Hyperhubeins A-I, Bioactive Sesquiterpenes with Diverse Skeletons from Hypericum hubeiense.

Nine new sesquiterpenes, hyperhubeins A-I (1-9), and 14 known analogues (10-23) were isolated from the aerial portions of Hypericum hubeiense. Their structures and absolute configurations were determined unambiguously via spectroscopic analysis, single-crystal X-ray diffraction, and electronic circular dichroism calculations. Compounds 1-3 possess an unprecedented sesquiterpene carbon skeleton. Further, a plausible biosynthetic pathway from farnesyl diphosphate (FPP) is proposed. The isolated phytochemicals were evaluated for neuroprotective and anti-neuroinflammatory properties in vitro. Compounds 1, 2, 5-8, 14, and 21 displayed notable neuroprotective activity against hydrogen peroxide (H2O2)-induced lesions in PC-12 cells at 10 µM. Additionally, compounds 1, 2, 12, and 13 exhibited inhibition of lipopolysaccharide (LPS)-induced nitric oxide (NO) production in BV-2 microglial cells, with their IC50 values ranging from 4.92 to 6.81 µM. Possible interactions between these bioactive compounds and inducible nitric oxide synthase (iNOS) were predicted via molecular docking. Moreover, Western blotting indicated that compound 12 exerted anti-neuroinflammatory activity by suppressing LPS-stimulated expression of toll-like receptor-4 (TLR-4) and inhibiting consequent activation of nuclear factor-kappa-B (NF-κB) signaling.

Identification of Forsythia suspensa (Thunb.) Vahl in different harvest periods using intelligent sensory technologies, HPLC characteristic fingerprint coupled with chemometrics.

INTRODUCTION: Forsythia suspensa (Thunb.) Vahl (FS), the fruit of Oleaceae plants, as a large part of traditional Chinese medicine, is classified as "Qingqiao (Q)" and "Laoqiao (L)" based on the harvest time. Because the maturation of FS is a gradual process, its accurate identification based on different maturity levels is an important issue. OBJECTIVES: We suggest colorimetric, electronic tongue, and high-performance liquid chromatography (HPLC) characteristic fingerprints to discriminate FS in different harvest periods. MATERIAL AND METHODS: First, FS fruits from different harvest times were collected, and then, their colour parameters, E-tongue sensory properties, HPLC characteristic fingerprints, and contents of nominal ingredients were determined. Finally, multivariate statistical analyses, including three-dimensional scatter plots, hierarchical cluster, principal component, linear discriminant, similarity, and partial least squares discriminant analyses were performed. RESULTS: The results demonstrated that the three experimental techniques could effectively discriminate FS based on different harvest times with 100% accuracy. Under the qualitative conditions, nine common peaks were identified in the HPLC fingerprints of 60 samples, among which, six peaks [variable importance in projection (VIP) > 1] could be used as index peaks for qualitative identification. In fact, the contents of quality marker components, including forsythin, phillygenin, rutin and forsythoside A, were significant different (P < 0.001) at different harvest times. Interestingly, the quality markers not only accurately reflected the maturity of FS but also showed close correlations with the colour parameters and sensory E-tongue responses. CONCLUSION: In our present investigation, bionic technologies, including a colorimeter, E-tongue analysis, and HPLC characteristic fingerprints, combined with chemometrics, were employed to develop a novel and accurate method for discriminating FS based on different harvest times.

Iridoid constituents from the male flower of Eucommia ulmoides and their promotion proliferation on ESF-1.

Four new iridoids (1, 2, 12, and 13), together with nine known iridoids (3-11), were isolated from the male flowers of Eucommia ulmoides Oliver and were characterized as 3ß-methoxyartselawnin C (1), 6ß-hydroxyl-1ß,3ß-dimethoxyartsclaenin III (2), 3,4-dihydro-3ß-ethoxyasperuloside (12) and 3,4-dihydro-3ß-ethoxydesacetylasperuloside (13) by extensive analyses of their 1D and 2D NMR spectroscopy and mass spectra. All of the isolated compounds were assayed for the promoting proliferation of skin fibroblasts cell (ESF-1) and compounds 4 and 7 (5 µM) significantly stimulated the proliferation of ESF-1 cells.

[Triterpenes constituents from male flowers of Eucommia ulmoides].

Nine triterpenes compounds were isolated from the male flowers of Eucommia ulmoides by recrystallization and chromatographic techniques over silica gel, Sephadex LH-20, and RP-18 gel. Their chemical structures were identified on the basis of spectral analysis and as 3-oxo-12-en-ursane-28-O-α-L-arabinofuranosyl (1 --> 6) -ß-D-glucopyranoside (1), 2α, 3ß-dihydroxyurs-12-en-28-oic acid(28 --> 1) -ß-D-glucopyranosyl ester (2), ursolic acid (3), α-amyrin (4), uvaol (5), ursolic acid acetate (6), 3-O-acetate oleanoic acid (7), betulinic acid (8), and betulinol (9). Compound 1 was a new compound, and compounds 2, 4-7 were isolated from the Eucommiu genus for the first time. Cytotoxic activity was tested for all the compounds against K562 and HepG2 cells. The results showed that only compound 3, exhibited cytotoxic activity.

[Effects of electroacupuncture on the expressions of neuroal nitric oxide synthase and astrocyte in dentate gyrus of rats with Parkinson's disease].

OBJECTIVE: To observe the changes in the expression of neuroal nitric oxide synthase (nNOS) and glial fibrillary acidic protein (GFAP) in dentate gyrus (DG) of rats with Parkinson's disease (PD) and effects of electroacupuncture (EA). METHODS: On the 7th day of stereotactic injection with 6-hydroxydopamine into right medial forebrain bundle, apomorphine-induced rotation was carried out to select the PD rats. The selected 12 rats were randomly divided into a model group (n = 6) and an EA group (n = 6). Moreover a normal group was established (n = 6). Then, the rats in EA group were treated with EA at "Hegu" (LI 4) and "Taichong" (LR 3), once a day for 21 days, and the other groups without any treatment. After EA treatment, the expressions of nNOS and GFAP in right DG were examined by immunohistochemistry method. RESULTS: The expression of nNOS in the right DG was weak in normal group, and the expression of nNOS in model group was significantly higher than that in normal group (P < 0.01), while in EA group, it was significantly lower than that in model (P < 0.01), with no difference between EA group and normal group (P > 0.05). The expression level of GFAP in model group was significantly higher than that in normal group (P < 0.01), while there was no difference in the number of GFAP positive cells between the above two groups (P > 0.05), and the number of GFAP positive cells in EA group was significantly increased compared with that in model group (P < 0.01), while with no significant difference in the expression level of GFAP between EA group and model group (P > 0.05). CONCLUSION: EA can reverse the increase of nNOS expression and promote the activation of astrocyte in DG on the injured side in rats with PD.

[Effect of electroacupuncture on expression of proliferating cell nuclear antigen and glial fibrillary acidic protein in subventricular zone of Parkinson's disease rats].

OBJECTIVE: To observe the effect of electroacupuncture (EA) on the expression of proliferating cell nuclear antigen (PCNA) and glial fibrillary acidic protein (GFAP) in the subventricular zone (SVZ) of Parkinson's disease (PD) rats, so as to explore its mechanisms underlying improvement of PD. METHODS: Eighteen SD rats were randomized into normal control, model and EA groups (6 rats/group). PD model was duplicated by microinjection of 6-hydroxydopamine (2.5 microg/microL, 6 microL) containing 0.02% antiscorbic acid into the right medial forebrain bundle region (P: -4. 0, L: 0.8, H: 8.0). On the 7th day after modeling, EA (100 Hz) was applied to bilateral "Hegu" (LI 4) and "Taichong" (LR 3) for 30 min, once a day for 21 days. The expression levels of tyrosine hydroxylase (TH), GFAP and PCNA in the right SVZ tissue were detected by immunohistochemistry. RESULTS: TH expression was strong in the normal control group, but no TH expression was found in the model and EA groups. In comparison with the normal control group, the expression levels of PCNA and GFAP proteins and the number of PCNA-IR-positive cells in the right SVZ were decreased significantly in the model group (P < 0.01); while compared with the model group, the expression levels of PCNA and GFAP proteins and the number of PCNA-IR-positive cells in the right SVZ were up-regulated considerably in the EA group (P < 0.01, P < 0.05). CONCLUSION: EA can up-regulate the expression of PCNA and GFAP proteins in the SVZ in PD rats without changes of TH expression, which may contribute to its effect in improving clinical symptoms of PD.