Total alkaloids of Coptidis Rhizoma combined with exercise inhibits tumor growth of orthotopically transplanted 4T1 breast cancer mice by blocking cell cycle G_1/S transformation / 中国中药杂志

Breast cancer is one of the leading causes for cancer-related death among women worldwide. Coptidis Rhizoma has antibacterial,anti-inflammatory,anti-tumor and other pharmacological activities,but whether exercise could synergistically promote the role of RC in the treatment of breast cancer has not been reported. In this experiment,the effects and mechanism of total alkaloids of Coptidis Rhizoma combined with exercise on the tumor growth of orthotopically transplanted 4 T1 breast cancer were systemically studied in mice. Balb/C mice transplanted with 4 T1 cells in situ were used as models. The total alkaloids of RC(145 mg·kg-1·d-1) alone or in combination with exercise(10 m·min-1,30 min/time,5 times/week) were given for 28 days,and then the changes in body weight and tumor volume,tumor weight,interleukin-1β(IL-1β),serum estradiol(E2) content,and expression levels of estrogen receptor α(ERα),cell cycle related proteins CDK4,CDK6,cyclin D1,CDK2,and cyclin E in tumor tissues. The results showed that total alkaloids of Coptidis Rhizoma could significantly inhibit the growth of 4 T1 breast cancer in mice(P< 0. 01),and exercise significantly promoted the anti-tumor activity of total alkaloids of Coptidis Rhizoma(P<0. 01),and reduced E2 and IL-1β levels in mice. Western blot and flow cytometry showed that the total alkaloids of Coptidis Rhizoma combined with exercise could down-regulate the protein expression levels of ERα,CDK4,CDK6,cyclin D1,CDK2 and cyclin E in cancer cells,block the transformation of G1/S in 4 T1 cell cycle,and inhibit DNA synthesis in breast cancer cells. The total alkaloids of Coptidis Rhizoma combined with exercise showed synergistic effect in inhibition of tumor growth in mice with orthotopically transplanted 4 T1 breast cancer.

Regulatory effect of coptisine on key genes involved in cholesterol metabolism / 中国中药杂志

To study the effect of cholesterol and 25-OH-cholesterol on cholesterol metabolism in HepG2 cells and the effect of coptisine (Cop) extracted from Coptidis Rhizoma (CR) in reducing and regulating cholesterol. In this study, TC, TG, LDL-c and HDL-c were measured by biochemical analysis; mRNA and protein expressions of LDLR, HMGCR and CYP7A1 were detected by qRT-PCR and Western blot. According to the results, cholesterol and 25-OH-cholesterol inducing could decrease in mRNA and protein expressions of LDLR and CYP7A1, so as to increase TC and LDL-c contents. However, Cop could up-regulate mRNA and protein expressions of LDLR and CYP7A1 and down-regulate that of HMGCR, so as to reduce TC and LDL-c levels. These findings suggested that Cop has potential pharmacological activity for reducing cholesterol, and may reduce cholesterol by regulating mRNA and protein expressions of key genes involved in cholesterol metabolism, such as LDLR, CYP7A1 and HMGCR. This study laid a firm theoretical foundation for developing new natural drugs with the cholesterol-lowering activity.

Study on membrane injury mechanism of total alkaloids and berberine from Coptidis Rhizoma on Aeromonas hydrophila / 中国中药杂志

To explore the antibacterial activity and mechanism of total alkaloids and berberine from Coptidis Rhizoma on Aeromonas hydrophila, and determine the effect of total alkaloids and berberine from Coptidis Rhizoma on minimum inhibitory concentrations, permeability and fluidity of cell membrane, conformation of membrane proteins and virulence factors of A. hydrophila. The results showed that both total alkaloids and berberine from Coptidis Rhizoma had antibacterial activities on A. hydrophila, with minimum inhibitory concentrations of 62.5 and 125 mg · L(-1), respectively. Total alkaloids and berberine from Coptidis Rhizoma could increase the fluidity of membrane, change the conformation of membrane porteins and increase the permeability of bacteria membrane by 24.52% and 19.66%, respectively. Besides, total alkaloids and berberine from Coptidis Rhizoma significantly decreased the hemolysis of exotoxin and the mRNA expressions of aerA and hlyA (P < 0.05, P < 0.01), the secretion of endotoxin and the mRNA expression of LpxC (P < 0.05, P < 0.01). The results suggested that the antibacterial activity of total alkaloids and berberine from Coptidis Rhizoma on A. hydrophila may be related to the bacteria membrane injury. They inhibited the bacterial growth by increasing membrane lipid fluidity and changing conformation of membrane proteins, and reduced the secretion of virulence factors of A. hydrophila to weaken the pathogenicity.

Efficient and rapid liquid reduction animal model / 中国中药杂志

To investigate the practicability of establishing zebrafish lipid-lowering drug screening model and the effect of berberine (BBR) on hyperlipidemic zebrafish. Three-month-old zebrafishes were fed with 4% cholesterol for 0, 2, 4, 8, 14, 20, 25, 30 days, and the level of total cholesterol in serum was measured. Zebrafish were randomly divided into four groups: the control group, the high cholesterol diet group, the 0.01% simvastatin-treated group, the 0.1% berberine-treated group and the 0.2% berberine-treated group. The levels of total cholesterol (TC), triglyceride (TC), low density lipoprotein cholesterol (LDL-c) and high-density lipoprotein cholesterol (HDL-c) in serum were measured; the expression of hepatic HMGCR, LDLR and CYP7A1a mRNA expressions were detected by real time PCR. Oil red O staining was performed to observe the changes in fat content in the liver. According to the result, the level of serum TC in the 4% cholesterol diet group significantly was higher than that of the normal control group in a time-dependent manner and reached a stable level at the 20th day. The BBR group showed significant decreases in the levels of TC, TG and LDL-c, HMGCR mRNA expression and fat content and increases in LDLR and CYP7A1a mRNA. The hyperlipidemia zebrafish model was successfully established by feeding with 4% cholesterol for 20 days. The findings lay a foundation for further screenings on lipid-lowering drugs.

Comparative study of pharmacokinetics and tissue distribution of 8-cetylberberine and berberine in rats / 药学学报

The concentrations of berberine (BBR) and 8-cetylberberine (8-BBR-C16) in rat plasma and tissue were determined by RP-HPLC. Both the plasma pharmacokinetics characteristic and tissue distribution differences of BBR and 8-BBR-C16 were compared to provide experimental data for the mechanism research and further drug development. After the oral administrations of BBR and 8-BBR-C16 at the dose of 80 mg x kg(-1) for rats, the pharmacokinetics result showed that compared with BBR, the C(max) and AUC(0-t), of 8-BBR-C16 increased by 2.8 times and 12.9 times respectively, t1/2 extended from 3.61 h to 11.90 h. The tissue distribution result showed that compared with BBR, the concentration of 8-BBR-C16 in various organizations increased and the retention time extended remarkably. The maximum concentration was achieved in lung and the highest concentration in it was 3 731.82 ng x g(-1). After being derived, the C(max) in plasma and bioavailability of 8-BBR-C16 increased remarkably and the circulation time in vivo extended. The drug concentration in tissue increased remarkably, and the distribution ratio changed too, with strong targeting selection in lung.

Effects of alkaloids from coptidis rhizoma on blood lipid metabolism and low-denstity lipoprotein receptor mRNA in golden hamsters / 中国中药杂志

To study the effects of alkaloids from Coptidis Rhizoma on low-density lipoprotein receptor (LDLR) mRNA expression and antihyperlipedemic levels. The LDLR mRNA expression were detected by real time fluorescence quantitative PCR, and the levels of total cholesterol (TC), triglyceride (TG), low density lipoprotein (LDL-c) and high-density lipoprotein cholesterol (HDL-c) in serum were measured at the first and last examination. The results show that, after the drug treatment, compared with the model group, each drug group showed a lipid-lowering effect. Especially, coptisine, palmatine, jatrorrhinze were significantly reduced TC, TG, LDL-c (P < 0.05, P < 0.01), and increased HDL-c (P < 0.01). In addition, they also increased mRNA expression of the LDLR in liver and HepG2 cells. The results showed that alkaloids from Coptidis Rhizoma can regulate lipid metabolism disorder, and coptisine have the best lipid-lowering effect.

Effect of 8-alkylberberine homologues on erythrocyte membrane.

8-alkylberberine homologues (Ber-C8-n, where n indicates carbon atom number of gaseous normal alkyl at 8 position, n =0, 2, 4, 6, 8, 10, 12, or 16) were synthesized and their effects on the hemolysis of rabbit erythrocyte, the fluidity of membrane and the fluorescence of membrane protein were investigated by fluorescence analysis technique. Ber-C8-n with mediate length alkyl (4<n<10) exhibited obvious hemolysis effect on rabbit erythrocyte when their concentration exceed 1.25×10-4 mol/L, and Ber-C8-8 displayed the highest hemolysis effect among all tested homologues. All of Ber-C8-n influenced the fluidity of erythrocyte membrane to different extents, which exhibited an obvious dose-effect relationship. The effect of Ber-C8-n on fluidity increased as the length of alkyl chain was elongated and decreased gradually when the alkyl carbon atoms exceeded 8. The fluorescence of erythrocyte membrane protein was quenched by Ber-C8-n, which showed a similar changing tendency on membrane fluidity. Experiments in vitro suggested that disturbing effects of Ber-C8-n on the conformation and function of membrane protein leaded to the changes of membrane fluidity and stability, and then the membrane was broken down.

Neuroprotective effects of ginsenoside Rg1 on 1-methyl-4-phenylpyridinum-induced apoptosis in PC12 cells / 第二军医大学学报

Objective To explore the neuroprotective effect of ginsenoside Rg1 against PC12 cell apoptosis induced by 1-methyl-4-phenylpyridinum (MPP++). Methods MPP++-induced apoptosis in PC12 cells, with the characteristics of dopaminergic neuron, were taken as the model of Parkinson disease in vitro. The cells were divided into control group, MPP++ group and 3 ginsenoside Rg1 pretreatment groups (concentrations 10, 20, and 50 μmol/L). MTT assay was used for detecting the cell viability, FCM for apoptosis ratio, TUNEL enzyme labelling for DNA fragment of the cell nuclear, and Western blotting analysis for cytochrome C protein. Results Ginsenoside Rg1 (10, 20, and 50 μmol/L) showed protective effect against MPP++-induced PC12 cells injury. Compared with MPP++-treated cells([52±4.7]%), pretreatment with 10, 20, and 50 μmol/ L ginsenoside Rg1 increased the cell viability to (64 ± 3. 4) %, (72 ± 5.2) % and (83±6.2)%, respectively (P<0.05 or P< 0.01). FCM analysis indicated that apoptosis rates decreased by ginsenoside Rg1 pretreatment, with the apoptosis rates in the control, MPP++ and 3 ginsenoside Rg1 groups (10, 20, 50 μmol/L) being 1.8%, 44.5%, 32.9%, 21.1% and 14.2%, respectively. We also found that ginsenoside Rg1 pretreatment greatly decreased DNA fragment of PC12 cells. Western blotting analysis indicated that the cytochrome C was depressed by the ginsenoside Rg1 pretreatment. Conclusion Ginsenoside Rg1 can protect PC12 cells against MPP++-induced apoptosis in a concentration-dependent manner, which may be closely related to down- regulation of cytochrome C over-expression in the mitochondria.

Neuroprotective effects of ginsenoside Rgl on l-methyl-4-phenylpyridinum-induced apoptosis in PC12 cells / 第二军医大学学报

Objective To explore the neuroprotective effect of ginsenoside Rg1 against PC12 cell apoptosis inducedby 1-methyl-4- phenylpyridinum (MPP+). Methods MPP+-induced apoptosis in PC12 cells, with the characteristics of dopaminergic neuron, were taken as the model of Parkinson disease in vitro. The cells were divided into control group, MPP+ group and 3 ginsenoside Rg1 pretreatment groups (concentrations 10, 20, and 50 μmol/L). MTT assay was used for detecting the cell viability, FCM for apoptosis ratio, TUNEL enzyme labelling for DNA fragment of the cell nuclear, and Western blotting analysis for cytochrome C protein. Results Ginsenoside Rg1 (10, 20, and 50 μmol/L) showed protective effect against MPP+-induced PC12 cells injury. Compared with MPP+-treated cells([52±4. 7]%), pretreatment with 10, 20, and 50 |imol/ L ginsenoside Rg1 increased the cell viability to (64 ± 3. 4) %, (72 ± 5. 2) % and (83±6.2)%, respectively (P<0. 05 or P< 0. 01). FCM analysis indicated that apoptosis rates decreased by ginsenoside Rg1 pretreatment, with the apoptosis rates in the control, MPP+ and 3 ginsenoside Rg1 groups (10, 20, 50 μmol/L) being 1. 8%, 44. 5%, 32. 9%, 21. 1% and 14. 2%, respectively. We also found that ginsenoside Rg1 pretreatment greatly decreased DNA fragment of PC12 cells. Western blotting analysis indicated that the cytochrome C was depressed by the ginsenoside Rg1 pretreatment. Conclusion Ginsenoside Rg1 can protect PC12 cells against MPP+-induced apoptosis in a concentration-dependent manner, which may be closely related to down-regulation of cytochrome C over-expression in the mitochondria.

Synthesis and anti-inflammatory activities of methylhesperetin-7-alkyl ether analogues / 药学学报

To investigate the relationship between the structures of methylhesperetin-7-alkyl ether analogues and their anti-inflammatory activities, nine new compounds, methyl-hesperetin (2), methylhesperetin-7-ethyl ether (3), 7-n-butyl ether (4), 7-n-hexyl ether (5), 7-n-octyl ether (6), 7-n-decyl ether (7), 7-n-dodecyl ether (8), 7-n-tetradecyl ether (9) and 7-n-hexadecyl ether (10), were synthesized with the lead compound of methylhesperidin (1). Their structures were confirmed by UV, 1H NMR, MS and HR-MS spectral data. The in vivo antiinflammatory activities of these compounds were tested on mouse paw edema induced by Freund's complete adjuvant (FCA) and mouse capillary permeability induced by acetic acid with po dose of 300 mg x kg(-1) x d(-1). The result indicated that the anti-inflammatory activities of the synthetic compounds increased firstly and then decreased with the elongating of the length of alkyl chain. After 25-day oral administration of compounds 6, 7 and 8, the inhibitory rates on mouse paw edema of adjuvant arthritis (AA) were 31.9%, 38.5%, 39.1%, respectively. They showed the concentrations of COX-2 in serum of AA mice respectively were 79.3, 75.4, 73.9 ng x L(-1) and the concentrations of PGE2 were in correspondence with 275.4, 258.9, 242.6 ng x L(-1). The inhibitory rates of compounds 6 and 7 on mouse capillary permeability induced by acetic acid were, respectively, 42.4% and 41.5% after 5-day oral administration. Compared with the lead compound of methylhesperidin, the anti-inflammatory activities of compounds 6, 7 and 8 were increased and showed an effective inhibition on the symptom of adjuvant arthritis and capillary permeability in mice.

Investigations on cordycepin production by solid culture of Cordyceps militaris / 中国中药杂志

An efficient method to produce cordycepin by solid culture using Cordyceps militaris was investigated in this study. Firstly, the changes of cordycepin during various growing periods of solid culture using 5 strains of C. militaris were detected, the best strain and optimal growing period for cordycepin production were determined. Then, by experiments of quadratic rotation-orthogonal combination design and orthogonal design, the medium composition and growth conditions for high yield of cordycepin were optimized. With the optimized method to produce cordycepin, the content of cordycepin in the medium was increased to 0.60%, which was nearly 2 times higher than the highest yield reported.

The Influence of Body Positions on Blood Pressure in Diabetic Patients / 中华高血压杂志

Objective To investigate the difference between the blood pressure readings between sitting and supine position,and to study the factors that associated with the sitting-supine blood pressure difference in patient with diabetes.Methods We measured the sitting blood pressure first then followed by the supine pressure in 356 diabetic patients,using a standard mercury sphygmomanometer.Patient's body weight,height and blood glucose levels were also measured.Results SBP and DBP were significantly higher in the supine position than in sitting position in diabetic patients(by 3.5?7.6/1.5?4.9 mm Hg,P