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The present study investigated the effect of active components of Descurainia sophia on allergic asthma and explored the underlying mechanism. SD male rats were randomly divided into a normal group(NC), a model group(M), a D. sophia decoction group(DS), a D. sophia fatty oil group(FO), a D. sophia flavonoid glycoside group(FG), a D. sophia oligosaccharide group(Oli), and a positive drug dexamethasone group(Y). The allergic asthma model was induced in rats by intraperitoneal injection of ovalbumin(OVA) and aluminum hydroxide gel adjuvant(sensitization) and atomization of OVA solution(excitation). After modeling, asthma-related indicators, tracheal phenol red excretion, inflammatory cell levels in the peripheral blood, lung permeability index(LPI), and oxygenation index(OI) of rats were detected. The pathological changes of lung tissues were observed by HE staining. Enzyme-linked immunosorbent assay(ELISA) was used to detect the content of inflammatory factors immunoglobulin E(IgE), interleukin-4(IL-4), and interferon-γ(IFN-γ) in the bronchoalveolar lavage fluid(BALF) and the content of endothelin-1(ET-1) and angiotensin-converting enzyme(ACE) in lung tissue homogenate. The serum content of nitric oxide(NO) was detected by colorimetry. Western blot was employed to determine the protein expression of Toll-like receptor 4(TLR4), nuclear factor κB-p65(NF-κB-p65), phosphorylated NF-κB-p65(p-NF-κB-p65), myosin light chain kinase(MLCK), vascular endothelial cadherin(VE cadherin), connexin 43, and claudin 5, and the mechanism of active components of D. sophia on allergic asthma was explored. As revealed by the results, the M group showed extensive infiltration of inflammatory cells around the bronchus of the lung tissues of the allergic asthma rats, thickened bronchial wall, severely deformed alveolar structure, increased number of wheezes, the content of IgE, IL-4, ET-1, and ACE, inflammatory cells, and LPI, and reduced latency of asthma, tracheal phenol red excretion, IFN-γ, NO content, and OI. After the intervention of the active components of D. sophia, the DS, FO, FG, Oli, and Y groups showed improved asthma-related indicators, tracheal phenol red excretion, and lung tissue lesions in allergic asthma rats, and the effects in the FO and Oli groups were superior. The content of inflammatory factors in BALF was recovered in the DS, FO, and Y groups and the FG and Oli groups. The number of inflammatory cells in rats was reduced in the DS and FO groups, and the FG, Oli, and Y groups to varying degrees, and the effect in the FO group was superior. DS, FO, Oli, and Y reduced ET-1, ACE, and LPI and increased NO and OI. FG recovered NO, ET-1, ACE, LPI, and OI to improve lung epithelial damage and permeability. Further investigation of inflammation-related TLR4/NF-κB pathways, MLCK, and related skeleton protein levels showed that TLR4, NF-κB-p65, p-NF-κB-p65, and MLCK levels were increased, and VE cadherin, connexin 43, and claudin 5 were reduced in the M group. DS, FO, FG, Oli, and Y could reduce the protein expression related to the TLR4 pathway to varying degrees, and regulate the protein expression of MLCK, VE cadherin, connexin 43, and claudin 5. It is inferred that the active components of D. sophia improve lung permeability in rats with allergic asthma presumedly by regulating the TLR4/NF-κB signaling pathway to improve airway inflammation, mediating MLCK and connexin, and regulating epithelial damage.
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Animals , Male , Rats , Asthma/drug therapy , Bronchoalveolar Lavage Fluid , Inflammation/metabolism , Lung , PermeabilityABSTRACT
OBJECTIVE@#To investigate the effects of methanol-ethyl acetate partitioned fractions from (MEDS) on the proliferation and apoptosis of human non-small cell lung cancer H1975 cells.@*METHODS@#The systemic solvent extraction method was used to preliminary separation of the effective fractions in the methanol extract of . The cytotoxicity of each extract (5, 10, 20, 40, and 80 μg/mL) was tested using MTT assay. Colony cloning method was used to assess the effect of different concentrations of methanol-ethyl acetate partitioned fractions from MEDS (5, 10, 20, 40, and 80 μg/ mL) on the proliferation of H1975 cells. Flow cytometric analysis with Annexin V-FITC/PI staining was performed to detect the apoptosis of the cells after treatment with different concentrations of MEDS fractions (10, 20, and 40 μg/mL). Western blotting was used to evaluate the effects of MEDS fractions on the expressions of apoptosis-related proteins Akt, Bax, and Bcl-2. The anti-tumor activity of 100 mg/kg MEDS fractions was tested in a nude mouse model bearing H1975 cell xenografts.@*RESULTS@#MTT assay and colony forming experiment showed that MEDS fractions significantly inhibited the proliferation of H1975 cells in a dose-and time-dependent manner ( < 0.05). The results of flow cytometry showed that MEDS fractions induced obvious apoptosis of H1975 cells in a concentration-dependent manner ( < 0.05). MEDS fractions also significantly decreased the expressions of Bcl-2 and Akt protein and increased the protein expression of Bax ( < 0.05). In the tumor-bearing nude mouse model, MEDS fractions showed potent anti-tumor effects with a low toxicity to affect the body weight and organs of the mice.@*CONCLUSIONS@#The methanol-ethyl acetate partitioned fractions from MEDS show potent anti-tumor activity both and , suggesting their value as promising therapeutic agents against lung cancer.
Subject(s)
Animals , Humans , Mice , Acetates , Antineoplastic Agents, Phytogenic , Pharmacology , Apoptosis , Carcinoma, Non-Small-Cell Lung , Pathology , Cell Line, Tumor , Cell Proliferation , Heterografts , Lung Neoplasms , Pathology , Methanol , Mice, Nude , Plant Extracts , PharmacologyABSTRACT
Objective To study the protective effect of aqueous extract from Descurainia Sophia (DS) on H2O2-induced H9c2 cardiomyocyte injury and to initially explore the potential mechanism. Methods The peaks of main components in DS were analyzed and identified by HPLC-MS. H9c2 cell injury model was established by H2O2. H9c2 cells were cultured in vitro and divided into control group, model group, probucol group, and DS at 100, 200, 400 μg/mL groups. In order to reveal the possible molecular mechanisms, the viability of H9c2 cells was measured by MTT assay; The apoptosis rate, autophagy rate, mitochondrial membrane potential, and reactive oxygen species (ROS) level were detected by flow cytometry; The relative indicators of cell oxidative stress were determined by biochemical kit; The expression levels of apoptosis-related protein and the autophagy-related protein were evaluated by Incell-western method. Results Seven components with the highest content were identified in DS through the results of mass spectrometry. Compared with the model group, DS can improve the cell viability (P < 0.05, 0.01) and survival rate of H9c2 cells (P < 0.01); At the same time, apoptosis was attenuated (P < 0.01), mitochondrial membrane potential was upregulated (P < 0.01), apoptosis related proteins Caspase-3, Bax/Bcl-2 were obviously downregulated (P < 0.01), autophagy phenomenon was attenuated (P < 0.01), autophagy related proteins LC3B and p62 were upregulated (P < 0.01). In addition, ROS level was decreased (P < 0.01), T-SOD and GSH-PX were upregulated and the levels of LDH and MDA were significantly decreased (P < 0.01). Conclusion This study suggests that DS can effectively protect H2O2-induced H9c2 cells injury, and the mechanism may be associated with improving oxidative stress in cells, inhibiting cell apoptosis and autophagy, which may be related to flavonoid glycosides.
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Objective: To explore the mechanism of QGG from Descurainia sophia on the airway inflammation and remodeling in COPD rats. Methods: SD Rats were randomly divided into six groups, including control, model, QGG high (QGGH, 0.20 g/kg), medium (QGGM 0.15 g/kg), low (QGGL 0.10 g/kg) groups and aminophylline group (0.09 g/kg). Rats with COPD in the stable phase we established by exposure to tobacco smoking combined with persistent bacterial infections for eight weeks. Rat in each group were ig administrated for twenty-eight days successive five weeks after smoking. Quantitative RT-PCR was applied to determine the mRNA expression of Th17 and Treg specific transcription factors RORγt and FOXP3. Ratio of Th17/Treg was tested by flow cytometry. Apoptosis cells of lung tissue were assayed by TUNEL. The expression of apoptotic related proteins cyctoC, Caspase-9, Caspase-3, bax, bcl-2, nucleus NF-κBp65, and cytoplasm NF-κBp65 in protein extracts of lung tissue were determined by Western blotting. Results: The growth rate of control group increased obviously with weeks, however, that of model group increased slowly in COPD establishment stage and the following smoking stage. The growth rate in QGGH and aminophylline groups increased quickly from the fifth week but in QGGL and QGGM groups increased slowly. The expression of RORγt and FOXP3 was increased in model group (P < 0.05). Compared with the model group, the expression of RORγt and FOXP3 was decreased in the QGGH and positive groups (P < 0.05). The expression of CD4+IL-17+/CD4+, FOXP3+CD4+/CD4 and the ratio of CD4+IL-17+/FOXP3+CD4+ in peripheral blood was significantly increased in the model group (P < 0.05). Compared with that of model, the expression of CD4+IL-17+/CD4+, FOXP3+CD4+/CD4 and the ratio of CD4+IL-17+/FOXP3+CD4+ were decreased in the QGGH group (P < 0.05). The expression of CD4+IL-17+/CD4+, FOXP3+CD4+/CD4 was decreased in the positive group, but the ratio of CD4+IL-17+/FOXP3+CD4+ was not changed. Compared with the control group, the apoptosis of lung tissues in the model group was significantly increased. Compared with the model group, apoptosis in the QGGH and positive groups was decreased (P < 0.05). The expression of bax, cytoC, Caspase-9, Caspase-3, and nucleus NF-κBp65 were significantly increased in the lung tissues of model rats, while that of bcl-2 and cytoplasm NF-κBp65 were decreased (P < 0.05). Compared with the model group, the expression of bax, cytoC, Caspase-9, Caspase-3, and nucleus NF-κBp65 in QGGH and positive groups were obviously decreased (P < 0.05), but that of bcl-2 and cytoplasm NF-κBp65 increased (P < 0.05). Conclusion: QGG can significantly improve the survival conditions, growth ratio and pulmonary functions of rats. QGG can reduce the ratio of CD4+IL-17+/FOXP3+CD4+ in peripheral blood. It can modulate Treg and Th17 specific transcriptional factors FOXP3 and RORγt, balance the ratio of Th17/Treg. QGG can reduce the apoptosis of lung tissues, repair damaged tissue, and maintain the integrity of organ.
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OBJECTIVE: To study the protective effects and mechanisms of aqueous extract from Descurainia sophia on doxorubicin(Dox)-induced cardiomyocyte injury.METHODS: The Dox induced H9c2 cell apoptotic model was established, then the cells were divided into normal group (NC), model group (Dox), positive group (resveratrol,RSV), and different doses of aqueous extract of Descurainia sophia (DS). The viability of H9c2 cells was detected by MTT assay. The apoptosis rate, mitochondrial membrane potential and reactive oxygen species (ROS) level were detected by flow cytometry. The levels of T-SOD, LDH, MDA and GSH-PX were measured. And the protein expression levels of caspase-3, Bcl-2, Bax and p53 were detected by western blot, HPLC-MS identification of aqueous extract from Descurainia sophia.RESULTS: After treating with DS products, the survival rate of H9c2 was increased (P<0.01), the apoptosis rate was significantly decreased (P<0.01),mitochondrial membrane potential was significantly increased (P<0.01), the level of ROS was significantly decreased (P<0.05), T-SOD and GSH-PX activities were significantly increased (P<0.01), the levels of LDH and MDA content were significantly decreased(P<0.01). Moreover, DS reduced the expression of caspase-3 (P<0.01), regulated the expression of Bax/Bcl-2 (P<0.01), decreased the expression of p53 (P<0.05). Seven components were identified from DS by HPLC/MS analysis.CONCLUSION: DS can effectively protect cardiomyocyte, and its mechanism is probably associated with correcting functional disorders of oxidative stress of cardiomyocytes, and inhibit mitochondrial apoptotic pathway.
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OBJECTIVE:To analyze the volatile components in Descurainia sophia and Lepidium apetalum and compare its dif-ferences. METHODS:HS-SPME was conducted for extracting volatile components in D. sophia and L. apetalum,GC-MS was used for detecting components,and area normalization method was adopted for calculating relative content of each component. RE-SULTS:The volatile components in D. sophia and L. apetalum were 25 and 18,accounting for 75.76% and 64.29% of total vola-tile components,respectively,and chemical components with the highest contents were β-caryophyllene and O-tolunitrile. CON-CLUSIONS:The method is simple,reliable,and can be used for the analysis of volatile components in D. sophia and L. apetalum. The volatile components show great differences in the kinds and contents,the study can provide basis for rapid identification of D. sophia and L. apetalum.
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This article was aimed to study the immunomodulatory effect of ethanol sediments of the seeds of Descurainia sophia(L.) Webb. ex Prantl. both in vitro and in vivo. The lymphocyte proliferation test in vitro was carried out to explore the effect of the ethanol sediments on the proliferation of T cell and B cell in the spleen of normal mice. And, the carbon clearance test, serum hemolysin test, and delayed-type hypersensitivity test were used to investigate the influence of fraction on non-specific immunity, humoral immunity and cellular immunity in the immunosuppressive mice induced by cyclophosphamide. Besides, the immunosuppressive model was used to evaluate the effect of fraction on immune organs and content of cellular factors in blood serum. The results showed that the ethanol sediments promoted Concanavalin A (Con A) induced T cell and Lipopolysaccharides (LPS) induced B cell (P < 0.01). It increased the carbon clearance index K, phagocytic index α, half value hemolysis (HC50), and swelling degree of auricula (P < 0.05 or P < 0.01). It reduced the body weight and atrophy of thymus and spleen index (P < 0.05 or P < 0.01). It increased the contents of IL-2, IFN-γ and IL-4 in serum in immunosuppressive mice (P < 0.05 or P < 0.01). It was concluded that ethanol sediments of the seeds of D. sophia(L.) Webb. ex Prantl. can boost the lymphocyte proliferation, protect the immune organs, and enhance the non-specific and specific immunity in immunosuppressive mice, which indicated that it had immune-promotion effect.
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This study was aimed to establish the method for fraction-splitting of seeds of Descurainia sophia, in order to study the stability and unoverlapping property of fractions of seeds of D. sophia. The fraction-splitting of seeds of D. sophia were obtained through the combination of various methods including decoction, distillation, extraction, ethanol precipitation and gradient elution of macroporous adsorption resin. HPLC and chemometrics software were used to analyze the stability and unoverlapping property of the fractions of D. sophia. The results showed that the chemical components from seeds of D. sophia was divided into six fractions. HPLC data and chemometrics analysis showed good stability of technology. The fraction-splitting of seeds of D. sophia was unoverlapping. It complied with the research model of chemical constituents of seeds of D. sophia which can be split and combinatorial. It was concluded that the established method for splitting fractions of seeds of D. sophia had good stability and repeatability. Each splitting fraction uncrossed others.
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OBJECTIVE:To identify Descurainia sophia and Semen Plantaginis by high performance capillary electrophoresis (HPCE). METHODS: HPCE was conducted with a fused silica capillary tube(60 cm?75 ?m) taking 30 mmol? L-1 sodium dihydrogen phosphate as running buffer (pH 8.2). The sample injection was performed by high pressure for 5 s at a separation voltage of 12 kV, an UV detection wavelength of 330 nm. RESULTS: The differences of HPCE electrophoregrams between Descurainia sophia and Semen Plantaginis were significant and easily to be differentiated. CONCLUSION: This method is simple, rapid and sensitive, and it can be applied to identify Descurainia sophia and Semen Plantaginis.
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OBJECTIVE: To establish an HPCE and polymerase chain reaction (PCR) assay for identification of Descurainia sophia and Plantago asiatica.METHODS: PCR products of D. sophia and P.asiatica were analyzed by capillary tube (60 cm?75 ?m), taking 30 mmol?L-1 sodium tetraborate decahydrate (pH 9.2) as run buffer. The injection sample was performed by high pressure of 5 s at a separation voltage of 12 kV and with UV detection wavelength of 254 nm. RESULTS: PCR products of D. sophia and P. asiatica were separated by HPCE rapidly and accurately. The significant differences were noted in electrophoregram between PCR products of D. sophia and P. asiatica.CONCLUSION: This method is simple, rapid and sensitive for identification of D.sophia and P. asiatica.