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Objective: To establish the HPLC fingerprint for effective quality control and scientific evaluation of Erigeron breviscapus. Methods: Separation was performed on a Zorbax SB-C18 column (150 mm × 4.6 mm, 5 μm) and the mobile phase was methanol-0.1% phosphoric acid with gradient elution. The flow rate at 1 mL/min, the column temperature at 30 oC, and the detection wavelength at 335 nm. A total of 19 batches of E. breviscapus and its related species were analyzed. Similarity evaluation combined with hierarchical clustering analysis (HCA) and principal components analysis (PCA) were used to evaluate the quality of herbs from different batches. Results: The HPLC fingerprint of E. breviscapus was established with 11 common peaks, and five peaks were identified. Similarities of the 19 batches of samples were 0.873-0.978. Two batches of samples from its related species were high similarity. These 19 batches of samples could be classified into three clusters. The PCA result was consistent with HCA. The comprehensive score of S5 was the highest and the quality was the best. There was possibility for using E. multiradiatus as herbs instead of E. breviscapus. Conclusion: The establishment of HPLC fingerprint and the recognition of chemical pattern can provide a more comprehensive reference for the quality control of herbs.
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This paper summarized the chemical structures and amounts of caffeoylquinic acids in Erigeron breviscapus, as well as its prevention and cure effects on ischemic stroke in clinical and possible pharmacological mechanism. The results showed 22 caffeoylquinic acids reported from E. breviscapus, accounting for 29% of all compounds from this herb; The average content of total polyphenols was 36.93%, and more than 95% of components in Erigeron Breviscapus Injection are caffeoylquinic acids, higher than that of scutellarin. Several high quality clinical studies confirmed that Erigeron Breviscapus Injection enhanced treatment performance and improve the neurological score in the treatment after ischemic stroke and had good safety. In pharmacological research, caffeoylquinic acid compounds display anti-oxidant, anti-free radical, anticoagulation, and anti-fibrosis effects, which can protect neuro, vascular endothelial cells, glial cells, and astrocytes. They are also able to inhibit inflammatory, suppress cytokines IL-1, TNF-α, and enhance SOD & GSH-Px, which play a role in different treatment stages of ischemic stroke. So, caffeoylquinic acid is a kind of important chemical in E. breviscapus.
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Objective To study the role of MYB transcription factor in phenylpropane biosynthesis pathway and the response to biotic or abiotic stress, a full length sequence of putative MYB gene was cloned in Erigeron breviscapus. Methods Based on a partial sequence of putative MYB gene from transcriptome we have previous reported, the full length cDNA was cloned by RACE method in E. breviscapus. According to the sequence of cloning cDNA, the nucleotide sequence similarity, physicochemical properties, hydrophobicity, transmembrane structure, secondary structure and tertiary structure were predicted and analyzed by various softwares. Meanwhile, multiple sequence alignment of cloned MYB gene was performed, and the phylogenetic tree was constructed. In addition, the fusion expression vector of this gene and green fluorescent protein was constructed. Results The MYB gene was cloned and named eBMYB06. The open reading frame was 783 bp, encoding 260 amino acid residues with a relative molecular mass of 63.80 kDa and a theoretical pI of 5.18 which proved to be a stable protein. The secondary structure of the protein is mainly composed of irregular coil, alpha helix and beta fold. According to the result of phylogenetic tree alignment of E. breviscapus with R2R3MYBs from Arabidopsis thaliana, the cloned MYB gene was clustered to two subsets of R2R3-MYB genes from A. thaliana, which suggested that the cloned MYB gene would be similar with the two groups in structure and function, which would be involved in response to biotic and abiotic stress or phenylpropane biosynthesis pathway, respectively. Further experimental results show that the constructed expression can be used for the efficient transformation in E. breviscapus. Conclusion For the first time, a MYB gene was cloned may be involved in the phenylpropanoid metabolism or gene metabolism or gene regulation in response to the environment in E. breviscapus.
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Objective To observe the effects of superfine powder of Erigeron breviscapus on chronic renal failure (CRF) in rats induced by adenine. Methods SD rats were randomly divided into control group, model group, superfine powder and common powder of E. breviscapus groups, 10 rats in each group. The control group received routine feeding, while the other three groups received intragastrically (ig) with adenine (250 mg/kg) for 21 d to establish rat CRF models. Then, the rats in the control group and model group were received ig with equal volume of distilled water. Also, the rats in the superfine powder and common powder of E. breviscapus group were administered ig with E. breviscapus (3 g/kg) for 42 d. During the experiment, the general condition of each group of rats were observed and the levels of serum creatinine (Scr) and blood urea nitrogen (BUN) were determined. Expressions of transforming growth factor-β1 (TGF-β1) and plasminogen activator inhibitor-1 (PAI-1) at mRNA level were determined by qRT-PCR. Concentrations of kidney tissue inflammatory cytokines tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β) were determined by ELISA. Moreover, the kidney tissue was taken for histopathological observation. Results The rats in CRF model group after treatment with superfine and common powder of E. breviscapus appeared different levels of decrease of Scr, BUN, TNF-α, IL-1β, and mRNA expression of TGF-β1 and PAI-1 (P < 0.05). The pathologic changes of kidney tissue were also alleviated in different degrees. The treatment effect of superfine powder of Erigeron breviscapus was better than that of common powder group. Conclusion E. breviscapus has a good renal protective effect in rats with CRF. Furthermore, the superfine powder is more effective than the common powder in the protective effects of renal.
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Objectives: To obtain the variety of Erigeron breviscapus with high yield, high content of active constituents, and stable agronomic characteristics. Methods: The group bulk selection breeding and individual selection breeding methods were applied respectively, and two varieties were evaluated partially by DUS testing and the fingerprint was analysed by AFLP markers. Results: The variety "Airuijie No. 1" had the yield of 3780 kg/hm2, the scutellarin content of 1.84%, and the total caffeic acid esters content of 1.79%; For the other variety "Airuijie No. 17", the yield was 3075 kg/hm2, the content of scutellarin was 1.71%, and the content of total caffeic acid esters was 1.50%. Compared with the wild local E. breviscapus after one-year acclimatization, the two new varieties showed better economic traits with higher uniformity, more stability, and unique AFLP fingerprint. Conclusion: The two varieties of E. breviscapus have high value of application and popularization.
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Background: Retinal ganglion cells (RGCs) are protected in rats with acute elevated intraocular pressure (IOP) by Erigeron breviscapus (vant.) hand-mazz (EBHM). However, it is unclear whether EBHM has neuroprotective effect on RGCs in animal with chronic elevated IOP. Objective: Investigate the protective effect of EBHM extract on RGCs in rabbits with chronic elevated IOP. Methods: Unilateral chronic elevated IOP was produced in rabbits by repeated injection of 2% methylcellulose into the anterior chamber. Secondary degeneration was measured with and without EBHM extract treatment for 60 days. At 60 days, the cells density of the RGCs layer, the thickness of retinal nerve fiber layer (RNFL), and the optic nerve axons were observed and analyzed using an image analysis system. The ultrastructural changes of RGCs and optic nerve axons were observed using transmission electron microscopy. Results: Compared with their contralateral control eyes with normal IOP, in the retinas of 3-4 mm from the optic disc, the cells density of the RGCs layer in the eyes with chronic elevated IOP was 23.2±6.5 cells (n = 6) and 36.0±8.9 cells (n = 10) per three 400x fields at 60 days in untreated and EBHM-treated group, respectively. The RNFL thickness in eyes with chronic elevated IOP was 3.4±0.4 ?m (n = 6) and 5.0±1.0 ?m (n = 10) at 60 days in untreated and EBHM-treated group, respectively. The axons number per 15057.8 ?m2 in eyes with chronic elevated IOP was 370.4±41.0 (n = 6) and 439.0±50.8 (n = 10) at 60 days in untreated and EBHM-treated group, respectively. The number of the organelles in RGCs plasm appeared decreased and mitochondrion vacuolated in the elevated IOP eyes of EBHM-treated group, while some dispersive mitochondrion and rough surfaced endoplasmic reticulum and ribosome still existed in the RGCs plasm. The myelin sheath plates condensed and degenerated, and the microfilaments and microtubules decreased or disappeared in the elevated IOP eyes, but the axons degeneration in the chronic elevated IOP with EBHM treatment was less than that in the chronic elevated IOP without treatment. Conclusion: EBHM extract provided a neuroprotective effect on retinal ganglion cells in rabbits with chronic elevated IOP.
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Objective: To breed Erigeron breviscapus triploid and evaluate the important biological properties of triploid Erigeron breviscapus. Methods: Tetraploid doubled by colchicine was hybridized with diploid cultivar to get the E. breviscapus triploid; The ploidy of E. breviscapus was identified by the chromosome number. The morphological charcteristics of E. breviscapus triploid in the field were observed and scutellarin was tested by HPLC. Results: Through hybirdization of diploid cultivar and tetraploid, E. breviscapus triploid (2n=3x=27) was obtained successfully. Compared with its parents, the plant height, leaf length and width, number of petals, stoma size of E. breviscapus diploids are larger than those of E. breviscapus diploids, but less than those of E. breviscapus tetraploids. However, leaf number and flower diameter of E. breviscapus triploids are better than those of the parents. The plant of E. breviscapus triploids grew vigorously, and the content of scutellarin (1.38%) is lower than that of E. breviscapus tetraploids (1.51%) and higher than that of E. breviscapus diploids (1.22%). However, the yield of E. breviscapus triploid is obvious higher than that of E. breviscapus diploid with 2.14 times and E. breviscapus tetraploid with 1.84 times. Conclusion: The content of scutellarin in E. breviscapus triploid bred by hybridizing E. breviscapus diploid and E. breviscapus tetraploid is lower than that of E. breviscapus tetraploid, while is much higher than that of E. breviscapus diploid. E. breviscapus triploid is an ideal and important one with the development and utilization value based on the characteristics mentioned above.
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Objective To investigate the effects of erigeron breviscapus (Vant.) Hand-Mazz (erigeron breviscapus) pretreatment on pathology and oxyradical level in the spinal cord after ischemia-reperfusion (I/R) injury in rabbits. Methods A total of 40 New Zealand white rabbits were randomly divided into three groups: sham-operation group with 10 rabbits treated with only abdominal aorta exposure without occlusion, control group with 15 rabbits that underwent ischemia for 50 minutes and treated with matched saline, and experimental group with 15 rabbits that underwent ischemia for 50 minutes and treated with erigeron breviscapus (9mg/kg) injection before ischemia. Malondialdehyde (MDA) level and superoxide dismutase (SOD) activity in the spinal cord were examined at 6 and 24 hours after I/R, respectively. The morphological changes and the number of the spinal cord anterior horn motor neurons were observed and counted under the light microscope and electron microscope, respectively. Results The level of MDA was markedly decreased and SOD activity was increased in the experimental group compared with those in the control group (P<0.01). Compared with that in the control group, the number of motor neurons in the experimental group significantly increased at 24h after I/R (P<0.01) and the morphous of the motor neurons improved. Conclusion Erigeron breviscapus can reduce oxyradical production and the apoptosis of nerve cells, and protect nerve tissue structure and function after spinal cord I/R.
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Objective: To investigate the effects of erigeron breviscapus (Vant.) Hand-Mazz (erigeron breviscapus) pretreatment on pathology and oxyradical level in the spinal cord after ischemia-reperfusion (I/R) injury in rabbits. Methods: A total of 40 New Zealand white rabbits were randomly divided into three groups: sham-operation group with 10 rabbits treated with only abdominal aorta exposure without occlusion, control group with 15 rabbits that underwent ischemia for 50 minutes and treated with matched saline, and experimental group with 15 rabbits that underwent ischemia for 50 minutes and treated with erigeron breviscapus (9 mg/kg) injection before ischemia. Malondialdehyde (MDA) level and superoxide dismutase (SOD) activity in the spinal cord were examined at 6 and 24 hours after I/R, respectively. The morphological changes and the number of the spinal cord anterior horn motor neurons were observed and counted under the light microscope and electron microscope, respectively. Results: The level of MDA was markedly decreased and SOD activity was increased in the experimental group compared with those in the control group (P<0.01). Compared with that in the control group, the number of motor neurons in the experimental group significantly increased at 24 h after I/R (P<0.01) and the morphous of the motor neurons improved. Conclusion: Erigeron breviscapus can reduce oxyradical production and the apoptosis of nerve cells, and protect nerve tissue structure and function after spinal cord I/R.
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Objective To observe the protective effect of Erigeron Breviscapus(Vant.)Hand-Mazz(EBHM) on retinal ganglion cells(RGCs) and optic nerve in rabbits with elevated intraocular pressure(IOP). Methods Twenty rabbits with chronic elevated IOP were divided into two groups randomly: EBHM treated group(the ocular hypertension with EBHM treated subgroup and the normal IOP with EBHM treated subgroup) and untreated group(the simple ocular hypertension subgroup and the simple normal IOP subgroup). EBHM was irrigated into the stomachs to the treated group after IOP elevated continuously for 7 d.For light and electron microscopy studies,the rabbits' eyes were made into eyeball samples and optic nerve samples after 60 d.The density of RGCs,thickness of retinal nerve fiber layer(RNFL),and optic nerve axons were observed and quantitated by computer image analysis system.The ultra-microstructure changes of RGCs and optic nerve axons were observed by electron microscope.Results(①The RNFL) thickness,RGCs density and number of axons of the two ocular hypertension subgroups were decreased,compared with those of the two normal IOP subgroups(P
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·AIM: To investigate whether Erigeron Breviscapus (vant) Hand-Mazz (EBHM) EBHM has neuroprotective effect against N-methyl-D-aspartate (NMDA)-induced neuron death in retinal ganglion cell layer (RGCL).· METHODS: 60 healthy SD rats were randomly divided into four groups. 6 animals were normal control group (group A). The others were divided as group B (EBHM group), group C (normal saline+NMDA group) and group D (EBHM + NMDA group). Each group had 18 rats.10nmol NMDA was intravitreally injected to induce partial damage of the neurons in RGCL in the right eyes of Groups C and D. Same volume PBS was intravitreally injected into the left eyes as self-control. Groups B and D were pre-treated intraperitoneally with 6g/L EBHM solution at a dose of 150mg/kg body weight/day seven days before and after NMDA treatment. Group C were administrated intraperitoneally with 9g/L normal saline at the same time of EBHM injection. Rats were sacrificed at 4,7,14d after NMDA treatment. Flat whole retinas were stained with 5g/L cresyl violet and neuron counting in RGCL from both eyes were observed. Each subgroup had 6 rats.· RESULTS: There was no significant difference of neuron counting in RGCL between the right eye and the left eye in group A (P=0.200). There was no significant difference between normal control group and EBHM group either in the right eyes or in the left eyes at 4, 7 and 14 d respectively after intravitreal injection of 10nmol NMDA in group C and group D. (P=0.636, P=0.193). Neuron counting of RGCL in group C and D was significantly decreased in the NMDA-treated eyes at 4, 7 and 14d after intravitreal injection (P＜0.001). There was no significant difference between self-control eyes group and normai control group(P＞0.05). However, neuron counting was significantly higher in the EBHM+NMDA group than normal saline +NMDA group at 14days after intravitreal injection (P=0.044), but was lowered than normal control group (P＜0.05).· CONCLUSION: EBHM has no effect on neuron counting of RGCL when administered alone in normal rats.The results indicates that EBHM plays a partial protective role in NMDA-induced neuron loss in RGCL in the rats.
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Objective To study the effects of N,P,and K fertilizers on the growth and accumulation of secondary metabolites,to reveal the accumulation relationship between the biomass and the secondary metabolites in medicinal plants in the plantation,and to establish the basic theory of fertilization in medicinal plant culture.Methods Orthogonal design was used to study the effects of nine combinations of N,P, and K on the biomass and active constituents in three levels of three factors.While plants flowered,they were harvested,weighed,and smashed.Then contents of N,total caffee esters,and scutellarin were anaylzed.Results High amount of N,P,and K fertilizers was optimum to the accumulation of biomass, but low amount of N,P,and K fertilizers was optimum to the accumulation of total caffee esters and scutellarin.The N fertilizer significantly increased the biomass and decreased the content of total caffee esters and scutellarin in Erigeron breviscapus,but did not affect the active constituents yield.The effect of P on the accumulation of active constituents was higher than that of K,but its effect on the biomass accumulation was lower than that of K.The effect of K on the active constituents yield was the highest.Conclusion N,P,and K fertilizers could increase biomass and active constituents yield,nevertheless,decrease the contents of flavonoids in medicinal plants.
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Object To select quality species of Erigeron breviscapus (Vant.) Hand.-Mazz. and study its growing practice and cultural habit. Methods The yield and content of breviscapinun of six species of E. breviscapus, the process of germinating and sprouding of its seeds, growing in the field, and content of breviscapinun of dry E. breviscapus in different growth duration were observed. Results The yield and content of breviscapiniun of QS-3, QS-6, and QS-1 of E. breviscapus species were higher than thatin others, and their yield in plastic green house is higher than that on open plots; with other crop, the percent of germinating and sprouding were lower, this process needs longer time; biomass of E. breviscapus under culturing is larger than that under wild; the content of breviscapinun of its dry grass in different growth duration is different, it is the highest of all flowering time. At last, methods of culture technology are put forward with above characteristics. Conclusion It will be optimistic to plant E. breviscapus artificilly.
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Object To study the variation of total flavonoids in Erigeron breviscapus (Vant.) Hand.-Mazz. with the development periods, the months, the affection of the environment and genetical factors on the total flavonoids. Methods The total flavonoids in organs of E. breviscapus during the different deve-lopment periods and the months were determined. The total flavonoids of E. breviscapus in the sunny and shady habitats and the total flavonoids of E. breviscapus with different colors were compared. Results The total flavonoids of E. breviscapus mainly existed in the ground part. The total flavonoids of E. breviscapus are the highest in its flourishing period, and the lowest in its withering period. The variety of the total flavonoids in every organs of E. breviscapus in flourishing period from April to June was not significant. Illumination intensity had little affection to the total flavonoids of E. breviscapus. The colors of E. breviscapus flowers could not be the selected markers for the total flavonoids. The genetic factor played an important role in deciding the difference of the total flavonoids among the individuals. Conclusion It will be optimistic to improve the total flavonoids in E. breviscapus by genetic breeding.
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Object To develop a method for the determination of scutellarin in Erigeron breviscapus (Vant.) Hand. -Mazz. and its extract (dengzhanhuasu) by capillary electrophoresis. Methods Separation was carried out in an uncoated fused silica capillary 57 cm ?50 ?m (ID). Meanwhile, a running voltage 20 kV, 40 mmol/L borax (pH 8.50 with H 3PO 4) buffer and a UV detector at 280 nm were adopted. Results The calibration curve showed good linearity over the range of 0.1-4.0 mg/mL (r=0.998 5). The average recovery was higher than 98.0%. Conclusion The method is proved to be simple, rapid and accurate, and can be used for quality control of E. breviscapus and its extract (dengzhanhuasu).
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Object To study the regular change of total flavonoids in Erigeron breviscapus (Vant.) Hand-Mazz (TFE) in different parts of the plant, different individual plant and population from different habitat. Methods Samples of E. breviscapus from eight districts around Qiubei, Kunming and at altitude on Cangshan mountain, Dali were collected and their flavonoids determined.Results TFE showed different values in samples from different origin, with the tendency to show an increase with increase of altitude. The aerial parts including stem, leaf, and flower were higher than that of the underground root.E. breviscapus grown in the same ecological conditions also showed different TFE values from individual to individual.Conclusion Variation in TFE was influenced by the combined effects of genotype and ecological conditions.
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Objective To study the effect of light intensity and light quality on the growth and total flavonoid accumulation of Erigeron breviscapus. Methods Young plants of E. breviscapus were planted under various color films and light intensities, their biomass and total flavonoid content were determined when plants flowered. Results The biomass and total flavonoid content of individuals under 100% and 80% sunshine were higher than those under 50% sunshine. The biomasses of plants under yellow, red, purple, or blue film were lower than those under white film. Under the blue film, the total flavonoid content of the plant was the highest; while under white film, the total flavonoid yield was the highest. Conclusion Light intensity and light quality significantly affect the growth and total flavonoid accummulation of E. breviscapus. The biomass and total flavonoid yield are the highest when under full sunshine.
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Objective To get genetic transformated Erigeron breviscapus plantlet with the antiinsect and antifungi effects.Methods The plasmid with gene RBBI2-3 was produced by PCR,middle carrier PBS and plant carrier YY46.By using the particle gun to bombard E. breviscapus,genetic transformation was realized.It was done with the control experiment on bombarded materials,bombarding pressures,and Gent.sensitivity.Results The result of genetic transformation by using true leaves as bombarded material was the best;Ⅱ-type callus were the next in order;the roots,stems,and Ⅰ-type callus were the last in the result.The bombarding pressure of 8.96?10~6 Pa was the best to induce true leaves forming adventitious bud.The result of using the pressure of 7.58?10~6 Pa and 8.96?10~6 Pa to bombard Ⅱ-type callus did not show any difference and was next to true leaves.The best result is using 50 ?g/mL Gent to choose the E.breviscapus of genetic transformation.Conclusion The paddy RBBI2-3 transformed into the genetic transformation on E.breviscapus by particle gun bombarding is the best.The genetic transformated E.breviscapus plantlet has been obtained.