ABSTRACT
ObjectiveTo clarify the therapeutic effect of Huashi Baidu prescription on pneumonia in mice caused by influenza A （H1N1） virus and explore its mechanism based on the transcriptome. MethodA mouse influenza viral pneumonia model was built by intranasal infection with influenza A virus， and mice were continuously administered the drug for five days， so as to investigate the general condition， lung index， viral load， pathological morphology of lung tissue， survival time， and prolongation rate of survival time of mice and clarify the therapeutic effect of Huashi Baidu prescription on influenza viral pneumonia. Transcriptome technology was used to detect the differentially expressed genes in the lung tissue of mice in the model group and the normal group， as well as the Huashi Baidu prescription group and the model group， and the potential core target of the Huashi Baidu prescription for the treatment of influenza viral pneumonia was screened. Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） was used to verify the effect of Huashi Baidu prescription on the mRNA expression level of core target genes. ResultCompared with the normal group， the lung index and viral load in the lung tissue of the model group were significantly increased （P<0.05， P<0.01）. Compared with the model group， the high-dose group of Huashi Baidu prescription significantly prolonged the survival time of mice infected with influenza A virus （P<0.05） and significantly reduced the lung index value of mice （P<0.05） and the viral load of lung tissue. The high-dose， medium-dose， and low-dose groups of Huashi Baidu prescription could significantly reduce lung tissue inflammation， blood stasis， swelling， and other pathological changes in mice （P<0.05， P<0.01）. Transcriptome analysis of lung tissue showed that core genes were mainly enriched in the nuclear transcription factor-κB （NF-κB） signaling pathway， interleukin-17 （IL-17） signaling pathway， cytokine-cytokine receptor interaction， and other pathways after the intervention of Huashi Baidu prescription. TRAF6， NFKBIA， CCL2， CCL7， and CXCL2 were the top five node genes with combined score values. Real-time PCR validation showed that Huashi Baidu prescription significantly downregulated the mRNA expression of key genes TRAF6 and NFKBIA in the NF-κB signaling pathway， as well as chemokines CCL2， CCL7， and CXCL2 （P<0.05， P<0.01）. ConclusionHuashi Baidu prescription has a therapeutic effect on influenza viral pneumonia， possibly by inhibiting the expression of key nodes TRAF6 and NFKBIA in the NF-κB signaling pathway and that of chemokines CCL2， CCL7， and CXCL2， reducing the recruitment of inflammatory cells and viral load， and exerting anti-influenza viral pneumonia effects.
ABSTRACT
Aim The TRPV1 plasmid was transiently transfected into human embryonic kidney HEK 293T cells to establish the heterologous expression system of TRPV1-channel. Methods The transfection efficiency was confirmed under fluorescence mi-croscope and the TRPV1 protein expression was identified by u-sing Western blot, and the functional characteristics of the chan-nel were studied by using the method of confocal microscopy. Results The transfection rate could reach 40% ~50%; the transfected cells were found to have a clear band at the corre-sponding position that TRPV1 should be, which indicated that TRPV1 channel protein was expressed in the transfected cells. The confocal microscopy imaging result showed that the trans-fected HEK 293T cells were activated by TRPV1 channel ago-nist. Conclusion Transient transfection of HEK 293T cells with TRPV1 channel is successfully constructed and the heterol-ogous TRPV1 channel is verified to have normal calcium-media-ting function.
ABSTRACT
Objective To study the effect of black granule capsules(BGCs) on growth of transplanted mouse hepatocarcinoma cells, cell proliferation cycle and apoptosis.Methods KM mouse were subcutaneously inoculated with Hepatocarcinoma cells (H22) and randomly divided into the model control group, the positive control group, the low, medium and high does of BGC group after 24h. The positive control group received intraperitoneal injection with 30 mg/kg cyclophosphamide. Mice of BGC groups were intragastricaly with different dosage of BGC (400, 800, 1 600 mg/kg). The model control group received intragastricaly with normal saline. The drugs were administrated once a day for seven days. The tumor inhibition rate was calculated at 24 h after the last administration. Flow cytometry was used to detect changes of cell cycle and apoptosis in harvested H22 tumor cells.Results The group of high does showed significant inhibitory effect on the growth of transplanted H22 tumor cells withthe inhibitory rate 38.78% (male) and 43.57% (female). Compared with model control group, groups of different dosages decreased time of G0-G1 phase (58.06% ± 9.65%, 55.10% ± 5.89%, 61.36% ± 15.95%vs. 74.47% ± 2.63%), increased tiem of Sphase (33.96% ± 11.90%, 32.67% ± 4.04%, 33.89% ± 9.82%vs. 14.37% ± 4.92%), and increased the apoptosis rate (31.12% ± 1.85%, 31.89% ± 2.16%, 40.64% ± 0.55%vs.21.75% ± 2.64%).Conclusion BGC has antitumor effect on mouse hepatocarcinoma H22 tumor cells, and its mechanism was to regulate cell proliferation cycle and induce the apoptosis.
ABSTRACT
Aim To further study the molecular mecha-nism of the herbs with hot nature on the regulational action on TRPV1 channel based on the 7900 Real-time PCR instrument. Methods 7900 PCR instrument was applied to detect the intracellular flurescence of TRPV1 channel in the dorsal root ganglions ( DRG ) neurons and the effect on the TRPV1 ’ s thermo-sensational functions of the selected 11 ingredients from hot herbs was explored. Results TRPV1 channels could be ac-tivated by gradually elevated temperature; the activa-tion process could be blocked by the TRPV1 specific blocking agent capsaizepine. Most of the ingredients from hot-nature herbs had the potential to up-regualate TRPV1 channel function. Conclusions The estab-lished TRPV1 channel detection system based on PCR instrument is suitable for the analysis of regulational functions of drugs on the heat-activated TRPV1 chan-nel;the functions of hot herbs may be related to the up-regualtional effects of its active ingredients on the TRPV1 channel which will further up-regulate energy metabolism.
ABSTRACT
The epoxy chloropropane Kelch sample related protein-1(Keap1)-nuclear factor erythroid-2 related factor(Nrf2)/antioxidant response element(ARE)signal pathway is of critical importance in cellular antioxidant response. The induction of down?stream phaseⅡmetabolic enzymes and antioxidant protein/enzyme offers cellular protection under oxidative stress. Research of Nrf 2 protein has gained much attention in recent years. This article reviewes Keap1-Nrf2/ARE signal pathway in three perspectives:the ba?sic structure including the structure of Nrf2,Keap1 and ARE,their involvement in anti-oxidative actions including the basic function of Keap1-Nrf2/ARE signal pathway and the downstream activation of phaseⅡmetabolic enzymes and antioxidant protein/enzyme,as well as their regulation mechanisms,such as decoupling,degradation,the latch and the hinge.
ABSTRACT
ObjectiveTo investigate the effects of Saposhnikovia divaricata extract combined with arsenic trioxide (ATO) on the proliferation and apoptosis in chronic myelogenous leukemia K562 cells. MethodsSaposhnikovia divaricata extract was prepared.Cultured K562 cells were treated with different concentration of Saposhnikovia divaricataextract or/and ATO for 48h. Cell proliferation was determined using the MTT assay. Apoptosis and cell cycle were detected using flow cytometry.ResultsThe MTT assay showed that Saposhnikovia divaricata extract of 750,1 000,1 250,1 500 μg/ml had a significantly proliferation inhibitory effect compared with control group, the inhibitory rates were 23.29% ± 3.31%, 48.30% ± 2.50%, 79.62% ± 3.41% and 88.94% ± 0.06%, respectively (allP<0.05); Saposhnikovia divaricata extract of 500 μg/ml combined with ATO of 1.0, 0.5 μg/ml significantly increased inhibitor rates compared with ATO of 1.0, 0.5 μg/ml (64.99% ± 5.18%vs. 44.48% ± 3.31%,38.59% ± 3.88%vs.26.30% ± 5.03%; allP<0.05). FCM showed that Saposhnikovia divaricata extract of 500 μg/ml combined with ATO of 2.0, 1.0, 0.5 μg/ml significantly increased apoptotic rate compared with ATO group of 2.0, 1.0, 0.5 μg/ml (33.97% ± 0.59%vs.20.97% ± 2.17%, 13.53% ± 0.47%vs.9.77%±0.64%、6.63%±&0.40%vs.4.00%±0.46%; allP<0.05 ). Cell cycle results showed that Saposhnikovia divaricata extract of 500μg/ml combined with ATO of 2.0,1.0, 0.5μg/ml significantly increased the rate of S phase compared with ATO group of 2.0, 1.0, 0.5 μg/ml (60.25 ± 2.59%vs.55.61 ± 1.28%, 60.89 ± 1.53%vs.37.96 ± 1.02%, 47.76 ± 0.87%vs.39.90 ± 0.92%; allP<0.05).ConclusionsSaposhnikovia divaricataextract could obviously inhibit the proliferation of K562 cells and enhance the apoptotic effect of ATO. ATO could induce a G2/M phase arrest, while Saposhnikovia divaricata extract combined with ATO could induce a S phase arrest in K562 cells.
ABSTRACT
To further uncover the scientific significance and molecular mechanism of the Chinese herbs with pungent hot or warm natures, endogenous and exogenous expression systems were established by isolation of dorsal root ganglion (DRG) neurons and transfection of HEK293 cells with TRPV1 channel gene separately. On this basis, the regulation action of capsaicin, one main ingredient from chili pepper, on TRPV1 channel was further explored by using confocal microscope. Besides, the three-sites one-unit technique and method were constructed based on the brown adipose tissue (BAT), anal and tail skin temperatures. Then the effect of capsaicin on mouse energy metabolism was evaluated. Both endogenous and exogenous TRPV1 channel could be activated and this action could be specifically blocked by the TRPV1 channel inhibitor capsazepine. Simultaneously, the mice's core body temperature and BAT temperature fall down and then go up, accompanied by the increase of temperature of the mice's tail skin. Promotion of the energy metabolism by activation of TRPV1 channel might be the common way for the pungent-hot (warm) herbs to demonstrate their natures.
ABSTRACT
Bitter taste receptors (BTRs) belong to the G protein-coupled receptors, which included 25 subtypes. BTRs, which were distributed in the oral cavity, mediated the bitter taste of mammalian. Furthermore, BTRs were also existed in the extra-oral digestive system such as the stomach and intestine to influence the digestion, absorption and energy regulation. It also can relax airway smooth muscles in the respiratory system and decrease blood pressure in the cardiovascular system. While being strikingly conformed to the latest advancements of BTRs, the efficacy of bitter taste property of Chinese materia medica (CMM) such as“excretion”,“dryness” and“strengthening” have been used in ancient theories of property and flavor in traditional Chinese medicine (TCM) for thousands of years in the treatment of multisystem diseases such as digestive, endocrine, respiratory and cardiovascular system. Therefore, applying modern techniques and new methods in the interpretation of the scientific connotation of bitter taste property of CMM based on BTRs should be a feasible way and a new research pattern. It played an important role in the enriching of the property theory in CMM, as well as enhancing the modernization and objectivization of CMM.
ABSTRACT
Acute promyelocytic leukemia (APL) is a distinctive subtype of acute myeloid leukemia with a distinct biology and clinical presentation. Its molecular biology characteristic is a aberrant chromosomal translocation of the promyelocytic leukemia (PML) gene on chromosome 15 and the retinoic acid receptor α(RARα) gene on chromosome 17. This translocation generates PML-RARα fusion protein, which plays an important role in the genesis, development, diagnosis and therapy of APL. The PML protein has a close relationship with PML-RARαfusion gene. This article mainly summarizes the character, the function of PML protein and the degradation pathway of PML-RARα.
ABSTRACT
<p><b>OBJECTIVE</b>To study the effects of the ingredients from Chinese herbs with the nature of cold or hot on the expression of TRPV1 and TRPM8.</p><p><b>METHOD</b>The effects of ingredients from herbs on primary culture DRG neurons are observed in vitro. The expression quantity of gene is detected by the method of real time PCR. the 2 (-deltadeltaCT) method is applied to analyze the data.</p><p><b>RESULT</b>Ingredients from herbs with the nature of cold up-regulate the expression level of TRPV1 and down-regulate that of TRPM8, especially under the temperature condition of 39 degrees C; while ingredients from herbs with the nature of hot up-regulate the expression level of TRPM8 and down-regulated that of TRPV1, which is more significant under the temperature condition of 19 degrees C.</p><p><b>CONCLUSION</b>The regulatory changes of TRPV1 and TRPM8 mRNA expression induced by the chemical ingredients might be related to the cold and hot natures of the herbs from which the ingredients are extracted. And this could be one of the therapeutic mechanisms for the treatment of Chinese herbal medicines to cold- and heat-related diseases.</p>
Subject(s)
Animals , Male , Rats , Drugs, Chinese Herbal , Gene Expression , Rats, Sprague-Dawley , TRPM Cation Channels , Genetics , Metabolism , TRPV Cation Channels , Genetics , MetabolismABSTRACT
AIM: To observe the changes of adengl cyclase(AC) and phosphodiesterase(PDE) activities of at different time point in hypothalamus of rats with fever and hypothermia. METHODS: Radioisotope method was used to measure the activity of AC and PDE. RESULTS:The fresh yeast caused rats fever after subcutaneous injection 4h( P
ABSTRACT
Increasing evidence suggests that a complex net-work of fever induction pathways in mammalian exists. In this article,the overview of recent studies on the mechanism of fever induced by different pyrogens using IL-1, IL-1R, ICE, IL-1ra, IL-1RacP, IL-6,IL-10,TNFR,cPLA 2,COX,EP,AT 2,iNOS and D 2/3 knockout mice is presented. Hyperthermia respond to localized infection/inflammation(e.g.,sc injection of turpentine) is mediated by IL-1? and IL-6 in turn.While fever induced by systemic infection/inflammation(e.g.,treatment with LPS intraperitoneally) varies with the different doses of pyrogens administered .Fever caused by a low dose of LPS administered ip is IL-6 dependent ,but the IL-6 independent pathway is crucial for the fever evoked by a high dose of LPS.Febrile responses during both local and systemic infection/inflammation develop totally through central PGE 2 dependent mechanism, but some stress induced hyperthermia otherwise.
ABSTRACT
The effect of Sin on the transmission of the impulses through isolated rabbit superior cervical sympathetic ganglia in comparison with that of curare by the sucrose - gap method was studied.Inhibi -tory effect of Sin on the action potential was dose-dependent but no effect on the excitability and conductivity of preganglionic nerves. The ID50 of Sin and curare were 1.2 mM and 0.2 mM, respectively. The effect of Sin was reduced in high Ca2+ concentration solution, and enhanced under Ca2+ free solution. Neostigmine antagonized the effect of Sin.
ABSTRACT
On the isolated rat phrenic nerve-diaphragm preparation Sin blocked the neuromuscular transmission without affecting the excitability & conductivity of phrenic nerve. Inhibitory effect of Sin on indirect contraction of diaphragm was dose-dependent, increasing with increasing dose & was antagonized by high Ca2+ solution. When the transmission was blocked by Sin, the ACh sensitivity of the chick biventer cervicis muscle was reduced. Like succinylcholine Sin induced the contractile response of the toade rectus abdominis. Neostigmine did not antagonize, but enhanced the blocking effect of neuromuscular transmission of Sin. These results indicate that Sin has some characteristics of depolarizing muscular relaxants.
ABSTRACT
AIM: To observe the changes of AC activity and content of cAMP at different time point in hypothalamus of rats with fever and hypothermia. METHODS: Radioisotope method was used to measure the enzymatic activity of AC and the content of cAMP. RESULTS:(1)The fresh yeast caused fever after making model 4 h( P