Study on nephrotoxicity of the extracts from different parts of Miao medicine Wikstroemia indica / 中国药房

OBJECTIVE To study the nephrotoxicity of the extracts from different parts o f Miao medicine Wikstroemia indica in healthy rats ，and to provide reference for the study of its toxicity mechanism and clinical drug use. METHODS Using 70％ ethanol as solvent ，total ethanol extract of W. indica was extracted with diacolation method. After dispersing the above extract with water，the fractions of corresponding fractions were obtained with petroleum ether ，ethyl acetate and n-butanol，and the rest was the extract of water fraction. SD rats were randomly divided into total ethanol extract group ，petroleum ether fraction group ，ethyl acetate fraction group ，n-butanol fraction group ，water fraction group and blank group ，with 12 rats in each group （half male and half female ）. The rats in the administration groups were given the corresponding dose of drug solution intragastrically （total ethanol extract 317.520 mg/kg，petroleum ether fraction 7.875 mg/kg，ethyl acetate fraction 78.435 mg/kg，n-butanol fraction 53.865 mg/kg and water fraction 76.545 mg/kg），once a day ，for conse- cutive 2 weeks，and then stopped taking drug for 2 weeks； rats in the blank group were given equal volume of 1.0％ . sodium carboxymethyl cellulose solution intragastrically. Duringthe experiment ，the general conditions of rats were observed. The samples of urine （on the 14th and 28th day ），serum and bilateral renal tissues （on the 15th and 29th day ）were taken respectively，the renal index was calculated ，the levels of @qq.com renal function indexes in serum and urine were detected ，and the pathomorphological changes of renal tissues were observed. RESULTS During administration ，compared with blank group ，the rats in the total ethanol extract group and ethyl acetate fraction group showed poisoning behavior and activity characteristics such as mental depression ，decreased activity and diet ，thin stool and decreased body mass. The mental state of the rats in the petroleum ether fraction group ，n-butanol fraction group and water fraction group were slightly worse than that in blank group，and slightly decreased activity and diet as well as thin stool ，and slowly increased body mass were found ；however，there was no significant difference in anal temperature in each group. After 2 weeks of administration ，the renal index in total ethanol extract group ，the serum levels of N-acetylglucosaminidase（NAG），urea nitrogen （BUN）and creatinine （Cr）in total ethanol extract group and ethyl acetate fraction group ，serum level of NAG in n-butanol fraction group and serum level of Cr in water fraction group ，as while as NAG levels in urine of rats in total ethanol extract group and petroleum ether fraction group ，NAG and urinary protein levels in urine of rats in ethyl acetate fraction group were increased significantly （P＜0.05 or P＜0.01）. In the pathomorphological observation ，renal tubules showed different degrees of unclear structure ，cell swelling and a few cell necrosis in the total ethanol extract group ，petroleum ether fraction group and ethyl acetate fraction group ，accompanying by glomerular pyknosis，renal tubular sclerosis and inflammatory cell infiltration ，compared with blank group. After drug withdrawal ，the mental state of rats in the administration groups were significantly improved ，the amount of activity and diet increased ，and the stool tended to be normal. Two weeks after drug withdrawal and recovery ，the levels of above indexes in serum and urine of rats in administration groups returned to be close to that in blank group （P＞0.05）；the glomerular structure of rats in each administration group gradually recovered clearly ，and cell swelling and inflammatory cell infiltration were rare in total ethanol extract group ， petroleum ether fraction group and ethyl acetate fraction group. CONCLUSIONS The total ethanol extract ，petroleum ether fraction and ethyl acetate fraction of Miao medicine W. indica have certain nephrotoxicity and reversibility. The toxic component may

Effects of “Sweat Soaking Method ”on the Content of Genkwanin in Wikstroemia indica and Its Anti-oxidation Ability / 中国药房

OBJECTIVE：To compare the content changes of active/toxic ingredient genkwanin in ethanol extract from Wikstroemia indica before and after processing with “sweat soaking method ”and the effects of processing method on its anti-oxidation ability. METHODS ：HPLC method was adopted to determine the content of genkwanin in W. indica before and after processing with “sweat soaking method ”. The separation was performed on Diamonsil C 18 column with 0.2％ phosphoric acid solution-methanol as mobile phase （gradient elution ）at the flow rate of 1 mL/min. The column temperature was 30 ℃ and detection wavelength was set at 346 nm. The sample size was 20 µL. SD rats were randomly divided into blank group ，W. indica raw product ethanol extract group （317.52 mg/kg，called“raw-product group ”as short ）and W. indica processed product ethanol extract group （317.52 mg/kg，called“processed-product group ”as short ），with 6 rats in each group. Blank group was given constant volume of 1.0％CMC-Na solution intragastrically ，and administration groups were given relevant medicine suspension intragastrically；all of them were given 20 mL/kg，once a day ，for consecutive 14 days. The contents of serum oxidant stress indexes（MDA，CAT，SOD）in rats were determined by ELISA. RESULTS ：The linear range of genkwanin were 0.147-27.360 μg （r＝0.999 9）；RSDs of precision ，reproducibility and stability tests were all lower than 3％ ；average recoveries were 98.64％-98.92％（RSD＜1％，n＝3）. Before and after processing with “sweat soaking method ”，average contents of genkwanin in W. indica were 0.377 6 and 0.234 0 mg/g. Compared with blank group ，the serum content of SOD in raw-product group was increased significantly ，while CAT content was decreased significantly （P＜0.05 or P＜0.01）；the serum content of MDA was decreased significantly in processed-product group ，while SOD content was increased significantly （P＜0.05 or P＜0.01）. MDA content of processed-product group was significantly lower than that of raw-product group ，while SOD content was significantly higher than raw-product group （P＜0.05）. CONCLUSIONS ：After proce ssing with “sweat soaking method ”，the content of genkwanin in W. indica is decreased ，and antioxidant activity is increased .“Sweat soaking method ”processes certain function of “reducing toxicity and increasing efficiency ”.

Correlation Study of “Dose-effect-toxicity”of Miao Medicine Wikstroemia indica on Anti-immune Inflammation of Mice before and after Processed by “Sweat Soaking Method ” / 中国药房

OBJECTIVE：To investigate the “dose-effect-toxicity”correlation of Miao medicine Wikstroemia indica before and after processed on anti-immnue inflammation of mice . METHODS ：Mice were divided into blank group ，model group ，ethanol extract of W. indica raw product groups 1-6，ethanol extract of W. indica processed product by “sweat soaking method ”groups 1-6 （hereinafter called “raw groups 1-6”“processed groups 1-6”for short ，drug dosage were 0.13，0.20，0.26，0.52，1.04，2.6 g/kg）， positive group （cyclophosphamide，36.4 mg/kg），with 10 mice in each group. Except for blank group ，other groups were given 1％ 2，4-dinitrofluorobenzene-acetone-sesame oil mixed solution to induce delayed type hypersensitivity model. After modeling， blank group and model group were given constant volume of 1.0％ CMC-Na solution intragastrically ，and administration groups were given relevant medicine 20 mL/kg intrag astrically，oncea day ，for consecutive 5 d. A fter last medication ，ear swelling degree of mice were recorded ；the inhibition rate of swelling degree， half effective dose （ED50） and 95％ confidence 158-02-32）； interval（CI）of raw and processed products were calculated ； the weight of heart ，liver，spleen，lung and kidney were measured and the indexes of organs were calculated ；ELISA 1161472062@qq.com and modified chemical oxidation method were used to determine the serum levels of inflammatory factors （TNF-α， IL-10） and liver and renal function indexes （ALT，AST， TBIL，BUN，CREA）. RESULTS：Compared with blank group ，the degree of ear swelling in the model group was significantly increased（P＜0.05）. Compared with model group ，ear swelling degree of mice were decreased significantly in different doses groups of ethanol extract of raw and processed products of W. indica （P＜0.05）. The inhibition rate of swelling increased with the increase of dose ，ED50 and 95％CI of delayed hypersensitivity ear swelling were 0.239 6（0.129 0，0.445 2）g/kg and 0.147 3（0.076 8，0.282 7）g/kg，respectively. Compared with blank group ，liver index and serum TNF-α level of mice were increased significantly in model group ，while lung index and serum IL- 10 level were decreased significantly （P＜0.05）. Compared with model group ，the levels of liver indexes （positive group ，raw group 3，processed groups 1-6）and serum TNF-α levels（positive group ，raw groups 1-3，processed groups 1-4） were decreased significantly in different administration groups ；while the levels of lung indexes （positive group ，raw groups 3-6 and processed groups 3-6），serum IL- 10 levels（raw groups 1，2，4 and 5，processed groups 2-6），ALT，AST，BUN and CREA levels （raw groups 4-6），TBIL levels （raw groups 3-6 ）were increased significantly （P＜ 0.05）. CONCLUSIONS ：The ethanol extract of raw product of W. indica has certain anti-inflammatory activity ，and has certain hepatorenal toxicity to mice ，with certain “dose-effect-toxity”correlation. The ethanol ectract of processed product of W. indica has certain anti-inflammatory activity too ，but its hepatorenal toxicity was lower than raw product. The “sweat soaking method ” possesses the function of “retaining efficiency and reducing toxicity ”for processing W. indica .

Chemical profiling from water extract of Wikstroemia indica by UPLC-Q-TOF-MS/MS / 中国中药杂志

In this study,a method using ultra performance liquid chromatography tandem quadrupole time-of-flight mass spectrometry( UPLC-Q-TOF-MS/MS) was established to identify complicated chemical constituents of Wikstroemia indica. Chromatographic separation was performed on an AcclaimTMRSLC 120-C18 column( 2. 1 mm×100 mm,2. 2 μm) using gradient elution with 0. 2% ammonium formate buffer salt solution( A)-0. 2% ammonium formate buffer salt solution methanol( B) as mobile phase. The column temperature was maintained at 30 ℃. The analytes were determined by positive and negative ion modes with electro-spray ionization source. A total of 52 compounds( including eleven coumarins,thirteen flavonoids,ten lignans,two amides,four phenolic acids,six sesquiterpenes and six other compounds) were identified or tentatively characterized from the water extract of W. indica by comparing their retention times and MS spectra with those of authentic standards or literature datas. Three compounds were found for the first time from W.indica namely isomer of indicanone,β-hydroxypropiovanillone and epiprocurcumenol. Furthermore,the fragmentation rules of some compounds were speculated and summarized. In addition,the cleavage pathways of guaiane sesquiterpenes were described for the first time,which can provide reference for studying the fragmentation pathways of similar compounds. This study provides an easy way to identify chemical constituents of traditional Chinese medicine and a basis for the further study on chemical fundamentals of W. indica.

Research progress on chemical constituents and biological activities of Wikstroemia indica / 中草药

Wikstroemia indica as a Chinese herbal medicine, belonging to family Thymelaeaceae, was the root and root bark of the plant. Studies found that W. indica had acids, alcohols or esters, quinones, steroids, coumarins, lignans, flavonoids, terpenes, diarylheptanes, and fat-soluble ingredients, and has the effects of antibacterial, anti-virus, cytotoxic, and anti-inflammatory activities etc. The contents of coumarins, flavonoids, quinines, and lignans were measured and applied quality evaluation. In addition, as a toxic Chinese herbal medicine, the toxicity of W. indica could be reduced by processing methods, so further research is needed to confirm the relationship between the compound and its pharmacological activity. In this paper, the researches from 2010 to 2017 on chemical constituents, biological activities, and components detection of W. indica were collected and summarized, aiming to provide reference for further research and development on W. indica.

Comparison of Acute Toxicity Effects of Ethanol Extract from Different Processed Products of Miao Medi-cine Wikstroemia indica on Mice / 中国药房

OBJECTIVE:To compare the acute toxicity of ethanol extract from raw product and different processed products of Wikstroemia indica on mice,and provide basis for optimizing the processing technology of perspiration method for W. indica and medication safety. METHODS:Perspiration method was used to process the W. indica pieces for 30 d to get processed product 1 and process its coarse powder for 14,7 d to get processed product 2,processed product 3,respectively. Then using 70％ ethanol as solvent,percolation method was used to extract the raw W. indica and its different processed products,and acute toxicity test was conducted on mice for different ethanol extracts. RESULTS:The median lethal dose(LD50)of ethanol extracts from raw W. in-dica and processed product 1 were 4.05 and 6.65 g/kg,equivalent to 19,32 times of the clinical daily dose of a 70 kg adult,re-spectively. While the LD50 of ethanol extract from processed product 2 and processed product 3 can not be measured,the maximum tolerated dose(MTD)were measured as 20.0,15.0 g/kg,equivalent to 95,71 times of the clinical daily dose of a 70 kg adult,re-spectively;the maximum dose(MLD)were measured as approximately 30.0,20.0 g/kg,equivalent to 143,95 times of the clini-cal daily dose of a 70 kg adult,respectively. CONCLUSIONS:The toxicity of processed products of W. indica is obviously lower than that of raw products,and its toxicity after processing the coarse powder for 14 d is lower than that after processing the coarse powder for 7 d.

Content Determination of 7 Constituents in Wikstroemia indica and Its Principal Component and Cluster Analysis / 中国药房

OBJECTIVE:To establish a method for simultaneous determination of 7 constituents in Wikstroemia indica,and to conduct principal component analysis and cluster analysis. METHODS:HPLC method was adopted. The determination was per-formed on Diamonsil Platisil ODS column with mobile phase consisted of acetonitrile-0.15% triethylamine solution(pH adjusted to 6.0 with phosphoric acid,gradient elution)at the flow rate of 1.0 mL/min. The detection wavelength was set at 280 nm,and col-umn temperature was 40 ℃. The sample size was 10 μL. The principal component analysis and cluster analysis were conducted for the results of content determination by SPSS 22.0 statistical software. RESULTS:The linear ranges were 2.688-53.76μg/mL for nar-ingin(r=0.9998),5.052-101.00 μg/mL for myricetin(r=0.9999),2.052-41.04 μg/mL for arctiin(r=0.9999),2.108-42.16 μg/mL(r=0.9999),5.112-102.20 μg/mL(r=0.9999),0.820-16.42 μg/mL(r=0.9999),2.070-41.40 μg/mL(r=0.9999),respec-tively. The limits of quantitation were no higher than 1.0720 μg/mL,the limits of detection were no higher than 0.3318 μg/mL. RSDs of precision,stability and reproducibility tests were all lower than 2.0%. The recoveries were 97.8%-102.5%(RSD=1.8%, n=6),97.2%-102.0%(RSD=2.0%,n=6),95.2%-100.1%(RSD=1.7%,n=6),95.2%-99.3%(RSD=1.6%,n=6), 97.0%-100.8%(RSD=1.3%,n=6),95.5%-98.6%(RSD=1.1%,n=6),95.0%-99.3%(RSD=1.8%,n=6),respectively. Three main components were belong to the samples of 10 batches of medicinal materials. The samples of medicinal materials from 10 pro-ducing area could be divided into 2 categories. The quality of W. indica from Qingyuan Guangdong and Guiyang Guizhou were bet-ter than others. CONCLUSIONS:The method is simple,precise,stable and reproducible,and it can be used for simultaneous de-termination of 7 constituents in medicinal material. The quality of W. indica from different regions are quite different.

Study on Antibacterial Effect in Vitro of Different Extract of Wikstroemia indica / 中国中医药信息杂志

Objective To research the antibacterial effect in vitro of different extract of Wikstroemia indica.Methods Double dilution method was used to determine the MIC of 11 species bacterium.Results The acetic ether extract had much obvious effect than n-butanol extract,both of them had high effect on staph.,but the aqueous extract had little antibacterial effect.Conclusion The acetic ether extract has obvious antibacterial effect,also has broad-spectrum antibacterial activity.