ABSTRACT
By reviewing ancient materia medica， prescription and medical books， combined with modern literature， the paper made textual research on the name， origin， producing area， quality evaluation， harvesting and processing methods of Angelicae Pubescentis Radix and Notopterygii Rhizoma et Radix， so as to provide a basis for the selection and use of these two herbs in the development of famous classical formulas. Through textual research， it can be found that Angelicae Pubescentis Radix and Notopterygii Rhizoma et Radix were mixed together in the early history of China， but the distinction was first made during the Southern and Northern dynasties， and since then there have been constant controversies， and it is not until contemporary times that they are distinguished clearly. In the past dynasties， Duhuo and Qianghuo were used as the rectification of names， some aliases and trade names were also seen. Angelica biserrata is the mainstream origin of Angelicae Pubescentis Radix in the past dynasties， and there are many plants belonging to Angelica， Heracleum and Aralia， which are also used as this medicine. However， the origin of Notopterygii Rhizoma et Radix used in the past dynasties is mostly Notopterygium incisum or N. franchetii， which is relatively uniform. The producing areas of Angelicae Pubescentis Radix and Notopterygii Rhizoma et Radix are mostly concentrated in the western and northwestern regions of China， among which Angelicae Pubescentis Radix is mainly produced in Hubei， Chongqing， Sichuan， Shaanxi and other places， and the border area between Hubei and Chongqing is the geo-authentic area. Notopterygii Rhizoma et Radix is mainly produced in Sichuan， Gansu， Qinghai， Shaanxi and others with the western and northern Sichuan and southern Gansu as the geo-authentic areas. Angelicae Pubescentis Radix and Notopterygii Rhizoma et Radix in the past dynasties were harvested in spring and autumn， especially in February and August of the lunar calendar. Angelicae Pubescentis Radix with strong main roots， few branches， firm texture and strong aroma is superior， and Notopterygii Rhizoma et Radix with strong rhizomes， tightly raised knots， purple-brown skin， tight cross-section， strong aroma and silkworm-like shape is superior. The processing methods of Angelicae Pubescentis Radix and Notopterygii Rhizoma et Radix are mostly cut after cutting the reeds， and the raw product is used as medicine. Based on the above research results， it is recommended that the roots of A. biserrata should be used for Angelicae Pubescentis Radix and the roots of N. incisum should be used for Notopterygii Rhizoma et Radix in the development of famous classical formulas， and raw products should be used in the formulas that do not specify processing requirements.
ABSTRACT
ObjectiveTo explore the efficacy and mechanism of the alcohol extract DH50 of Angelicae Pubescentis Radix in treating gouty arthritis induced by monosodium urate （MSU） crystals in vivo and in vitro. MethodFifty male SD rats were randomly assigned into five groups （n=10）： a normal group， a model group， a dexamethasone （DXMS， 0.07 mg·kg-1） group， and low- （DH50-D， 9 mg·kg-1） and high-dose （DH50-G， 18 mg·kg-1） DH50 groups. The rats in the normal group and model group were administrated with the same amount of pure water. On day 5， the gouty arthritis model was established by injecting MSU into the right ankle joint of rats. The toe volume and joint inflammation index were measured 4， 8， 24， and 48 h after modeling. The pathological changes of the synovial tissue were detected by hematoxylin-eosin （HE） staining. Enzyme-linked immunosorbent assay （ELISA） was employed to measure the levels of tumor necrosis factor-alpha （TNF-α）， interleukin （IL）-1β， and IL-6 in the synovial tissue. Western blot was employed to measure the protein levels of NOD-like receptor protein 3 （NLRP3）， cysteine-aspartic protease-1 （Caspase-1）， apoptosis-associated speck-like protein containing a C-terminal caspase recruitment domain （ASC）， IL-1β， and cyclooxygenase-2 （COX-2） in the synovial tissue. Furthermore， the cell inflammation model was established with RAW264.7 cells stimulated with MSU （75 mg·L-1）. The cell experiments were carried out with 6 groups： a normal group， a model group， a positive drug （DXMS， 100 μmol·L-1） group， and low- （DH50-D， 25 mg·L-1）， medium- （DH50-Z， 50 mg·L-1）， and high-dose （DH50-G， 100 mg·L-1） DH50 groups. Methyl thiazolyl tetrazolium （MTT） assay was employed to determine the cell viability， ELISA to determine the content of TNF-α in the supernatant of cell culture， and Western blot to determine the protein levels of NLRP3， cleaved Caspase-1， IL-1β， TNF-α， and COX-2. ResultCompared with the normal group， the rat model group showed increased toe swelling degree and joint inflammatory index （P<0.01）， serious infiltration of the synovium， elevated levels of inflammatory cytokines in the tissue homogenate （P<0.01）， and up-regulated protein levels of NLRP3， Caspase-1， ASC， IL-1β， and COX-2 （P<0.05， P<0.01）. Compared with the rat model group， low- and high-dose DH50 mitigated the toe swelling degree， decreased the joint inflammatory index， alleviated the inflammatory infiltration， lowered the levels of inflammatory cytokines in the tissue homogenate （P<0.01）， and down-regulated the expression of related proteins （P<0.05， P<0.01）. Compared with the normal group， the cell model group showed elevated level of TNF-α in the supernatant （P<0.01） and up-regulated protein levels of NLRP3， cleaved Caspase-1， IL-1β， TNF-α， and COX-2 （P<0.05）. Compared with the model group， low， medium， and high doses of DH50 lowered the level of TNF-α in the supernatant of cell culture in a dose-dependent manner and down-regulated the expression of related proteins （P<0.05， P<0.01）. ConclusionDH50 can mitigate gouty arthritis both in vitro and in vivo by inhibiting the activation of NLRP3 inflammasomes and the production of inflammatory cytokines.
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Objective To establish HPLC analysis methods and fingerprint of different proportions of Angelicae Pubescentis Radix in Angelicae Sinensis Radix; To use HPLC fingerprint method to identify the adulteration of Angelicae Pubescentis Radix in Angelicae Sinensis Radix. Methods Agilent TC-C18 column (5 μm, 4.6 mm × 250 mm) was adopted. Acetonitrile (A) - 0.05% phosphoric acid (B) was the mobile phase, and the gradient elution (0–20 min, 95%–75%B; 20–60 min, 75%–65%B; 60–80 min, 65%–5%B) was used; The flowrate was 1 mL/min; Detection wavelength was 254 nm; Column temperature was 30 ℃. The HPLC spectra of Angelicae Sinensis Radix, Angelicae Pubescentis Radix and adulterated samples were compared and analyzed by using TCM chromatogram fingerprint similarity evaluation system. Results Angelicae Sinensis Radix and Angelicae Pubescentis Radix showed good chromatographic peaks in chromatogram separation. 11 peaks in HPLC fingerprint could be used as the mark, including 7th peak for ferulic acid, 32th peak for osthole, and 34th peak for columbianadin. Conclusion This method is rapid, simple and feasible, can fully reflect the quality of Angelicae Sinensis Radix and clearly identify whether Angelicae Pubescentis Radix is adulterated in Angelicae Sinensis Radix.
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Aim To investigate the neuroprotective effects of osthole(Ost)on the primary cultured cortical neurons transfected with APP595/596 gene and its underlying mechanism.Methods Neonatal mouse cortical neurons were transfected with APP595/596 gene to establish AD cell models for the further study.Then,the cell viability was detected by CCK-8 assay,and the leakage of lactate dehydrogenase(LDH)was assayed by LDH kit to evaluate the injury degree.Transferase-mediated nick end labeling(TUNEL)was used to evaluate the cell apoptosis.The expression of β-amyloid peptide(Aβ)and β-site APP cleaving enzyme 1(BACE1)was measured by immunofluorescence,while the miRNA-107 was measured by RT-PCR.Results Compared to model group,Ost could significantly improve the neurons viability,decrease the LDH release and prevent the apoptosis.Ost also inhibited the expression of Aβ and BACE1 at protein level,while enhanced the expression of miRNA-107 at gene level.Conclusion Ost plays a neuroprotective role in neurons transfected with APP595/596 gene in part through up-regulating miRNA-107.
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Objective: To provide the evidence for the molecular identification of Angelicae Pubescentis Radix (APR) and discuss the scientific classification standard through ITS analysis on 26 samples of APR from 17 species. Methods: ITS of 26 populations was amplified and sequenced. The differences among the different samples were compared and K2P genetic distances of ITS sequence were calculated. NJ tree was constructed and haplotype network map was obtained by using the Network 4.2.0.1 software. Results: NJ tree and haplotype network map suggested that 26 populations were clustered into five groups, and the 17 APR could be sorted into four types after comprehensive analysis. Angelica pubescens was different from other species due to the distinct base sequence in the ITS sequence. All species were identified by ITS analysis except for the three species in Heracleum L. Conclusion: The ITS sequence is powerful for the identification and classification of medicinal APR. Tetrataenium-type plant could be the first choice as succedaneum and followed by Heracleum-type plant, then Aralia-type plant be the last candidate as well.