Scutellarin Alleviates Ischemic Brain Injury in the Acute Phase by Affecting the Activity of Neurotransmitters in Neurons.

Cerebral ischemic stroke is a common neuron loss disease that is caused by the interruption of the blood supply to the brain. In order to enhance the CIS outcome, both identifying the treatment target of ischemic brain damage in the acute phase and developing effective therapies are urgently needed. Scutellarin had been found to be beneficial to ischemic injuries and has been shown to have potent effects in clinical application on both stroke and myocardial infarction. However, whether scutellarin improves ischemic brain damage in the acute phase remains unknown. In this study, the protective effects of scutellarin on ischemic brain damage in the acute phase (within 12 h) were illustrated. In middle cerebral artery occlusion and reperfusion (MCAO/R) modeling rats, the Z-Longa score was significantly down-regulated by 25% and 23.1%, and the brain infarct size was reduced by 26.95 ± 0.03% and 25.63 ± 0.02% when responding to high-dose and low-dose scutellarin treatments, respectively. H&E and TUNEL staining results indicated that the neuron loss of the ischemic region was improved under scutellarin treatment. In order to investigate the mechanism of scutellarin's effects on ischemic brain damage in the acute phase, changes in proteins and metabolites were analyzed. The suppression of scutellarin on the glutamate-inducing excitatory amino acid toxicity was strongly indicated in the study of both proteomics and metabolomics. A molecular docking experiment presented strong interactions between scutellarin and glutamate receptors, which score much higher than those of memantine. Further, by performing a parallel reaction monitoring-mass spectrometry (PRM-MS) study on both the cortex and hippocampus tissue of the ischemic region, we screened the scutellarin-regulating molecules that are involved in both the release and transportation of neurotransmitters. It was found that the aberrant levels of glutamate receptors, including EAAT2, GRIN1, GRIN2B, and GRM1, as well as of other glutamatergic pathway-involving proteins, including CAMKK2, PSD95, and nNOS, were significantly regulated in the ischemic cortex. In the hippocampus, EAAT2, GRIN1, nNOS, and CAM were significantly regulated. Taken together, scutellarin exerts potent effects on ischemic brain damage in the acute phase by regulating the activity of neurotransmitters and reducing the toxicity of excitatory amino acids in in neurons.

[Effect of Valeriana jatamansi extract on fecal UPLC-MS/MS metabolomics in rats with diarrheal irritable bowel syndrome].

The purpose of this study was to understand the pharmacodynamic effect of Valeriana jatamansi extract in diarrhea predominant irritable bowel syndrome(IBS-D) rat model induced by maternal separation combined with three kinds of stress, and observe the changes of endogenous metabolites in feces after intervention to find potential biomarkers and related metabolic pathways. The animal model of IBS-D was established by maternal separation combined with restraint, ice swimming and tail clamping. The therapeutic effect of each dose group of V. jatamansi extract was evaluated in terms of abdominal withdrawal reflex pressure threshold, fecal water content and immobility time of forced swimming test. In addition, rat feces were collected for detection of metabolic profiles of small molecular metabolites with UPLC-LTQ-Orbitrap MS platform, so as to find the biomarkers of differential metabolism with multivariate statistical analysis methods such as principal component analysis(PCA) and orthogon partial least squares discrimination analysis(OPLS-DA). The results showed that as compared with the normal group, the threshold of abdominal withdrawal reflex pressure was decreased, the fecal water content was increased, and the immobility time of forced swimming test was prolonged in the model group. The results of fecal metabonomics showed that the levels of 39 metabolites were down-regulated and those of 37 metabolites were up-re-gulated. Further analysis showed that these metabolites were related to bile acid metabolism, unsaturated fatty acid metabolism, amino acid metabolism, ceramide metabolism and other metabolic pathways. This study proved that the extract of V. jatamansi had definite pharmacodynamic effect on IBS-D model rats, and the mechanism was discussed from the perspective of fecal metabonomics.

[Effects of baldrinal of Valeriana jatamansi on expression of CRF, TPH1 mRNA and 5-HT in rats with irritable bowel syndrome].

This study aimed to investigate the effects of baldrinal of Valeriana jatamansi on the expression of corticotropin releasing factor (CRF) and tryptophan hydroxylase 1 (TPH1) mRNA and levels of 5-hydroxytryptamine (5-HT) in colon of rats with irritable bowel syndrome (IBS), and to explain its therapeutic mechanism on IBS through 5-HT pathway. Fifty-four male SD rats were randomly divided into 6 groups: blank group, model group, baldrinal high, medium and low dose groups, and pinaverium bromide group, n=9 in each group. The IBS rat models were established by using unpredictable chronic stress for 3 weeks followed by 1-hour acute restraint stress (CAS) after 7 days of rest and independent feeding. CRF expression was detected by IHC-P; TPH1 mRNA expression was detected by using RT-PCR and the 5-HT level was measured by high performance liquid chromatography(HPLC). The results indicated that the method of chronic stress with acute restrain stress method and independent feeding could lead to the increase in expressions of CRF and TPH1 mRNA and levels of 5-HT in IBS rats(P<0.05). The expressions of CRF, TPH1 mRNA and 5-HT in baldrinal groups were significantly lower than those in model group(P<0.05). The experimental results showed that IBS could result in increase in the expressions of CRF, TPH1 mRNA and levels of 5-HT, and the baldrinal of V. jatamansi could improve the symptoms of IBS by reducing the expressions of CRF, TPH1 mRNA and levels of 5-HT in colon of rats.

Phenolic constituents from Rheum nobile and their antioxidant activity.

Although the rhizomes of Rheum nobile Hook. f. et Thoms (Polygonaceae) are widely used in Tibetan medicine, no previous investigations regarding the biological activities and rarely chemical constituents of this plant have been reported. As part of an ongoing search for novel bioactive agents, a phytochemical investigation of R. nobile led to the isolation of two new compounds Rheumone B (1) and piceatannol-4'-O-ß-D-(6″-O-acetyl)-glucoside (2), together with 15 known compounds by gel filtration over Sephadex LH-20 and preparative HPLC. Their structures were determined by combined spectroscopic methods. Compounds 1-10 were evaluated for their ability to scavenge 2,2-diphenyl-1-picrylhydzyl (DPPH) radical and compounds 7-10 showed relatively strong scavenging abilities with IC50 values from 2.76 µM to 11.80 µM. In conclusion, naphthalene glycosides, stilbene glycosides, flavanols, especially anthraquinones are main chemical constituents of this plant. The ability to scavenge DPPH radical of compound 8 was the highest among compounds 1-10.

[Textual research for Tibetan medicine Qumazi].

Qumazi is a commonly used Tibetan medicine. With a long history, it can be found in the Four Medical Tantras written by gYu-thog rNying-ma Yon-tan mGon-po since the 8th century AD. Qumazi grows in mudflats and fields, including species growing in highlands, lowlands, mountains and farmlands. According to records in Crystal Beads Materia Medica, it features green sword-shaped leaves, thin stems with red veins, inserted panicles, white chicken-like flowers and copper needle row-like roots. However, there are many inconsistent morphological descriptions for Qumazi plants in many Chinese versions of Tibetan medicine books. In this article, after studying ancient and modern Tibetan medicine books, consulting experts and conducting surveys, the authors confirmed that Qumazi belongs to Rheum of Polygonaceae, including Rheum nobile Hook. f. et. Thoms, R. globulosum Gage, R. alexandrae Hook. f. et. Thoms, R. pumilum Maxim and R. delavayi Franch. In some regions, Qumazi is substituted by R. spiciforme Royle and R. przewalskyi Losinsk. After the Chinese version of Qinghai-Tibet Plateau Drug Illustrations was published in 1972, Qumazi has been miswritten as P. sibiricum Laxm in many Chinese versions of Tibetan medicine books, perhaps because P. sibiricum Laxm has many similar features with Qumazi as described in Crystal Beads Materia Medica and then is mistranslated from Tibetan to Chinese versions. According to records, Qumazi can reduce edema and is mainly applied to treat the minamata disease in clinic.

[Sequence analysis and identification of a chloroplast matK gene in Rhei Rhizoma from different botanical origins].

Rhei Rhizoma is a Chinese medicine with multiple botanical origins. There is a problem to identify it with conventional methods. To compare the characteristics of chloroplast matK gene sequences of different Rheum species and authenticate inspected species, the matK gene sequences of different species from different origins were amplified, cloned, and sequenced. Genomic DNA of Rheum plants was extracted using modified DNA extracted Kit and matK gene sequences were analyzed by ContingExpress, DNAman and MEGA5.0. The length of matK gene sequences of Rheum palmatum, R. tanguticum and R. officinale were 1 518 bp containing 57 variable loci. According to the mutation sites, R. palmatum, R. tanguticum and R. officinale were divided into different genotypes separately. Based on the established method according to the loci 587, 707, 838, we successfully identified the genuine Rheum species from its adulterants.

[Influence of iridoid from Valeriana jatamansi on 5-HT and 5-HIAA in rats with irritable bowel syndrome].

OBJECTIVE: To investigate the mechanism of iridoid from Valeriana jatamansi treating irritable bowel syndrome. METHOD: Sixty male SD rats were equally divided into 6 groups (2 controls, 1 model and 3 treatment doses) with 10 rats per group. The test groups were administered with iridoid (24.92, 12.46, 6. 23 mg x kg(-1)) while the control groups were administered with fluoxetine (2.5 mg x kg(-1), positive control) or distilled water (negative control). The model was established by chronic stress and independent feeding. The influence of iridoid from V. jatamansi on 5-HT and 5-HIAA in colon, serum and hypothalamic were observed in all groups. RESULT: In the model group, the content of 5-HT in colon and serum increased significantly, but the content of 5-HT in hypothalamic decreased significantly. The content of 5-HIAA and the value of 5-HT/5-HIAA had no significant change. In three iridoid-treated groups, the content of 5-HT in colon and serum decreased, but the content of 5-HT in hypothalamic increased. The content of 5-HIAA had no significant change. The value of 5-HT/5-HIAA in colon and serum reduced. CONCLUSION: The mechanism of iridoid from V. jatamansi treating irritable bowel syndrome may be related to the regulation effect to the levels of 5-HT from Gastrointestinal to central nervous system.

[Study on quality specification of Rhizoma et Radix Valeriana Jatamansi].

OBJECTIVE: To provide scientific basis for the utilization and development of Valeriana jatamansi by setting up the quality control specification of V. jatamansi. METHOD: The pharmacognostical methods were applied. The extract of V. jatamansi was examined. Moisture and ash were determined. And the bioactive constituents were analyzed by TLC and HPLC. RESULT: The morphological and histological characters of V. jatamansi were observed. Content of total ash, acid-insoluble ash, and moisture of 15 samples from different habitats and times were determined. The qualitative and quantitative analysis of valtrate and acevaltrate by TLC and HPLC were preformed respectively. CONCLUSION: The established method can be used for the quality control of V. jatamans.

[Determination of complanatoside A in semen Astragali complanati by HPLC].

OBJECTIVE: To establish the determination method for complanatoside A in seeds of Astragalus complanatus. METHOD: An HPLC method has been developed to separate complanatoside A on ZORBAX EXTEND-C18 (4.6 mm x 250 mm, 5 microm) column with acetonitrile-water-phosphoric acid (20:80:0.2) as mobile phase and UV detection at 267 nm. RESULT: The good linearity of complanatoside A ranged 0.086-0.430 microg, r = 0.9999. An average recovery of 99.8% (n = 5) was obtained with a RSD of 1.0%. CONCLUSION: The established method is proved to be stability, fast, accurate and can be used for quantification of Complanatoside A in Semen Astragali Complanati.

[Determination of quercetin-3-O-beta-D-glucopyranosyl-7-O-beta-D-gentiobioside in semen descurainiae by HPLC].

OBJECTIVE: To establish a method for the determination of quercetin-3-O-beta-D-glucopyranosyl-7-O-beta-D-gentiobioside in Semen Descurainiae. METHOD: HPLC was used with self-made quercetin-3-O-beta-D-glucopyranosyl-7-O-beta-D-gentiobioside as reference substances. RESULT: The average collection was 99.78%, RSD 2.4%. CONCLUSION: The method is appropriate for quality control of Semen Descurainiae.

[Identification and evaluation of TCM shayuanzi].

OBJECTIVE: To understand the varieties of Shayuanzi on sale and establish the method for evaluating its quality. METHOD: 22 samples collected from all over the country were identified by TLC and the contents of total flavone in them were determined with UV. RESULT: All samples could be classified into three varieties, and they could be identified by TLC. The contents of total flavone in them were 0.3%-1.0%. CONCLUSION: Shayuanzi on sale mainly includes Astragalus complanatus, and there are A. adsuragen and A. sinicus as well. TLC is a better method for the identification of shayuanzi. The total flavone in A. complanatus is 0.33%-0.82%.

[Identification on genuine and pseudo shayuanzi collected from different place by HPLC fingerprint].

OBJECTIVE: To establish the identification method for genuine and pseudo Shayuanzi by HPLC fingerprint. METHOD: The flavone, a kind of active composition in genuine (Astragalus complanatus) and analysed species was analysed by RP-HPLC. RESULT: The fingerprints of genuine Shayuanzi collected from different places showed the similar characteristics, but they were different from those of pseudo species A. adsuragen and A. sinicus. CONCLUSION: The HPLC fingerprint can be an identifing method for Shayuanzi.

[Study of cerebral protective effects of naosaitong in animals].

OBJECTIVE: To investigate the influence of Naosaitong (NST) on the cerebral blood flow (CBF), the infarct areas and blood rheology in animals. METHOD: NST's cerebral protective effects were investigated by using middle cerebral artery occlusion (MCAO), bilateral common carotid artery ligation, and carrogeenin-induced thrombus model rats, being administrated with medicine for seven days. RESULTS: Three dosage groups of NST increased CBF in anesthetized rabbits, reduced the infarct areas in MCAO rats, decreased the physical sign indexes, and water quantities. They increased the activities of Glutathione peroxidase (GPX) and Catalase (CAT), decreased the contractions of Lipid peroxidase (LPO) and Lactate (LD) in the cerebral ischemia-reperfusion rats; shortened the length of thrombus and improved the blood rheology in the carrogeenin-induced thrombus model rats, and prolonged hypoxia-resisting time in mice. CONCLUSION: NST can evidently increase CBF in rabbits, improve the cerebral edema brain tissues' injure and nervous physical sign indexes in the cerebral ischemia-reperfusion rats, reduce the infarct areas in MCAO rats, postpone thrombosis course and have antioxidation effects, which show that NST can obviously protect the brain tissues in the experimental cerebral infarct model rats.