To explore the effect of geniposidic acid (GPA) on the signal pathway of small heterodimer dimer receptor (SHP) and liver receptor homologue 1 (LRH-1) in cholestasis rats induced by alpha-naphthalene isothiocyanate (ANIT).
 Methods: Fifty SD rats were randomly divided into five groups: a blank group, an ANIT group, an ANIT+GPA (100 mg/kg) group, an ANIT+GPA (50 mg/kg) group, and an ANIT+GPA (25 mg/kg) group (n=10 in each group). The GPA were intragastrically given to rats for 10 days, and the control group and the ANIT group were given normal saline. At the eighth day of administration, all rats except the blank group were given 65 mg/kg ANIT once until the tenth day. After the last administration, serum total cholesterol (TC), triglyceride (TG) and total bile acids (TBA) were measured. The primary hepatocytes (RPH) were isolated from normal rats and cultured. The cells were divided into a blank group, an ANIT (40 μmol/L) group, an ANIT (40 μmol/L)+GPA (4.00 mmol/L) group (A4.00G group), an ANIT (40 μmol/L)+GPA (1.00 mmol/L) group (A1.00G group), and an ANIT (40 μmol/L)+GPA (0.25 mmol/L) group (A0.25G group). The mRNA transcription levels of SHP and cholesterol 7 alpha hydroxylase (CYP7A1) in RPH were detected by real-time-PCR, and the protein levels of SHP and CYP7a1 were detected by Western blotting. In the LRH-1 silence experiment, the RPH were divided into a blank group, a negative transfection group, a siRNA-LRH group (ZR group), a siRNA-LRH+GPA (4.00 mmol/L) group (ZR4.00G group), a siRNA-LRH+GPA (1.00 mmol/L) group (ZR1.00G group) and a siRNA-LRH+GPA (0.25 mmol/L) group (ZR0.25G group). The protein and mRNA levels of SHP, CYP7a1, LRH-1 were detected. In the over-expression experiment, the RPH were also divided into a blank group, a negative transfection group, a LRH-1 over-expression plasmid group (OE group), a LRH-1 over-expression plasmid+GPA (4.00 mmol/L) group (OE4.00G group), a LRH-1 over-expression plasmid+GPA (1.00 mmol/L) group (OE1.00G group), and a LRH-1 over-expression plasmid+GPA (0.25 mmol/L) group (OE0.25G group). The protein and mRNA levels of SHP, CYP7a1 and LRH-1 were detected.
 Results: Compared with the blank control group, TC and TBA were significantly increased (both P<0.01) in the ANIT group, but there was no difference in TG; compared with the ANIT group, the contents of TC and TBA in the AG100 and AG50 groups were significantly reduced (all P<0.01). Compared with the blank control group, the proteins and mRNA levels of SHP were significantly decreased (P<0.01), while CYP7a1 were dramatically increased (P<0.01) in the ANIT group; compared with the ANIT group, the proteins and mRNA levels of SHP in the A4.00G group and the A1.00G group were significantly increased (both P<0.01), while the levels of CYP7a1 proteins and mRNA levels were evidently decreased in the A4.00G and A1.00G groups (both P<0.01). Compared with the negative transfection group, the proteins and mRNA levels of CYP7a1 and LRH-1 were dramatically restrained (all P<0.01), while there was no change in SHP in the ZR group; compared with the ZR group, the proteins and mRNA levels of SHP were significantly increased (all P<0.01), while LRH-1 and CYP7a1 were not changed in the ZR4.00G, ZR1.00G and ZR0.25G groups. Compared with the negative transfection group, the protein and mRNA levels of CYP7a1 and LRH-1 were significantly suppressed in the OE group (all P<0.01). Compared with the OE group, the protein and mRNA levels of SHP were evidently increased in the OE4G and OE1G groups (all P<0.01), while LRH-1 and CYP7a1 were not changed in the OE4G, OE1G and OE0.25G groups.
 Conclusion: The over-expression of LRH-1 in RPH can up-regulate the mRNA and protein levels of CYP7a1. GPA can improve the biochemical and liver pathology of ANIT-induced cholestasis rats, which may be related to the decrease of CYP7a1 by activating SHP through LRH-1 in RPH.
Animals ; Cholestasis ; Iridoid Glucosides ; Rats ; Rats, Sprague-Dawley ; Receptors, Cytoplasmic and Nuclear ; Signal Transduction

Objective To observe the damage effect of borneol on rabbit corneal epithelial cells. Methods After the treatment with borneol at 100, 200, 400 μg·mL-1 respectively, the viability of rabbit corneal epithelial cells was determined by methyl thiazolyl tetrazolium ( MTT) assay, cell apoptosis was determined by flow cytometry with Annexin V- fluorescein isothiocyanate/propidium iodide ( FITC/PI) staining, and Caspase-3 mRNA expression was detected with real-time reverse transcription polymerase chain reaction ( RT-PCR) . Results Borneol at the concentrations of 100, 200, 400 μg·mL-1 inhibited the activity of rabbit corneal epithelial cells. Compared with the normal control group, borneol increased the rate of apoptosis, and enhanced the Caspase-3 mRNA expression in rabbit corneal epithelial cells ( P<0.05 or P<0.01) . Conclusion Borneol can inhibit the proliferation of rabbit corneal epithelial cells, induce cell apoptosis through enhancing the expression of apoptosis-related gene Caspase-3 mRNA.

Objective To explore the optimal method of extraction and determination of rhynchophylline and isorhynchophylline in Ramulus Uncariae cum Uncis. Methods A RP-HPLC was performed on a Phenomenex C18 column (4. 6 mm× 150 mm, 5m) at 30℃. The mobile phase consisted of methanol-0. 01 mmol/L triethylamine solution (70: 30, pH 7.0 adjusted by glacial acetic acid) at the flow rate of 0. 5 mL/min. The automatic sample injector was set at 5℃ and the ultraviolet detector was operated at 245 nm. And then, the effect of different extraction condition on the contents of rhynchophylline and isorhyn-ehophylline in Ramulus Uncariae cum Uncis was investigated. ResultsA good linearity of rhynchophylline was in the range of 2. 5μg/mL to 80. 0μg/mL (Y=76. 7170X-0. 0727,r=0. 999 8),the average recoveries were from 99. 84 ％ to 116. 91％, and the RSD (n=6) were less than 8.8 ％. A good linearity of isorhynchophylline was in the range of 2. 0 μg/mL to 80. 0 μg/mL (Y=87. 4729X-0. 3666, r=0.999 7), the average recoveries were from 87.08 ％ to 104. 97 ％, and the RSD (n=6) were less than 7.3 ％. The contents of rhynchophylline and isorhynchophylline in the methanol-extracted solu- tion were approximately ten times as much as those in water-decocted solution. The main factors which had effect on the ex-traction of rhynchophyUine and isorhynchophylline in methanol solution were supersonic time,methanol amount and cold-dousing time, in a decreasing sequence. The best condition selected by orthogonal experiment wasb as follows: extracting the medicinal material with 20-fold volumes of methanol, cold maceration for 24h and supersonic extraction for 1 h. Conclusion The extraction percentage of rhynchophylline and isorhynchophylline in the extraction condition of cold maceration with methanol and supersonic extraction is superior to that in the water-decocting condition. The method is simple, fast and accurate, and it can be used for the quality control of Ramulus Uncariae cum Uncis.

Objective To establish a HPLC fingerprint analysis method of salt-prepared Cortex Phellodendri. Methods C18 column was used, with gradient elution by the mobile phase consisted of acetonitrile-0. 1% phosphoric acid (contained 0. 2 % triethylamine). The detection wavelength was at 230 nm, and the flow rate was at 0. 8 mL/min. Results Seventeen char-acteristic peaks were selected and the fingerprints on HPLC was set up. Conclusion HPLC fingerprint method is repro-ducible, accurate and stable, and can be used for the quality control of salt-prepared Cortex PheUodendri.

BACKGROUND: Borneol can open blood-brain barrier (BBB) but the mechanisms are not very clear. Histamine and 5-hydroxtryptamine can take part in regulation of permeability of BBB. There is not report on the rela tion between the effect of opening BBB of borneol and the regulation of permeability of BBB of histamine (HA)/5-hydroxytryptamine (5-HT). OBJECTIVE: To study the relation between the effect of opening BBB of bornool and the regulation of permeability of BBB of histamine (HA)/5-hydroxytryptamine (5-HT).DESIGN: A completely randomized and controlled study.SETTING: Clinical Pharmacological Institute of University of Traditional Chinese Medicine of Guangzhou.MATERIALS: The experiment was carried out at the Clinical Pharmacological Institute of University of Traditional Chinese Medicine of Guangzhou from September 2003 to February 2004. Totally 104 healthy male SD rats, weighting 230-27.0 g, supplied by Guangdong Medical Experimental Animal Center, were selected.METHODS: The rats were randomly divided into 13 groups: pre-medicine group and post-medicine group (high, moderate and low dosage group and 5, 20, 45 and 60 minutes group in each dosage group) with 8 in each group. Borneol was mixed as 10% millet oil suspension. Rats were fasted before experiment for whole night, and medicine was perfused on the next morning with the high, moderate and low dosage of 0.15, 0.12 and 0.09 g/kg respectively. Hypothalami of rats was selected at various time points to make biological samples. Contents of HA and 5-HT were assayed with HPLC system electrochemical detector.MAIN OUTCOME MEASURES: Changes of level of histamin (HA) and 5-HT in hypothalamus of rats after administration of borneol.RESULTS: Data of totally 104 rats was entered the final analysis. ① Contents of HA and 5-HT in hypothalami were 2.07±0.54 μg/g and 1.45±0.14 μg/g respectively. ② The level of HA in hypothalamus of rats after different doses of borneol were higher than that of before administration. Comparing the level of HA in 20 minutes after moderate dose with before administration, the level of HA in 20 minutes was increased significantly [(3.36±0.21) μg/g, P < 0.01], the others of moderate dose, the 45 minutes after high dose, the 20 and 45 minutes after low dose were also increased significantly than before administration (P < 0.05). ③ After administration of different doses of borneol, the level of 5-HT after high dose were higher than that of before administration [5, 20, 45 and 60 minutes after medicine: (1.90±0.32), (3.28 ±0.25), (2.66±0.46), (2.80±0.34) μg/g, respectively; (P < 0.05 or P < 0.01)]; the level of 5-HT after 5, 20 and 45 minutes of moderate and low dose were increased significantly too (P < 0.05 or P < 0.01).CONCLUSION: Borneol could open BBB by increasing levels of HA and 5-HT in hypothalamus of rats.Borneol mediates opening of BBB by increasing levels of HA and5-HT in rats.

OBJECTIVE: To observe the therapeutic effect of Wuling Powder extract on rats with renal hypertension and to evaluate the influence of it on the volume of urine and the concentrations of Na(+), K(+), Cl(-). METHODS: Reformed Gold-blatt hypertension rat model (G-2K1C) was established. The rats were divided into 6 groups as follows: sham-operation group; model group, Wuling Powder high dosage group (80 g/kg), Wuling Powder middle dosage group (40 g/kg), Wuling Powder low dosage group (20 g/kg), and hydrochlorothiazide (HCT) group (25 mg/kg). Urine volume of the rats was measured during the experiment. Tail arterial pressure and [Na(+)], [K(+)], [Cl(-)] in serum of the rats were detected after 30 days of treatment. RESULTS: The blood pressure of the G-2K1C rats was decreased in the three Wuling Powder groups (P<0.05 or P<0.01), but higher than that of the false-operation group (P<0.01), and there was no difference between each of the Wuling Powder groups and the HCT group (P>0.05). Diuretic effect of the three dosages of Wuling Powder was weaker than that of the HCT (P<0.05 or P<0.01). The effects of the three dosages of Wuling Powder and HCT on [Na(+)] and [Cl(-)] in the serum were not obviously different (P>0.05), but [K(+)] of the HCT group was significantly decreased compared with that of the false-operation group and the three Wuling Powder groups (P<0.01). CONCLUSION: Wuling Powder extract had satisfying therapeutic effects in increasing the discharge of urine, decreasing the blood pressure and keeping the balance of the serum electrolyte contents in rats with renal hypertension.

Objective To observe the analgesic action and anti-inflammation effect of borneol. Methods The analgesic action and anti-inflammation effect of borneol were observed by mouse hot-plate test,mouse acetic-acid-induced twisting test,mouse abdominal cavity capillary permeability increase test induced by acetic acid,and rat pedal swelling test induced by carrageenin. Results Borneol reduced the pedal swelling of rats,ED50=0.3242 g?kg-1,increased pain threshold in mice ED50=0.3870 g?kg-1,inhibited twisting response of mice obviously,ED50=0.5813 g?kg-1. In the same dose,the analgesic action of borneol was stronger than the anti-inflammatory effect. Conclusion Borneol has certain analgesic action and anti-inflammation effect.

Objective: To study the effect of Fructus Psoraleae on antipyrine elimination in rats. Methods:After the medication of Fructus Psoraleae decoction once daily for 6 days in a dose of 5. 3 g/kg, antipyrine was fed to rats in a dose of 400mg/kg. Concentration of antipyrine in various time was measured by HPLC, and the pharmacokinetics parameters of antipyrine elimination was caculated. Results: The elimination rate constant (Kel) of antipyrine in rats fed with Fructus Psoraleae decoction was 1.55 times more than that in the control group, and the elimination half - life (t1/2) was shortened by 60. 7 % . Conclusion: Fructus Psoraleae decoction accelerate the elimination of antipyrine in rats by inducing the activities of hepatic oxygenases.

Objective To investigate the effects of Rhizoma Zingiberis Extract (RZE) on the cardiac functions of rabbits with heart failure. Methods Forty pure-bred New-Zealand rabbits were randomly allocated to 4 groups: control group, low-dosage RZE group, moderate-dosage RZE group and high-dosage RZE group. The rabbit model of heart failure was established by intravenous dripping of 20 g/L pentobarbital sodium. Oral tube perfusion of RZE was given after model establishment.The hemodynamic changes were observed before and after the modeling and 5,10,15,20,30,45,60,90,120,150 minutes after treatment by a RM-6000 4-graph physiological monitor. Results After treatment, LVSP, lv?dp/dtmax,lv +dp/dtmax,lv-dp/dtmax showed an ascending tend in all the groups, particularly in RZE groups; the differences between the control group and RZE groups were significant 30 minutes after treatment (P

Objective To determine the intestinal absorption characters of zingerone in rats.Methods The concentration of zingerone in the intestine perfusate was determined by HPLC.The absorption kinetics were obtained by the in-situ perfusion method in rats.Results The absorption rate constants(Ka) of zingerone were 0.0137,0.0135 and 0.0139 min-1 at the concentration of 25.0,50.0 and 100.0 ?g?mL-1,respectively;Ka of zingerone in the duodenum,jejunum,ileum and colon were 0.0070,0.0031,0.0032 and 0.0026 min-1,respectively.Conclusion Concentration of zingerone has no significant effect on its absorption kinetics.The best absorption segment of the intestine was duodenum,ileum,jejunum and colon by turns.The intestinal absorption of zingerone has the first-order kinetic characters with passive diffusion mechanism.