Analysis of Chemical Composition in Sancao Baogan Decoction Based on UPLC-ESI-HRMSn / 中国实验方剂学杂志

Objective::To analysis the chemical constituents in Sancao Baogan decoction by ultra-high performance liquid chromatography coupled with hybrid quadrupole-orbitrap mass spectrometry (UPLC-ESI-HRMSn). Method::The separation was performed on an ACQUITY UPLC BEH C18 column (2.1 mm×100 mm, 1.7 μm) by a gradient elution of methanol (A)-0.1% formic acid solution (B) (0-2 min, 5%A; 2-20 min, 5%-12%A; 20-35 min, 12%-40%A; 35-38 min, 40%A; 38-48 min, 40%-80%A; 48-50 min, 80%A). The flow rate was 0.3 mL·min-1 and the column temperature was set at 30 ℃, the injection volume was 10 μL. Electrospray ionization was applied and the data were collected via positive and negative ion modes. By using Xcalibur 3.0 software, the chemical constituents were analyzed based on the relative retention time, excimer ion peak and fragment ion peak of the compounds, as well as comparison with human metabolome database (HMDB), reference substances and literature data. Result::A total of 40 chemical components were identified from Sancao Baogan decoction, including 16 phenolic acids, 19 flavonoids, 2 anthraquinones, 1 triterpenoid, 1 sterol, and 1 monoterpenoid. Six compounds (dansensu, α-pinene, epigallocatechin, 2, 5-dihydroxybenzoic acid, naringenin and emodin) were reported for the first time from Sancao Baogan decoction. Conclusion::The established UPLC-ESI-HRMSn can quickly, accurately and comprehensively analyze the chemical constituents of Sancao Baogan decoction, which lays a foundation for the basic research of pharmacodynamic substances and quality control of this formula.

CD11c+DCs promote the occurrence and development of psoriasis-like morbidity in K14-VEGF transgenic mice / 基础医学与临床

Objective To investigate the auxoaction and mechanism of CDllc+DCs on psoriasis. Methods CDllc+ DCs in the psoriasis-like lesions were confirmed by immunofluorescence double staining experiments. The CD1lc+ DCs were subcutaneously injected into the neck skin of transgenic mice without disease. The clinical features were observed and assessed with PASI score every day. The ear and back skin were checked by HE stained. At the same time, the T lymphocyte and DCs of bone marrow, spleen, submandibular lymph nodes and blood were analyzed by flow cytometry, and the T lymphocyte in the skin lesions were analyzed by immunofluorescence. The heart, liver, spleen, lungs and kindey were HE stained. Results In the psoriasis-like lesions, about 90％ CDllc+ cells expressed CDllc. HE test showed that the thickness of the skin layer was significant different between the experimental group (ear: 29±4 μm; back: 25±3 μm) and the control group(ear: 11±2 μm; back: 9±1 μm) (P＜0.01). CD3 + CD4+ T lymphocyte cells in the blood of the experimental mice (17.87％) were decreased significantly (P＜0.01) compared with the control group mice (31.77％). The numbers of Thl and Th17 cells from bone marrow, spleen, submaxillary lymph nodes and blood, as the same as CD1lc+DCs from bone marrow and submaxillary lymph nodes, were decreased in the experimental mice compared with the control group mice (P＜0.01). The number of CD4﹢T cells, CD8+T cells, IL-17a+cells and IFN-γ+cells in the skin lesions from the experimental group all were higher than that from the control group (P＜0.01). And the vessels in the lesions of the experimental group were higher than that in the control group (P＜0.01). Conclusions CD1 lc+CD1 lc+DCs may play an important role in the occurrence and development of psoriasis by increasing the T lymphocyte and the blood vessel in the skins of K14-VEGF transgenic mice.

Effect of tetramethylpyrazine on the responses of respiration and expression of nNOS in brainstem to hypoxia in rats / 生理学报

The aim of the present study was to investigate the effect of tetramethylpyrazine (TMP) on the changes of respiration and expression of neuronal nitric oxide synthase (nNOS) in brainstem induced by hypoxia in the rats. Hypoxia was induced by inhalation of 8% O2-balanced N2.The electromyogram (EMG) of diaphragm was monitored to evaluate the respiratory response of the rats to hypoxia. The immunohistochemical staining technique was used to study the change of the expression of nNOS in the brainstem during hypoxia. In the rats of hypoxia group, a successive process of response, excitatory followed by inhibitory, was produced. Twenty min after hypoxia, a significant inhibition of respiration occurred, which was characterized with a marked decrease in the inspiratory duration, the respiratory frequency, and the amplitude of inspiration and a prolongation of expiratory duration (P<0.05). In the rats of pretreated with TMP, the respiratory activity was not obviously depressed (P>0.05). In the rats of hypoxia group, the level of nNOS immunoreactivity was enhanced remarkably in the lateral reticular nucleus, nucleus of trapezoid, hypoglossal nucleus and the facial nucleus compared with the control group (P<0.05). In the rats of pretreated with TMP, the nNOS level increased further in the nuclei mentioned above (P<0.05). The results obtained indicate that TMP can reverse the inhibitory effect of hypoxia on respiration in the rats and that nNOS may be involved in the respiratory protective action of TMP.

Effect of stimulation of the facial nucleus on discharge of respiratory neurons in the pre-Bözinger complex and its neurotransmitter mechanism in rats / 生理学报

The experiments were carried out on adult Sprague-Dawley rats. We investigated the discharge response of respiratory neurons (RNs) in the pre-Bözinger complex (PBC) to electrical stimulation of the facial nucleus in which the motor neurons were retrogradely degenerated and the antagonistic effects of microiontophoresis of CNQX, bicuculline (BIC), strychnine (Stry) and atropine on the discharge responses of the neurons. In 12 rats with retrograde degeneration of the facial motor neurons, 116 RNs in the PBC ipsilateral to the facial nerve sectioned were extracellularly recorded. The response of pre-inspiratory (Pre-I) (24 / 26) and inspiratory (I) (30 / 35) neurons to the electrical stimulation of the facial nucleus was mainly excitatory, and the response of expiratory (E) (20 / 22) and inspiratory-expiratory phase-spanning (I-E) (28 / 33) neurons was mainly inhibitory. CNQX partially or completely block the excitatory effect of the stimulation on Pre-I (18 / 24) and I (23 / 27) neurons. Stry could partially or completely block the immediate transient inhibition on Pre-I (12 / 18) and I (14 / 23) neurons and the inhibitory effect on I-E (20 / 28) and E (9 / 16) neurons induced by the stimulation. BIC partially or completely blocked the inhibitory effect on I-E (22 / 25) and E (9 / 9) neurons induced by the stimulation. Atropine did not have obvious influence on the response of RNs to the stimulation. These results suggest that non-motoneurons in the facial nucleus may participate in the modulation of respiration by affecting the activities of RNs in the PBC and that Glu, GABA and Gly serve as neurotransmitters or modulators to regulate the activities of the RNs in the PBC and hence the rhythmic respiratory movement.