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Objective:To explore the improvement effect of total flavonoids of Mori Cortex combined with total saponins of Anemarrhena Asphodeloide on hyperlipidemia rats with osteoporosis and its possible mechanism. Method:The 40 SPF male SD rats were adaptively fed for 7 days， and then randomly divided into normal group， model group， calcitriol group （45 ng·kg<sup>-1</sup>）， total flavonoids of Mori Cortex and total saponins of Anemarrhena Asphodeloide 1∶2 group （0.6 g·kg<sup>-1</sup>+0.4 g·kg<sup>-1</sup>） and 2∶1 group （1.2 g·kg<sup>-1</sup>+0.2 g·kg<sup>-1</sup>）. Except for the normal group， rats in the other groups were fed with high fat for 9 weeks， the normal group and the model grouotal flavonoids of total flavonoids of Mori Cortex and total saponins of Anemarrhena Asphodeloip were given normal saline by gavage， and the other groups were given corresponding drugs by gavage， after 12 weeks of administration， except for the normal group ， the other groups were given intramuscular injection of glucocorticoids at the same time. After 22 weeks of administration， the weight of rats with total flavonoids from Mori Cortex combined with total saponins of Anemarrhena Asphodeloide was measured. Serum levels of total cholesterol （TC）， triglyceride （TG）， low density lipoprotein cholesterol （LDL-C）， osteocalcin （BGP） and bone alkaline phosphatase （BALP） were determined by biochemical assay. Hematoxylin-eosin （HE） staining to observe the pathological changes of rat tibia. Real-time fluorescent quantitative polymerase chain reaction （Real-time PCR） was used to detect the expression levels of peroxisomal proliferators activate the receptor gamma（PPAR<italic>γ</italic>） and Runt-related transcription factor 2 （Runx2） mRNA in rat bone tissue， immunofluorescence was used to detect the expression of PPAR<italic>γ</italic> and Runx2 in rats. Result:Compared with normal group， the body mass of rats in model group was significantly increased （<italic>P</italic><0.01）， and the contents of TC， TG， and LDL-C in the serum were significantly increased （<italic>P</italic><0.01）. Compared with model group， the body weight of rats in thet total flavonoids of Mori Cortex and total saponins of Anemarrhena Asphodeloide 1∶2 group and 2∶1 group were significantly reduced （<italic>P</italic><0.01）， and the contents of TC， TG， and LDL-C in the serum were significantly reduced （<italic>P</italic><0.01）， the content of BGP and BALP increased （<italic>P</italic><0.01）. HE staining results showed that compared with the normal group， the tibia fat vacuoles of the model group increased， and the number of osteoblasts decreased， compared with the model group， the total flavonoids of the Mori Cortex and the flavonoids-total saponins of Anemarrhena Asphodeloide 1∶2 group and 2∶1 group decreased in tibia fat vacuoles and increased the number of osteoblasts， the results of immunofluorescence and Real-time PCR showed that， compared with normal group， the expression of Runx2 in the model group decreased and the expression of PPAR<italic>γ</italic> increased （<italic>P</italic><0.01）. Compared with model group， the total flavonoids of Mori Cortex-total saponins 1∶2 group and the total flavonoids of Mori Cortex-total saponins 2∶1 Group up-regulated the expression of Runx2 and down-regulated the expression of PPAR<italic>γ </italic>（<italic>P</italic><0.05，<italic>P</italic><0.01）. Conclusion:The total flavonoids of Mori Cortex combined with the total saponins of Anemarrhena Asphodeloide up-regulated Runx2 and down-regulated the expression of PPAR<italic>γ</italic> mRNA and protein， thereby affecting the metabolism of TG and TC in the blood， achieving a therapeutic effect on osteoporosis， provides experimental basis for the clinical prevention and treatment of hyperlipidemia with osteoporosis.
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Objective::To observe the effect of sanggenone C (SanC) on the proliferation and differentiation of mouse MC3T3-E1 osteoblasts induced by dexamethasone (DEX), and to explore its mechanism. Method::Molecular docking was conducted between SanC and Runt-associated transcription factor 2(Runx2) protein structure obtained by homologous modeling. MC3T3-E1 cells were jointly treated by different concentrations of SanC (8, 16, and 32 μmol·L-1) and 1 μmol·L-1 DEX, and then cell counting kit-8(CCK-8) method was used to detect the effect of SanC on the proliferation of MC3T3-E1 osteoblasts. The alkaline phosphatase (ALP) activity of MC3T3-E1 osteoblasts was determined by reagent kit and the formation of mineralized bone nodules were detected by alizarin red staining. Real-time fluorescent quantitative polymerase chain reaction (Real-time PCR) was used to detect the mRNA expression of Runx2, ALP and Osterix. The protein expression of Runx2 was detected by Western blot. Result::The docking score of SanC and Runx2 was -9.78.As compared with the normal group, DEX group significantly reduced the cell survival rate (P<0.01), and the greatest difference occurred on the seventh day. As compared with DEX group, SanC could significantly promote the cell proliferation of MC3T3-E1 (P<0.01), in which 32 μmol·L-1 SanC had the largest difference in proliferation rate on seventh day. As compared with the normal group, the expression of Runx2, ALP and Osterix mRNA increased to a certain extent in DEX group(P<0.01). As compared with DEX group, the expression levels of Runx2, ALP and Osterix mRNA were up-regulated in different concentration groups of SanC in a dose-dependent manner (P<0.01). As compared with the normal group, the expression of Runx2 protein in DEX group decreased significantly (P<0.05), and as compared with DEX group, the expression of Runx2 protein in cells under the intervention of SanC increased significantly (P<0.01). Conclusion::SanC can promote the proliferation, differentiation and mineralization of MC3T3-E1 osteoblasts, and the mechanism may be related to the up-regulation of Runx2 expression.
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Objective:To explore the effect of anemarrhena asphodeloside BⅡ (TBⅡ) on the expressions of nuclear transcription factor-κB receptor activator factor ligand （RANKL）, RANK and C-FOS genes during osteoclast differentiation. Method:Molecular docking software LeDock was used to score the docking of TBⅡ with RANKL, RANK and C-FOS. RAW264.7 was treated with soluble RANKL（sRANKL） and divided into control group, sRANKL group (model group), Icariin (Ica) group, low-dose TBIⅡ group (2 μmol·L-1), medium-dose TBⅡ group (4 μmol·L-1), and high-dose TBⅡ group (8 μmol·L-1). The corresponding kit was used to detect iconic enzyme (TRAP) of osteoclast differentiation. Total RNA was extracted by trizol method, Real-time fluorescence quantitative polymerase chain reaction (Real-time PCR) was used to detect the expressions of C-FOS, upstream RANKL/RANK and downstream nuclear factor of activated T-cells cytoplasmic 1 (NFATC1), and osteoprotegerin OPG. Result:The molecular docking score were -11.86, -11.38, -12.34 kcal·mol-1, and there might be multiple binding sites between TBII as well as RANKL, RANK and C-FOS. Compared with the control group, the content of TRAP in model group increased significantly (P<0.01), and compared with model group, the content of TRAP in each administration group decreased significantly (P<0.01), and TBⅡ decreased the content of TRAP in a dose-dependent manner. Compared with the control group, the expressions of RANKL, RANK, C-FOS and NFATC1 increased (P<0.01), whereas the expression of OPG decreased (P<0.01) in model group. Compared with model group, the expressions of RANKL, RANK, C-FOS and NFATC1 decreased (P<0.01), while the expression of OPG increased (P<0.01) in each administration group. Conclusion:TBⅡ may inhibit the differentiation of osteoclast precursors into osteoclasts, inhibit osteoclast activity, reduce bone resorption and improve osteoporosis by regulating RANKL/RANK/C-FOS signal pathway.
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Gambogic acid (GA), the main active ingredient in gamboge, has been reported to have good anti-tumor activity with excellent selectivity. However, its clinical application is limited by the poor water solubility. GA nanosuspensions were designed in this study in order to solve this problem. GA nanosuspensions were prepared by microprecipitation method based on pH adjustment. Suitable stabilizer was screened according to the size and polydispersity index (PDI) of the resultant nanosuspensions. Dynamic light scattering method was used to measure the particle size and transmission electron microscopy was used to observe the morphology. The stability was studied in different medium. The drug release was evaluated using a dialysis method. MTT assay was used to assess their cytotoxicity in vitro against cancer cell line. Anti-tumor effect in vivo was investigated on H22-bearing mice. In result, Poloxamer (P188) was found to be a good stabilizer. The resultant GA nanosuspensions (GA-NSps) were 135.9 ±5.1 nm in diameter, with PDI value being 0.26 ±0.01 and the zeta potential being −35.1 ±1.36) mV. GA-NSps were nearly spherical. They were quite stable in various physiological media. GA-NSps exhibited a sustained drug release pattern, with the cumulative release reaching 90.26% within 312 h. In MTT assay, GA-NSps had a stronger cytotoxicity against HepG2 cells than the free drug (IC50, 0.851 8 μg·mL−1 vs 2.104 μg·mL−1, P vs 66.80%, P < 0.01). In summary, we prepared GA-NSps with high drug loading capacity, small particle size and good stability, and provided a solid basis for the effective dosage form of gambogic acid.
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Honokiol (HK) have extensive pharmacological activities, but its poor solubility and instability restricted its clinical application and efficacy exertion. HK nanosuspensions (HK-NSps) were designed in this study in order to solve the problems. HK-NSps were prepared by antisolvent precipitation method, using poly-vinylpyrrolidone (PVP) and bovine serum albumin (BSA) as a combined stabilizer. The particle size was measured using dynamic light scattering method, the morphology was observed by transmission electron microscopy. The size change and drug content of HK-NSps in various physiological media during the storage at ambient temperature was examined to evaluate their storage stability. Dialysis method was used to study their drug release in vitro. MTT assay was used to assess their in vitro cytotoxicity against 4T1 breast cancer cell line. Anti-tumor effect in vivo was also investigated in 4T1 tumor-bearing mice. HK-NSps were prepared with high drug loading content of 48.62%, nearly spherical shape and good storage stability. The average particle size was (83.40 ±1.042) nm, the polydispersity index (PDI) value was 0.223 ±0.011, the zeta potential was (-42.2 ±1.2) mV. HK-NSps showed sustained in vitro drug release and enhanced cytotoxicity in contrast to free HK against 4T1 cells (IC50, 8.36 μg·mL-1 vs 37.58 μg·mL-1, Pin vivo study on 4T1 tumor-bearing mice demonstrated that HK-NSps showed good dose-dependent tumor inhibition rate (TIR). In contrast to 4 mg·kg-1 of PTX injection (TIR, 47.9%), medium and high dose of HK-NSps displayed improved therapeutic efficacy (TIR, 55.67% for 40 mg·kg-1, 67.28% for 60 mg·kg-1, P-1) had TIR of only 54.13% even administrated every day. In conclusion, HK-NSps were prepared with small size, high drug-loading capacity, and good stability. The improved in vitro and in vivo antitumor efficacy demonstrated that HK can be a promising antitumor drug in combination with nanosuspensions technology.
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To investigate the mechanism of the treatment of hyperlipidemia rats induced by Huangqi San. The 40 male SD rats were randomly divided into normal group, model group, Huangqi San low and high dose group (1, 2 g·kg⁻¹), and positive lipitor group (2 mg·kg⁻¹). The normal group feeds on base feed, and other groups feed on high-fat feed. After 8 weeks, the hyperlipidemia model was successful. After intervention by drugs for 13 weeks, fasting blood glucose, total cholesterol, triglycerides and LDL cholesterol content of all rats were measured. The pathological changes of liver and skeletal muscle of rats were observed in rats. Real-time PCR and Western blot were used to detect the mRNA and protein expression levels of AMPK signaling pathway in the liver and skeletal muscles (AMPK, ACC, CPT1A, SREBP2, HMGCR). The degree of FPG, TC, TG and LDL-C were the highest in the model group, and the liver and skeletal muscle pathology were the most obvious. After intervention by Huangqi San and lipitor, a significant reduction in the blood sugar blood fat, liver, and skeletal muscle injury has improved significantly, except SREBF2 and HMGCR mRNA and protein expression of this enzyme is reduced, other AMPK pathway related mRNA and protein expression increased significantly. Huangqi San effect is superior to lipitor. Huangqi San may improve hyperlipidemia by regulating the AMPK signaling pathway, increasing the oxidation of fatty acids and inhibiting cholesterol synthesis.
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<p><b>OBJECTIVE</b>To assess the value of an intracavitary convex array probe in detecting internal carotid artery (ICA) disease.</p><p><b>METHODS</b>Eighty-six carotid arteries in 43 cases were examined with intracavitary convex array probe, low-frequency convex array probe and high-frequency linear probe to collect the data including the ICA visible length, peak systolic velocity (PSV), internal diameter, blood vessel shape; common carotid artery (CCA) intimae-medial thickness (IMT), PSV, and internal diameter.</p><p><b>RESULTS</b>Significant differences were noted in the visible length, PSV of ICA, and internal diameter detected by different frequency ultrasound probes. Intracavitary probe and high-frequency probe produced significantly different findings of the blood vessel shape.</p><p><b>CONCLUSION</b>Intracavitary convex array probe has important clinical value in detecting of ICA disease.</p>
Subject(s)
Adult , Aged , Female , Humans , Male , Middle Aged , Carotid Artery Diseases , Diagnostic Imaging , Ultrasonography , MethodsABSTRACT
<p><b>OBJECTIVE</b>To observe the effects of phlegm-dispelling and blood stasis-resolving traditional Chinese drugs on the cell cycle of cardiac myocytes and left ventricular reconstruction in hypertensive rats.</p><p><b>METHODS</b>Bilateral renal artery stenosis was conducted to induce hypertension in rats, which were randomly divided into hypertensive model group (n = 10), sham-operated group (n = 8), high-dose drug group (n = 11) and low-dose drug group (n = 11), with 8 normal untreated rats as the normal control group. The systolic blood pressure (SBP) was measured in the tail artery of the rats. Two months after the operation, the left ventricular mass (LVM) and LVM index (LVI) were calculated in all the rats. The cell cycle changes in the left ventricular cardiac myocytes were evaluated using flow cytometry.</p><p><b>RESULTS</b>The mean blood pressure and LVI of the hypertensive model group were significantly higher than those of the normal control (P < 0.05) and sham-operated group (P < 0.01). After treatment with preparation of the traditional Chinese drugs at either high or low dose, the mean blood pressure and LVM of the rats showed obvious reduction, and LVI was decreased significantly compared with that of the model group (P < 0.05). Compared with the hypertensive model group which showed obviously decreased cell percentage in G0/G1 phase and increased S phase cells, the treatment at both doses significantly increased the cells in G0/G1 phase (P < 0.05) and decreased the S-phase cells (P < 0.05) to levels comparable to those in the normal control and sham-operated groups (P > 0.05). The percentage of G2/M-phase cells showed no significant difference between the groups (P > 0.05).</p><p><b>CONCLUSION</b>The traditional Chinese drugs can significantly decrease blood pressure and LVI in hypertensive rats, and induce cell cycle arrest in G0/G1 phase to reverse left ventricular hypertrophy by regulating the cell cycle and inhibiting the division and proliferation of the cardiac myocytes.</p>
Subject(s)
Animals , Male , Rats , Cell Cycle , Cells, Cultured , Drugs, Chinese Herbal , Therapeutic Uses , Hypertension , Drug Therapy , Hypertrophy, Left Ventricular , Drug Therapy , Pathology , Myocytes, Cardiac , Pathology , Phytotherapy , Random Allocation , Rats, Sprague-Dawley , Ventricular RemodelingABSTRACT
The relationship between adjunctive drug (AD) and other drugs in a compound prescription and the actions of AD are very complex. From historical view, it showed the adjunctive action of AD was only be recognized with emphasis on its action as corrigent and effect on complications. In modern medicine, the actions of AD were reduced to 3 aspects: the assistant action, the restrained action and the corrigent action, with new contents added in each aspect on the basis of the ancient understanding. The authors hold that to master the various actions of AD and its relationship with other drugs in the prescription would be helpful in choosing them accurately and scientifically, thus to contribute the treatment in accordance with disease.
Subject(s)
Humans , China , Drug Combinations , Drug Prescriptions , Drug Synergism , Drugs, Chinese Herbal , History, 16th Century , History, 17th Century , Medicine, Chinese Traditional , HistoryABSTRACT
Researches in recent close to dozen years concerning Tripterygium wilfordii (TW) toxicity-reducing and efficacy-enhancing by combined use of Chinese medicine in treating intractable diseases such as rheumatoid arthritis, nephropathy, psoriasis etc. were summarized in this paper. Furthermore, the therapeutic mechanisms and adverse reaction of TW were elaborated and how to arrange properly the TCM hebal drugs used in combination was analyzed.