited Col Ⅰ obviously in the concentration of 2?10~(-2) g/mL and could inhibit Col Ⅲ in all the concentrations.Shengdi Solution for injection could inhibit Col Ⅲ in all the concentrations. CONCLUSION: Some constitutents of Radix Rehmanniae can inhibit the expression of Col Ⅰ and Col Ⅲ of pulmonary fibroblast which is one of mechanisms that Radix Rehmanniae takes effect on fibrosis disease in pulmonary interstitial tissue.
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AIM: To establish the quality standard of Agui Yangxue Tablets (Radix Angelicae sinensis, Radix Codonopsis, Rhizoma Atractylodis macrocephalae, etc.) METHODS: Radix Astragali, Radix et Rhizoma Glycyrrhizae and Radix Rehmanniae praeparata were identified by TLC. The content of ferulic acid and paeoniflorin was determined by HPLC. The HPLC condition of ferulic acid was as follows: column was Diamonsil~ TM C_ 18 (4.6 mm?200 mm,5 ?m), the mobile phase consisted of acetonitrile -0.085% phosphoric acid (22 ∶ 78), the UV detection wavelength was at 322 nm, the flow rate was 1.0 mL/min with column temperature at 35 ?C . The HPLC condition of paeoniflorin was as follows: column was Diamonsil~ TM C_ 18 (4.6 mm?200 mm,5 ?m), the mobile phase consisted of acetonitrile -0.1% phosphoric acid (15 ∶ 85), the UV detection wavelength was at 229 nm, the flow rate was 1.0 mL/min with column temperature at 25 ?C . RESULTS: The developed TLC spots were quite clear. The calibration curve of ferulic acid was linear with 0.012 9-0.154 8 ?g(r=0.999 7). The average recovery was 99.32% with RSD of 1.74% (n=9). The calibration curve of paeoniflorin was linear with 0.106 8-1.602 0 ?g(r= 0.999 9). The average recovery was 98.68% with RSD of 0.98 (n=9). CONCLUSION: The established TLC and HPLC methods are exclusive, reproducible and suitable for the quality control of Agui Yangxue Tables.