ABSTRACT
Objective To optimize the method of combining azomethane oxide(AOM)and dextran sodium sulfate(DSS)to create a colitis-associated colon cancer(CAC)model,and to explore the pathogenesis of the intestinal flora in CAC.Methods Model groups A and B were established by one and two injections of AOM,respectively,combined with free drinking of DSS,and a normal control group was injected intraperitoneally with normal saline combined with purified water(n=10 mice per group).The better modeling scheme was selected by comprehensive evaluation of the disease activity index score,colon length,tumor rate,and mortality.Serum levels of interleukin-6(IL-6),tumor necrosis factor-α(TNF-α),and tumor markers CA199,CEA,and CA724 were detected by enzyme-linked immunosorbent assay.Colon lesions were evaluated by hematoxylin and eosin(HE)staining.Changes in the intestinal microbiota in CAC mice were detected by 16S rDNA high-throughput gene sequencing analysis of mouse feces.Results Both single and enhanced AOM injections combined with DSS induced CAC mice;however,colon growths were larger,more closely arranged,and their morphological size was more consistent in group B compared with group A,with a tumor-formation rate of 100%.IL-6 levels were increased in the model group compared with the normal group(P<0.05).TNF-α levels were increased in the model group compared with the normal group(P>0.05).The CA199 and CEA levels were also significantly increased(P<0.05),but CA724 levels were not.Infiltration of inflammatory cells in the colon detected by HE pathology was accompanied by high-grade intraepithelial tumor-like changes on the surface of the lumen.The diversity and abundance of intestinal bacteria were decreased in CAC mice compared with normal mice:phyla Verrucomicrobiota and Actinobacteriota were significantly increased(P<0.05),Bacteroidota and Campilobacterota were significantly decreased(P<0.05).Akkermansia,Prevotellaceae,Ruminococcus,and Bifidobacterium were significantly increased(P<0.05),and Roseburia,Rikenellaceae_RC9_gut_group,Anaeroplasma,and Muribaculaceae were significantly decreased(P<0.05).Conclusions Two injections of AOM combined with 1.5%(1.5 g/100 mL)DSS induced CAC model mice with a high colon-tumorigenesis rate,uniform tumor morphology,and low mortality,and may thus be the preferred modeling scheme for pharmacodynamic experiments.Disorders or dysfunction of the intestinal flora may lead to increased permeability,loss of intestinal mucosal barrier function,and the release of enterogenic endotoxins,Resultsing in a sustained inflammatory response,as an indirect or direct cause of CAC pathogenesis.
ABSTRACT
ObjectiveTo induce the rat model of ulcerative colitis (UC) with spleen-kidney Yang deficiency and liver depression, and explore the efficacy and mechanism of Sishenwan combined with Tongxie Yaofang (SSW&TXYF) based on the therapeutic principles of tonifying spleen, soothing liver, warming kidney, and astringing intestine. MethodSixty male SD rats were randomized into normal, model, mesalazine, and high-, medium-, and low-dose SSW&TXYF groups. The rats in other groups except the normal group were administrated with Sennae Folium decoction and hydrocortisone and received tail clamping for 14 days. On day 14, rats received enema with TNBS-ethanol solution to induce UC. The rats were administrated with corresponding drugs from day 15 of modeling, and the body weight and mental state were observed and recorded. The sucrose preference test was performed from day 25. On day 28, the rectal temperature was measured, and the rats were administrated with 3% D-xylose solution at a dose of 10 mL·kg-1 by gavage. Blood was sampled 1 h later, from which the serum was collected for measurement of the D-xylose content. The serum, hippocampus, and colorectum samples of rats were collected on day 29. The levels of gastrin (GAS), adrenocorticotropic hormone (ACTH), corticosterone (CORT), cyclic adenosine monophosphate (cAMP), cyclic guanosine monophosphate (cGMP), interleukin (IL)-4, tumor necrosis factor (TNF)-α, and interferon (IFN)-γ in the serum and 5-hydroxytryptamine (5-HT) in the hippocampus were determined by enzyme-linked immunosorbent assay. Hematoxylin-eosin staining was employed to reveal the colonic lesions. The mRNA and protein levels of p38 mitogen-activated protein kinase (MAPK), extracellular signal-regulated kinase (ERK), and c-Jun N-terminal kinase (JNK) in the colon tissue were determined by Real-time PCR and Western blot, respectively. ResultCompared with the normal group, the model group showed decreased body weight, anal temperature, and D-xylose content in the serum and increased GAS content (P<0.01). The modeling led to cAMP/cGMP unbalance and decreased the ACTH and CORT content in the serum (P<0.01), the preference for sucrose water, and the 5-HT content in the hippocampus (P<0.01). Moreover, it shortened the colorectal length and caused massive infiltration of inflammatory cells and severe structural damage in the colon tissue. High, medium, and low doses of SSW&TXYF improved above indicators (P<0.05, P<0.01), reduced inflammatory infiltration, and repaired the pathological damage of the tissue. Compared with the normal group, the model group showed lowered IL-4 level (P<0.01) and elevated TNF-α and IFN-γ levels (P<0.05, P<0.01) in the serum, as well as up-regulated expression of p38 MAPK, ERK, and JNK (P<0.05, P<0.01). Compared with the model group, SSW&TXYF elevated the IL-4 level (P<0.01), lowered the TNF-α and IFN-γ levels (P<0.05, P<0.01), and down-regulated the mRNA and protein levels of p38 MAPK, ERK, and JNK (P<0.05, P<0.01). ConclusionA rat model of UC with spleen-kidney Yang deficiency and liver depression was successfully established. SSW&TXYF can significantly mitigate this syndrome by reducing the inflammatory response in the colon and inhibiting the MAPK pathway.
ABSTRACT
ObjectiveTo investigate the efficacy of Huangqintang on mouse models of colitis-associated colon cancer (CAC) and explore the mechanism of Huangqintang in regulating immune function and inflammatory response, inhibiting abnormal cell proliferation, and delaying or inhibiting CAC formation in CAC. MethodC57BL/6J mice were randomly divided into a normal group, model group, mesalazine group, and high- and low-dose Huangqintang groups according to body weight, with 12 mice in each group. Except for the normal group, the rest of the mice were given two intraperitoneal injections of 10 mg·kg-1 azomethane (AOM) and allowed to drink 1.5% dextran sodium sulfate (DSS) freely for seven days and water normally for two weeks. Then, two cycles of ''DSS-drinking water'' were repeated. During the administration of DSS, mice in the normal group and model group were given gavage in equal doses of pure water. Mice in the mesalazine group were given 150 mg·kg-1·d-1 mesalamine suspension for gavage, and mice in the high- and low-dose Huangqintang groups were given 18 and 9 g·kg-1·d-1 Huangqintang for gavage, respectively. Each group was given one dose daily until the end of three cycles. After the intervention, the body weight, colon length, and number of colon tumors in each group were measured, and disease activity index (DAI) scores were performed. The serum contents of interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), interleukin-4 (IL-4), interleukin-10 (IL-10), and gastrointestinal tumor marker carbohydrate antigen-199 (CA199) were detected by enzyme linked immunosorbent assay (ELISA). The colonic lesions were observed by hematoxylin-eosin (HE) staining. The expression of proliferative cell-associated antigen (Ki67) was observed by immunohistochemistry. The expression of T lymphocyte subsets (CD3+, CD4+, CD8+, and CD49b+) in mouse plasma was detected by flow cytometry. Fluorescein isothiocyanate-D (FITC-D) content in mouse serum was detected by fluorescent labeling method. The Western blot method was used to detect the expression of Cyclin D1, cyclin-dependent kinase 2 (CDK2), cyclin-dependent kinase 4 (CDK4), and tightly junction-related Occludin and Claudin-1. ResultCompared with the normal group, the body weight of mice in the model group decreased. DAI score increased significantly, and the colon became shorter. Pro-inflammatory factors such as IL-6, TNF-α, and IL-1β increased, and IL-6 and TNF-α were significantly increased (P<0.05). The inflammatory factor IL-4 (P<0.05) and IL-10 were significantly reduced, and the tumor marker CA199 was significantly increased (P<0.01). HE staining showed that colon lesions, intestinal mucosal epithelial defects with a large number of inflammatory infiltrates, serious crypt structure damage, and glandular arrangement disorder were observed in the model group. Ki67 positive granules were expressed in large areas of colonic tissue. The serum CD4+ and CD4+/CD8+ of mice in the model group decreased significantly (P<0.05), and CD8+ increased significantly (P<0.05). The plasma content of FITC-D in the model group was significantly increased (P<0.05), and the expression of Cyclin D1, CDK2, and CDK4 proteins in colon tissue was significantly increased (P<0.05, P<0.01). In addition, the expression of Occludin and Claudin-1 was significantly decreased. Compared with the model group, the body weight of mice in the mesalazine group and the high- and low-dose Huangqintang groups increased. DAI score decreased, and the colon became longer. IL-6, TNF-α, and IL-1β expression decreased (P<0.05, P<0.01), but there was no significant change in IL-4 and IL-10. The content of CA199 was significantly reduced (P<0.05), and the colomatoid lesions and inflammatory infiltrates were reduced in the mesalazine group and the Huangqintang group. The crypt structure damage was lighter, and the positive expression of Ki67 was reduced. CD4+, CD4+/CD8+, and CD49b+ increased, and the difference was not statistically significant. FITC-D content decreased (P<0.05). The expression of Cyclin D1, CDK2, and CDK4 decreased (P<0.05, P<0.01), and Claudin-1 and Occludin protein expression increased in the high-dose Huangqintang group (P<0.05). ConclusionHuangqintang has a certain delay and inhibitory effect on AOM/DSS-induced inflammatory cancer transformation, and its mechanism of action may be related to regulating immune function and inflammatory response, inhibiting the release of pro-inflammatory factors, repairing damaged intestinal barriers, inhibiting abnormal proliferation of colon cells, and intervening in the formation and development of CAC colon tumors.
ABSTRACT
At present, major depressive disorder (MDD) is highly prevalent with advanced neurological disorders as the main pathological manifestations. As the physiological function bearer of higher neural activity, gray matter has become the focus of MDD treatment. However, recent research has shown that white matter and gray matter are independent of each other in the central nervous system (CNS), and their functions are integrated and linked. In addition to gray matter damage, white matter damage is also the core driving event of disease progression and determines the outcome of MDD. At the treatment level, the current drug treatment of MDD mainly focuses on gray matter repair, while ignoring the importance of white matter integrity for the treatment of the disease, which has become the weakness of the current treatment of MDD. Traditional Chinese medicine (TCM) has good application potential in white matter repair. This paper elaborated on the following three aspects. ① The roles of white matter damage in the occurrence and development of MDD were summarized. ② The key link of white matter repair in MDD was elaborated with microglia microenvironment regulation as the entry point. ③ The application value of TCM in white matter repair in MDD was analyzed. This review aims to highlight the importance of white matter integrity in the treatment of MDD and is expected to expand the understanding dimension of the activity of related Chinese medicines in MDD from the perspective of white matter repair and analyze its potential application value.
ABSTRACT
Ulcerative colitis (UC) is a chronic intestinal disease with unknown etiology, with main symptoms of abdominal pain, diarrhea, mucus, pus, and blood in the stool. It can be accompanied by various complications and has a high risk of developing to colon cancer. In recent years, the incidence of UC and related colon cancer has been increasing, which seriously affects human health and quality of life. The operation, immunosuppressant, etc. are the main approaches in the modern clinical treatment of UC and related colon cancer, but these methods all have different toxic and side effects, and the therapeutic effect is not ideal. For many years, traditional Chinese medicine (TCM) has attracted much attention in the treatment of UC and related colon cancer due to its slightly toxic side effects and remarkable curative efficacy. Huangqintang, derived from the Shang Han Lun (伤寒论), is composed of Scutellariae Radix, Paeoniae Radix Alba, Glycyrrhizae Radix et Rhizoma, and Jujubae Fructus with the functions of clearing heat, checking diarrhea, harmonizing the middle, and relieving pain, and has a significant effect on the treatment of UC. Huangqintang has complex compositions and plays roles with multiple targets and pathways. According to the literature and the research results of this research group for many years, it was found that the mechanism of Huangqintang in the treatment of UC and related colon cancer was presumably related to the protection of the intestinal mucosal barrier, inhibition of inflammatory response, promotion of mitophagy, inhibition of oxidative stress, regulation of intestinal flora, cell cycle, and gene expression, suppression of cell proliferation, and promotion of apoptosis. To provide theoretical references for an in-depth study of the mechanism and clinical use of Huangqintang, this paper reviewed the research advances in recent years.
ABSTRACT
ObjectiveTo evaluate the pharmacodynamic effect of Huangqintang (HQT) on ulcerative colitis (UC) model mice and investigate its protective effect against UC by regulating intestinal flora. MethodMale Balb/c mice were randomly divided into control group,model group, high-, medium-, and low-dose HQT groups (20, 10, 5 g·kg-1), flora interference group, flora interference model group, and flora interference-drug treatment group (HQT, 20 g·kg-1). The flora interference model was constructed through intragastric administration of antibiotics (200 mg·kg-1 bacitracin and 200 mg·kg-1 vancomycin) for 8 d, and the UC model was constructed by allowing mice with free access to 3% dextran sulfate sodium (DSS) solution for 7 d. HQT was administered for 7 d. After the experiments, the mice were sacrificed, and blood, colon, and feces were collected. Hematoxylin-eosin (HE) staining was performed to observe the colonic lesions. The serum levels of interleukin (IL)-4, IL-6, IL-10, and tumor necrosis factor (TNF)-α were detected by enzyme-linked immunosorbent assay (ELISA). The mRNA and protein expression of Claudin1, MUC1, Occludin, and zonula occludens-1(ZO-1) in colon tissues was detected by Real-time fluorescence quantitative polymerase chain reaction (Real-time PCR) and Western blot, respectively. The fecal DNA of mice was extracted and analyzed by high-throughput sequencing. ResultCompared with the normal group, the model group showed increased serum content of IL-4, IL-6, and TNF-α (P<0.05, P<0.01) and decreased IL-10 (P<0.05). Compared with the model group, the HQT groups displayed decreased serum levels of IL-4, IL-6, and TNF-α (P<0.05, P<0.01), increased IL-10 content (P<0.01), increased mRNA and protein expression levels of Claudin1, MUC1, Occludin, and ZO-1 (P<0.05, P<0.01). After flora interference, the diversity and abundance of intestinal bacteria decreased. To be specific, Proteobacteria increased (P<0.01), and Firmicutes and Bacteroidetes decreased (P<0.01). After UC induction by DSS, Bacteroidetes and Tenericutes decreased (P<0.05). The high-, medium-, and low-dose HQT groups showed increased Bacteroidetes and Tenericutes (P<0.05, P<0.01) and decreased Firmicutes (P<0.05). Additionally, the abundance of Lactobacillus, Lachnospiraceae NK4A136 group, Escherichia-Shigella, and Helicobacteris was positively proportional to the dose of HQT. ConclusionHQT can inhibit the inflammatory response of UC mice, restore the imbalance of intestinal flora, and repair the damaged intestinal mucosal barrier.
ABSTRACT
ObjectiveTo explore the anti-inflammatory mechanism of Huangqintang based on the inflammation model in RAW264.7 cells. MethodHuangqintang was prepared and the safe dose to RAW264.7 cells was screened out. The RAW264.7 cells were seeded in 24-well plates and incubated with Huangqintang and lipopolysaccharide (LPS), successively. The concentrations of nitric oxide (NO), interleukin (IL)-6, tumor necrosis factor (TNF)-α, and prostaglandin E2 (PGE2) were measured by Griess assay and enzyme-linked immunosorbent assay (ELISA), respectively. Meanwhile, RAW264.7 cells were inoculated in 6-well plates, and normal group, LPS group, LPS+Huangqintang group, nuclear factor-κB (NF-κB) p65 inhibitor PDTC group, p38 mitogen-activated protein kinase (MAPK) inhibitor SB203580 group, extracellular signal-regulated kinase (ERK) inhibitor PD98059 group, c-Jun N-terminal kinase (JNK) inhibitor SP600125 group, and Janus kinase (JAK) inhibitor AG490 group were set up. After the cells were incubated with corresponding inhibitors and Huangqintang and stimulated by LPS, RNA and protein were extracted. The mRNA and protein expression levels of NF-κB p65, p38 MAPK, ERK, JNK, and JAK were detected by Real-time fluorescence-based quantitative polymerase chain reaction (Real-time PCR) and Western blot, respectively, to explore the anti-inflammatory mechanism of Huangqintang by regulating the NF-κB, MAPK, and JAK/signal transducer and activator of transcription protein (STAT) signaling pathways. ResultAfter stimulation with LPS, the concentrations of NO, IL-6, TNF-α, and PGE2 in the cells of the model group increased significantly(P<0.05,P<0.01). Compare with the model group, after incubation with Huangqintang, the secretion of NO, IL-6, TNF-α, and PGE2 showed a downward trend (P<0.05,P<0.01). Compared with the normal group, the model group showed increased mRNA expression of p38 MAPK, ERK, JNK, JAK, and NF-κB p65 and total protein expression in cells after stimulation with LPS (P<0.05,P<0.01). Compare with the model group,after incubation with Huangqintang, the total protein and mRNA expression of p38 MAPK, ERK, JNK, JAK, and NF-κB p65 in inflammatory cells decreased (P<0.05,P<0.01). Meanwhile, the expression of NF-κB p65 total protein and mRNA in each inhibitor group showed a downward trend (P<0.05,P<0.01). ConclusionHuangqintang can inhibit the inflammatory response through the NF-κB, MAPK, and JAK-STAT signaling pathways.
ABSTRACT
ObjectiveTo verify the anti-oxidative stress effect of Huangqintang based on the nuclear factor E2-related factor 2 (Nrf2) signaling pathway by using Caco-2 cells as a carrier and RNA interference (RNAi) technology with in vitro experiments. MethodThe Caco-2 cells in the logarithmic growth phase were transfected with siRNA to construct siRNA Caco-2 cells. After normal Caco-2 cells and siRNA Caco-2 cells were incubated with Huangqintang of different doses, RNA and protein were extracted. Real-time fluorescence quantitative polymerase chain reaction (Real-time PCR) and Western blot were used to detect the mRNA and protein expression of heme oxygenase-1 (HO-1), NAD(P)H quinone oxidoreductase 1 (NQO1), glutathione S-transferase (GST), Kelch-like ECH-associated protein 1 (Keap1), and Nrf2. Meanwhile, the activities of superoxide dismutase (SOD) and GSH-Px, as well as the expression levels of malondialdehyde (MDA) and reactive oxygen species (ROS), were detected by the colorimetric method and the probe method. ResultCompared with the results in the normal group, only the 400 mg·L-1 Huangqintang group and the sulforaphane (SFN) group could reduce the content of ROS and MDA in Caco-2 cells (P<0.01), while the activities of SOD and GSH-Px in the cells of the Huangqintang groups and the SFN group showed an upward trend. Furthermore, there were significant differences in the 400 mg·L-1 Huangqintang group/the SFN group and the normal group (P<0.01). Meanwhile, the protein and mRNA expression levels of HO-1, GST, Keap1, NQO1, and Nrf2 showed an upward trend in all groups (P<0.05, P<0.01). After transfection, compared with the normal group, the model group showed increased content of MDA and ROS, blunted activities of GSH-Px and SOD, and reduced protein and mRNA expression of HO-1, GST, Keap1, and NQO1 (P<0.05, P<0.01). After drug incubation, compared with the model group, the SFN group showed potentiated SOD activity, and the SFN group and the Huangqintang groups showed enhanced GSH-Px activity (P<0.01). Moreover, the activities of SOD and GSH-Px in the 400 and 200 mg·L-1 Huangqintang groups and the SFN group showed an upward trend (P<0.01), and the content of MDA in the 400 mg·L-1 Huangqintang group and the SFN group showed a downward trend. ROS decreased in all groups with drug intervention (P<0.01), and the protein and mRNA expression of HO-1, GST, Keap1, NQO1, and Nrf2 increased to varying degrees (P<0.05, P<0.01). ConclusionHuangqintang can play an anti-oxidative stress role by regulating the Nrf2 pathway.
ABSTRACT
Objective To investigate the death of MHD patients with different blood phosphorus variability.Methods The clinical data of seventy-four cases of MHD patients with more than 3 months dialysis age in the hemodialysis center of Chaozhou Central Hospital from January 2014 to December 2016 were retrospectively analyzed.The general data and laboratory biochemical indexes were collected and according to the difference in coefficient of variation (CV),the patients were divided into two groups:high blood phosphorus variability group (CV≥0.226 mmol/L) and low blood phosphorus variability group (CV <0.226 mmol/L).The relationship between the coefficient of variation and all-cause mortality and cardiovascular mortality in MHD patients was analyzed.Results Among all selected patients,all-cause mortality rate was 22.9% (17/24),12.2% (9/74) died of cardiovascular disease,and the all-cause mortality 34.2% (13/38)in the high blood phosphorus variability group was significantly higher than that in the other one1 1.1% (4/36) (x2=5.574,P<0.05);but there was no significant difference between the two groups in cardiovascular disease mortality (X2 =0.962,P>0.05).And there was no significant difference between the serum phosphorus standard group and the substandard group in the all-cause mortality and the cardiovascular disease mortality (x2 =0.389,0.065,P > 0.05).There was no significant difference in the cumulative survival rate between the patients in the serum phosphorus standard group who experienced all-cause death and cardiovascular disease death and the patients in the substandard group (P>0.05).In the high blood phosphorus variability group,patients who reached serum phosphorus standard had higher total mortality rate and cardiovascular disease mortality rate,compared with patients who achieved serum phosphorus standard or experienced low phosphorus deficiency in the low blood phosphorus variability group (x2=0.211,0.197,P >0.05).Kaplan/Meier analysis showed that the cumulative survival rates of all patients with all-cause death and cardiovascular death in the high blood phosphorus variability group were significantly lower than those in the low level group (P< 0.05).Conclusion All-cause mortality and cardiovascular disease mortality are high in MHD patients with large variability in blood phosphorus,and the degree of blood phosphorus variation is helpful to predictthe death of MHD patients.
ABSTRACT
OBJECTIVE:To study the protective effects of Baimai ointment on sciatic nerve injury in rabbit. METHODS:The rabbit sciatic nerve injury model was induced by forceps operation. 180 rabbits were randomly divided into 6 groups,with 30 rab-bits in each group (10 rabbits of 7,14,28 d),including sham operation group (blank matrix of Baimai ointment for external use),model group(same as sham operation group),positive group [ig Mecobalamine tablet solution 1.25×10-4 g/(kg·d)] and Bai-mai low-dose,medium-dose and high-dose groups [Baimai ointment 0.33,0.67 and 1.34 g/(kg·d)for external use],once a day, for consecutive 7,14 and 28 d. Gait instrument was used to test the foot-touch-land force of rabbit;pathology examination was conducted for sciatic nerve resection;immunohistochemistry was used to detect the levels of NOS and NMDA in nerve tissue. RE-SULTS:Baimai ointment can significantly relieve sciatic nerve injury,edema and inflammatory reaction;7,14 and 28 d after med-ication,compared with model group,foot-touch-land force decreased significantly in rabbits with sciatic nerve injury (P<0.01);14,28 d after medication,foot-touch-land force of rabbits increased significantly in positive group,Baimai ointment groups,while the levels of NOS and NMDA decreased in nervous tissue(P<0.05). CONCLUSIONS:Baimai ointment can protect sciatic nerve with function injury in rabbits.
ABSTRACT
The quality control processes for herbal medicines have been problematic. Flavonoids are the major active components of Huangqin Tang (HQT, a traditional Chinese medicine formula). In this study, we used a combinative method approach consisting of chromatographic fingerprinting (high performance liquid chromatography; HPLC), quantitative methods and a pharmacodynamic evaluation model to analyze the flavonoids of HQT obtained from different sources. Ten batches of HQT were analyzed by the HPLC fingerprinting method and 26 common peaks were detected, of which 23 peaks corresponded with the chemical profile of HQT. In addition, 11 major compounds were identified by LC-MS analysis (liquid chromatography-tandem mass spectrometer; LC-MS (n) ) and quantified by the HPLC quantitative method approach. The studied 10 batches of HQT were found to be homogeneous in their composition with a similarity between 0.990 and 1.000. The distribution of the 11 identified compounds was found to be very similar among the batches. Only slight pharmacodynamic differences were detected between the different batches, confirming the homogeneity of HQT. The results of this study prove that the combination of chromatographic fingerprinting and quantitative analysis can be readily used for comprehensive quality control of herbal medicines.
ABSTRACT
To investigate the effect of huangqin tang on expression of cytokines and NF-κB p65 in rats with ulcerative colitis (UC), and to probe into its underlying mechanisms of action. The mode of UC rats with cell immunoreactivity was made using compound method (trinitrobenzene sulfonic acid and ethanol). Rats were randomly divided into control group, model group, SASP group and high dose, middle dose and low dose of huangqin tang group. The food intake, body weight and microscopic damage of rats in each group were evaluated after being treated for five days. The blood and colon tissue were also collected. Production of NO was detected by Griess assay, the expression levels of IL-6, TNF-α, PGE2 were detected by ELISA. ICH method was undertaken to determine the expression of NF-κB p65 protein in colon tissue. The food intake and body weight of model group rats were lower than that of control group. The expression levels of NO, IL-6, TNF-α, PGE2 in serum and NF-κB p65 protein of colon tissue in model group were higher than that of control group. The above indexes were ameliorated in high and middle dose of huangqin tang groups. But there was no significant difference with SASP group. NF-κB p65 may be involved in the pathogenesis of UC, and huangqin tang can inhibit the relative activity of NF-κB p65, and decrease the expression levels of NO, IL-6, TNF-α and PGE2.
ABSTRACT
A simple and selective HPLC method for simultaneous determination and quantification of anthraquinones, lignans and flavonoids in Xiao-Cheng-Qi Tang (XCQT), Hou-Po-San-Wu Tang (HPSWT) and Hou-Po-Da-Huang Tang (HPDHT) was developed and validated. An Agilent Zorbax SB-C 18 (4.6 mm x 250 mm, 5 µm) column with the mobile phase of acetonitrile and 0.5% acetic acid aqueous solution in gradient elution mode was used. The flow rate was 1.0 mL · min(-1) at 30 °C, and injection volume was 10 µL. The detection wavelength was set at 254 nm and 294 nm simultaneously for the quantitative analysis. The current HPLC assay was validated for linearity, intra-day and inter-day precisions, accuracy, recovery and stability. The method was applied to the content comparison of the gallic acid, cinnamic acid, sennoside A, sennoside B, rhein, emodin, aloe-emodin, chrysophanol, physcion, magnolol, honokiol, narirutin, naringin, hesperidin, neohesperidin, hesperetin, naringenin and nobiletin in XCQT, HPSWT and HPDHT. The good linear equations of eighteen constituents were obtained within the investigated ranges (r > 0.998). The recovery of the method was 94.28%-99.89% and the precision was less than 5%. The sample was stable within 16 h. There were some differences between the contents of anthraquinones, lignans and flavonoids in analogous formulae about XCQT. XCQT contained the greatest abundance of anthraquinones and flavonoid, HPSWT contained the greatest abundance lignans. In conclusion, the methods are simple, low-cost, precise, accurate and reliable for the determination of eighteen constituents in analogous formulae about XCQT, and these results provide methodological support for its quality control.
ABSTRACT
The pharmacodynamic (PD) and pharmacokinetic (PK) properties of Huangqin Tang (HQT) were investigated in yeast-induced febrile rats. Blood sample and rectal temperature data of the rats were collected at different times after single oral administration of HQT at 20 g x kg(-1). The plasma concentrations of paeoniflorin, baicalin, wogonoside, baicalein, wogonin, oroxylin A, glycyrrhizic acid and glycyrrhetinic acid were quantified by a sensitive liquid chromatography-tandem mass spectrometric (LC-MS) method. The blood concentrations of PGE2, 1L-1β and TNF-α were detected by radioimmunoassay (RIA). All pharmacokinetic parameters were processed by non-compartmental analysis using WinNonlin software. The potential relationship between the mean concentration of eight constituents and the antifebrile efficacy was investigated by calculating Pearson correlation coefficients. It was found that HQT had significant antifebrile efficacy in yeast-induced febrile rats, but had no effect to normal rats. The antifebrile effect of HQT can be attributed to the inhibition of PGE2, 1L-1β and TNF-α. The constituents (baicalin, wogonoside, baicalein, wogonin, oroxylin A, glycyrrhizic acid and glycyrrhetinic acid) in febrile rats had delayed absorption and elimination, a longer residence time in the body, and higher C(max) and AUC than those in normal rats. Febrile condition could affect the pharmacokinetic behaviour of HQT in vivo; the flavonoids with the same backbone showed the similar fate in the body; baicalein and wogonin had a strong positive correlation (R > 0.66, P ≤ 0.02) with the antifebrile efficacy determined. Together, these constituents demonstrated different pharmacokinetic properties in the febrile body.
ABSTRACT
Objective To investigate the effects of Saussurea involucrate by cell culture on increasing the calcium content and bone density after 90 days feed in ovariectomized rats. Methods Osteoporosis model rats were induced by ovariectomy, treated with 16.7、33.3、100 mg/(kg?d)cell culture of Saussurea involucrate for 90 days. Body weight was observed, calcium content was measured through atomic absorption method and the bone density was detected by bone sonometers. Results cell culture of Saussurea involucrate(16.7、33.3、100 mg/kg)can increase the calcium content[(380.1 ± 1.9)mg/g、(370.7 ± 1.1)mg/g、(363.5 ± 2.4)mg/g] significantly(P<0.01)than the model group; increase distal femoral bone mineral density[(0.041 ± 0.017)g/cm2、(0.042±0.023)g/cm2、(0.040±0.008)g/cm2] significantly(P<0.01 or 0.05) than the model group(0.022±0.014)g/cm2; and also increase the middle femoral bone mineral density, the low-dose group (0.305±0.030)g/cm2 significantly (P<0.05) than the model group(0.259±0.061)g/cm2. Conclusion The cell culture of Saussurea involucrate has the effect of increasing the bone density.
ABSTRACT
Objective To explore the influence on anti-inflammatory efficacy and bitter cold properties of Coptis processed by Evodia juice. Methods Auricle swelling model induced by croton oil in mice was used in the anti-inflammatory efficacy experiment. The mice were assigned randomly into distilled control group, dexamethasone group, crude drug group, 3 processed drug groups that Euodia juice which plant origin was Euodia rutaecarpa (Juss.) var. bodinaieri (Dode) Huang (SMYHL), Evodia rutaecarpa (Juss.) Benth and Euodia rutaecarpa (Juss.) Benth var. officinalis (Dode) Huang, respectively. The crude drug group and the 3 processed drugs groups were orally administrated one time and the dosages were all 6 g/kg. The control group was orallly administrated the same volume water. Dexamethasone group was intraperitoneally injected dexamethasone 30 g/kg. The swelling degree of each group was observed. In the bitter cold properties experiment, rats were assigned randomly into control group, crude drug group and 3 processed drug groups. The crude drug group and the 3 processed drug groups were orally administrated for 14 days and the dosages were all 3 g/kg. The control group was orally administrated the same volume water. The body weight, capacity for eating and drinking, body temperature, heart rate and blood viscosity of rats were measured. Meanwhile, TSH, IL-2, IL-8 in serum and plasma were determined by radioimmunoassay. Results The degree of mouse auricle swelling was reduced by the crude and processed drugs. Fourteen days after medication, the weight, capacity of eating and drinking of rats were declined to different degrees in crude drug group and in processed drugs group as compared with control group. The blood viscosity was increased and IL-2 was decreased in SMYHL group as compared with control group (P<0.05). TSH was decreased in crude drug group as compared with control group (P<0.05). Conclusion There is no obvious change in anti-inflammatory function of Coptis before and after processed. The cold properties of Coptis can be reduced after processed by Evodia Juice.
ABSTRACT
The current study aims to investigate the pharmacokinetic properties of Huangqin Tang on different oral doses. An LC-MS method for simultaneous determination of flavonoids and terpenoids in rat plasma was developed and validated. Plasma samples were treated with hydrochloric acid (containing 1% ascorbic acid), precipitated with acetonitrile, separated on a Zorbax SB-C18 column, detected by single quadruple mass spectrometry with an electrospray ionization interface, and quantified using selected ion monitoring mode. All pharmacokinetic parameters were processed by non-compartmental analysis using WinNonlin software. The results of specificity, linearity, intra-day and inter-day precisions, accuracy, and stability for LC-MS assay were suitable for the quantification of paeoniflorin, baicalin, wogonoside, baicalein, wogonin, oroxylin A, glycyrrhizic acid and glycyrrhetinic acid in rat plasma. The concentration-time profiles of baicalin, wogonoside, baicalein, wogonin, oroxylin A and glycyrrhizic acid showed double-peak phenomenon after Huangqin Tang was orally administered at 40 g x kg(-1) dose; all eight constituents in rat plasma showed good dose-exposure relationship within the dosage of 10-40 g x kg(-1); although plasma concentrations were different, the flavonoids with the same backbone showed the similar fate in the body with the corresponding dosage. In conclusion, the LC-MS assay was successfully applied for the pharmacokinetic study of multi-constituents of Huangqin Tang with different doses. Additionally, these constituents demonstrated good pharmacokinetic properties in the body.
ABSTRACT
To illustrate the compability rule of Jinlingizi powder, by investigating the effects of Jinlingzi Powder with different compatibility on the enzymatic activity of cytochrome P1 A2 (CYP1A2) from rat liver microsome. The different compability of Jinlingizi powder is designed, based on the orthogonal array L9 (3(4)). In vitro test, rat liver microsomes incubation system is applied to detect the 50% inhibitory concentraton of Jinlingzi powder with different compatibility to cytochrome P1A2 (CYP1A2) enzyme. In vivo experiments, rats is treated orally with the different compability of Jinlingizi powder for 5 days, then be injected with probe drug phenacetin. The biosample from liver tissue is obtained by microdialysis probe, then analysisd by HPLC. The concentration-time data are modulated by software WinNonlin. IC50 data show no significant inhibitory activty to cytochrome P1 A2. Acetaminophen and phenacetin PK parameters indicate that the different compability of Jinlingizi powder can modulate the CYP 1A2 mediated metabolism, which is associate with the compatibility of Jinlingzi powder.
Subject(s)
Animals , Male , Rats , Cytochrome P-450 CYP1A2 Inhibitors , Drugs, Chinese Herbal , Pharmacology , Medicine, Chinese Traditional , Microsomes, Liver , Powders , Rats, WistarABSTRACT
<p><b>OBJECTIVE</b>To observe the impact on absorption of berberine and palmatine in different compatibilities of Wuji pill by the perfused rat intestine-liver preparation.</p><p><b>METHOD</b>Use L9 (3(4)) orthogonal design table, establish the perfused rat intestine-liver preparation, the twelve Wuji pill compatibilities duodenal administrated, collect the perfusate at different times points for LC-MS detection, calculate the absorbed score, Ka.</p><p><b>RESULT</b>Evodiae Fructus and the absorption score, Ka of berberine and palmatine are inverse correlated. The most superior portion which promote the absorption is Coptidis Rhizoma-Evodiae Fructus-Paeoniae Radix Alba 3:1:3.</p><p><b>CONCLUSION</b>Evodiae Fructus suppressed the absorption of berberine and palmatine. With the different portion the absorption also have big different.</p>
Subject(s)
Animals , Male , Rats , Berberine , Metabolism , Pharmacokinetics , Berberine Alkaloids , Metabolism , Pharmacokinetics , Drugs, Chinese Herbal , Pharmacology , Evodia , Chemistry , Intestinal AbsorptionABSTRACT
<p><b>OBJECTIVE</b>To research the representative ingredient excretion in bile using the different compatibilities of Wuji Wan, in order to indicate the regularity of compatibility.</p><p><b>METHOD</b>L9 (3(4)) orthogonal design table was used, in addition 9 simples, altogether 18 compatibilities. After duodenal administration bile at different time spot was collected for LC-MS detection.</p><p><b>RESULT</b>The excretion of Evodiae Fructus was negative correlated with that of berberine and palmatine in bile. The excretion of Paeoniae Radix Alba was positive correlated with that of berberine and palmatine in bile. The excretion of Coptidis Rhizoma, Paeoniae Radix Alba was negative correlated with that of evodiamine, rutaecarpine; meanwhile, the most superior proportion was also caculated which promote the representative ingredient excrete from bile.</p><p><b>CONCLUSION</b>Evodiae fructus suppresses representative ingredients of Coptidis Rhizoma excrete through bile. Paeoniae Radix Alba promote suppresses representative ingredients of Coptidis Rhizoma excrete through bile. Coptidis Rhizoma, paeoniae Radix Alba suppresses representative ingredients of Evodiae Fructus excrete through bile. Coptidis Rhizoma, Evodiae Fructus suppresses representative ingredients of Paeoniae Radix Alba excrete through bile.</p>