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ObjectiveTo investigate the anti-inflammatory effect of the component compatibility of Gentianae Macrophyllae Radix and Clematidis Radix et Rhizoma on the rat model of rheumatoid arthritis （RA） and the mechanism. MethodSeventy-two SPF-grade SD rats （male and female） aged 5 to 6 weeks were selected. Except the blank group， the rat model of collagen-induced arthritis （CIA） was replicated by the type Ⅱ collagen induction method. The 64 rats after successfully modeling were randomly divided into model group， methotrexate group （0.375 mg·kg-1）， gentianoside with magnoflorine group （150.454 1 mg·kg-1+5.061 8 mg·kg-1）， gentianoside with clematichinenoside AR group （150.454 1 mg·kg-1+16.433 1 mg·kg-1）， sweroside with magnoflorine group （3.455 8 mg·kg-1+5.061 8 mg·kg-1）， sweroside with clematichinenoside AR group （3.455 8 mg·kg-1+16.433 1 mg·kg-1）， swertiamarin with magnoflorine group （9.303 2 mg·kg-1+5.061 8 mg·kg-1）， and swertiamarin with clematichinenoside AR group （9.303 2 mg·kg-1+16.433 1 mg·kg-1）， with 8 rats in each group. Each group was given the corresponding medicinal solution or normal saline by gavage for 15 d. During the experiment， the general status， of rats in each group were observed and recorded. Tumor necrosis factor-α （TNF-α）， interleukin-1β （IL-1β）， rheumatoid factor （RF）， C reactive protein （CRP）， prostaglandin E2 （PGE2）， and anti-cyclic peptide containing citrulline antibody （anti-CCP Ab） in the serum of rats were measured by enzyme-linked immunosorbent assay （ELISA）. The histopathological changes in rat ankle joints were observed by hematoxylin-eosin （HE） staining. Immunohistochemistry （IHC） and Western blot were used to detect the protein expression of nuclear factor-κB （NF-κB） and vascular endothelial growth factor （VEGF） in rat ankle joints. Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） was used to detect the mRNA expression of NF-κB and VEGF in rat ankle joints. ResultCompared with those in the blank group， rats in the model group were in poor general conditions with significant foot-plantar swelling， and the content of CRP， anti-CCP Ab， and IL-1β in the rat serum was significantly increased （P<0.01）. In the model group， the tissue structure of the ankle joint was severely damaged， and the protein and mRNA expression of NF-κB and VEGF in the rat ankle joints were significantly up-regulated （P<0.01）. As compared with the model group， the general status of rats in each administration group was significantly improved. The levels of serum TNF-α， IL-1β， RF， CRP， PGE2， and anti-CCP Ab were reduced to different degrees in these administration groups， among which the effects of the gentianoside with clematichinenoside AR group on down-regulating serum TNF-α and IL-1β， the gentianoside with magnoflorine group on down-regulating serum RF and CRP， the sweroside with magnoflorine group on down-regulating serum PGE2， and the swertiamarin with clematichinenoside AR group on lowering serum anti-CCP Ab were better than those of administration groups. The histopathological changes in the ankle joint were improved to different degrees. The protein and mRNA expression of NF-κB and VEGF in rat ankle joints in the administration groups was significantly down-regulated （P<0.05， P<0.01）， and the swertiamarin paired with clematichinenoside AR group had the most significant effect. ConclusionThe component compatibility of Gentianae Macrophyllae Radix and Clematidis Radix et Rhizoma exerts a good therapeutic effect on the rat model of RA， and the compatibility of components from the two medicines has a multi-channel， multi-target， and synergistic effect. The five component compatibility patterns， namely gentiobioside with magnoflorine， gentiobioside with clematichinenoside AR， sweroside with clematichinenoside AR， swertiamarin with magnoflorine， and swertiamarin with clematichinenoside AR， all have potential advantages. The mechanism may be related to the reduction of inflammatory factor secretion and the inhibition of abnormal protein and mRNA expression of NF-κB and VEGF.
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Objective:To evaluate the compatibility laws of effective-component compatibility of Bufei Yishen formula Ⅲ (ECC-BYFⅢ) in regulating mucus hypersecretion of chronic obstructive pulmonary disease (COPD).Methods:According to the efficacy of the original Chinese medicine, the components of ECC-BYFⅢ were divided into four categories: Buqi (Ginsenoside Rh1+Astragaloside), Bushen (Icariin), Huatan (Nobiletin), and Huoxue (Paeonol). The four categories were divided into 14 groups based on the method of mathematical permutation. ① The rats were divided into control group, model group, ECC-BYFⅢ, and different components compatibility groups according to the random number table, totaling 17 groups. COPD rat model in stable phase was established by cigarette smoke exposure combined with repeated bacterial infections. The corresponding drugs were given by gavage at the 9th week of modeling, and the samples were collected at the end of the 16th week. The levels of matrix metalloproteinase-9 (MMP-9) and tissue inhibitors of metalloproteinase 1 (TIMP-1) in serum and bronchoalveolar lavage fluid (BALF), and the levels of mucin (MUC) 5AC in lung tissue and BALF were detected by enzyme linked immunosorbent assay (ELISA). ② Human lung epithelial cells BEAS-2B were divided into blank group, model group, and different components compatibility groups. Hypoxia-induced mucus hypersecretion model of human lung epithelial cells BEAS-2B was established 4 hours after corresponding drug pretreatment. The mRNA expressions of MUC5AC, MUC5B, and MUC1 were detected by quantitative polymerase chain reaction (PCR). The mucus secretion indexes of rats and BEAS-2B cells were evaluated by Region (R) value comprehensive evaluation method.Results:① Compared with the control group, MMP-9 in serum and BALF from the model group were significantly increased, the level of TIMP-1 was significantly decreased, and MUC5AC in lung tissue and BALF were significantly increased. The results of R value comprehensive evaluation showed that except for the Buqi and Bushen groups, ECC-BYFⅢ and other components compatibility groups significantly corrected mucus hypersecretion in COPD rats, ECC-BYFⅢ, Bushen Quxie, Fuzheng Huatan, and Quxie groups were much better (R values were 2.15±0.42, 2.11±0.23, 2.16±0.23 and 2.16±0.55, respectively), compared with the model group (R value: 3.00±0.00), the differences were statistically significant (all P < 0.05). ② Compared with the blank group, the mRNA expressions of MUC5AC, MUC5B, and MUC1 increased in the model group. But different components compatibility groups had no significant effects on the mucus secretion of BEAS-2B cells. ③ The comprehensive evaluation results of R value about each in vivo and in vitro index showed that ECC-BYFⅢ, Huoxue, Quxie, Bushen Huoxue, Fuzheng Huatan, Buqi Quxie groups significantly corrected the mucus hypersecretion (R values were 2.30±0.43, 2.33±0.44, 2.12±0.68, 2.27±0.64, 2.24±0.27 and 2.29±0.47, respectively), compared with the model group (R value: 3.00±0.00), the difference was statistically significant (all P < 0.01). The order was: Quxie > Fuzheng Huatan > Bushen Huoxue > Buqi Quxie > ECC-BYFⅢ > Huoxue. Conclusions:Different components compatibility of ECC-BYFⅢ had different effects on COPD mucus secretion. The components containing Huatan (Nobiletin) or Huoxue (Paeonol) showed a better inhibitory effect on mucus secretion.
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Objective:This study intends to study the regulatory effect and mechanism of the effective components of Salviae Miltiorrhizae Radix et Rhizoma and Carthami Flos on inflammatory factors related to cerebral ischemia-reperfusion injury in rats through multiple levels of neuropathology， molecular neurobiology and functional behavior. Method:The 32 male rats were randomly divided into four groups： sham group， model group， Danhong components compatibility group（720 mg·kg-1）， nimodipine （0.5 mg·kg-1）groups，each group of eight male rats.Cerebral ischemia was established by middle cerebral artery occlusion （MCAO） approach. The treatment was performed immediately and at 6 hour after MCAO.Hematoxylin-eosin （HE）staining was used to check the changes of brain histopathology， immunohistochemistry and Real time polymerase chain reaction （Real-time PCR） were used to check the expression of IL-1β and Nrf2 in brain tissue，Western blot was used to detect the protein expression of Nrf2 in brain tissue. The aim is to investigate the treatment mechanism of Salviae Miltiorrhizae Radix et Rhizoma and Carthami Flos components in a rat model of cerebral ischemic-reperfusion injury. Result:HE staining results showed， compared with sham group， the surviving neurons amount in the model group was significantly reduced（P<0.01），compared with the MCAO group，the number of surviving neurons in the brain tissue of Salviae Miltiorrhizae Radix et Rhizoma and Carthami Flos component compatibility group and nimodipine group was significantly increased（P<0.05，P<0.01）.The results of immunohistochemistry and Real-time PCR showed that，compared with normal group，IL-1β and Nrf2 expression in model group were significantly increased （P<0.01），compared with MCAO group， the expression of IL-1β and Nrf2 in Salviae Miltiorrhizae Radix et Rhizoma and Carthami Flos component compatibility group and the nimodipine group was significantly decreased （P<0.05，P<0.01）. Western blot results showed that， compared with sham group， Nrf2 positive expression in model group was much more increased （P<0.01）， compared with MCAO group， the expression of Nrf2 in Salviae Miltiorrhizae Radix et Rhizoma and Carthami Flos component compatibility group and the nimodipine group was significantly decreased （P<0.01）. Conclusion:The combination of effective components of Salviae Miltiorrhizae Radix et Rhizoma and Carthami Flos can significantly down-regulate the expression of IL-1β and Nrf2 proteins.The mechanism is to activate the protein expression of inflammatory pathways， reduce the apoptosis of nerve cells， and finally inhibit the inflammatory response in the process of ischemic stroke injury.
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OBJECTIVE: To explorethe effect of total flavonoids from Cudrania tricuspidata Bun on Lewis lung cancer mice and its new component compatibility on the autophagy of LLC cells. METHODS: A mouse model of Lewis lung cancer xenografts was constructed. The body weight, tumor weight, tumor volume and organ index were measured before and after taking total flavonoids of Cudrania tricuspidata Bun. The HE staining of the xenograft pathological sections were also observed. The TNF-α, IL-2, IL-6 and IL-12 levels in the serum were calculated by ELISA. The content of the main active ingredient and its ratio in the flavonoid extract were measured by UPLC. Western blot was used to detect the effect of the new component compatibility on the expression of autophagy protein in LLC cells, and the ultra structural changes of LLC cells were observed by transmission electron microscopy. Flow cytometry was used to detect the effect of the new component compatibility on autophagy of LLC cells. RESULTS: The high-dose group of total flavonoids from Cudrania tricuspidata Bun can significantly inhibit the growth of tumor in mice and enhance the organ index of tumor-bearing mice, and the survival rate of mice could be improved in all groups of total flavonoids from Cudrania tricuspidata Bun. The serum levels of TNF-α, IL-2, IL-6 and IL-12 in the tumor-bearing mice of each group were higher than those in the model group, and there was significant difference in the high dose group of total flavonoids (P<0.01). The main active ingredients were taxifolin, kaempferol and naringenin by UPLC. The ratio of taxifolin to kaempferol was 60:1. Western blot assay showed that its new component compatibility significantly increased the expression of autophagic protein in LLC cells (P<0.01). The autophagosomes in the cytoplasm were observed by transmission electron microscopy. The results of flow experiments showed that the average fluorescence intensity of its new component compatibility was significantly higher than that of the control group. CONCLUSION: Total flavonoids of Cudrania tricuspidata Bun can effectively inhibit tumor growth and have less harm to immune organs. Its mechanism may be related to up-regulation of cytokines TNF-α, IL-2, IL-6 and IL-12 levels and improve immune function in the body. Moreover, its main active ingredient promotes the increase of autophagy protein expression in LLC cells and induces autophagy in LLC cells.
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Objective: To study antibreast cancer activity and the mechanism of appropriate components of polysaccharides (P), triterpenes (T), the essential oil (V), and different compatibilities (PT, PV, TV, and PTV) of Prunella vulgaris in vivo. Methods: 4T1 breast cancer model was established to evaluate anti-breast cancer activity. The appropriate components of P. vulgaris were screened. The tumor volume and the shaded areas in breast cancer mice were detected by living small animal Micro-CT scan. The structural changes of each organ were detected by HE staining. TUNEL staining was used to detect apoptotic rate of tumor. The expression of PCNA, CD-31, and E-cadherin were detected by immunohistochemistry (IHC). Serum estradiol was detected by Elisa kit. Results: T and PTV had significant anti-breast cancer activity. There were inflammatory cells infiltration, degeneration and necrosis, tumor cell apoptosis in the T and PTV groups. T and PTV could inhibit cell proliferation by reducing the estradiol level and downregulating the overexpression of PCNA proteins. T and PTV could also reduce tumor angiogenesis by downregulating the protein expression of CD-31. T and PTV also inhibited the metastatic process by upregulating the protein expression of E-cadherin. Conclusion: T and PTV showed significant anti-breast cancer activities, suggesting that T and PTV can be a potential drug for breast cancer.
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To reveal the relationship between components and components, components and syndromes based on data mining. Analyzing the component compatibility regularity in treatment of cardiovascular diseases from the component level provides the basis for treating the diseases with multi-component Chinese medicine based on the combination of syndrome and symptom in Chinese medicine clinic. Through the collection and structural processing of the key points in the literature data in recent 20 years, the mining results of Chinese material medica (Chinese medicinal materials/decoction pieces), Chinese patent medicine and related components for the treatmet of cardiovascular diseases were analyzed by association rules mining algorithm. The component compatibilities for treating cardiovascular diseases with a higher correlation were screened out. New component compatibility forms were found. The effective substances and/or possible component compatibility of single herb and Chinese herbal compound were confirmed or speculated. The relationship between components compatibility and diseases, components and syndromes were revealed. Based on association rules mining, it can provide new ideas and methods for component compatibility and optimization, and provide technical support for the research and development of component Chinese medicine.
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To explore the anti-atherosclerotic mechanism of active component compatibility of Danshen and Shanzha (SC121) based on network pharmacology and in vitro research validation with cell model. On one hand, according to the chemical structures and pharmacological activities of the compounds reported in Danshen and Shanzha, 5 compounds, i.e., salvianolic acid B, tanshinone ⅡA, tanshinol, epicatechin and procyanidin B2 were chosen and used for network pharmacology analysis. Then the TCMSP（http://lsp.nwsuaf.edu.cn/tcmsp.php）was used for finding the network targets for 5 compounds from SC121. The signaling pathway associated with cardiovascular disease was analyzed by KEGG mapping, the biological process associated with cardiovascular disease was analyzed by Uniprot. And, the mechanism of SC121 was predicted by network pharmacology. In vitro cell model was subsequently performed for validation. HUVEC and RAW264.7 cell injuries and foam cell formation were constructed by ox-LDL, and the intervention effects of SC121 were assayed. The result showed that SC121 not only alleviated the damage of HUVEC and RAW264.7, lowered the ROS level, but also decreased the area of foam cell in a dose-dependent manner, which indicated that SC121 could inhibit the damage of endothelial cells and lower the oxidative stress. The experimental data validated the prediction of network pharmacology, and elucidated the mechanism of SC121's effect on AS.
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Objective: To investigate the regulation of in vitro myocardial cell L type calcium channel of rats with active component of Shenfu Decoction and its combination. Methods: With Langendorff cardiac infusion, the single ventricular myocardial cell with acute enzymylosis approach was acquired and the voltage current of L type calcium channel was recorded with whole cell patch technique. Results: The active ginsenosides Re (3, 10, 30, and 100 μmol/L), and Rg1 (10 and 100 μmol/L) can reversibly reduce L type calcium current in a dose dependant manner, Rb1 (10 and 100 μmol/L) has no effect on L type calcium current; the active component aconine (10 and 100 μmol/L) of aconite could irreversibly enhance L type calcium current in a dose dependant manner; The combination of ginsenosides Re (20 μmol/L) and Rg1 (80 μmol/L) also reduced the L type calcium current, which is more significant than either ginsenosides Rg1 and Re; The inhibitory rate of the combination of ginsenosides Rg1, Re and aconine (100 μmol/L) has no difference from the combination of ginsenosides Rg1 and Re. Conclusion: The active component ginsenosides Rg1 and Re of Shenfu Decoction can reduce the L type calcium current, while aconine can irreversibly enhance the L type calcium current; The combination of ginsenosides Rg1 and Re can obviously increase the reduction of L type calcium current, while the combination of ginsenosides Rg1, Re and anonine can not change the reduction of L type calcium current.
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Objective: To explore the research method suitable for the component compatibility and optimization design of Chinese materia medica (CMM) by taking the blood pressure-lowering effect of the effective components in Ramulus Uncariae cum Uncie total alkaloids (RUCUTA) and Semen Raphani total alkaloids (SRTA) as the research objects. Methods: According to the procedure of "orthogonal design-evaluation on drug effect-partial least-squares regression (PLSR)", using contractive pressure as index, the ratio of RUCUTA and SRTA was optimized by orthogonal design method. The data were analyzed by range analysis, variance analysis, multiple regression analysis, and PLSR analysis. Results: According to the result of the data analysis and considering the economic costs, the optimal ratio of the two components was 25 μg/g RUCUTA and 30 μg/g SRTA. Conclusion: The compatibility of RUCUTA and SRTA could effectively reduce blood pressure. The orthogonal design method combined with PLSR could eliminate the interaction among the various factors and be suitable for the component compatibility in CMM and optimization design.