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Anthraquinones,dianthrones and tannins are the main active ingredients of Rheum tanguticum. In this study the three components were determined by HPLC,and the results were analyzed by multiple comparisons,principal components analysis(PCA)and correspondence analysis(CA). The results showed that the contents of components in different growing areas and types(wild and cultivated) reached a significant level(P<0. 05). Baiyu county,Xiaojin county and Ruoergai county had obvious advantages in the accumulation of catechin hydrate,rhien and sensenoside A respectively. The principal component was different in two growing type and the wild environment was conducive to combined anthraquinones accumulation. For active components,normalized planting was better than retail cultivating. Therefore,the effect on the accumulation of chemical components in Rh. tangusticum,should be taken into full account in the selection of the cultural base of Rh. tanguticum. The standardized cultivating is superior to retail cultivating in terms of the accumulation of active ingredients,and standardized planting is inferior to the wild.
Subject(s)
Anthraquinones , Chromatography, High Pressure Liquid , Drugs, Chinese Herbal , Phytochemicals , Plants, Medicinal , Chemistry , Rheum , Chemistry , TanninsABSTRACT
Objective: To improve the survival status of wild Rheum tanguticum, which is threatened to meet the market demand, Illumina high-throughput sequencing technologies is used to bring new directions for relevant research. Methods The complete chloroplast genome of Rheum tanguticum was constructed with Illumina high-throughput sequencing technologies in this paper. Results: The genome was 161 054 bp in length, and exhibited a typical quadripartite structure of the large (LSC, 86 441 bp) and small (SSC, 12 745 bp) single-copy regions, separated by a pair of inverted repeats (IRs, 30 934 bp each). The chloroplast genome contained 132 genes, including 88 protein-coding genes, 36 transfer RNA genes, eight ribosomal RNA genes and two pseudogenes, 19 genes located in each IR area. Conclusion: The phylogenetic tree constructed with 11 sequences of seven Polygonaceae species and four other family species demonstrated a close relationship between R. tanguticum and R. palmatum in Polygonaceae, which was coincided with their morphological similarity, in addition, there were certain SNP sites in rpl32 and other genes, which provided a new basis for the effective identification of related species.
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Objective To develop an HPLC method to determine the contents of chrysophanol, emodin, physcion, aloe-emodin, sennoside A, and sennoside B simultaneously in the aerial part of Rheum tanguticum Maxim.ex Balf. Methods The analysis was achieved with an Agilent ZORBAX SB-C18 analytic column (250 mm × 4.6 mm, 5 μm) by gradient elution of methanol-0.1％ phosphoric acid in water gradient elution system (0 min, 70％ B; 5 min, 65％ B;8–16 min, 60％ B; 18–23 min, 55％ B; 25–30 min, 45％ B; 32–39 min, 35％ B; 49–57 min, 24％ B; 65–72 min, 20％ B). The flow rate was 1.0 mL/min; detection wavelength was 254 nm; the column temperature was room temperature; the quantification used external standard method. Results The peak areas and concentrations of chrysophanol, emodin, physcion, aloe-emodin, sennoside A, and sennoside B showed good linear relationship within a certain concentration range (r≥0.9995); the RSD of precision was 0.75％–1.17％; the RSD of repeatability was 0.99％–2.06％; the RSD of stability was 0.97％–1.76％; the average recoveries were 99.7％–100.4％. The results showed that there were differences in content between the root and aerial part of Rheum tanguticum Maxim.ex Balf. Conclusion HPLC method for simultaneous determination of contents of 6 contents of the aerial part of Rheum tanguticum Maxim.ex Balf can be used as the references for quality control.
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This study was aimed to investigate the effect of Rheum tanguticum on expression of serum endothelial nitric oxide synthase (eNOS), endothelin (ET) and 6-keto-prostaglandin F1α (6-Keto-PGF1α) in uric acid nephropathy rat model. After 3 weeks of treatment with different extractions of Rheum tanguticum on uric acid nephropathy rat model, levels of serum BUN, Cr and UA were measured by biochemical analyzer. ELISA method was used to measure the expression of serum eNOS, ET and 6-Keto-PGF1α. Meanwhile, pathological changes of renal tissues were observed by light microscope. The results showed that compared with the normal control group, the levels of serum BUN, Cr, UA and renal index were higher in the model group (P< 0.05). Compared with the model group, the concentration of eNOS in the model group was obviously higher in the rhubarb ethanol extraction group, rhubarb water extraction group, and allopurinol group (P< 0.05). Concentration of ET was obviously reduced in the rhubarb water extraction group and allopurinol group (P< 0.05). There was no statistical difference with the rhubarb ethanol extraction group. There was no significant difference in the concentration of 6-Keto-PGF1α among groups. Lots of inflammatory cells and uratic crystallization were observed by light microscope in renal tissues among rats in the model group. Damages of renal tissues in medication groups were alleviated than the model group. It was concluded that Rheum tanguticumcan alleviate the damage of uratic crystallization and inflammatory cell infiltration, increase the concentration of serum eNOS, and decrease the concentration of serum ET. Thus, it can reduce the damage of renal endothelial cells and protect renal functions. The effect of rhubarb ethanol extraction on reducing ET expression was less than the water extraction.
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AIM: To explore the effects of rheum tanguticum polysaccharides(RTP) on TNBS-induced colitis in mice and its probable mechanisms.METHODS: Mice colitis model was induced by 2,4,6-trinitrobenzene sulfonic acid(TNBS).The mice were randomly divided into 5 groups,normal group,model group,RTP-100,200 and 400 group.The macroscopical and histological changes of the colon were evaluated and the cytokines IL-8,IL-10,TNF-?and IL-4 produced by splenocyts were analyzed with ELISA.RESULTS: Compared with the model group,both symptoms and the lesions of colonic mucosa of RTP-200 and 400 group were slighter on ulcerative colitis induced by TNBS in mice. Furthermore,the expressions of TNF-?and IL-8 in colon tissue of mice with TNBS-induced colitis were higher and the IL-10 and IL-4 were lower than that of normal control(P
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Aim To investigate the protective effect of RTP1,one of the polysaccharide isolated from Rheum tanguticum,on H2O2 induced apoptosis in IEC-6 cells and its possible mechanism.Methods H2O2(100 mmol?L~-1)was used to induce IEC-6 cell apoptosis.Different doses(10,30,100 mg?L~-1)of RTP1 were administrated before H2O2 was added into IEC-6 cell culture.Cell viability was observed by MTT assay.Reactive oxygen species were measured with laser scanning confocal microscopy(LSCM).DNA content and percentage of apoptosis were assayed by DNA agarose gel electrophoresis,acridine orange staining and flow cytometry.The activation of Caspase-3 was detected with Western blot analysis.Results Following treatment with H2O2 for 2 h,H2O2 induced a significant decrease in cell viability,while DNA ladder was observed and apoptosis percentage was as high as 31.3%.Accumulation of intracellular ROS and increase in Caspase-3 activity were also detected.Pretreatment with RTP1 for 24 h exhibited cytoprotective effects in a dose-dependent manner.RTP1 obviously enhanced cell viability,reduced formation of DNA ladder and significantly reduced the number of cells labeled with Annexin V.The percentage of apoptosis/necrosis cells was markedly decreased to 24.4% and 21.5%,respectively.LSCM showed that RTP1 attenuated the accumulation of ROS.The significant decrease in Caspase-3 activity was detected.Conclusion RTP1 has cytoprotective capacity to antagonize H2O2-induced IEC-6 cell apoptosis and injury,and this effect may be related to decrease ROS and inhibit Caspase-3 activity
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Objective To establish the HPLC fingerprints of Rheum tanguticum from Qinghai Region.Methods By RP-HPLC,acetonitrile and water including 0.04% H_3PO_4 as mobile phase,gradient elution,flow rate was 1.0 mL/min,the detection wavelength was 280 nm,and the temperature was 40 ℃.Results The RSD of peak areas and retention time of common peak in the test of precision,repeatability,and stability has not reached 5%,respectively.Average similarity was 0.925 in R.tanguticum collected from different habitats.Conclusion All above results show that this method is simple,practicable,and reliable.It can be used as a standard analysis method in the quality control of R.tanguticum from Guoluo Region in Qinghai Province.It also provides a theory foundation on which the species cultivated could replace the wild ones.