Mechanism of Alisma plantago-aquatica Linn. improving chronic glomerulonephritis based on network pharmacology and experimental verification / 药学学报

This study aims to explore the improvement and the mechanism of the Alisma plantago-aquatica Linn. (ApL) on chronic glomerulonephritis (CGN). All animal experiments were followed the regulation of the Experimental Animal Ethical Committee of Shanghai University of Traditional Chinese Medicine. CGN mouse model was established by a single tail-vein injection of doxorubicin (Dox) (20 mg·kg-1). One week after Dox administration, the mice received water extract of ApL (85 and 255 mg·kg-1) by gavage once a day for 14 days. At the end of experiment, the urine albumin-to-creatinine ratio (ACR), serum albumin (ALB), blood urea nitrogen (BUN) and serum creatinine (SCr) were detected, kidney histopathological H&E staining was analyzed. Active ingredients and action targets of ApL were collected from TCMSP database, and CGN-related targets were obtained from Genecards database. STRING platform was employed to perform protein-protein interaction (PPI), and Metascape platform was used for KEGG pathway and GO enrichment analysis. The results of experiments demonstrated that ApL (85 and 255 mg·kg-1) could reduce the ACR and the content of SCr and BUN, and increase the content of ALB in mice. Network pharmacology results predicted that nuclear factor kappa-B (NF-κB)-related pathway and biological process of oxidoreductase activity regulation may be involved in the ApL-provided amelioration on CGN. The verification results showed that ApL could inhibit the activation of NF-κB and the expression of inflammatory factors in mice, and reduce the activity of renal myeloperoxidase (MPO). Meanwhile, ApL promoted the activation of nuclear factor erythroid 2-related factor 2 (Nrf2) and increased the expression of its downstream gene mRNA, and reduced the level of renal malondialdehyde (MDA) and reactive oxygen species (ROS), and further elevated renal glutathione (GSH) level. Based on network pharmacology combined experiments, this study found that ApL may improve CGN in mice through multiple targets and multiple pathways, in which the inhibition of NF-κB signaling and the activation of Nrf2 signaling may be important mechanisms involved.

Effects of Zhibitai capsule combined with pitavastatin calcium tablets on coronary heart disease complicated with diabetes mellitus / 中国基层医药

Objective:To investigate the effects of Zhibitai capsule combined with pitavastatin calcium tablets on blood lipids, blood glucose, and glycated hemoglobin in patients with coronary heart disease complicated by diabetes mellitus. Methods:A total of 100 patients with coronary heart disease and diabetes mellitus who received treatment in The Third Affiliated Hospital of Jinzhou Medical University from January 2017 to June 2020 were included in this study. They were divided into a control group ( n = 50) and an observation group ( n = 50) according to different treatment methods. Both groups were given conventional treatment such as pitavastatin calcium tablets. The control group was given pitavastatin calcium tablets based on conventional treatment. The observation group was given Zhibitai capsule combined with pitavastatin calcium tablets based on conventional treatment. After 6 months of treatment, serum levels of triacylglycerol, total cholesterol, low-density lipoprotein cholesterol, high-density lipoprotein cholesterol, fasting plasma glucose, and glycated hemoglobin were compared between the two groups. Results:After treatment, serum levels of triacylglycerol, total cholesterol, low-density lipoprotein cholesterol, fasting plasma glucose, glycated hemoglobin in the observation group were (4.26 ± 0.67) mmol/L, (1.85 ± 0.38) mmol/L, (3.16 ± 0.27) mmol/L, (8.29 ± 1.07) mmol/L, and (8.20 ± 0.77)%, respectively, and they were (4.50 ± 0.39) mmol/L, (1.99 ± 0.19) mmol/L, (3.28 ± 0.27) mmol/L, (8.80 ± 0.66) mmol/L, (8.54 ± 0.74)%, respectively in the control group. After treatment, these indices in each group were decreased compared with those before treatment (control group: t = 19.56, 14.60, 10.66, 8.60, 10.18; observation group: t = 15.04, 14.68, 11.36, 12.36, 12.89, all P < 0.05). After treatment, these indices in the observation group were significantly lower than those in the control group ( t = -2.12, -2.23, 2.26, -2.84, -2.44, all P < 0.05). After treatment, the level of high-density lipoprotein cholesterol in the observation and control groups was (1.16 ± 0.18) mmol/L and (1.09 ± 0.13) mmol/L, respectively. After treatment, the level of high-density lipoprotein cholesterol in each group was increased compared with that before treatment (control group: t = -11.10, observation group: t = -11.07, P < 0.05). After treatment, the level of high-density lipoprotein cholesterol in the observation group was significantly higher than that in the control group ( t = 2.11, P < 0.05). Conclusion:Zhibitai capsule combined with pitavastatin calcium tablets can greatly improve the level of blood lipids and blood glucose in patients with coronary heart disease complicated by diabetes mellitus.

Textual Research on Origin of Alismatis Rhizoma in China Pharmacopoeia / 中国实验方剂学杂志

This paper origin studies the origin of Alismatis Rhizoma in Chinese pharmacopoeia， and puts forward some suggestions for modification. Through the changes in the records of the source of Alismatis Rhizoma in the various versions of the Chinese Pharmacopoeia and the records of Flora of China and Materia Medica of China，it is found that the source of Alismatis Rhizoma in the Chinese Pharmacopoeia is confused. Specifically， the Chinese name of Alismatis Rhizoma does not correspond to the Latin name. As a common Chinese herbal medicine，Alismatis Rhizoma has a large market circulation. Many classic Chinese medicine prescriptions released by China Food and Drug Administration contain Alismatis Rhizoma. The development of the classic Chinese medicine prescriptions will further increase the market circulation of Alismatis Rhizoma. As a major national move to promote the development of traditional Chinese medicine， the study for classic Chinese medicine prescriptions requires defining the origin of the medicinal materials used，and the confused origin of Alismatis Rhizoma recorded in the Chinese Pharmacopoeia seriously hinder the development of the classics. Therefore，in order to regulate the origin of Alismatis Rhizoma， ensure the clinical efficacy and promote the development of classic Chinese medicine prescriptions，the confused origin of Alismatis Rhizoma in the Chinese Pharmacopoeia has to be resolved as soon as possible. Based on the analysis of the changes of Alismatis Rhizoma's producing areas in the past dynasties， it is found that the producing areas of Alismatis Rhizoma have continuous changed from Wei and Jin dynasties to present， and finally formed the current situation of Sichuan as the main producing area. In comparison of chemical composition，origin and market circulation of Alismatis Rhizoma in Sichuan Province that is the most productive， and Fujian Province that is the best quality， it is found that the two species are different in every aspects. Nowadays，Alisma plantago-aquatica occupies the majority of the market， which doesn't conform to Alisma orientale as specified in the 2015 edition of the Chinese Pharmacopoeia. Therefore， through textual research and analysis， it is suggested that both A. plantago-aquatica and A. orientale. Shall be used as the origin of Alismatis Rhizoma. In the 2015 edition of Chinese Pharmacopoeia，Cassiae Semen，Schizonepetae Herba，Aisaematis Rhizoma，Fibraureae Caulis and Ajugae Herba have the same problem. This paper provides ideas for the revision of sources of traditional Chinese medicine.

Novel C-17 spirost protostane-type triterpenoids from with anti-inflammatory activity in Caco-2 cells

Twenty-one protostane-type triterpenoids with diverse structures, including nine new compounds (-), were isolated from the of Linn. Structurally, alisolides A‒F (-), composed of an oxole group coupled to a five-membered ring, represent unusual C-17 spirost protostane-type triterpenoids. Alisolide H () is a novel triterpenoid with an unreported endoperoxide bridge. Alisolide I () represents the first example of 23,24-acetal triterpenoid. Their structures were elucidated based on spectroscopic analysis, wherein the absolute configurations of ‒, were further confirmed by the Mo(OAc)-induced ECD method. Furthermore, all isolates were evaluated for their inhibitory effects on LPS-induced NO production in Caco-2 cells, and all the compounds showed remarkable inhibitory activities, with IC values in the range of 0.76-38.20 μmol/L.

Correlation study between accumulation of triterpenoids and expression of relative genes in Alisma orientale / 中国中药杂志

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.

Improvement Effects of Alisma orientalis Polysaccharide on Insulin Resistance and Lipid Metabolism Disorder in Type 2 Diabetes Mellitus Rats and Its Mechanism Study / 中国药房

OBJECTIVE:To investigate the effects ofAlisma orientalis polysaccharides on the insulin resistance and lipid metabolism disorder in type 2 diabetes mellitus (T2DM) rats and its possible mechanism.METHODS:T2DM rat model was induced by high fat diet combined with intraperitoneal injection of streptozotocin (30 mg/kg).50 model rats were randomly divided into model group (normal saline),glibenclamide group (positive control,25 mg/kg) and A.orientalis polysaccharide high-dose,medium-dose and low-dose groups (400,200,100 mg/kg),with 10 rats in each group.Other 10 normal rats were included in normal control group (normal saline).All groups were given drugs ig,once a day,for consecutive 6 weeks.After 5 weeks of administration,the glucose tolerance test was conducted.Fasting blood glucose (FBG) levels were measured in rats 0,30,60,90,120 min after intraperitoneal injection of 2 g/kg Glucose solution,respectively.After 6 weeks of administration,serum levels of FINS and FBG were measured,and HOMA-IR and insulin sensitivity index (ISI) were calculated;serum levels of FFA,LDL-C,HDL-C,TG and TC were detected,and the levels of SOD,GSH-Px and MDA in liver tissue were detected.RESULTS:Compared with normal control group,FBG levels of rats in model group at 0-120 min were increased significantly in glucose tolerance test.After 6 weeks of administration,the serum levels of FINS,LDL-C and ISI,and the levels of SOD and GSH-Px in the liver tissue of the model group were decreased,and the differences were statistically significant (P＜0.05).Compared with model group,each index of other administration group were improved significantly (P＜0.05),except that HOMA-IR,the serum levels of LDL-C and HDL-C were not significantly improved in A.orientalis polysaccharide low-dose group (P＞0.05).CONCLUSIONS:A.orientalis polysaccharide can improve insulin resistance and lipid metabolism disorder in T2DM rats through antioxidant effect.

Effect of pretreatment with Rhizoma Alismatis extract on cardiac function and cysteinyl aspartate specific protease-3 in ischemia-reperfusion injury of rats / 中华老年医学杂志

Objective To observe the effect of pretreatment with Rhizoma Alismatis extract on cardiac function and cysteinyl aspartate specific protease-3 (caspase-3)in ischemia-reperfusion injury of rats.Methods Male Sprague-Dawley(SD)rats were randomized into a sham-operated group (S group),an ischemia-reperfusion group(IR group),Rhizoma Alismatis water extract group(S1 group),Rhizoma Alismatis alcohol extract group(S2 group)and Rhizoma Alismatis polysaccharide group(S3 group).At the end of 14-day of intragastric gavages,rats were subjected to 40 min(T1)of LAD(left anterior descending)coronary artery ligation (ischemia)and 120 min (T2)of ligation loosening (reperfusion),called as myocardial ischemia-reperfusion(IR)models.Then,at the end of T1 and T2,the left ventricular end diastolic pressure (LVEDP),left ventricular systolic pressure (LVSP),and maximum rise/fall rate of left intraventricular pressure(± dp/dtmax)were recorded respectively.The level of creatine kinase(CK)and lactate dehydrogenase(LDH)were determined.The area of myocardial infarction and the expression level of caspase-3 protein were tested.Results At end of T2,compared with the index values of IR group as a non-treatment control[LVEDP(6.70 ±0.22)mmHg,LVSP (86.16±15.11)mmHg,+dp/dtmax(997.99±151.03)mmHg,-dp/dtmax(663.71±68.55)mmHg,CK(10.54±2.04)U/L,LDH(296.51±7.00)U/L,the size of myocardial infarction(39.82±11.80)％and expression level of caspase-3(123.42±14.77)],the treatment groups of S1,S2,and S3 showed a lower levels of LVEDP [(5.89 ± 0.47) mmHg,(5.89 ± 0.67) mmHg,(6.07 ±0.51) mmHg],of activity of CK[(8.60± 1.67)U/L(8.90±1.27)U/L,(9.39±0.83) U/L],of LDH[(239.33±30.81) U/L,(223.63 ± 20.26) U/L,(241.19 ± 45.56) U/L],of size of myocardial infarction[(30.39 ± 5.44) ％,(32.18±5.90)％,(33.12±8.16)％],and of expression level of caspase3 protein[(73.44± 15.28),(65.47±12.53),(65.05± 10.45)],(all P＜0.01 or＜0.05);but showed a higher levels of LVSP[(99.24±12.00)mmHg,(97.05±12.45)mmHg,(97.06±7.61) mmHg],and of ±dp/dtmax [(1 137.33±85.70)mmHg,(1 147.24±118.07)mmHg,(1 124.50±141.47)mmHg];[(604.77± 68.37)mmHg,(616.61±46.73)mmHg,(708.76±81.44)mmHg],(all P＜0.01 or＜0.05).Conclusions Pretreatment with Rhizoma Alismatis extract can effectively improve the cardiac function of ischemic repeffusion injury in vivo in rats,and reduce myocardial infarction size and myocardial enzyme release.The mechanism may be related to the down-regulation of apoptotic protein caspase-3 in myocardial tissue.

Prokaryotic expression, function verification and immunoassay activity of farnesyl pyrophosphate synthase of Alisma orientale (Sam.) Juzep / 药学学报

Farnesyl pyrophosphate synthase of Alisma orientale (Sam.) Juzep. (AoFPPS) is considered as one of the important rate-limiting enzymes in the biosynthetic pathway of protostane triterpenes. In order to investigate the expression and function of AoFPPS, the gene (accession No. HQ724508) was cloned into a bacterial expression vector pCzn1, then the combined plasmid pCzn1-AoFPPS was transformed into Escherichia coli BL21, and a fusion protein was obtained after induction. The fusion protein was purified by Ni resin, and the function was verified through in vitro enzymatic reaction. High performance liquid chromatography (HPLC) analysis revealed that the products were able to catalyze the synthesis of farnesyl pyrophosphate (FPP). High purity recombinant protein was used to immunize New Zealand rabbits to generate a polyclonal antibody. The titer of the antibody was determined by enzyme linked immunosorbent assay (ELISA), and Western blot results demonstrated that the antibody could specifically recognize the AoFPPS protein in A. orientale (Sam.) Juzep. So,the method of rapid immunoassay to detect AoFPPS was established. This study lays the foundation for further study of the AoFPPS gene expression, regulation and mechanism of action in A. orientale (Sam.) Juzep., and it also provides a scientific basis on improving the quality of Alismatis Rhizoma using the plant genetic engineering.

Prokaryotic expression, functional identification of squalene synthase in Alisma orientale and its immunoassay study / 中国中药杂志

Squalene synthase of Alisma orientale catalyzes farnesyl diphosphate (FPP) to form squalene, which is the key regulatory enzyme of the carbon source flow to protostane triterpenes biosynthesis. For further research on the function and expression of AoSS gene, the open reading frame (ORF) of squalene synthase gene (accession no. JX866770) from A. orientale was subcloned into a prokaryotic expression vector pCzn1 and induced the expression of AoSS gene in Escherichia coli BL21(Roseta). The fusion protein was mainly in the form of inclusion bodies and purified to obtain high purity protein. By verifying its functionality through vitro enzymatic reaction, the results showed that the catalytic protein had the catalytic activity of FPP into squalene. In order to research the expression of AoSS in A. orientale, the purified protein was used to immunized rabbits to prepare polyclonal antibody which was then purified, the titer of the antibody was greater than 1∶51 200 by ELISA detection, and displayed good specificity by Western blotting. The prepared antibody was used for immunoassay of AoSS in different organs of A. orientale, and the results showed that the AoSS expression level was the highest in tubers, followed by leaves, and lowest in root. Successful construction of prokaryotic expression vector, validation of gene functions and establishment of rapid immunoassay lay the foundation for further researches on the function and regulation of AoSS gene, and also provide scientific basis on the application of the protostane triterpenes of A. orientale in the field of synthetic biology.

Research on effective components compatibility of Alismatis Rhizoma resisting calcium oxalate calculus in vitro based on uniform design / 中草药

Objective: To investigate the inhibitory effect of six active triterenoids and their compatibility from Alisma orientale on the formation of urinary calcium oxalate calculus in vitro. Methods: A uniform design method was used to design the different compatibilities from six triterenoids. The inhibitory effects of the six triterenoids along with their different compatibility groups were evaluated in vitro using a standard seeded crystallization technique. Results: The six active triterenoids and their compatible preparations could significantly inhibit calcium oxalate crystal formation in vitro (P < 0.05), and the group of alisol A-alisol A 24-acetate-alisol B-alisol B 23-acetate-alisol F-alisol F 24-acetate (2.2∶3.8∶1∶3.5∶2.2∶1) was the most efficient with an inhibitory index of 188.29%. Conclusion: The triterenoids in A. orientale play a key role in the inhibition of calcium oxalate calculus formation and compatibility of alisol A, alisol A 24-acetate, alisol B, alisol B 23-acetate, alisol F, and alisol F 24-acetate can inhibit the calcium oxalate calculus well in vitro especially cooperated with each other, and the best compatibility proportion is 2.2∶3.8∶1∶3.5∶2.2∶1, respectively.

Simultaneous determination of four active components in Alisma orientale (Sam.) Juz. by HPLC-DAD using a single reference standard / 药物分析学报

A rapid, simple and practical high-performance liquid chromatography method coupled with diode array detector (HPLC–DAD) was developed to evaluate the quality of Alisma orientale (Sam.) Juz. through a simultaneous determination of four major active triterpenes using a single standard to determine the multi-components (SSDMCs). Alisol B 23-acetate was selected as the reference compound for calculating the relative response factors. All calibration curves showed good linearity (R240.9998) within test ranges. RSDs for intra- and inter-day of four analytes were less than 3.6% and 2.3%; the overall recovery was 92.1–110.2%(SSDMC). The proposed method was successfully applied to quantify the four components in 20 samples from different localities in China. Moreover, significant variations were demonstrated in the content of these compounds. In addition, hierarchical clustering analysis (HCA) and principal components analysis (PCA) were performed to differentiate and classify the samples based on the contents of Alisol C 23-acetate, Alisol A, Alisol A 24-acetate and Alisol B 23-acetate. This simple, rapid, low-cost and reliable HPLC–DAD method using SSDMC is suitable for routine quantitative analysis and quality control of A. orientale (Sam.) Juz.

Effect of Rhizoma Alismatis extracts on oxidative stress induced by cerebral ischemia-reperfusion injury in rats / 中华老年医学杂志

Objective To investigate the effects of Rhizoma Alismatis extracts on oxidative stress induced by cerebral ischemia-reperfusion injury in rats,and to explore its protective mechanism in cerebral ischemia-reperfusion injury.Methods A total of 60 male SD rats were randomly divided into sham operation group,model group,Alisma orientalis group and Nimodipine positive control group (n=15,each).Cerebral ischemia-reperfusion injury model was prepared by suture method after 14 days of intragastric administration.After 24 hours,scores of neurological dysfunction,the infarct size,the water content of the brain,the malondialdehyde (MDA),superoxide dismutase (SOD),nitric oxide (NO) levels in serum and brain tissues,and the activity of inducible NO synthase (iNOS)were detected.Results As compared with the model group,Alisma orientalis group showed that the scores of neurological dysfunction,cerebral water content,cerebral infarction size,contents of MDA and NO,and the activity of iNOS were significantly reduced,and the activity of SOD was significantly increased in respectively [(2.21 ± 0.38) vs.(2.78 ± 0.43),(81.18 ± 2.09)％ vs.(88.33±4.15)％,(0.26±0.07) ％ vs.(0.35±0.04)％,(5.92±1.64) μmol/L vs.(8.21±1.47)μmol/L,(115.48±18.65) mU/L vs.(75.52±20.78) mU/L,(28.23±4.32) μmol/L vs.(41.73±3.85) μmol/L,(15.31±1.68) mU/L vs.(23.49±3.53) mU/L,(5.41±0.68) μmol/L vs.(7.58±1.49) μmol/L,(168.57±10.65) mU/L vs.(150.11±13.62) mU/L,(14.37±0.77) μmol/L vs.(22.08±1.57) μmol/L,(9.83±0.75) mU/L vs.(13.28±1.84) mU/L,respectively,all P＜0.05]Conclusions Alisma orientalis extract has the protective effect on focal cerebral ischemia reperfusion injury,and the mechanism may be related to antioxidant and scavenging free radicals.

Progress on chemical components and diuretic mechanisms of traditional Chinese diuretic medicines Poria cocos,Cortex Poriae, Polyporus umbellatus and Alisma orientalis / 中国药理学与毒理学杂志

Poria cocos,Cortex Poriae,Polyporusumbellatusand Alisma orientalisare common tra-ditionaI Chinese diuretic medicines. According to reported Iiterature,P.cocostriterpenes and poIysaccha-rides,steroids and tetracycIic triterpenes are the main chemicaI components of P.cocos,its epidermis, Pol.umbellatusand A.orientalis,respectiveIy. most of these diuretic drugs contain tetracycIic triterpenes and steroids,which have a simiIar structure to aIdosterone nucIeus structure. Therefore,this characteris-tic may reveaI their diuretic mechanisms. The tetracycIic triterpenes and steroids may exert diuretic effect through competitive inhibition of aIdosterone receptors in different parts of tubuIar reabsorption to increase urine output. The present articIe reviewed the chemicaI components of these diuretic Chinese medicines. Furthermore,their bioactive components and action mechanisms were aIso anaIyzed and discussed.

Extraction and isolation of 23-acetate alisol B from Alisma orientalis by supercritical fluid extraction and high-speed counter-current chromatography / 中草药

Objective: To develop an effective and rapid method for the preparation of 23-acetate alisol B from Alisma orientalis. Methods: The SFE-CO2 extract from A. orientalis was injected into high speed counter current chromatography (HSCCC) directly, and eluted with difierent solvent systems. The crystalline purity was detected by HPLC. The structure of the target compound was identified by UV, IR, MS, and NMR. Results: The solvent system composed of n-hexane-ethylacetate- methanol-water (3∶2∶3∶2) was the best. The lower phase was used as the mobile phase and performed at a flow rate of 2 m/min, while the apparatus rotated at 800 r/min, and detected at 254 nm. The prepared alisol B 23-acetate was identified with infrared spectrometry (IR), mass spectrometry (MS), and nuclear magnetic resonance (NMR) detection, and its purity was 99.8% analyzed by HPLC. Conclusion: The established method is relatively simple, fast, and suitable for the fast isolation and separation of alisol B 23-acetate.

Chemical constituents from Alisma orientalis extracts with hypoglycemic effect / 中草药

Objective: To study the chemical constituents in Alisma orientalis extracts with hypoglycemic effect. Methods: To study the in vivo hypoglycemic effects of A. orientalis extracts, high fat diet (HFD)-induced insulin resistance male C57BL/6J mice were treated with water and ethanol extracts of A. orientalis in diet, and glucose tolerance test was carried out following the intervention. Silica gel, ODS, and preparative HPLC were used to isolate the compounds. Their chemical structures were elucidated on the basis of NMR and MS spectral data. Results: Sixteen compounds were identified as sitosterol (1), palmitic acid (2), heptadecanoic acid (3), eicosanoic acid (4), 11-deoxy-alisol B (5), 23-acetate alisol B (6), 23-acetate alisol C (7), alisol B (8), 24-acetate alisol A (9), alisol G (10), 24-acetate alisol F (11), alisol L (12), alisol C (13), alisol F (14), alisol A (15), and 16-oxo-24-acetate alisol A (16), and nine of the triterpenes could improve glucose uptake in HepG2 cells. Conclusion: Compounds 3 and 4 are isolated from A. orientalis for the first time. The water and ethanol extracts of A. orientalis could improve glucose tolerance test. Triterpenes may be one of the therapeutic material basis in hypoglycemic activities in A. orientalis.

Effect of reproductive growth on source-sink relationship of Alisma plantago-aquatica / 中草药

Objective: The effects of the reproductive growth on the source-sink relationship and source-sink characteristic indexes of Alisma plantago-aquatica were studied in order to provide a potential reference for the cultural techniques of high yield and quality and breeding. Methods: The dry weight in each part and the content of non-structure carbohydrates and nitrogen compounds in the leaves and tubers from the wiping off bolting and bolting plant were tested; The accumulative process of physiological indexes was compared. Results: Reproductive growth could benefit the accumulation of reproductive sink, go against the increase of source of leaves and vegetative sink, and obviously increase the plant total storage capacity and the total biomass. Wiping off the reproductive growth could increase the C/N ratio in the leaves at the later stages of plant growth and the tubers during the yield formative period, and promote the production of the leaves and tubers. Conclusion: During the process of cultivation, it is supposed to promptly adjust the plant C/N ratio by artificial measures for increasing the tuber production. The strains with suitable amounts and high C/N ratio in the leaves and high nitrogen content in the tubers at the later stages of the plant growth should be chosen for breeding.

Gene cloning of squalene synthase in Alisma orientalis and its bioinformatic analysis / 中草药

Objective: To clone the full-length cDNA encoding squalene synthase (SS), a key enzyme of protostane type triterpenes biosynthesis, from Alisma orientalis and to perform bioinformatic analysis. Methods: With the total RNA as template, the full-length cDNA of SS in A. orientalis was cloned via homology-based cloning approach and rapid amplification of cDNA ends technique. The bioinformatics of the cloning SS gene was analyzed by DNAMAN and ExPASy online analysis. Results: The full-length cDNA (1 577 bp) of SS gene was obtained (GenBank accession number JX866770), with an open reading frame of 1 230 bp, encoding 409 amino acid polypeptides, which had higher homology with the known SS in other medicinal species. The calculated relative molecular mass was 4.68 × 104, the isoelectric point was 5.97, and there was no signal peptide in SS. The deduced protein sequence exhibited two conserved domains rich in Asp (DXXDD). Conclusion: The cDNA encoding SS from A. orientalis is cloned and reported for the first time. This work provides a foundation for exploring the biosynthetic pathway of protostane type triterpenes in A. orientalis and their applications in bioengineering.

The Effects of Alisma canaliculatum Butanol Fraction with Selenium on Glycogen Level, Lipid Metabolism and Lipid Peroxidation in Streptozotocin-Induced Diabetic Rats

The purpose of this study was to investigate the effect of butanol (BuOH) fraction of Alisma canaliculatum (Ac) and/or selenium (Se) treatment on glycogen level, lipid metabolism and lipid peroxidation in streptozotocin (STZ)-induced diabetic rats. Male Sprague-Dawley rats were assigned to one of the five groups: normal, STZ-control, and three experimental groups (Ac group, Ac-Se group, and Se group). Diabetes was experimentally induced by intravenous administration of 45 mg/kg of STZ in citrate buffer. The BuOH fraction of Ac (400 mg/kg bw) was orally administered for 3 weeks. The Se group were fed a AIN-93 recommended diet mixed with Na(2)SeO(3) (2 mg/kg diet). The liver glycogen level of Ac and Ac-Se groups were significantly higher, when compared with the STZ-control groups. The muscle glycogen level was not significantly differ among all groups. The levels of liver triglyceride were higher in Ac-Se group than the STZ-control group. Pancreas protein levels were significantly increased in Ac-Se group than STZ-control group. The concentration of liver malondialdehyde (MDA) was significantly decreased in Ac and Se groups and decreased in Ac-Se group. Administration of BuOH fraction of Alisma canaliculatum and selenium supplementation increased the liver glycogen and triglyceride levels, and reduced peroxidative liver damage in STZ induced diabetic rats. These results suggest that treatment with a BuOH fraction of Alisma canaliculatum in combination with selenium has no synergistic antioxidative effect. Selenium supplementation may lead a decrease MDA of liver in diabetic rats.

The Effects of Zhu Ling Decoction and Alisma Orientalis on Osteopotin mRNA Expression in the Experimental Rat Model of Renal Stone / 中国医师杂志

Objective To explore the effects of Zhu Ling decoction and Alisma orientalis on the osteopotin mRNA expression in the experimental rat model of renal stone. Methods The experimental rat model of renal stone was set up using glyoxylic acid. Osteopontin mRNA expression in the renal tissues in the experimental rat model of renal stone was detected by RT-PCR. Results Glyoxylic acid obviously increased osteopontin mRNA expression in the renal tissues. Zhu Ling decoction and Alisma orientalis remarkably decreased the expression of osteopontin mRNA in the renal tissues. Conclusion Zhu Ling decoction and Alisma orientalis could decrease the osteopotin mRNA expression in the renal tissues in the experimental rat model of renal stone. The results were helpful for prevention and treatment of renal stone.

Protective Effect of Extracts from Radix et Rhizoma Rhei and Alisma orientalis against Acute Kidney Injury Induced by Diethylene Glycol / 中国药房

OBJECTIVE:To explore the protective effect of the extracts from Radix et Rhizoma Rhei and Alisma orientalis against acute kidney injury induced by diethylene glycol (DEG) and its action mechanism. METHODS: The acute kidney injury model of mice was induced by intragastric administration of DEG. Then the model mice were administered intragastrically with the extracts of Radix et Rhizoma Rhei and Alisma orientalis. Serum levels of creatinine(Cr) and blood urea nitrogen (BUN), the activities of superoxide dismutase (SOD) and glutathione peroxidase(GSH-PX) and the content of malonaldehyde(MDA) were determined. RESULTS: After administration of DEG, the mice presents with obvious toxic reactions including the increase of the ratio of kidney to body weight, the increase of serum levels of BUN and Cr, the decrease of the activities of SOD and GSH-PX in kidney tissue, the increase of the content of MDA. However, after treatment with the extracts of Radix et Rhizoma Rhei and Alisma orientalis, the toxic symptoms in mice were attenuated markedly, the ratio of kidney to body weight decreased, serum levels of BUN and Cr decreased significantly, the activities of SOD and GSH-PX in renal tissues increased and the content of MDA decreased. CONCLUSION: The remarkable protective effect of the extracts from Radix et Rhizoma Rhei and Alisma orientalis against DEG-induced acute kidney injury in mice might be related to its actions of improving the activity of antioxidase of kidney while inhibiting lipid peroxidation.