RESUMO
The ethanol precipitation process of Nauclea officinalis extract was optimized based on the concept of quality by design(QbD). Single factor tests were carried out to determine the levels of test factors. The ethanol volume fraction, pre-ethanol precipitation drug concentration, and ethanol precipitation time were taken as critical process parameters(CPPs). With the comprehensive scores of strictosamide transfer rate and solid removal rate as the critical quality attributes(CQAs), Box-Behnken design was employed to establish the mathematical models and space design between CPPs and CQAs, and the obtained optimal operating space was validated. The optimal operating space included ethanol volume fraction of 65%-70%, pre-ethanol precipitation drug concentration of 22-27 mg·mL~(-1), and ethanol precipitation time of 12 h. Based on the concept of QbD, this study adopted the design space to optimize the ethanol precipitation process of N. officinalis extract, which provided a reliable theoretical basis for the quality control in the production process of N. officinalis preparations. Moroever, this study provided a reference value for guiding the research and industrial production of traditional Chinese medicines.
Assuntos
Medicamentos de Ervas Chinesas , Etanol , Medicina Tradicional Chinesa , Controle de Qualidade , Modelos TeóricosRESUMO
Pai-Nong-San (PNS), a prescription of traditional Chinese medicine, has been used for years to treat abscessation-induced diseases including colitis and colorectal cancer. This study was aimed to investigate the preventive effects and possible protective mechanism of PNS on a colitis-associated colorectal cancer (CAC) mouse model induced by azoxymethane (AOM)/dextran sodium sulfate (DSS). The macroscopic and histopathologic examinations of colon injury and DAI score were observed. The inflammatory indicators of intestinal immunity were determined by immunohistochemistry and immunofluorescence. The high throughput 16S rRNA sequence of gut microbiota in the feces of mice was performed. Western blot was used to investigate the protein expression of the Wnt signaling pathway in colon tissues. PNS improved colon injury, as manifested by the alleviation of hematochezia, decreased DAI score, increased colon length, and reversal of pathological changes. PNS treatment protected against AOM/DSS-induced colon inflammation by regulating the expression of CD4+ and CD8+ T cells, inhibiting the production of HIF-α, IL-6, and TNF-α, and promoting the expression of IL-4 and IFN-γ in colon tissues. Meanwhile, PNS improved the components of gut microbiota, as measured by the adjusted levels of Firmicutes, Bacteroidetes, Proteobacteria, and Lactobacillus. PNS down-regulated the protein expression of p-GSK-3ß, ß-catenin, and c-Myc, while up-regulating the GSK-3ß and p-ß-catenin in colon tissues of CAC mice. In conclusion, our results suggested that PNS exhibits protective effect on AOM/DSS-induced colon injury and alleviates the development of CAC through suppressing inflammation, improving gut microbiota, and inhibiting the Wnt signaling pathway.
Assuntos
Colite , Medicamentos de Ervas Chinesas/farmacologia , Via de Sinalização Wnt/efeitos dos fármacos , Animais , Azoximetano/toxicidade , Linfócitos T CD8-Positivos , Colite/induzido quimicamente , Colite/tratamento farmacológico , Colite/genética , Sulfato de Dextrana/toxicidade , Modelos Animais de Doenças , Glicogênio Sintase Quinase 3 beta , Camundongos , Camundongos Endogâmicos C57BL , RNA Ribossômico 16SRESUMO
Objective: To investigate the dynamic regulation of self-assembled aggregations (SAA) in Coptidis Rhizoma decoction on the permeability of intestinal tissue and the mechanism underlying. Methods: The effects of SAA on berberine (Ber) absorption were respectively analyzed in an in situ intestinal perfusion model and in an Ussing Chamber jejunum model with or without Peyer's patches (PPs). The expression levels of ZO-1, Occludin and Claudin-1 were detected by immunofluorescence to evaluate the tight junction (TJ) between intestinal epithelium cells. The expression levels of T-box-containing protein expressed in T cells, signal transducers and activators of tranion-6, retinoic acid receptor-related orphan receptor γt and forkhead box P3 in PPs were detected by the reverse transcription-polymerase chain reaction and the secretions of interferon-γ (IFN-γ), interleukin-4 (IL-4), interleukin-17 (IL-17) and transforming growth factor-ß (TGF-ß) in PPs were evaluated by immunohistochemistry, to reflect the differentiation of T lymphocyte in PPs to helper T (Th) cell 1, Th2, Th17 and regulatory T (Treg) cell. To confirm the correlation between SAA in Coptidis Rhizoma decoction, PPs-associated immunity and intestinal epithelium permeability, SAA were administrated on an Ussing Chamber jejunum model with immunosuppressed PPs and evaluated its influences on intestinal tissue permeability and TJ proteins expression. Results: SAA in Coptidis Rhizoma decoction could dose-dependently promote Ber absorption in jejunum segment, with the participation of PPs. The dose-dependent and dynamical regulations of SAA on permeability of intestinal tissue and TJ proteins expression level between intestinal epithelium cells occurred along with the dynamically changed T lymphocyte differentiation and immune effectors secretion in PPs. The administration of SAA on immunosuppressed PPs exhibited dose-dependent PPs activation, inducing dynamic promotion on intestinal tissue permeability and inhibition on TJ proteins expression. Conclusion: SAA can improve the Ber absorption in small intestine, through the PPs-associated immunity induced dynamic regulation on intestinal tissue permeability and TJ proteins expression. These findings might enlighten the research of traditional Chinese medicine decoction.
RESUMO
Ligustrazine was the active ingredient of the traditional Chinese medicine Chuanxiong Rhizoma. However, the content of ligustrazine is very low. We proposed a hypothesis that ligustrazine was produced by the mutual effects between endophytic Bacillus subtilis and the Ligusticum chuanxiong Hort. This study aimed to explore whether the endophytic B. subtilis LB5 could make use of Chuanxiong Rhizoma fermentation matrix to produce ligustrazine and clarify the mechanisms of action preliminarily. Ultra high performance liquid chromatography with quadrupole time-of-flight mass spectrometry analysis showed the content of ligustrazine in Chuanxiong Rhizoma was below the detection limit (0.1 ng/mL), while B. subtilis LB5 produced ligustrazine at the yield of 1.0268 mg/mL in the Chuanxiong Rhizoma-ammonium sulfate fermentation medium. In the fermented matrix, the reducing sugar had a significant reduction from 12.034 to 2.424 mg/mL, and rough protein content increased from 2.239 to 4.361 mg/mL. Acetoin, the biosynthetic precursor of ligustrazine, was generated in the Chuanxiong Rhizoma-Ammonium sulfate (151.2 mg/mL) fermentation medium. This result showed that the endophytic bacteria B. subtilis LB5 metabolized Chuanxiong Rhizoma via secreted protein to consume the sugar in Chuanxiong Rhizoma to produce a considerable amount of ligustrazine. Collectively, our preliminary research suggested that ligustrazine was the interaction product of endophyte, but not the secondary metabolite of Chuanxiong Rhizoma itself.
Assuntos
Bacillus subtilis/química , Medicamentos de Ervas Chinesas/análise , Pirazinas/análise , Rizoma/química , Bacillus subtilis/metabolismo , Cromatografia Líquida de Alta Pressão , Medicamentos de Ervas Chinesas/metabolismo , Fermentação , Espectrometria de Massas , Medicina Tradicional Chinesa , Pirazinas/metabolismo , Rizoma/metabolismo , Fatores de TempoRESUMO
OBJECTIVE: To study the effect of supplementing Qi and activating blood circulation method with danshen and huangqi on protein uptake in NRK-52E cells injured by high glucose. METHODS: The influence of combined use of huangqi and danshen on NRK-52E cells proliferation were determined by MTT method, FITC-BSA uptake were observed by fluorescence microscope and analyzed by fluorescence spectrophotometer, and megalin expression were detected by the method of immunohistochemistry. RESULTS: Combined use of huangqi and danshen could strengthen its protection effect. The FITC-BSA uptake and expression of megalin in NRK-52E cells were decreased in high glucose environment. Combined use of huangqi and danshen could increase FITC-BSA uptake and megalin expression in high glucose-injured NRK-52E cells. CONCLUSION: The method of supplementing Qi and activating blood circulation can protect high glucose injured NRK-52E cells and improve the function of protein uptake by increasing expression of megalin.
Assuntos
Medicamentos de Ervas Chinesas/farmacologia , Células Epiteliais/efeitos dos fármacos , Glucose/efeitos adversos , Túbulos Renais/citologia , Triazinas/metabolismo , Animais , Astragalus propinquus/química , Proliferação de Células , Células Cultivadas , Nefropatias Diabéticas/metabolismo , Nefropatias Diabéticas/patologia , Células Epiteliais/metabolismo , Células Epiteliais/patologia , Fluoresceína-5-Isotiocianato/metabolismo , Imuno-Histoquímica , Túbulos Renais/efeitos dos fármacos , Túbulos Renais/metabolismo , Substâncias Protetoras/farmacologia , Ratos , Salvia miltiorrhiza/química , Soroalbumina Bovina/metabolismoRESUMO
OBJECTIVE: To study the protective effect of medicated serum prepared with Taohong Siwu Tang on hydrogen peroxide-injured human umbilical vein endothelial cells (HUVECs). METHOD: Sprague Dawley rats were orally administered with 10 mL x kg(-1) extracts from Taohong Siwu Tang (1.75 g crude drug), twice a day for three days, in order to prepare medicated serum of Taohong Siwu Tang. The effect of medicated serum pre-treated with Taohong Siwu Tang (with concentrations of 5%, 10%, 15%) on reduction of H2O2-induced cell activity was detected MTT. Cell morphological changes were observed under microscope. The effect of medicated serum prepared with Taohong Siwu Tang on the apoptosis and antioxidant capacity of HUVECs was detected with the content of malondialdehyde (MDA) and dehydrogenase (LDH), the activity of superoxide dismutase (SOD) and AO/EB staining. The expression of Caspase-3 was determined by western blot. RESULT: Compared with the blank serum control group, cell were significantly less active after being damaged by H2O2 (400 micromol x L(-1)). Medicated serum could significantly improve the SOD activity, reduce the levels of LDH and MDA, and inhibite the expression of Caspase-3. AO/EB staining and inverted microscope also showed that medicated serum prepared with Taohong Siwu Tang could reduce cell apoptosis induced by H2O2. CONCLUSION: Medicated serum prepared with Taohong Siwu Tang has significant protective effect on HUVECs injured by H2O2.
Assuntos
Medicamentos de Ervas Chinesas/farmacologia , Células Endoteliais da Veia Umbilical Humana/efeitos dos fármacos , Peróxido de Hidrogênio/farmacologia , Soro/química , Animais , Apoptose/efeitos dos fármacos , Western Blotting , Caspase 3/metabolismo , Sobrevivência Celular/efeitos dos fármacos , Células Cultivadas , Medicamentos de Ervas Chinesas/química , Feminino , Células Endoteliais da Veia Umbilical Humana/citologia , Células Endoteliais da Veia Umbilical Humana/metabolismo , Humanos , L-Lactato Desidrogenase/metabolismo , Malondialdeído/metabolismo , Ratos , Ratos Sprague-Dawley , Superóxido DismutaseRESUMO
OBJECTIVE: To investigate the effect of Chinese herbal medicine with Supplement Qi and Activating Blood Circulation (huangqi and danshen) on urinary protein, kidney function and tubular reabsorption of diabetic nephropathy rats. METHODS: SD rats were randomly divided into a nondiabetic control group (normal group) and three groups in which diabetes were induced by a single intraperitoneal injection of freshly prepared streptozotocin( STZ,55 mg/kg body weight). Then the diabetes rats were randomly assigned to three groups: diabetic model group, Supplement Qi and Activating Blood Circulation traditional Chinese medicine group (huangqi and danshen group) and Gliquidone group (as a reference hypoglycemic drug). Each group was treated with corresponding drugs for 6 weeks. At the end of the study, the rats from each group were injected with FITC-labeled BSA through tail vein. The 24 h urinary protein excretion were measured and blood was collected for measuring plasma glucose levels, serum creatinine (Cr), blood urea nitrogen (BUN), triglyceride (TG) and total cholesterol (T-CHO). Renal tissue was used to measure the level of LPO,SOD,GSH-Px and AGEs and Paraffin-embedded sections were stained with HE, PAS and immunohistochemistry. RESULTS: The plasma glucose, the 24 h urinary protein excretion, the levels of serum Cr, BUN, TG and T-CHO in STZ-induced diabetic rats were higher than those of nondiabetic rats. Diabetic rats showed significantly increase in LPO and AGEs (P < 0.01) and decrease in antioxidant enzyme activity (both GSH-Px and SOD) (P < 0.05) as compared with non-diabetic control rats. Treatment with the Supplement Qi and Activating Blood Circulation traditional Chinese medicine for 6 weeks in diabetic rats significantly reduced the 24 h urinary protein excretion compared with model control (P < 0.01), and markedly decreased the levels of serum Cr,BUN,TG and T-CHO as compared with those of diabetic rat (P < 0.05). The levels of LPO and AGEs were decreased and the activity of GSH-Px was increased by Supplement Qi and Activating Blood Circulation treatment. The kidney proximal tubule lesions were improved and the reabsorption of FITC-BSA in tubular was increased in diabetic rats treated by huangqi and danshen, and the expression of megalin in proximal tubular was enhanced as compared with diabetic rats. CONCLUSION: Diabetic nephropathy rats treated with traditional Chinese medicine therapeutic principles "Supplement Qi and Activating Blood Circulation" can reduce the 24 h urinary protein excretion and improve the function of tubular reabsorption. These protect effects may be in correlation with enhancement the renal tissue activity of antioxidant and up-regulation the expression of megalin in renal tubular epithelial cells in diabetic rats.
Assuntos
Nefropatias Diabéticas/tratamento farmacológico , Medicamentos de Ervas Chinesas/uso terapêutico , Túbulos Renais/efeitos dos fármacos , Proteína-2 Relacionada a Receptor de Lipoproteína de Baixa Densidade/metabolismo , Absorção/efeitos dos fármacos , Animais , Astragalus propinquus/química , Biomarcadores/sangue , Biomarcadores/urina , Glicemia/metabolismo , Diabetes Mellitus Experimental/complicações , Diabetes Mellitus Experimental/tratamento farmacológico , Nefropatias Diabéticas/metabolismo , Nefropatias Diabéticas/fisiopatologia , Modelos Animais de Doenças , Medicamentos de Ervas Chinesas/administração & dosagem , Testes de Função Renal , Túbulos Renais/metabolismo , Túbulos Renais/fisiopatologia , Masculino , Fitoterapia/métodos , Ratos , Ratos Sprague-Dawley , Salvia miltiorrhiza/química , EstreptozocinaRESUMO
OBJECTIVE: To observe the effect of Taohong Siwu decoction (THSWD) on micro-vascular density (MVD) in rat uterus, the content of angiopoietin-1 (Ang-1) and angiopoietin-2 (Ang-2) in serum, and the expression of tyrosine kinasa receptor (Tie-2) in uterus. METHOD: Early pregnancy rats were intragastrically administrated with misoprostol (100 microg x kg(-1)) and mifepristong (8.3 mg x kg(-1)) to established the incomplete-abortion model. The incomplete-abortion rats were randomly divided into the model group (the same volume of distilled water), the positive control group (at the daily dose of 4.3 g x kg(-1) Motherwort Particles), and THSWD-treated groups (at the daily dose of 18.0, 9.0 and 4.5 g x kg(-1)). Pregnant rats were taken as the control group (the same volume of distilled water). After the successive oral administration for 7 days, blood was collected from aorta abdominalis, and rat uterine tissues were collected. The content of serum Ang-1 and Ang-2 were detected by ELISA; And the levels of Tie-2 and MVD in uterine tissues were detected by SP immunohistochemistry. RESULT: THSWD remarkably increased the levels of MVD in uterus of medicine-induced abortion rats, the content of Ang-1 and Ang-2 in serum, and the expression of Tie-2 in uterine tissues. CONCLUSION: THSWD has the effect in markedly promoting angiogenesis in incomplete-abortion rats. Its mechanism may be related to the regulation of concentrations of Ang-1 and Ang-2 in serum and Tie-2 in uterine tissues.
Assuntos
Aborto Incompleto/tratamento farmacológico , Aborto Incompleto/genética , Angiopoietina-1/genética , Angiopoietina-2/genética , Medicamentos de Ervas Chinesas/uso terapêutico , Receptor TIE-2/genética , Útero/irrigação sanguínea , Aborto Incompleto/sangue , Angiopoietina-1/sangue , Angiopoietina-2/sangue , Animais , Feminino , Expressão Gênica/efeitos dos fármacos , Humanos , Gravidez , Ratos , Ratos Sprague-Dawley , Receptor TIE-2/metabolismo , Útero/efeitos dos fármacos , Útero/metabolismoRESUMO
Atherosclerosis is a chronic inflammatory disease characterized by increased expression of adhesion molecules, which contribute to monocytes adhesion to vascular endothelial cells (VECs). Paeonol, an active compound isolated from cortex Moutan, has been shown to have therapeutic effects on atherosclerotic animals. The present study aims to investigate whether paeonol can inhibit monocyte adhesion to vascular endothelial cells induced by oxidized Low-Density Lipoprotein (ox-LDL) and its possible therapeutic molecular mechanism. Exposure to ox-LDL (50, 100 µg/mL) induced damaged to VECs leading to decreased survival rates (p<0.01). Paeonol (7.2-18.0 µM) partially restored survival and reduced lactate dehydrogenase (LDH) release in VECs in a concentration-dependent manner (p<0.01). Adhesion of monocytes to VECs was dramatically prevented by paeonol at 21.6 and 25.2 µM (p<0.01). In addition, paeonol (14.4-21.6 µM) repressed the expression of vascular cell adhesion molecule-1 (VCAM-1) and lowered the levels of phosphor-c-Jun N-terminal kinase (P-JNK)1/2, phosphor-extracellular signal-regulated kinase (P-ERK)1/2 and P-p38 in a dose-dependent manner. The molecular effects of paeonol were more pronouced when companied with mitogen activated protein kinases (MAPKs) inhibitors. These data suggest that paeonol (10.8-25.2 µM), at certain concentrations, prevents monocyte adhesion to VEC induced by ox-LDL, probably by means of blocking one or more target proteins on MAPKs signaling pathway. These results indicate that paeonol has potential protective effects on the development of atherosclerosis.
Assuntos
Acetofenonas/farmacologia , Aterosclerose/metabolismo , Células Endoteliais/efeitos dos fármacos , Lipoproteínas LDL/metabolismo , Proteínas Quinases Ativadas por Mitógeno/antagonistas & inibidores , Monócitos/efeitos dos fármacos , Paeonia/química , Acetofenonas/uso terapêutico , Animais , Aterosclerose/tratamento farmacológico , Adesão Celular/efeitos dos fármacos , Relação Dose-Resposta a Droga , Medicamentos de Ervas Chinesas/farmacologia , Medicamentos de Ervas Chinesas/uso terapêutico , Células Endoteliais/metabolismo , MAP Quinases Reguladas por Sinal Extracelular/metabolismo , Proteínas Quinases JNK Ativadas por Mitógeno/metabolismo , L-Lactato Desidrogenase/metabolismo , Monócitos/citologia , Fitoterapia , Ratos , Ratos Sprague-Dawley , Transdução de Sinais/efeitos dos fármacos , Molécula 1 de Adesão de Célula Vascular/metabolismo , Proteínas Quinases p38 Ativadas por Mitógeno/metabolismoRESUMO
AIM: To observe the effects of Kupffer cells on hepatic drug metabolic enzymes. METHODS: Kunming mice were i.p. injected with GdCl310, 20, 40 mg/kg to decrease the number and block the function of kupffer cells selectively. The contents of drug metabolic enzymes, cytochrome P450, NADPH-cytochrom C redutase (NADPH-C), aniline hydroxylase (ANH), aminopyrine N-demethylase (AMD), erythromycin N-demethylase (EMD), and glutathione s-transferase (mGST) in hepatic microsome and S9-GSTpi, S9-GST in supernatant of 9 000 g were accessed 1 d after the injection. The time course of alteration of drug metabolic enzymes was observed on d 1, 3, and 6 treated with a single dose GdCl3. Mice were treated with Angelica sinensis polysaccharides (ASP) of 30, 60, 120 mg/kg, i.g., qd x 6 d, respectively and the same assays were performed. RESULTS: P450 content and NADPH-C, ANH, AMD, and EMD activities were obviously reduced 1 d after Kupffer cell blockade. However, mGST and S9-GST activities were significantly increased. But no relationship was observed between GdCl3 dosage and enzyme activities. With single dose GdCl3 treatment, P450 content, NADPH-C, and ANH activities were further decreased following Kupffer cell blockade lasted for 6 d, by 35.7%, 50.3%, 36.5% after 3 d, and 57.9%, 57.9%, 63.2% after 6 d, respectively. On the contrary, AMD, EMD, mGST, and S9-GST activities were raised by 36.5%, 71.9%, 23.1%, 35.7% after 3 d, and 155%, 182%, 21.5%, 33.7% after 6 d, respectively. Furthermore, the activities of drug metabolic enzymes were markedly increased after 30 mg/kg ASP treatment, and decreased significantly after 120 mg/kg ASP treatment. No change in activity of S9-GSTpi was observed in the present study. CONCLUSION: Kupffer cells play an important role in the modulation of drug metabolic enzymes. The changes of drug metabolic enzyme activities depend on the time of kupffer cell blockade and on the degree of Kupffer cells activated. A low concentration of ASP increases the activities of drug metabolic enzymes, but a high concentration of ASP decreases the activities of drug metabolic enzymes.