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In this research, we collected information of eighty nine sampling points of Isatidis Radix nationwide through data query and field survey, and the medicinal component contents of samples were determined by HPLC. By using Maxent Model and ArcGIS, along with ecological factor data, the national habitat suitability distribution of Isatidis Radix was predicted. R-language was adopted to establish a model of the relationship between the medicinal component contents and ecological factors. The medicinal quality was divided by ArcGIS grid computing. The results indicated that the three main ecological factors affecting the distribution of Isatidis Radix were precipitation in the driest season, mean annual temperature and mean temperature in the wet season. The suitable cultivation region of Isatidis Radix is mainly distributed in the north of China, but the medicinal quality is quite different, Isatidis Radix in Xinjiang province has higher medicinal quality. This study provides a reference for rational selection of planting areas of Isatidis Radix.
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The method of quantitative nuclear magnetic resonance spectroscopy (qNMR) for determination of epigoitrin in Radix Isatidis was established based on solid phase extraction (SPE).The twice ultrasonic extraction method using pure water was used for fully extracting epigoitrin in sample, and then the extraction was enriched and concentrated by poly-Sery MCX SPE cartridge.The effect of sample pretreatment and qNMR experimental conditions was investigated.The qNMR experiment conditions were selected using DMSO as solvent, calibrated 2,3,5-triiodobenzoate as internal standard, and P1(pulse width)=14.1 μs, d1(pulse delay time)=5 s, NS(number of scan)=256.The .1H-NMR peaks of δ 5.365-5.399 (H-7b, d, 1H) of epigoitrin were chosen as the quantitative peaks.Method validation was performed including precision (intra-day precision RSD was 0.5%, and the inter-day precision was 0.8%), linearity (correlation coefficient r>0.9991), LOD (0.05 mg/g, standard curve method) and LOQ (0.19 mg/g, S/N≥150).The recoveries of the SPE-qNMR were 97.4%-101.7%.The result showed that the method was stable, accurate and reliable.With this method the epigoitrin in a real Radix Isatidis was determined to be <0.19-1.26 mg/g.SPE combining with qNMR could extend the application field of qNMR, especially in the detection of low-content component in complex samples.
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Aim To observe the effects of active com-ponent of Radix Isatidis ( ACRI ) on anti-anoxia and anti-fatigue in mice and investigate its possible mecha-nism. Methods Based on the weights, the mice were randomly divided into 5 groups: blank control group, ACRI 25, 50 and 100 mg·kg-1 groups, positive drug ( American ginseng liquid) control group 3 mL·kg-1 . Drugs were administered to the mice for about 14 con-secutive days, and during the experiment general situa-tions of mice were observed. The experiment of bearing hypoxia at normal pressure and the experiment of swim-ming while weight-bearing were conducted to study the effect of ACRI on anti-anoxia and anti-fatigue in mice. Then the superoxide dismutase ( SOD ) activities, the content of maleic dialdehyde ( MDA ) of mice serum and liver and blood urea nitrogen, blood lactic acid, liver glycogen were detected, in order to investigate its mechanism. Results ACRI decreased the growth rate of body weight in mice significantly, obviously pro-longed the survival time of anoxic mice at normal pres-sure and the swimming time of loaded mice, enhanced the SOD activities of mice blood and liver, decreased the MDA content of mice blood and liver, increased the content of liver glycogen, and decreased the blood urea nitrogen and blood lactic acid in mice after swim-ming. Conclusion ACRI has the anti-anoxia and anti-fatigue functions.
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Objective To evaluate the influence of Chinese patent medicine Banlangen Granules and Ganmao Qingre Granules on war‐farin anticogulation effect during the process for treating acute upper respiratory tract infection in the postoperative patients with mechanical valve replacement in order to provide the theoretical reference for clinical medication .Methods The patient receiving mitral valve replace‐ment ,aortic valve replacement or double‐valve replacement and long term postoperative oral warfarin anticoagulation treatment were select‐ed ,moreover the symptoms of complicating acute upper respiratory tract infection occurred .The patients were divided into 2 groups ,30 cases in each group .The group A adopted oral Banlangen Granules ,while the group B was treated by oral Ganmao Qingre Granules .The treat‐ment course was 3 d .The International Normalized Ratio (INR) in the two groups was monitored before medication ,at 72 h after medication and at 72 h after drug withdrawal .Complicating bleeding or embolism reaction was observed .The INR values at various time points were compared between the two groups and the INR values in each group were compared among 3 time points .Results The INR values in the Banlangen Granules group had statistical differences between 72 h and other two time points ,and the INR values at the same time point had statistical difference compared with the Ganmao Qingre Granules group (P0 .05) .Thirty cases had no complication occurrence .The Ganmao Qingre Granules group had no statistical difference among various time points (P>0 .05) ,thirty cases had no complication occur‐rence .Conclusion The warfarin combined with Banlangen Granules causes the INR value increase ,while warfarin combined with Ganmao Qingre Granules has no impact on the INR value ,therefore which suggests that the combination use of warfarin and Banlangen Granules should be cautious in clinic .
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OBJECTIVE To investigate the effect of Radix Isatidis and its constituents indigo and in?dirubin on two principal subtypes of organic cation transporters(OCT)OCT1,OCT2 in vivo in mice. METHODS Decoction of Radix Isatidis (DRI) 1.6 and 6.4 g · kg-1,granules of Radix Isatidis (GRI) 0.615 and 2.460 g·kg-1,indigo 0.008 and 0.640 mg·kg-1 and indirubin 0.0192 and 1.536 mg·kg-1 were ig given to NIH mice(60 mice per group),twice a day for 5 d. Four control groups were set up,including the vehicle of water,0.5% sodium carboxymethyl cellulose(CMC),additives of sucrose plus dextrin (1.5 g · kg-1)and positive control quinidine(0.025 g · kg-1). Sixty minutes after the last dosing,all the mice were iv given metformin(Met)5 mg·kg-1,and at 1.0,2.5,5.0,7.5,10.0 and 20.0 min after Met iv,10 mice in each group were sacrificed to collect whole blood and kidneys respectively. The right kidney was homogenized for Met accumulation test and the left one used to extract total RNA for analysis of OCT1 and OCT2 mRNA expressions by real-time PCR. The contents of Met in sera and kidneys were quantified by HPLC. Major pharmacokinetic parameters of Met in sera were analyzed by pharmacokinetic software(DAS 2.0). RESULTS There was no significant difference between water control group,0.5%CMC group and sucrose plus dextrin group in any examined item. Compared with vehicle control group (water and 0.5%CMC group),all the related pharmacokinetic parameters in DRI 6.4 g · kg- 1,GRI 2.46 g · kg-1,indigo 0.640 mg · kg-1 and indirubin 1.536 mg · kg-1 groups were changed significantly (P<0.05,P<0.01). The elimination half time (t1/2β) was prolonged 13%-97%,volume of distribution reduced by 13%-72%,clearance(Cl)reduced by 9%-65%,and the area under the concentration-time curve (AUC0-20 min) increased by 13%-135%. AUC0-20 min obtained from renal Met accumulations was significantly increased(P<0.01)while Met uptake by kidney slices was reduced(P<0.05,P<0.01). The expressions of OCT1 and OCT2 mRNA were obviously down-regulated(P<0.05,P<0.01). CONCLUSION The mouse renal OCT1 and OCT2 are significantly inhibited by DRI,GRI,indigo and indirubin. The inhibitory effect of Radix Isatidis on OCT1 and OCT2 probably arises from indigo and indirubin contained.
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Objective To investigate the effect of extraction of radix isatidis on the expression of transforming growth factor(TGF-β)in lung tissues of rats with radioactive lung injury.Methods 72 female Wistar rats were randomly divided into 3 groups,blank control group(groupA,n=24),simple irradiation group(group B,n=24),irradiation with Chinese medicine group(group C,n=24).Rats in group A and group B were given distilled water 2 mL/200g by gavage for one week,the rats in group C were given extraction of radix isatidis 2 mL/200g(1.6 g/200 g)per day for one week,then the rats received irradiation,during irradiation,the rats' weight was measured once a week,to illuminate the end.After 10-time irradiation(the 30th day), and at 60 days and at 120 days after irradiation,the expression levels of TGF-βin lung tissue were detected;In addition each rat lung tissue morphology was observed.Results The expression levels of TGF-βprotein in group B reached peak at the 30 days after irradiation,which at different time points in group A and group C was significantly lower than that in group B(P<0.01).Rats of group A had no significant change in lung tissue.Rats of group B were observed inflammatory cells in interstitial pulmonary alveolar,alveolar structural damaging,interstitial pulmonary edema,interstitial hemorrhage and inflammatory pulmonary with alveolar cavity;Rats of group C pulmonary interstitial fewer inflammatory cells in alveolar damage were not obvious, with no-pulmonary interstitial edema, hemorrhage and no inflammatory symptoms, comparison between group B and C was not obvious. Conclusion Extraction of radix isatidis could reduce the expression levels of transforming growth factor(TGF-β)in lung tissues of rats with radioactive lung injury ,has better preventive effect on radioactive lung injury,could protect lungtissue from damaging,inhibite alveolar permeability,and present the effect of pulmonary edema.
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Objective To explore the effect of Radix Isatidis( indigowoad root,RI) polysaccharide on NO and ET-1 in orthopotic liver autotransplantation of rats.Methods The rats used in the experiment were randomly divided into three groups:normal control group (NC), autotransplantation group (AT) and RI polysaccharide group (RIPS).At 1, 6, 12 and 24 hours after autotransplantation, the content of NO,ET-1, alanine aminotransferase ( ALT), and aspartate amin-otransferase ( AST) in serum was assayed.Morphological observation of hepatic tissues was also performed.Results The serum level of ET-1 in RIPS group was significantly lower than that of AT group (P<0.05), and NO content in serum of RIPS group was increased significantly compared with AT group ( P<0.05) at 1, 6, 12 and 24 hours after autotransplanta-tion.The serum level of ALT and AST in RIPS group was lower than that of AT group (P<0.05) at each measurement point.The morphology of hepatocytes changed abnormally under light microscopy.The blood stasis and swelling of hepatic tissues, denaturalization and necrosis of hepatocytes in RIPS group were milder than that in AT group.Conclusion RIPS has a protective effect on hepatic ischemia reperfusion injury after orthotopic liver autotransplantation, which may be related to increased NO levels and lower levels of ET-1.
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OBJECTIVE: To establish an HPLC-ELSD method to determine a new compound named Fructopyrano-(1 → 4)-glucopyranose in Radix Isatidis. METHODS: The HPLC method was conducted on an Agilent XDB-C18 column (4.6 mm × 150 mm, 5 μm), with methanol-water (5:95) as the mobile phase. The drift tube temperature was 115°C and flow fate of carrier gas was 3 L · min-1. RESULTS: The calibration curve was linear for FG in the range of 8.40 - 21.00 ng (r = 0.9990). The recovery was 97.27% - 102.72%. CONCLUSION: This method is accurate, reliable, rapid, sensitive, reproducible, and can be used for the determination of FG in Radix Isatidis. Copyright 2012 by the Chinese Pharmaceutical Association.
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OBJECTIVE: To study the effects of fructopyrano-(1 → 4)-glucopyranose (FG) extracted from Radix Isatidis on the tumor growth and immune activity in tumor-bearing mice. METHODS: The antitumor activity of FG in vivo was observed by implanted H22 and S180 mouse models. The effect of FG on immune function in tumor-bearing mice was investigated by using phagocytosis test, T plymphocyte proliferation test, and Elisa for determining the expression levels of TNF-α, IL-2, IL-6, and IL-12. RESULTS: At dosages of 50, 100 and 200 mg · kg-1, the inhibition rates of FG were 23.60%, 36.76% and 49.26% for S180, 22.61%, 36.52%, and 46.95% for H22, respectively. FG increased the spleen index and chest gland index, improved the phagocytosis function of the macrophage and enhance the T lymphocytes proliferation, and increased the expression levels of TNF-α, IL-2, IL-6, and IL-12. CONCLUSION: FG exhibits significant anti-tumor effect in vivo and the mechanism may be related to enhancing immune function. Copyright 2012 by the Chinese Pharmaceutical Association.
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OBJECTIVE:To isolate the chemical constituents from Radix isatidis.METHODS:The 95% ethanol extract of Radix Isatidis underwent adsorption by silica gel,and the portion eluted by different solvents was isolated and purified on silica gel column repeatedly.The physico-chemical constants and spectral data of the obtained compounds were determined and their chemical structure was identified.RESULTS:6 compounds were separated from chloroform and n-Butanol parts of Radix Isatidis and identified as:cholesterol(Ⅰ),indigotin(Ⅱ),indirubin(Ⅲ),p-Hydroxybenzaldehyde(Ⅳ),salicylic acid(Ⅴ),lariciresinol-4,4'-2-O-?-D-glucopyranoside(Ⅵ),respectively.CONCLUSION:For the first time,compound(Ⅰ)and compound(Ⅳ)were isolated from Radix Isatidis,this finding serves as reference.
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To find a component with the strongest anti-endotoxin effect from the four components (F021 , F022, F023, and F024 ) of chloroform extract (F02) of Radix Isatidis, the protective effects of the five components on endotoxin-challenged mice and their effects on the production of TNFα and IL-6 by macrophages from the endotoxin-challenged mice were compared. It was found that the five components all had protective effects on endotoxin-challenged mice in terms of mortality. The mortalities of the mice challenged by 2.42 mg/kg endotoxin were 30 %, 50 %, 20 %, 50 % and 60 %after treatment with F02, E021 , F022, F023, and F024. The 5 components all had inhibitory effects on the production of TNFα and IL-6 by macrophages from the mice. At a concentration of LPS of 50 ng/mL, the inhibitory rates of F02 , F021 1, F022 , F023 , and F024 for the production of TNFα and IL-6were 76.54 %, 30.86 %, 78. 60 %, 36.63 %, 2.06 % and 75.85 %, 18. 45 %, 77.68 %, 24.41%, 3.64% and at the dose of 100 ng/mL, the inhibitory rates were 49.65 %, 16.86 %, 66.97 %, 13.39 %, 7.16 % and 59.78 %, 1.28 %, 61.86 %, 2.08 %, 1.44 %, respectively. It is concluded that the component F022 of Radix Isatidis has the strongest anti-endotoxin effect.
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The effects of active antiendotoxin chemical fraction isolated from Radix Isatidis (fraction D) on TNF-α and IL-8 secretion in HL-60 cells induced by lipopolysaccharide (LPS) were studied. The appropriate densities of cell suspension and fraction D solution were determined by MTT colorimetric method. Fraction D and LPS were added to HL-60 cell suspension with three different methods respectively. The contents of TNF-α and IL-8 in the cultured supernatant induced by LPS were detected by using ELISA method. The results showed that the absorbance (A) was directly proportional to the number of cells and the linearity was good in the range from 0.25 × 105 to 2 ×105 cell/mL cell suspension. The fraction D significantly inhibited the oversecretion of TNF-α and IL-8 in HL-60 cells induced by LPS at the concentration of 7. 812 mg/mL which had no cytotoxicity. It was indicated that the antiendotoxin mechanism of the active fraction from Radix Isatidis was contributed to the inhibition of the oversecretion of cytokines induced by LPS.
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OBJECTIVE:To establish a GC-MS method to identify the volatile constituents in Radix Isatidis.METHODS:The method of wet distillation was applied to extracting volatile oil from Radix Isatidis,the chromatographic column was OV-1,and the flow rate was 1.0mL?min-1,EI ion source was used by MS.The volatile constituents in Radix Isatidis were identified and analyzed by NIS107 system.RESULTS:22 chromatographic peaks were separated,19 chemical compounds were identified,representing 90.51% of total amount,hexadecanoic acid had highest content,accounting for 38.52% of total amount.CONCLUSION:This experiment has provided initial usable data for comprehensive utilization of Radix Isatidis.
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Objective To observe the anti-pyretic effect of F022 from Radi x Isatidis.Methods The fever model was induced by intravenous injection of endo toxin into rabbits.Body temperature of the rabbits was measured at 0.5,1,2,3,an d 4h after administration in semi-vivo and in vivo.Results F022 subsided th e fever in rabbits induced by endotoxin in semi-vivo and vivo.Conclusion It is valuable for further research that F022 component is used as the main to scr een anti-pyretic active constituent.
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Objective To investigate the relationship between pharmacokinetics(PK) and pharmacodynamics PD of epigoitrin,a main component in total alkaloids of Radix Isatidis,in yeast-induced febrile rats with the combined PK-PD model.Methods The plasma concentration of epigoitrin after ig administration with total alkaloids of Radix Isatidis was determined by HPLC method and the body temperature was recorded by electronic thermometer.The individual PK parameters were fitted using one compartmental model.The PD parameters were fitted by three kinds of PK-PD binding models,such as indirect inhibition-Kin model,indirect stimulation KoutPD model,and Sigmoid-Emax model.Results The main PK parameters t1/2,Cmax,AUC were(4.94?0.84) h,(4.01?0.21) ?g/mL,(28.37?2.42) ?g?h/mL and(5.71?0.91) h,(4.15?0.25) ?g/mL,(30.35?2.58) ?g?h/mL in both normal and febrile rats,respectively.The relationship between pharmacological effects and effect compartment concentration was better fitted with the indirect inhibition-Kin PD model.The corresponding PD parameters were Kin=(0.70?0.10) h-1,Kout=(0.54?0.12) h-1,R0=1.33?0.16,IC50=(0.94?0.66) mg/L.ConclusionThe PK parameters of epigoitrin in total alkaloids of Radix Isatidis show that there is no significant difference in PD behavior in vivo in both normal and febrile rats.Relationship between in vivo PK and PD of epigoitrin in febrile rats is established using indirect inhibition Kin model.
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Object To study the anti endotoxic effect of syringic acid (SA) which was isolated from Radix Isatidis (Banlangen in Chinese, BLG). Methods SA was extracted and isolated from BLG and made to 1% solution. The content of SA pretreated ET was quantitatively determined using Limulus Test. Then the ability of fever induction of endotoxin (ET) pretreated with SA was measured using endotoxin induced fever test in rabbits. At last, the lipopolysaccharide (LPS) induced death in mice pretreated with and without SA was compared. Results Being pretreated with SA, 83.16% ET was destroyed, the ET induced fever in rabbits relieved markedly and the LPS induced death rate in mice dropped from 68% to 20%. Conclusion SA isolated from BLG has anti endotoxic effects.
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Objective To study the feasibility of chelating resins A and B removing heavy metals from the extracts of Radix Isatidis.Methods Heavy-metal Removal experiment was performed on automatic control platform in industry mode.With dry ointment yield,comparability of HPLC and removal rate of heavy metals as the indexes,the effects of two kinds of Chelating resins in removing heavy metal were compared.Results After finishing heavy metal removal step,the loss rate of the dry-extract was lower than 7 %,and the heavy metal contents in dry-extract were lower than national limits.HPLC similarity of the extracts before and after removing heavy metals by resin A was higher than 0.97,but that by resin B was very low.Conclusion Chelating resin A is suitable for removing heavy metals from extracts of Radix Isatidis.
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AIM:To improve the quality control standard based on 3 kinds of Radix Isatidis preparations-Radix Isatidis syrup,chewable tablet and granules. METHODS: A TLC method was established for iden tification of with Isatidis indigotica RS,Vleucine CRS and Arginine CRS as the references.A HPLC method was developed for the determination of adenosine and igoitrin in Radix Isatidis Preparations,with DIKMA Diamonsil C_(18) column(5 ?m,200 mm?4.6 mm id),a mixure of acetonitrile-water-triethylamine(10∶90∶0.2,using glacial acetic acid to adjust to pH 5.2)-water(85∶15) as the mobile phase.The detection wavelength was set at 245 nm. RESULTS: Isatidis indigotica,Vleucine and Arginine were detected in all the preparations.The contents of adenosine and igoitrin varied greatly in tested samples from different pharmaceutical enterprises. CONCLUSION: The above methods are simple,accurate,and suitable for the quality control of Radix Isatidis Preparations.
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AIM: To determine epigoitrin and 2,4(1H,3H)-quinazolinedione in Radix Isatidis and its preparation by HPLC-DAD. METHODS: HPLC was carried out with an Agilent 1200,equipped with ZORBAX SB-C_(18) column(4.6 mm?250 mm,5 ?m).The mobile phase of acetonitrile:(0.1% phosphoric acid+0.03% triethylamine)aqueous solution=12∶88.The flow rate was 0.7 mL/min.The detection wavelength was set at 224 nm.The column temperature was at 30 ℃. RESULTS: The linear ranges of epigoitrin was 0.040 9 ?g-1.103 8 ?g;2,4(1H,3H)-quinazolinedione was 0.010 6 ?g-0.169 6 ?g,The recoveries of epigoitrin and 2,4(1H,3H)-(quinazolinedione) were 98.44%(n=9),with RSD of 1.43% and 98.67%,with RSD of 1.65%,respectively. CONCLUSION: The method is simple,sensitive and reliable.It can be used for quantitative determination of Radix Isatidis and its preparation.
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AIM:To establish a method for determining guanosine, uridine,and andadenosine for Radix Isatidis. METHODS: The Prevail C_(18) analytical column(4.6 mm?250 mm,5 ?m)was used.The mobile phases consisted of water and acetonitrile with gradient.The flow rate was 0.5 mL/min and column temperature was at 25 ℃.The detection wavelength was at 254 nm. RESULTS: The method had good linear relationship within the range(1.92)-38.4 ?g/mL;1.75-35 ?g/mL; 2.12-42.4 ?g/mL of guanosine,uridine and andadenosine,respectively,with the correlation coefficients of 0.999 9.The average recoveries of the three nucleosides were 99.64%(RSD=0.38%),98.94%(RSD=0.32%) and 98.85%(RSD=0.48%),correspondingly.CONCLUSION: The method is fast,rapid,convenient,accurate and can be used for quality control of Radix isatidis.