This study explored whether Sagittaria sagittifolia polysaccharides(SSP) activates the nuclear factor erythroid-2-related factor2(Nrf2)/heme oxygenase-1(HO-1) signaling pathway to protect against liver damage jointly induced by multiple heavy metals. First, based on the proportion of dietary intake of six heavy metals in rice available in Beijing market, a heavy metal mixture was prepared for inducing mouse liver injury and HepG2 cell injury. Forty male Kunming mice were divided into five groups: control group, model group, glutathione positive control group, and low-and high-dose SSP groups, with eight mice in each group. After 30 days of intragastric administration, the liver injury in mice was observed by HE staining. In the in vitro experiment, MTT assay was conducted to detect the effects of SSP at 0.25, 0.5, 1, and 2 mg·mL~(-1) on HepG2 cell survival at different time points. The content of alanine transaminase(ALT) and aspartate aminotransferase(AST) in the 48-h cell culture fluid was measured using micro-plate cultivation method, followed by the detection of the change in reactive oxygen species(ROS) content by flow cytometry. The mRNA expression levels of Nrf2 and HO-1 in cells were determined by RT-PCR, and their protein expression by Western blot. HE staining results showed that compared with the model group, the SSP administration groups exhibited significantly alleviated inflammatory cell infiltration and fatty infiltration in the liver, with better outcomes observed in the high-dose SSP group. In the in vitro MTT assay, compared with the model group, SSP at four concentrations all significantly increased the cell survival rate, decreased the ALT, AST, and ROS content(P<0.05), and down-regulated Nrf2 and HO-1 mRNA and protein expression(P<0.05). SSP significantly improves inflammatory infiltration in the liver tissue of mice exposed to a variety of heavy metals and corrects the liver fat degeneration, which may be related to its regulation of the Nrf2/HO-1 signaling pathway, reduction of ROS, and alleviation of oxidative damage.
Animals ; Heme Oxygenase-1/metabolism* ; Liver ; Male ; Metals, Heavy/metabolism* ; Mice ; NF-E2-Related Factor 2/metabolism* ; Oxidative Stress ; Polysaccharides/pharmacology* ; RNA, Messenger/metabolism* ; Reactive Oxygen Species/metabolism* ; Sagittaria/metabolism*

To establish a quality constant evaluation system of Alismatis Rhizoma decoction pieces,in order to provide reference for regulating the market circulation of this decoction pieces. A total of 18 batches of Alismatis Rhizoma decoction pieces were collected from different pharmaceutical factories,and the morphological parameters of each sample were tested. The content of alisol B 23-acetate in Alismatis Rhizoma decoction pieces was determined by HPLC in the 2015 edition of Chinese Pharmacopoeia,and the parameters such as quality constant and relative quality constant were calculated. The quality constant range of 18 batches of Alismatis Rhizoma decoction pieces was 0. 390-2. 076. If 18 batches of Alismatis Rhizoma decoction pieces were divided into 3 grades,taking 80% of the maximum quality constant as first grade,50% to 80% as second grade,and the rest as third grade,then the quality constant of firstgrade samples was ≥1. 66,the quality constant of second-grade samples was ≥1. 04 and <1. 66,and the quality constant of third-grade samples was <1. 04. The established quality constant evaluation method is objective and feasible,which can be used to classify the grade of Alismatis Rhizoma decoction pieces and provide a reference method to control the quality of this decoction pieces.
Alisma ; chemistry ; Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; standards ; Quality Control ; Rhizome ; chemistry

To research the correlation between accumulation of triterpenoids and expression of key enzymes genes in triterpenoid biosynthesis of Alisma orientale,the study utilized UPLC-MS/MS method to detect eight triterpenoids content in the tuber of A. orientale from different growth stages,including alisol A,alisol A 24 acetate,alisol B,alisol B 23 acetate,alisol C 23 acetate,alisol F,alisol F 24 acetate and alisol G,and then the Real time quantitative PCR was used to analyze the expression of key enzymes genes HMGR and FPPS in triterpenoid biosynthesis. Correlation analysis showed that there was a significant positive relation between the total growth of these eight triterpenoids and the average relative expression of HMGR and FPPS(HMGR: r = 0. 998,P<0. 01; FPPS: r = 0. 957,P<0. 05),respectively. Therefore,the study preliminarily determined that HMGR and FPPS genes could regulate the biosynthesis of triterpenoids in A. orientale,which laid a foundation for further research on the biosynthesis and regulation mechanism of triterpenoids in A. orientale.
Alisma ; chemistry ; genetics ; Chromatography, Liquid ; Geranyltranstransferase ; genetics ; Hydroxymethylglutaryl-CoA-Reductases, NADP-dependent ; genetics ; Phytochemicals ; analysis ; Plant Extracts ; Plant Proteins ; genetics ; Plant Tubers ; chemistry ; Tandem Mass Spectrometry ; Triterpenes ; analysis

Alisma orientalis is a traditional herb medicine commonly used in clinical. With the increasing report of its toxicity in clinical, the renal toxicity of Alisma orientalis has got gradually attention. This paper systematically reviews the research on the chemical material basis of Alisma orientalis including its chemical composition and toxicity of ingredients; and also declares its toxic ingredients and targets according to Network toxicology. Based on the controversy on renal toxicity of Alisma orientalis, we analyzed the possible reasons that may be associated with renal toxicity. It might be associated with the differences of the material basis composition and regulatory toxicology network, differences in employed processing technology, the metabolic function leading to accumulation of compounds, dosage and duration of the experiment and compatibility. The review provides possible reference and ideas for the quality control and rational use of Alisma orientalis.
Alisma ; chemistry ; toxicity ; Animals ; Drugs, Chinese Herbal ; chemistry ; toxicity ; Humans ; Molecular Structure

OBJECTIVETo compare the roles of alisma and gliclazide in the treatment of diabetes in Goto-Kakizaki (GK) rats.

METHODSGK rats were randomly divided into alisma group, gliclazide group, and blank group, and Wistar rats were used as the normal group. After two weeks of treatment, body weight, food intake,fasting glucose, impaired glucose tolerance, and other indicators were measured.

RESULTSThe body weight increased after the treatment in the normal group,blank group,and gliclazide group [(241.3 ± 7.0)g vs.(263.5 ± 11.1)g, (242.8 ± 7.1)g vs.(267.9 ± 16.8)g, (243.9 ± 12.2)g vs.(277.9 ± 9.8)g, P<0.05] but decreased in alisma group [(244.6 ± 9.2)g vs.(227.9 ± 13.7)g, P<0.05]. The food intake showed no significant change before and after administration among different groups(P>0.05). Fasting glucose was significantly lower in normal group than in control group,alisma group,and gliclazide group [(4.8 ± 0.2) mmol/L vs.(8.2 ± 1.4) mmol/L,(8.1 ± 0.6) mmol/L, (8.1 ± 0.9)mmol/L, P<0.05] one week after drug administration; it was not significantly different among blank group,alisma group,and gliclazide group before drug administration (P>0.05); however, it significantly decreased in alisma group and gliclazide group two weeks after administration [(6.9 ± 0.7) mmol/L vs.(8.1 ± 0.6) mmol/L; (5.8 ± 0.5) mmol/L vs.(8.1 ± 0.9) mmol/L, P<0.05]; compared with the blank group, the fasting glucose was significantly lower in the alisma group and gliclazide group,and it was also significantly different between these two groups [(6.9 ± 0.7) mmol/L vs.(8.8 ± 0.6) mmol/L,(5.8 ± 0.5)mmol/L vs.(8.8 ± 0.6)mmol/L, (6.9 ± 0.7) mmol/L vs.(5.8 ± 0.5)mmol/L, P<0.05]. Compared with the normal group,glucose tolerance was abnormal in blank group,alisma group,and gliclazide group;after two weeks of treatment,glucose tolerance was significantly improved in alisma group (P<0.05); compared with the pretreatment level and that in the blank group,the glucose tolerance in gliclazide group showed no significant difference (P> 0.05).

CONCLUSIONSBoth alisma and gliclazide monotherapy is effective in lowering fasting blood glucose. As a single-target drug,gliclazide has stronger effecacy in lowering fasting glucose. However, alisma, as a mixture, can also control weight and improve glucose intolerance.

Alisma ; Animals ; Blood Glucose ; Body Weight ; Diabetes Mellitus, Experimental ; Gliclazide ; Rats ; Rats, Wistar

Climate in China has fluctuated greatly for last two thousand years. Also, the temperate-subtropical transition zone, as well as the distribution boundaries of subtropical biology and growth of suitable areas appear north-south lapse. In historical period, significant climate change will also be bound to the changes of the medicinal organism distribution suitable areas. The past dynasties herbal herbs have documented origin in detail, especially genuine medicinal materials producing areas. In this paper, Alisma orientale and Citrus aurtantium as examples, were used to elaborate the impact of climate change fluctuations on genuine producing area by evolution and change of genuine producing areas. The results showed that medicinal species were more sensitive to climate change such as A. orientale and C. aurtantium, its main producing areas or genuine producing area from north to south shifted in the Ming and Qing dynasties, consistent with the characteristics of climate change in China in last two thousand years. Factors prompted producing areas southward are not only low temperature and cold damage, and temperature and humidity have often synergistic effect. The human activities are more likely to exacerbate the impact.
Alisma ; growth & development ; Citrus ; growth & development ; Climate Change ; Time Factors

OBJECTIVETo sift and identify the nephrotoxic components in Zexie for controlling the quality of the herb.

METHODThe fractions of zexie were prepared by Pre-HPLC, then the nephrotoxicity of the fraction was sifted using LLC-PK1 labelled with fluorescein diacetate and MTT assay. Finally, the compounds in the most obvious nephrotoxic fraction were identified with LC-MS.

RESULTUsing MTT and FDA assay, similar results were obtained. Fraction C13 was found to be the most toxic with FDA assay, in which three compounds, alisol C, 16, 23-oxido-alisol B and alisol O, were detected and characterized by multi -stage mass spectrometric analysis.

CONCLUSIONAlisol C, 16, 23-oxido-alisol B and alisol O in Zexie may cause nephrotoxicity.

Alisma ; chemistry ; toxicity ; Animals ; Chromatography, High Pressure Liquid ; Kidney ; drug effects ; LLC-PK1 Cells ; Mass Spectrometry ; Swine

In the present study, A fluorescent imaging-based high-throughput screening method was developed for identifying anti-migratory compounds with 96-well Transwell plates. The correlation, precision and stability of this method were examined and the incubation time of dye Hoechst 33342 in addition to migration time was optimized. In addition, The inhibitory activity of anti-cancer drug paclitaxel on tumor cell migration was assayed and an IC50 value of 0.717 micromol x L(-1) was obtained. Using this method, 24 components from Rhizoma Alismatis were screened and one component with anti-migration activity was found. These results show that the new proposed method with good precision, stability and linear range has the potential to assay the inhibitory activity of anticancer compounds.
Alisma ; chemistry ; Antineoplastic Agents, Phytogenic ; pharmacology ; Cell Line, Tumor ; Cell Movement ; drug effects ; Drug Screening Assays, Antitumor ; methods ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Fluorescence ; High-Throughput Screening Assays ; methods ; Humans ; Inhibitory Concentration 50 ; Paclitaxel ; pharmacology ; Plants, Medicinal ; chemistry ; Rhizome ; chemistry

Triterpenes, which have large application potential in the treatment of cancer, are the main active components of genuine medicinal material Alisma orientale (Sam.) Juzep. Farnesyl pyrophosphate synthase (FPPS) is one of the important rate-limiting enzymes in the synthetic pathway of triterpenes. In this study the FPPS full length cDNA of the A. orientale, was cloned via homology-based cloning approach and rapid amplification of cDNA ends (RACE). The full length of the FPPS cDNA was 1 531 bp (accession no. HQ724508), which contained a full 1 032 bp ORF that encoded 343 amino acids. The deduced protein sequence exhibited five conserved motifs, two of which is riched of Asp (DDXXD). The result of real-time quantitative PCR (QRT-PCR) showed that FPPS gene was expressed in different organs of A. orientale. The expression increased from October to the first ten-day period of December, and then decreased. The FPPS gene expression was higher in leaves but lower in leafstalk, tuber and root. HPLC analysis of active components 23-acetyl-alismol B of A. orientale. during different periods indicated that its change trend should be consistent with FPPS gene expression. It can be primarily deduced that FPPS gene should be an important control point in the synthetic pathway of Alisma terpenes. This study may facilitate the quality of medicinal plants through gene engineering in the future.
Alisma ; enzymology ; genetics ; Amino Acid Sequence ; Base Sequence ; Cloning, Molecular ; Computational Biology ; Conserved Sequence ; DNA, Complementary ; genetics ; DNA, Plant ; genetics ; Gene Amplification ; Geranyltranstransferase ; genetics ; isolation & purification ; metabolism ; Molecular Sequence Data ; Phylogeny ; Plant Leaves ; enzymology ; genetics ; Plant Roots ; enzymology ; genetics ; Plants, Medicinal ; enzymology ; genetics ; RNA, Messenger ; metabolism ; Real-Time Polymerase Chain Reaction ; methods

OBJECTIVETo develop an HPLC method for the determination of alisol A, alisol F, alisol A 24-acetate and alisol B 23-acetate in the rhizome of Alisma orientalis.

METHODThe separation was performed on a Shim-Pack CLC-ODS a C18 column (6 mm x 150 mm, 5 microm) eluted with the mobile phases of acetonitrile (A)-water (B) in gradient elution. The absorbance was monitored at 210 nm. Orthogonal test was adopted to optimize the extraction conditions of the four alisols.

RESULTThe correlation coefficients of the four alisols were higher than 0.999 and the average recoveries were 99.23%, 96.67%, 97.30% and 99.61%, respectively. All the RSDs were less than 3%.

CONCLUSIONThe validation data demonstrated that the method was accurate and repeatable, and can be used to measure the four alisols in Rhizoma Alismatis.

Alisma ; chemistry ; Cholestenones ; analysis ; Chromatography, High Pressure Liquid ; methods ; Chromatography, Reverse-Phase ; methods ; Reproducibility of Results ; Rhizome ; chemistry ; Time Factors ; Triterpenes ; analysis