Table of Content

Volume 10 Issue 6
26 June 2005

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Animal models of neuropathic pain

LIU Guo-kai, HUANG Yu-guang, LUO Ai-lun

2005, 10(6):  601-603. 

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Although there are a number of shortcomings with these animal models, they provide important clues in understanding the underlying pathophysiology of neuropathic pain in humans. In these models, cutaneous sensory threshold of the hindlimb ipsilateral to nerve injury is measured. The presence of neuropathic pain in experimental animal models is mainly measured as allodynia or hyperalgesia, in which the normally nonnoxious or mildly noxious stimuli induce a nociceptive behavioral response. This paper mainly discusses the recent findings from the peripheral nerve injury model of neuropathic pain, as well as the different characteristics of these animal models of neuropathic pain.

Protective effects of pivanampeta on ischemia-reperfusion injury of rat heart

CAO Ze-ling, CHEN Kai, YANG Ting-shu, LONG Chao-liang, LI Xiao-wei, WANG Hai

2005, 10(6):  604-612. 

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AIM: To observe the effects of pivanampeta (Piv) on ischemia-reperfusion injury of rat heart in vivo. METHEDS: Sprague-Dawley rats were randomly divided into two groups. One was 30 min reperfusion group, which was subdivided into sham, control and Piv groups with 30 min ischemia followed by 30 min reperfusion to detect some data related to cardiac function and the area of myocardium infarction. Anotherwas 2 h reperfusion group, which was further subdivided into such sections as follows: sham, model, Piv 3, 6 and 9 mg·kg^-1 groups.Apart from the sham, Piv and NS (control) groups were administered intravenously 30 min before occlusion. Then hearts were excised, paraffined and cut into 4 μm think. Immunohistochemistry, in situ hybridization, TUNEL and DNA agarose gel electrophoresis were performed to detect the expression of Bax, Bcl-2, caspase-3, MMP-2 and PPAR γprotein and MMP-2, and PPARγmRNA. RESULTS: Compared with control group, the ratio of nec aar after Piv injected decreased 21 % (P <0.05), while nec lv reduced to 22 %(P < 0.05). Heart rate, +dp /dt_max and V_max representing the systolic function of heart as well as-dp /dt_max which was the indicator of diastole improved dramaticly at 1 min and 30 min after reperfusion, respectively (P <0.05). In dose-dependent manner, the expression of Bax and caspase-3 protein, MMP-2 protein and mRNA depressed, while the PPARγprotein and mRNA enhanced by Piv. The apoptotic index of subgroups injected Piv reduced with important significance by TUNEL compared with model(P <0.05). DNA ladder existed in model, Piv 3、6 mg·kg^-1, rather than Piv 9 mg ·kg^-1 by agarose gel electrophoresis. CONCLUSION: Piv shows the protective ability against I R injury of myocardium. Piv administration may reduce the area of MI and improve cardiac function as well as decrease apoptotic cardiomyocytes.

Comparative study on changes of glycogen metabolism in model mice of diabetes mellitus

XIAO Mei-fang, ZHANG Xue-mei, BAI Xiang, TAN Huan-ran

2005, 10(6):  613-616. 

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AIM: To explore the changes of glycogen metabolism in diabetic animal models and the role of glycogen metabolism in the pathogenesis of diabetes mellitus. METHODS: Firstly four types of diabetic animal models were generated including mice models of type 1 and type 2 diabetes mellitus. Then the liver glycogen and muscle glycogen contents of these diabetic mice were determined according to anthrone-reagent method and glucokinase expressions in liver of glucokinase gene knockout mice were assayed using western blot. RESULTS: Blood glucose levels were increased in all types of model mice. In particularly, model mice of type 1 diabetes mellitus displayed significantly higher blood glucose levels than those of model mice of type 2 diabetes mellitus. Liver glycogen contents in model mice significantly declined except neonatal STZ-induced model mice. In addition, muscle glycogen contents were decreased in all model mice. Western blot assay showed that glucokinase expression decreased in liver from glucokinase gene knockout mice, which might contribute to the diminished liver glycogen in this kind of diabetes model. CONCLUSION: Diabetic subjects show impaired glycogen metabolism with the diminished liver glycogen and muscle glycogen contents, which indicates the key role of glycogen metabolism in the pathogenesis of diabetes.

Effects of tubeimoside isolated from Bolbostemma paniculatum on microtubule in CNE-2Z cells

SONG Gang, MA Run-di, YU Li-jian

2005, 10(6):  617-621. 

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AIM: To examine the effects of tubeimoside isolated from Bolbostemma paniculatum on microtubule in low differentiated nasopharyngeal carcinoma CNE-2Z cells. METHODS: The interactions of tubeimoside with microtubule in CNE-2Z cells were investigated by immunofluorescence microscopy, and the interactions of tubeimoside with tubulin in CNE-2Z cells were detected by Western blotting. RESULTS: Tubeimoside (25 μmol·L^-1, 3 h) was sufficient to cause the microtubular network disruption. Western blot analysis revealed that the proportion of cytosolic tubulin of cells treated with tubeimoside increased in a time-dependent and concentration-dependent manner. CONCLUSION: Tubeimoside is an effective antimicrotubule agent.

Pharmacokinetics and tissue distribution of daurisoline in rabbits

SHI Shao-jun, LI Zhong-fang, GU Shi-fen, CHEN hui, ZENG Fan-dian

2005, 10(6):  622-626. 

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AIM: To investigate the pharmacokinetics and tissue distribution of daurisoline (DS)after administration in rabbits. METHODS: Single dose of 2.5, 5 or 10 mg·kg^-1 DS of body weight was administrated in rabbits. The tissue distribution of DS (10 mg·kg^-1)was investigated, and the concentrations in plasma and tissues were measured by a validated high performance liquid chromatography method. RESULTS: Plasma concentration-time profiles were adequately described by a twocompartment open model. The main pharmacokinetic parameters after iv DS 2.5, 5 and 10 mg·kg^-1 were as follows: T_1/2β were 3.0 ±0.6, 3.4 ±0.9 and 6.9 ±0.6 h, respectively, Cls were 3.1 ±0.6, 3.6 ±0.4 and 4.4 ± 0.3 L·h·kg^-1, respectively, V_d were 13.1 ±2.7, 18.0 ±6.2 and 43.6 ±4.4 L·kg^-1, respectively, and AUC_0~t were 0.84 ±0.13, 1.41 ±0.17 and 2.30 ±0.18 mg·h·L^-1, respectively. At dosages of 2.5 and 5 mg·kg^-1, no statistically significant difference existed in main pharmacokinetic parameters. However, T_1/2β and C₀ increased nonproportionally when the dosage was 10 mg·kg^-1. CONCLUSION: Across the dosages of 2.5-5 mg·kg^-1, DS demonstrates linear kinetics. However, a nonlinear kinetics is found at dosage of 10 mg·kg^-1. Assessment of tissue concentrations of DS reveals preferential distribution to the lung, kidney, spleen, and liver, and displays substantial penetration into tissues.

Effects of crocin on apoptosis of vascular endothelial cells induced by triol

LIU Juan, QIAN Zhi-yu

2005, 10(6):  627-632. 

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AIM: To investigate effects of Crocin on apoptosis of vascular endothelial cells injured by triol. METHODS: Apoptosis model induced by triol (cholestane-3β, 5α, 6β-triol) was adopted. Malondialdehyde (MDA) content was measured by colorimetric assay. The morphology and ultrastructure were detected by light microscope and electron microscope. DNA ladder was analyzed by DNA electrophoresis. The apoptosis rate was assessed with the flow cytometer assay. Bax mRNA expression of endothelial cells was measured by RT-PCR analysis. RESULTS: In Triol group, MDA content was 1.761 nmol·L^-1 (P <0.01). BAECs was shrinkage. The nucleus concentrated and dye deeply, mitochondria swelling and cavitations, mitochondrial cristae fragmentation, formation of apoptotic bodies, the DNA ladder in agarose gel and hypodiploid cusp was observed. Apoptosis rate was 30.62 %. Bax mRNA expression increased. In the Crocin +Triol group, MDA content decreased, majority of cells grown in spreading out like that in control and cholesterol group. Apoptosis rate of Crocin (10^-7 or 10^-6 mol·L^-1) +Triol group were 24.4 %, 6.3 %, respectively. Bax mRNA expression of endothelial cells in the crocin (10^-6 mol·L^-1) +Triol group decreased. CONCLUSION: Crocin maybe abnormally decrease apoptosis by anti-lipid peroxidation and adjust Bax mRNA expression.

Relaxation selectivity of three structurally diverse ATP-sensitive potassium channel openers, iptakalim, pinacidil and diazoxide on non-vascular smooth muscles

GAO Min, WANG Yu, WANG Hai

2005, 10(6):  633-636. 

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AIM: To investigate the relaxation effects of iptakalim (Ipt), a novel structural ATP-sensitive potassium channel opener (KCO), compare the relaxation characteristics of Ipt to the other structurally diverse KCOs, pinacidil (Pin)and diazoxide (Dia), and further understand the relaxation selectivity of Ipt. METHODS: Strips of stomach, ileum, and bladder isolated from rats were prepared and treated with different concentrations of drugs after contraction of 10^-5 mol·L^-1 acetylcholine. RESULTS: Ipt at the concentrations of 10^-8-10^-4 mol·L^-1 caused no relaxation effects on strips of stomach, ileum and bladder. The same results as Ipt were detected in diazoxide-treated tissue strips. Pin showed no effects on strips of stomach and bladder, however, Pin at concentrations of 10^-5 and 10-4mol·L^-1 induced significant relaxations in ileum strips compared with the former concentration, and the relaxation rates were 28.8 % and 51.9 %, respectively. CONCLUSION: Ipt shows no effect on the relaxation of stomach, ileum and bladder, which is different from Pin, but the same as Dia. Moreover, similarities and differences in relaxation are also found among the structurally diverse KCOs.

Effects of human tissue kallikrein gene A1789G polymorphism on plasma creatinine levels in patients with essential hypertension

HONG Zong-yuan, ZHANG Xiu-qing, HUANG Guo, LING Dai-jun, XU Xi-ping

2005, 10(6):  637-641. 

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AIM: To investigate the effect of the human tissue kallikrein gene (hKLK1) polymorphism on plasma creatinine levels in hypertensive subjects. METHODS: The hKLK1 A1789G polymorphism was genotyped by PCR-restriction fragment length polymorphism (RFLP) in 733 hypertensive subjects. The relationship between genotype and plasma creatinine level was performed by a multiple regression analysis. The interactive effect of genotype and blood pressure on the plasma creatinine level was accessed by ANOVA. RESULTS: Multiple regression analysis showed that the plasma creatinine level was significantly higher in the subjects with mutant allele G (AG or GG genotype) than in those with wild allele A (AA genotype) (P =0.009 and P = 0.046, respectively). ANOVA indicated that the AG and GG genotype individuals had high plasma creatinine levels in SBP and DBP Compared with AA genotype individuals, and the plasma creatinine level increased with blood pressure rises (P <0.05). CONCLUSION: The hKLK1 A1789G polymorphism influences plasma creatinine level, and the A1789→G variation is a risk factor in renal plasma creatinine clearance rate decline in hypertensive individuals.

Antiproliferative effects of pioglitazone involved of nitric oxide on cultured rabbit aortic smooth muscle cells

CHEN Lu-ying, XU Jiang-ping, PANG Jian-xin, SHI Dao-hua

2005, 10(6):  642-645. 

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AIM: To investigate the antiproliferative effects of pioglitazone (PIO) and its possible mechanisms on the cultured rabbit aortic smooth muscle cells (VSMCs). METHODS: Proliferation of VSMCs was induced by high concentration of glucose and insulin to mimic insulin resistance (IR) status. Cells proliferation and viability were measured by MTT assay. NO content in culture solution and NOS activities of VSMCs were determined by assay kits, respectively. RESULTS: Proliferation of VSMCs were markedly decreased by PIO in all groups (P <0.01). NO content in culture medium and the activities of total NOS and iNOS of VSMCs were significantly increased by PIO in both normal and mimic IR culture condition (P <0.01). The effects of PIO on VSMCs were stronger in mimic IR culture condition than that in normal culture condition, and these effects of PIO were partially blocked by the NOS inhibitor L-NAME. CONCLUSION: The antiproliferative effects of PIO were partially involved in NO pathway on cultured VSMCs both in normal and mimic IR culture condition.

Inhibition effects and mechanisms of anisodamine on calcium transient and contractile function in isolated adult rat ventricular myocytes

LIU Zhen-guo, YAO Xiu-juan, LI Xiao-qiang, YANG Zhi-fu, WANG Ru-tao, MEI Qi-bing

2005, 10(6):  646-650. 

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AIM: To study the effects and mechanisms of anisodamine on calcium transient and contractile function in isolated adult rat ventricular myocytes. METHODS: Single cell edge motion detection system and dual-excitation fluorescence photomultiplier system were used for measuring the intracellular calcium transient properties and contractile function. ΔFR, TTP, T₅₀ D, PS, TPS, TR₉₀ and ± d_L /d_t were recorded by the ionoptix system. Antagonist of IP₃ receptor, VDCC and muscarinic receptors were used for study the mechanism of anisodamine. RESULTS: Anisodamine (1 ×10^-9, 1 × 10^-7, 1 ×10^-5 mol·L^-1) directly inhibited intracellular calcium transient and contractile function. Carbachol (1 ×10^-6 mol·L^-1) abolished the effect of anisodamine. Heparin (1 ×10^-6 mol·L^-1) and verapamil (1 ×10^-6 mol·L^-1) decreased the effect of anisodamine. CONCLUSION: The effect of anisodamine on intracellular calcium transient and contractile function may be related to inhibition of calcium releasing from intracellular calcium pool and VDCC.

Study of chloride channel ClC-3 gene expression in patient with atrial fibrillation

WU Jia-bin, XU Chun-xuan, ZHANG Jian-cheng, CHEN Lin, LIN Li-fang, HU Xi-zhong

2005, 10(6):  651-654. 

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AIM: To investigate alterations in gene expression of chloride channel ClC-3 in patients with atrial fibrillation (AF). METHODS: Right atrial appendages specimen were obtained from 33 patients with chronic AF(CAF group), 7 with paroxysmal AF(PAF group) and 31 matched controls in sinus rhythm (SR group) who underwent open-heart surgery. In this study, the semi-quantitative polymerase chain reaction (RT-PCR) was used to examine the mRNA expression of ClC-3 channel in atrial tissue. RESULTS: The content of ClC-3 mRNA in CAF group was higher than that in SR group(P <0.01), and it was correlated with AF duration (r =0.376, P = 0.001). There was no significant difference in the gene expression of ClC-3 between PAF group and SR group, although the content of ClC-3 mRNA was slightly higher than that in SR group (P >0.05). CONCLUSION: The increased gene expression of chloride channel ClC-3 may be served as the molecular basis of I_Cl.VOL remodeling in patients with atrial fibrillation (AF).

Effects of furosemide, piretanide and azathioprine on thiopurine methyltransferase activity in red blood cells from patients with chronic inflammatory bowel disease

XIN Hua-wen, FISCHER Christine, SCHWAB Matthias, KLOTZ Ulrich

2005, 10(6):  655-658. 

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AIM: To study the possible effects of furosemide, piretanide and azathioprine on thiopurine methyltransferase (TPMT) activity in red blood cells (RBC) from patients with chronic inflammatory bowel disease (IBD). METHODS: The inhibitory potential of furosemide, piretanide and azathioprine on TPMT activity in RBC was assessed in vitro by HPLC in three groups of patients with very high, normal and intermediate TPMT activity (n =6 in each). Individual concentration response curves IC₅₀-values were determined. RESULTS: Independent of the basal TPMT activity, lowest IC₅₀-values were calculated for furosemide (15-19 μmol·L^-1), followed by piretanide (300-313 μmol·L^-1) and azathioprine (430-532 μmol·L^-1). Compared with reported plasma concentration achieved during treatment, only furosemide showed the potential to inhibit TPMT in vivo, whereas the IC₅₀-values of the other agents were far above the corresponding plasma levels. CONCLUSION: Only furosemide has the potential to inhibit TPMT activity in patients with IBD. Clinically relevant drug interactions should be considered in patients treated simultaneously with this diuretic and thiopurines.

Effects of arsenolite on genic expression of cytoplastic Phospholipases A₂ and leukotriene C₄ in asthmatic mice

SONG Ze-qing, CHEN Hen-hui, YAO Wei-min, LIANG Biao, WANG Hui

2005, 10(6):  659-663. 

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AIM: To study the effects of arsenolite on the leukotriene C₄ and the expression of cPLA₂ mRNA in ovalbumin challenged asthmatic mice. METHODS: 56 mice were randomly divided into 7 groups. Establishing the ovalbumin sensitized asthmatic model, RT-PCR was employed to detect the expression of cPLA₂ mRNA in the lung. The TLC₄ and PLA₂ in BALF were measured by ELISA. Expiration time /respiratory total time and expiration maximum pressure were detected. RESULTS: The expression of PLA₂ mRNA in the lung and the TLC₄, PLA₂ activity in BALF in asthmatic group were significantly higher than those in normal group. After therapy with dexamethasone and different dosage of arsenolite (0.625, 1.25, 2.and 5.00 mg·kg^-1), the expression of PLA₂ mRNA and the TLC₄, PLA₂ activity were significantly lower than those in asthmatic group. Expiration time /respiratory total time and expiration maximum pressure in asthmatic group were significantly increased compared with normal group. Expiration time /respiratory total time and expiration maximum pressure of groups treated with 4 doses arsenolite were significantly decreased compared with asthmatic group. CONCLUSION: PLA₂ mRNA expression increases in asthma group, and arsenolite can efficiently treat asthma. Its mechanism maybe partly relates to decreasing TLC₄ in airway and the inhibition of PLA₂ mRNA expression.

Protective effects of tanshinone on brain slices injury induced by oxygen glucose deprivation

SUN Zhen-rong, ZHU Jian-qian, DIN Feng, DA Yi-dong, CHEN Hua

2005, 10(6):  664-669. 

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AIM: To investigate the protective effects of tanshinone on the brain slices injury induced by oxygen glucose deprivation(OGD). METHODS: Rat brain slices were made and randomly assigned to four groups (n = 10 in each): control group, OGD group, tanshinone 20 mg ·L^-1 group and tanshinone 200 mg·L^-1 group. Changes of the neuron injury and apoptosis were observed with TTC staining, LDH releases, TUNEL staining, immunohistochemistry and electromicroscope. In addition, changes of intracellular calcium were measured by confocal laser-scanning microscopy. RESULTS: Tanshinone 20 mg·L^-1 and 200 mg·L^-1 attenuated the decrease of TTC staining and the increase of LDH release induced by OGD in brain slices. Neuronal apoptosis and changes of neuronal ultrastructures were attenuated by different concentrations of tanshinone. Bcl-2 and Bax protein expression was increased after OGD. Bax protein expression was decreased by different concentrations of tanshinone, while Bcl-2 protein expression was increased by different concentrations of tanshinone. Intracellular calcium concentration was increased by OGD and then it was lowered by different concentrations of tanshinone. The protective effects of 200 mg·L^-1 tanshinone were more obvious better than those in the 20 mg·L^-1 tanshinone group. CONCLUSION: Different concentrations of tanshinone have neuronprotective effects against OGD injury in rat brain slices, and 200 mg·L^-1 tanshinone shows more obvious protective effects than those in the 20 mg·L^-1 tanshinone.

Effects of D-galactose on lumber vertebra density and serum testosterone concentration in male rats

LUO Hong-mei, CUI Liao, WU Tie

2005, 10(6):  670-673. 

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AIM: To observe the effects of D-galactose on the lumber vertebra body and investigate the reasons. METHODS: 12 rats at age of 6 months were divided into two groups, control group and D-galactose group (n =6 in each). The control group were administered saline solution sc, and the D-galactose group were administered 5 %D-galactose solution sc at dose of 100 mg·kg^-1. After 3 month, the rats were killed by exsanguination from heart. The fourth lumber vertebra was taken and immerged in formalin. The testicle were taken and immerged in formalin at the same time. The blood serum was collected by centrifugating the collected blood after resting for a while, and it was preserved in refrigeratory at the degree of-70 ℃.The vertebra body were embedded in plastic and sliced up after being dehydrated step by step with different concentration ethanol. The slices were analyzed under the image analysis apparatus. The testicle were made into paraffin slices and observed under the common microscope. The concentration of serum testosterone and luteinizing hormone were measured by radio-immunity assay. RESULTS: The lumber vertebra body in D-galactose group appeared osteoporosis. The serum testosterone hormone concentrations of D-galactose rats were significantly decreased. And the microstructure of testicle present aging change, but no change of serum LH concentration was observed. CONCLUTION: D-galactose can cause the osteoporosis in male rats, which may be related to affect the function of thalamus-pituitary-testicle axis, decrease the content of testosterone of D-galactose.

In vivo study on apoptosis and cell proliferation induced by adriamycin in mice with MCF-7 breast cancer

WANG Wen-wu, OU-YANG Xue-nong, JIANG Hao

2005, 10(6):  674-676. 

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AIM: To observe the apoptosis and cell proliferation induced by adriamycin in mice with breast cancer. METHODS: Therapeutic effect of 1.25 mg·kg^-1 adriamycin (ADR)on mice MCF-7 breast cancer was observed in vivo. Apoptosis and PCNA of tumor cells were also studied by TUNEL and immune his to chemical methods. RESULTS: In the control group, the apoptotic cell index (AI)in tumor cell was 1.11 %and PCNA index (PI)was 56.38 %.In the ADR group, the rate of tumor inhibition was 43 % and the AI was 1.93 % (P <0.05).The 35.75 % of PI in ADR group was lower than that in the control group (P <0.01). CONCLUSION: Adriamycin may induce apoptosis and reduce cell proliferation.

Effects of uroacitides on proliferation ability of breast cancer cells

WANG Yan-hong, ZHENG Wei

2005, 10(6):  677-681. 

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AIM: To investigate the growth inhibition effects of CDA-II on breast cancer cells. METHODS: The effects of CDA-II on growth curve and morphology of breast carcinoma cell lines MCF-7 and MDA-MB-231 were observed in vitro cultures. RESULTS: CDA-II reduced the growth speed and proliferation ability. CONCLUSION: CDA-II has remarkable effects on anti-cellproliferation and shows good prospects of application in the treatment of patients with breast carcinoma.

Effects of angiotensin type-1 receptor blockade on expression of angiotensin-converting enzyme 2 in renal of spontaneously hypertensive rats

YANG Liu, LIU Chang-hui, WANG Yi, BAI Yu-peng

2005, 10(6):  682-686. 

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AIM: To investigate the expression of angiotensin-converting enzyme 2 (ACE₂) in the renal of spontaneously hypertensive rats and the effects of angiotensin type-1 receptor blockade. METHODS: Twenty of 14-week-old male SHR were randomly divided into two groups (n =10 in each): SHR group and irbesartan group. Ten of 14-week-old male WKY group were served as control. Irbesartan group were given a daily dose of 50 mg·kg^-1 irbesartan for 12 weeks by gavage and the other two groups were fed with a normal diet. Angiotensin-converting enzyme 2 was examined by immunohistochemistry and RT-PCR. RESULTS: ACE₂ in the renal was significantly down-regulated in the SHR group compared with the WKY group (0.72 ±0.11 vs 1.11 ±0.15). After 12 weeks treatment with irbesartan, the attenuated ACE₂ expression in irbesartan group was markedly enhanced compared with the SHR group (1.03 ±0.13 vs 0.72 ± 0.11). CONCLUSION: Hypertension induces the reduction of ACE₂ in the renal in rats and irbesartan markedly increases ACE₂ expression. It suggests that up-regulation ACE₂ which is one of counter-regulatory particular components in the renin-angiotensin system may be a new mechanism of curing hypertension beyond blockade angiotensin type-1 receptor.

Study of fibroblast apoptosis selectively induced by artesunate

KONG Heng, YU Qing-sheng

2005, 10(6):  687-690. 

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AIM: To study the specificity of the antiproliferative effects of artesunate on fibroblast. METHODS: Human embryo lung fibroblasts cell line (HLF), human skin scar fibroblasts cell (HSF) and human umbilical vein endothelial cells (HUVEC) were selected in this trial. Cells were verified by immune his to-chemical method. A quantitative cytotoxicity was measured by MTT assay. Cell morphology change was identified by light microscopy. The apoptosis was evaluated by flow cytometry analysis and agarose gel electrophoresis. RESULTS: Artesunate inhibited the proliferation of HLF and HSF cells after cells were treated with artesunate for 24 hours in the dose range of 30-240mg·L^-1 (P <0.05). It had little inhibitory effects on HUVEC(P >0.05). Their IC₅₀ were 64, 60 and 600 mg·L^-1, respectively. Under light microscope, fibroblast treated with artemisinin assumed smaller, ellipse rather than fusiform in shape with increase of cytoplasmic particles, even with cell apoptosis. Ladder pattern of DNA were observed in artesumate-treated cells in the dose of 240 mg·L^-1.However, DNA of HUVEC cells was not affected in expropriate condition. CONCLUSION: The inhibitory effect of artesunate is specific on fibroblast coming from skin and lung. Artesunate can induce the apoptosis of the two fibroblasts, but it has no effect on HUVEC cells.

Effects of budesonide on signal pathway of IKK /NF-κB in asthma rats

LIN Yi-ping, LI Chang-chong, HU Ye, LI Men-grong, Chen Xiao-fang, MAO Yu-fei

2005, 10(6):  691-696. 

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AIM: To study the role of budesonide in regulating asthma inflammation and the activity of IKKβmRNA and NF-κBp65 protein. METHODS: Thirtysix Sprague-Dawley (SD) rats were divided into three groups: normal group, asthmatic model group and budesonide (Bud)group (n =12 in each).The asthmatic model was established by ovalbumin (OVA)injection and inhalation. The rats in Bud group were administered budesonide atomized inhalation within 14 days. The mRNA expression of IKKβ and the protein NF-κBp65 in lung tissue were assessed by situ hybridization with oligonucleotide probe and immunohistochemisty, respectively. The concentration of interleukin 5 (IL-5)in serum and BALF were measured by sandwich ELISA. RESULTS: Twentyfour hours after the last antigen chenllenge, the mRNA expression of IKKβ(0.203 ±0.038)and the protein expression of NF-κBp65(0.256 ±0.063)in asthmatic group increased compared with control group (0.015 ±0.006, 0.034 ±0.008, P <0.01, respectively).The mRNA expression of IKKβ (0.101 ±0.010)and the protein expression of NF-κBp65(0.062 ±0.014)in Bud group decreased compared with asthmatic group (P <0.01, respectively). CONCLUSION: In asthmatic rats, the expression of IKKβmRNA and the protein of NF-κBp65 were increased significantly, budesonide atomized inhalation can inhibit the expression of IKKβmRNA and the protein of NF-κBp65 and hold down the activity of the signal pathway IKK /NF-κB.

Antitumor effects of novel thymidylate synthase inhibitor ZD1694 in vitro and in vivo

LI Ling, YANG Su-rong, YAO Ming-hui

2005, 10(6):  697-700. 

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AIM: To observe the inhibitory effects of ZD1694 on the growth of S180 sarcoma cells in mice and investigate the effects of ZD1694 on the proliferation of gastric gland carcinoma SGC-7901 cell lines in vitro. METHODS: The tumor mice models were administered (ip) with ZD1694 at the different dose levels in one day or five consecutive days. The tumor weights were measured after the mice were slaughtered and the antitumor rates were calculated. In addition, MTT assay and flow cytometry were used for testing the effects of ZD1694 on SGC-7901 cell lines. RESULTS: The growth of S180 sarcoma cells in mice was inhibited by ZD1694.The order of the antitumor rates in different groups were listed as 200 mg·kg^-1 × 1 d > 100 mg·kg^-1 × 1 d > 40 mg·kg^-1·d^-1 × 5 d > 50 mg·kg^-1 × 1 d > 20 mg·kg^-1·d^-1 ×5 d >10 mg·kg^-1·d^-1 ×5 d. The in-hibitory rate of ZD1694(0.001-1 000 mmol·L^-1) on SGC-7901 cells were increased in concentration and timedependent manner. The cell numbers in G₀-G₁ phase in SGC-7901 cells were raised by ZD1694 as well. CONCLUSION: ZD1694 can take obvious antitumor effects on S180-bearing mice. The antitumor rate of ZD1694 is in a dose-dependent manner, and it is higher when the total dosage is administered in one day than that the total dosage was divided into 5 consecutive days. ZD1694 can inhibit the growth of SGC-7901 cells and block the cells in G₁ phase in vitro.

Expression of antibacterial peptide gloverin in vitro

LUO Ping, XU Bo, LU Yong-ling, ZHOU Hong

2005, 10(6):  701-704. 

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AIM: To construct a recombinant vector containing gloverin and express it in vitro by Rapid Translation System(RTS500). METHODS: The gloverin cDNA was amplified by PCR and inserted into the prokaryotic expression vector pIVEX2.3.The recombinant product was identified by PCR and enzyme digestion. The positive reconstructed expression plasmid pIVEX2.3-G was expressed by RTS500 in vitro. The expressed protein was identified by SDS-PAGE and western blotting. RESULTS: Positive recombinant plasmid pIVEX2.3-G was successfully constructed. Target protein of 13.8 Kda with 6-his tag was detected by SDS-PAGE and western blotting. CONCLUSION: Gloverin polypeptide is successfully expressed in inactive E. coli in vitro.

Research of regulating Bccap-37 cell treated with baicalin on TIMP2 expression in vitro

FENG Zheng-quan, SHEN Min-he, WU Liang-cun

2005, 10(6):  705-708. 

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AIM: To study the effects of baicalin on the growth and TIMP2 expression of bcap-37 cells. METHODS: The effects of baicalin at different concentration on the growth curve of bcap-37 cells were tested by MTT method and TIMP2 expression were analyzed by RTPCR and western blot. RESULTS: Baicalin inhibited the growth of the bcap-37 cells in a concentration relying way. Baicalin up-regulated TIMP2 expression in the bcap37 cells at both Mrna and protein levels at low dose, but down-regulated it at high dose. CONCLUSION: Baicalin can inhibit the growth of human mammary cancer cell line bcap37, and this effect maybe related to downregulate TIMP2 expression in the cancer cells.

Intracellular metabolism and bioconversion of β-L-D4A

WU Jin-ming, LIN Ju-sheng, ZHANG Jin-yan, LIANG Kuo-huan

2005, 10(6):  709-712. 

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AIM: To investigate the metabolism of β-L-D4A in 2.2.15 cells for further clarifying its anti-HBV mechanism and establishing a background on the studies of its anti-HBV effect and pharmacokinetics in animal model and human body. METHODS: 2.2.15 cells were treated with [³H] β-L-D4A or [³H] β-D-D4A at 2 μmol·L^-1 concentrations for 2, 4, 8, 12 and 24 hours, then the cells were extracted by adding 0.4 mol·L^-1 perchloric acid containing 0.08 mol·L^-1 triethylammonium phosphate. Then it centrifuged at 1 000 ×g for 5 min, the acid-soluble supernatant were directly isolated immediately by HPLC and monitored by connected ultraviolet detector, then the peaks were analyzed. RESULTS: Both of the two compounds present 4 peaks of metabolites, and the emergence time of each corresponding metabolite peak were similar. The retention times for β-LD4A, mono-, di and triphosphates were 6, 10, 19 and 28 min, respectively. Peaks for β-L-D4A metabolites were significantly higher than that for β-D-D4A metabolites in 2.2.15 cells after treated with each of the compounds for 24 h. Rapid conversion of β-L-D4A to its phosphorylated forms could be seen, especially for triphosphorylated form.With the concentrations used, maximal metabolite formation was observed at 8 h, then the metabolites began to reduce gradually. After 24 h treatment, when β-L-D4A was withdrawn, the levels of mono-, di, and triphosphates dropped rapidly in the first 8 h. In the subsequent 16 h, the triphosphate was removed at a lower rate, with 35.6 %of the triphosphate still present at 24 h in comparison with the amounts at 8 h. CONCLUSION: β-LD4A can be more easily phosphated metabolism than β-DD4A in 2.2.15 cells. Monophosphorylation may be the rate-limiting step. Triphosphate of β-L-D4A is degraded at a lower rate in 2.2.15 cells, and it may have a longer half-life.

Preventive effects of ginseng fiber on hepatic fibrosis induced by bone loss in mice

FENG You-hui, HE Kang, ZOU Li-yi, XU Bi-lian, LIU Yu-yu

2005, 10(6):  713-716. 

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AIM: To study the relationship between hepatic fibrosis and osteoporosis, and observe the preventive effects of ginseng fiber on bone loss. METHODS: Forty PCR micewere randomly divided into 4 groups. The mice in control group were treated by daily oral gavage with vehicle. The mice in other three groups were given SC injection of 40 %CCl₄ 10 ml·kg^-1 and treated by either daily oral gavage with vehicle, colchicine, or ginsen hair for 42 d. The liver injury indexes were measured and the mineral elements and hydroxyproline of femur were determined. RESULTS: Compared with the control group, the serum enzyme activity of alanine aminotransferase (ALT), aspartic acid aminotransferase (AST) markedly increased and serum albumin (Alb)and A /G distinctly decreased in CCl₄ group whose liver slides also showed typical liver cirrhosis. The dried weight of femur markedly reduced and the bone calcium content and bone hydroxyproline content significantly decreased in CCl₄ group. Bone copper and bone magnesium increased in CCl₄ group. Ginseng fiber markedly decreased the serum enzyme activity of ALT and increased the bone calcium content and bone hydroxyproline content. The preventive effects of ginseng fiber was similar to that of colchicine. CONCLUSION: The bone mass is lost in mice with chronic hepatic injury induced by CCl₄. Ginseng fiber can prevent bone loss and hepatic fibrosis induced by CCl₄ in mice.

Influence of rosiglitazone and metformin on blood pressure in treatment of patients with type 2 diabetes mellitus

LI Hong-hui, LIU En-bo, XIE Qun, DING Su-jie, HE Tao, LI Jia-fu

2005, 10(6):  717-720. 

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AIM: To investigate the influence of insulinsensitizer (Rosiglitazone) and metformin on blood pressure in treatment of patients with type 2 diabetes mellitus. METHODS: 58 patients with type 2 diabetes mellitus were randomly assigned to two groups.Patients orally received rosiglitazone 4-8 mg, qd, or metformin 250-500 mg, tid, for 12 weeks. The patients did not take any hypotensive drug during the investigation. BP, blood glucose and insulin were measured before and after treatment. The insulin resistance index and insulin sensitive index were calculated. RESULTS: SBP and DBP decreased after treatment by rosiglitazone and metformin (P <0.05). The decrease of blood pressure in rosiglitazone group were more than that in metformin group. FBG, PBG and FINS and PINS decreased after treatment (P < 0.05). ISI increased and IR decreased in the two groups (P <0.05). The extents of ISI and IR in rosiglitazone group were higher than that in metformin group. CONCLUSION: Rosiglitazone and metformin can effectively decrease IR and blood pressure, and the effect of rosiglitazone is better than that of metformin.