Chinese Journal of Clinical Pharmacology and Therapeutics

Advance research of effect of Ligustrazine on cardiovascular system

JIANG Hong, SHI Guang-fei, ZHU Zhu

2007, 12(5): 484-487.

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Ligustrazine is an active alkaloidal monomer which is extracted from the Chinese traditional herb Chuanxiong (Ligusticum chuanxiong Hort. ). With the in-depth research of Ligustrazine, its clinical use is widen day by day, especially its remarkable efficiency to the Cardiovascular Disease. The chemical structure of Ligustrazine is tetramethyl pyrazine, its four methyl can show different efficiency, when they are replaced by different functional units. It is found that the effect of Ligustrazine in the cardiovascular system has different ways and mechanisms of action. The pharmacological development of Ligustrazine in the cardiovascular system is reviewed.

Research advances of osteopontin in pathogenesis of rheumatoid arthritis

ZHANG Ge, WU Yu-lin

2007, 12(5): 488-492.

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Rheumatoid arthritis is one of the most critical diseases that impair the quality of patients’life,but its pathogenesis has not yet been fully understood. Osteopontin (OPN) is an extracellular matrix protein containing Arg-Gly-Asp (RGD) sequence and has been recently recognized as a potential inflammatory cytokine. It is discovered that OPN plays an important role in the pathogenesis of rheumatoid arthritis. It is a bridge between bone and the immune system. This paper reviewed the pivotal role that OPN plays in the pathogenesis of rheumatoid arthritis.

Drug interactions with antianginal drugs

LI Xiao-min, GUO Xin, CHENG Ze-neng

2007, 12(5): 493-498.

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Angina pectoris is the major clinical symptom of myocardial ischemia. Angina pectoris can be managed by using drugs that either improve perfusion of the myocardium or reduce its metabolic demand. Many of the drugs used in angina pectoris are the substrate of cytochrome P450, drug interactions are easily to occur when antianginal drugs co-administrated with other drugs and foods. In this article, the drug interaction of main antianginal drugs in pharmacokinetics were reviewed, and it will help the rational use of antianginal drugs in clinical medication.

Progress in treatment of multiple sclerosis

ZHANG Liu-fu, HUANG Qiong, CHU Lan

2007, 12(5): 499-503.

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Multiple sclerosis is a demyelinating disease affecting central nervous system induced by a chronic autoimmune disorder. Its exactly etiological mechanism remains unknown. Viral infection and autoimmunity functional disorder may be one of the reasons. In this article, we mainly reviewed the progress in the treatment of the disease including immune depressant, gene therapy, transplant therapy of stem cells and other therapies.

Neuroprotective effects of paeoniflorin against cerebral ischemia through pharmacological preconditioning

WANG Guo-feng, CHEN Dong-mei

2007, 12(5): 504-511.

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AIM: To investigate the neuroprotective effects of paeoniflorin (PF) , the principal component of Paeoniae Radix prescribed in traditional Chinese medicine against the damage of rat middle cerebral artery occlusion (MCAO) and reperfusion through pharmacological preconditioning (PPC) and its molecular mechanisms, in order to offer novel drug targets and therapeutic strategies against ischemic stroke. METHODS: In rat MCAO and reperfusion model, the effects of PF pretreatment through PPC on the infarct volume and neurological deficits were examined. Furthermore, the mRNA or protein expression levels of cyclooxygenase-2 (COX-2) and 5-lipoxygenase (5-LOX) have been examined after PF preconditioning using RT-PCR and Western Blot analyses. Finally, the protein expressions of MAPK signaling effectors were examined after PF preconditioning. RESULTS: At a dosage of 20 or 40 mg kg, PF preconditioning reduced the infarct volume dose-dependently and reversed the neurological deficits caused by ischemia. Similarly, the ameliorative effects on infarct size and neurological impairment induced by MCAO emerged as well at a time-dependent manner when PF was administered 24, 48 h or 5 d before MCAO at the dose of 20 mg kg. The over expressions ofCOX-2 mRNA and protein and 5-LOX protein in ischemia were reversed by PF preconditioning. The activation of MAPK signaling pathway was also inhibited by PF preconditioning. CONCLUSION: The PPC could be induced by PF, mimicking the effects of ischemic preconditioning. The molecular mechanisms underlying its ameliorative effects are through inhibiting the inflammation involved in arachidonic acid metabolism and the activation of MAPK signaling pathway.

Suppressing effect of gastrin-releasing peptide DNA vaccine on EMT6 breast cancer growth

OUYANG Ke-dong, GUO Wei, WU Guo-jun, ZHANG Shu-ya, LIU Jing-jing

2007, 12(5): 512-515.

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AIM: To observe the inhibiting effect of the GRP DNA vaccine on EMT6 breast cancer tissue. METHODS: Female BALB c mice were immunized intramuscularly with GRP DNA vaccine pCR3. 1-VS-HSP65- TP-GRP6-M2 5 times at 2-weekly intervals. The specific anti-GRP antibody was detected by ELISA method. Two weeks after the last immunization, tumor challenge experiments were performed by using EMT6. After 14 d of challenge experiments, all mice were killed and tumors were weighted. Histological analysis of tumor tissue was carried out with HE staining. RESULTS: The specific anti-GRP antibodies were detected in the antiserum of the female BALB c mice immunized with pCR3. 1-VS-HSP65-TPGRP6- M2 DNA vaccine. It showed that EMT6 tumor growth in mice of GRP DNA vaccine group was obviously suppressed (P<0.01) compared with that in pCR3. 1- VS-HSP65-TP or saline control group, with tumor inhibitory rate of 46. 53 %. Histological analysis showed that GRP DNA vaccine successfully induced anti-tumor immune responses in vivo, and, the invasiveness of EMT6 tumor tissues was markedly decreased in mice of GRP DNA vaccine group compared with that in pCR3. 1-VSHSP65- TP control group. CONCLUSION: GRP DNA vaccine can significantly suppress the growth of EMT6 breast cancer in vivo, which lays a basis for further research.

Effect of tetrahydroxystilbene-glucoside on lipid and inflammatory factor levels of experimental atherosclerosis in rats

ZHANG Wei, LI Feng, WANG Yu-qin,WANG Chun-hua, SHEN Yan

2007, 12(5): 516-520.

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AIM: To investigate the preventive effect of tetrahydroxystilbene-glucoside (TSG) on experimental atherosclerosis rats. METHODS: Sixty one male rats were randomly divided into six groups. normal control; model control ;TSG low dose (30 mg∙kg^-1∙d^-1) ;TSG middle dose (60 g∙kg^-1∙d^-1 ) ;TSG high dose (120mg∙kg^-1∙d^-1) ;simvastatin(2 mg∙kg^-1∙d^-1). The AS model of rats was made by feeding high grease food and injecting VitD3. All the rats were fed for 12 weeks, blood samples were drawn from carotid artery of rats, the levels of TC, TG, HDL-C, LDL-C , CRP, IL-6 and TNF-αin serum were measured with biochemical method. After blood samples were collected , the aorta samples were separated from the bodies, then they were placed 4 % paraformaldehydea and were through Sudan Ⅳ and HE staining. STATA7. 0 software was used to evaluate the differences between groups. RESULTS: Data of the study demonstrated that the levels of TC 、TG 、LDL-C 、TNF-α、IL-6 and CRP were decreased remarkably, the level of HDL-C was increased by TSG 60, 120 mg∙kg^-1∙d^-1group in the high cholesterol-fed rats,which showed a dose-dependent effect. The result of SudanⅣ and HE staining suggested that the lipid deposits in aortic endothelium in TSG group were less than those in model group. CONCLUSION: TSG has preventive effect on the experimental atherosclerosis among the high cholesterol-fed rats. The anti-atherogenic effect of TSG seems to be closely related to regulating plasma lipid profile, and inhibiting inflammation.

Effect of simvastatin on regression of pressure overload-induced rat’ s myocardial fibrosis and its relationship with monocyte chemoattractant protein-1

ZHANG Li-juan, ZHAO Lian-you, ZHENG Qiang-sun, SHANG Fu-jun, YANG Run-tao, LIU Hui, LIU Shao-wei

2007, 12(5): 521-525.

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AIM: To study the effect of 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase inhibitor simvastatin on MCP-1 expression and macrophages infiltration in pressure overload-induced myocardial fibrosis in rats with hypertension induced by abdominal aortic coarctation and further clarify the underlying mechanism of the treatment. METHODS: Pressure overload-induced rat model was established by abdominal aorta constriction (AC). Eighteen SD rats were divided randomly into sham group, AC group and simvastatin group. At the ending of observation, mean arterial blood pressure (MBP) was measured by carotid artery intubation, and ratio of ventricle mass to body weigh (LVW BW) was calculated. Myocardial interstitial fibrosis and perivascular fibrosis were evaluated by PAS. Immunohistochemistry was used on myocardium for ED-1 which was a marker of macrophage. MCP-1 mRNA and protein expression levels in myocardium were determined with reverse transcription polymerase chain reaction (RT-PCR) and ELISA, respectively. RESULTS: MBP, LVW BW, MCP-1 protein and mRNA expression in both AC group and simvastatin group were significantly higher than those in sham group (P<0.05 or P<0.01). Collagen volume fraction (CVF) and perivascular circumferential area (PCVA) , ED-1 positive cells in AC group were increased significantly compared with sham group (P<0.01). LVW BW, CVF, PCVA, ED-1 positive cells, MCP-1 protein and mRNA expression in simvastatin group were significantly lower than those in the AC group (P<0.01). CONCLUSION: Simvastatin prevents pressure overload-induced rat’ s myocardial fibrosis, which is associated with the reduction of MCP-1 in myocardium and interstitial macrophage infiltration.

Experimental study of effect of Shuanghuanglian liquor on scalded or burnt rats

LI Li-hua, HA Na, YANG Meng-huan, SUN Zhao-jun

2007, 12(5): 526-529.

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AIM: To clarify the healing effect of Shuanghuanglian liquor on the wounded surface of the scalded or burnt rats. METHODS: We made rats’scalded or burnt models on backs or feet, respectively. Those rats were divided into three groups :the model group, the sulfadiazine silver treated group and the Shuanghuanglian liquor treated group, in order to observe the healing process of the wounded surface in the three experimental groups. RESULTS : The cold and wet application of Shuanghuanglian liquor could not only shorten the time for the healing of the wounded surface of scalded or burnt rats, but also reduce the size of the wound. The healing rate of Shuanghuanglian liquor was similar to that of sulfadiazine silver group, but was much higher than that of the modeled group. Under the microscope, we observed that the skin appendages in dermis were still in integrity. No edema or inflammatory cell infiltration was found. CONCLUSION: The cold and wet application of Shuanghuanglian liquor shortens the healing course of wounded skin in scald or burnt rats.

Effect of polystyrene sulphur lanthanum on hyperphosphatemia in rats with chronic renal failure induced by adenine

LI Jie, WANG Peng, XIE Yan-ying, ZHANG Qiang, ZHANG Xiu-qin, LIN Xia

2007, 12(5): 530-534.

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AIM: To observe the effect of polystyrene sulphur lanthanum on hyperphosphatemia in rat model with chronic renal failure induced by adenine. METHODS: Chronic renal failure of rats was induced by Yokozawz method through infusing adenine (0.3g·kg^-1·d^-1 ) for 25 days, and then model rats were given polystyrene sulphur lanthanum (1.500, 0.750, 0.375g·kg^-1·d^-1 ) and positive drug for 22 days, and the adenine was given continuously for another 10 days. During the experiment courses, the body weight, general status,appetite, urination and defecation were observed. And levels of BUN, Cr, P3 +, Ca2 +, K+, RBC, Hb, PLT,and HCT in blood were monitored after the drug was given for 10 and 21 days. Rats were killed for kidney pathology and a quotient calculation. RESULTS: The polystyrene sulphur lanthanum at a dose of 1.500 g·kg^-1 ·d^-1 could reduce the phosphor level significantly, and there was obviously different from that of control model group (P<0.05). CONCLUTION: The polystyrene sulphur lanthanum can significantly reduce the phosphor level of rat with chronic renal failure induced by adenine.

Effect of cilostazol on neuronal injury after focal cerebral ischemia in mice

CHEN Li-ping, YE Yi-lu, SHI Wen-zhen, WANG Meng-ling, ZHANG Wei-ping, WEI Er-qing

2007, 12(5): 535-539.

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AIM: To determine the protective effect of cilostazol on neuronal injury after persistent focal cerebral ischemia in mice. METHODS: Persistent focal cerebral ischemia was induced by middle cerebral artery occlusion. Cilostazol (3-30 mg/kg) and pranlukast (0.1mg/kg) were i. p. injected 30 min before ischemia. The changes in the morphology and the densities of neurons and degenerated neurons were determined 24 h after ischemia. RESULTS: After ischemia, neuron density was reduced and the degenerated neurons were increased. Cilostazol (3,10 mg/kg) and pranlukast significantly increased the neuron density and reduced the degenerated neurons in the ischemic hemisphere. CONCLUSION: Cilostazol has protective effect on neuronal injury after persistent focal cerebral ischemia in mice.

Protective effect of ligustrazine on acute injury fatty liver in mice

SUN Yu-qin, GAO Tian-yun, ZHOU Juan, HONG Bei-bei, DING Hong

2007, 12(5): 540-543.

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AIM: To investigate the protective effect of ligustrazine on fatty liver induced by carbon tetrachloride in mice and study the possible mechanisms. METHODS: The fatty liver model was established by 0.5 % carbon tetrachloride (CCl4)-oil solution i. p. on the first day of the experiment. One hour before the model was made, mice in high-dose, middle-dose and low-dose groups were given ligustrazine 50, 25 and 12.5 mg/kg i.p., respectively. Furthermore, the model group, Tiopronin group and different dosages of ligustrazine groups were taken normal saline, Tiopronin 200 mg/kg, and ligustrazine 50,25 and 12.5 mg/kg i.p., respectively, twice a day for 7 days. The indexes of liver function such as levels of GOT and GPT in serum were evaluated. The liver lipid and lipid superoxidation were measured, and pathology was examined. RESULTS: Compared with the model group, the activities of ALT and AST significantly decreased in serum (P<0.05) ;Ligustrazine markedly decreased the contents of triglycerides (TG) (P<0.05) and free fatty acid (FFA) (P<0.01) in liver tissue ;the activities of lipase and SOD were higher (P<0.05) , and the content of malondialdehyde (MDA ) decreased markedly (P<0.05 or P<0.01) in liver tissue. CONCLUSION: Ligustrazine improves the protective effect on injury liver in mice and decreases the deposition of triglycerides in liver. The possible mechanism is that ligustrazine can reduce the TG deposition, promote the β-oxidation of FFA and decrease the lipid peroxidation injury.

Effect of D-aminoglucose derivatives on reactive oxygen species and mitochondrial membrane potential in human esophageal cancer cell line Eca-109

YANG Guo-dong, WU Jing, QIANG Zhan-rong, ZHOU Yong-ning, WANG Ai-qing, XUE Qun-ji

2007, 12(5): 544-547.

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AIM: To explore mechanism of apoptosis in Eca-109 cells induced by 2-(3-carboxy-1-oxoprogy1)amino-2-deoxy-D-glucose (COPADG ). METHODS: Eca-109 cells were cultured with different concentrations of COPADG for 24 h. Then we examined cell growth inhibitory rate , apoptosis rate , reactive oxygen species(ROS)and ΔΧm of Eca-109 cells. RESULTS: The apoptosis rate of Eca-109 cells and the concentration of COPADG presented a positive correlation (rs=1.0 , P<0.01). The ΔΧm and the apoptosis rate of Eca-109 cells were related (rs=1.0 , P<0.01). The ROS and the apoptosis rate showed a positive relationship (rs=1.0 ,P<0.01). The levels of ROS and the ΔΧm were negatively related (rs=1.0 , P<0.01). CONCLUSION: COPADG promotes apoptosis of Eca-109 cells , raises ROS level in Eca-109 cells and lows level of ΔΧm. The experimental results suggest COPADG increases ROS level and lows level of ΔΧm triggering the thoroughfare of cell apoptosis to induce apoptosis in Eca-109 cells. The exaltation of ROS results in a decrease in ΔΧm.

Effects of cinobufacine on intracellular adriamycin accumulation in human breast cancer cells

WANG-Ling, LIU Shi-kun, ZHOU Yu-lu , PEI Qi, YANG Ning

2007, 12(5): 548-551.

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AIM: To study the effects of cinobufacine on intracellular adriamycin accumulation in human breast cancer MCF-7/ADM cells, and further explore the mechanism of cinobufacine reversing the multidrug resistance of human breast cancer MCF-7/ADM cells. METHODS: The cytotoxic effect was determined by MTT assay. Intracellular ADM concentration was examined by HPLC. RESULTS: 15 mg/L cinobufacine decreased the IC₅₀ of ADM to MCF-7/ADM cells from 38.14 mg/L to 12.93 mg L, but increased intracellular adriamycin accumulation in MCF-7/ADM cells. CONCLUSION: Cinobufacine can increase intracellular ADM accumulation and partly reverse MDR of MCF-7/ADM cell line.

Protective effects of bendazac lysine against development of diabetic nephropathy

YIN Xiao-xing, ZHANG Yin-di, YU Jun-xian, ZHANG Bei, SHEN Jian-ping, QIU Jun

2007, 12(5): 552-561.

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AIM: To investigate the protective effects of bendazac lysine (BDL) on diabetic nephropathy (DN) in vitro and in vivo experiments. METHODS: After rat mesangial cells were cultured in 3 concentrations of BDL for 36 h, the percentages of S phase of cells were determined by flowcytometry ;the transforming growth factor β₁(TGF-β₁ ) mRNA level was assayed by reverse transcription PCR;and two main components of extracellular matrix (ECM) , collagen Ⅳ and laminin, were determined by radioimmunoassay. Streptozotocin (STZ) induced diabetic rats were administered BDL at doses of 100, 200, 400 mg/kg for 8 weeks. The physical behavior andHbAlC levels of rats were observed. RESULTS: In the presence of high glucose and H₂O₂ , the percentages of S phase of cells were lowered, and TGF-β₁ mRNA level, collagen Ⅳ and laminin level were significantly increased. When compared with those in the high glucose group, the percentages of S phase of cells were significantly raised, and the levels of TGF-β₁mRNA, collagen Ⅳ and laminin were statistically decreased. The physical behavior of high BDL treated rats restored to be vibrant, vigorous and weight gaining, and the HbAlC level was significantly reduced. CONCLUSION: BDL has the protective effects against damage caused by DN, and is a potential drug candidate worth further study in preventing and treating DN.

Protective effect of medicinal serum of Jiangtang compound recipe on oxidative-injured endothelial cells

HUANG Wei-jia, WU Guan-zhong

2007, 12(5): 562-565.

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AIM: To study the protective effect of medicinal serum of Jiangtang compound recipe on endothelial cells which were damaged by H2O2. METHODS: The effects of medicinal serum of Jiangtang compound recipe on cell viability, concentration of malondialdehyde (MDA) , NO, endothelin-1 (ET-1) and activity of superoxide dismutase (SOD) were determined for human umbilical vein endothelial cells (ECV304) damaged by H2O2. RESULTS: The medicinal serum of Jiangtang compound recipe significantly improved the structural change of ECV304 cells damaged by H2O2 , suppressed the production of MDA and the content of ET-1, increased activity of SOD and the rate of animate cells, and promoted secretion of NO. CONCLUSION: The medicinal serum of Jiangtang compound recipe can resist the injury of ECV304 caused by H2O2 , playing a role in the protective effect.

Induction of apoptosis by matrine in RPMI8226 cells and its effect on Bcl-2 and proliferating cell nuclear antigen

ZHANG Sheng-hui, YU Kang, WU Jian-bo, HAN Yi-xiang, XIONG Shu-dao, TAN Ying-xia

2007, 12(5): 566-570.

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AIM: To explore whether the apoptosis was induced by matrine and its effect on the expression of Bcl-2 and proliferating cell nuclear antigen (PCNA) in human mutiple myeloma cell line RPMI8226 cells. METHODS: RPMI8226 cells were incubated with indicated concentrations of matrine. The growth of RPMI8226 cells was observed by CCK-8 colorimetric assay, and apoptosis rates were detected by flow cytometry using Annexin V-FITC PI staining. The cell cycles were analyzed by PI staining. Flow cytometry was used to detect the expression of Bcl-2 and PCNA. RESULTS: RPMI8226 cells viability in presence of matrine decreased markedly in a dose and time-dependent manner. The apoptosis could be induced by matrine and its level increased following the augmentation of the drug concentration. After treated by matrine for 48 h, a concentration-dependent increase of cells in G0 G1 phase and a decrease in S phase were detected, but there was no obvious change at G2 M phase. Treatment of RPMI8226 cells with matrine for 48 h provoked a decrease in the level of expression of Bcl-2 and PCNA. CONCLUSION: Matrine can significantly inhibit the growth of RPMI8226 cells, its function is performed by inducing apoptosis and arresting cell cycles. Bcl-2down-expression may probably function as an important regulator in the process of apoptosis induced by matrine.

Effect of garlicin on protecting nonalcoholic fat liver in SD rats induced by high fat diet

SHAO Liang, HAN Zhen, WU Wan-chun, HE Chi-yi

2007, 12(5): 571-574.

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AIM: To study the effect of garlicin on protecting nonalcoholic fat liver in rats induced by high fat diet and explore the pathogenesis involved. METHODS: According to the dosage of garlicin and diet, fifty SD rats were randomly divided into five equal groups:normal control group, model control group and garlicin (10, 20, 30 mg/kg) groups. Apart from the rats in normal control group, the rats were all fed with high fat and high cholesterol diet. After 12 weeks, the levels of serum endotoxin (ETX) , total cholesterols (TC) , triglyceride (TG) , superoxide dismutase (SOD) , malondialdehyde (MDA ) , transaminase and free fatty acids (FFA) were detected. The levels of MDA, SOD, glutathione hormone (GSH) in hepatic tissue were also detected. Then the features of live pathology were observed. RESULTS: The levels of ETX, TC and TG in garlicin groups were significantly lower than those in model control group (P<0.05 or P<0.01). The SOD and GSH in serum and hepatic tissue of rats treated with garlicin were significantly higher than those in model control group (P<0.05 or P<0.01) ,while the serum MDA and FFA as well asMDA in hepatic tissue were significantly lower than those in model control group (P<0.05, or P<0.01). High dosage of garlicin could alleviate adipose degeneration of hepatic cells compared with low-dose garlicin group (P<0.05). CONCLUSION: Garlicin has high effect of preventing nonalcoholic fat liver disease in a dose-dependent manner. The anti-lipid peroxidation and lowering serum endotoxin effect of garlicin may be the mechanism.

Therapeutic effects of benthiacthiactzine against circulatory failure induced by organophosphate insecticides and hemorrhagic shock in rats

LIU Nian, CAO Jie-wei, WANG Ru-huan, LONG Chao-liang, WANG Hai

2007, 12(5): 575-581.

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AIM: To investigate the therapeutic effects of benthiactzine against circulatory failure induced by organophosphate insecticides DDV and hemorrhagic shock in rats. METHODS: 42 healthy Wistar male rats, weighing 320 ±20 g, were divided into six groups randomly :DDV poisoning rats treated with benthiactzine 0.5, 1. 0 or 2.0 mg/kg, and hemorrhagic shock rats treated with saline or benthiactzine 0.5, 2.5 mg/kg (n= 7). Rats were treated with organophosphate insecticides (i.p. ) to induce circulatory failure shock in which the mean blood pressure (MBP) dropped to 45 mm Hg as an index, then treated with benthiactzine (i.m). The others were exsanguinated untill MBP decreased to 35 -40 mm Hg within 15 min, and models of hemorrhagic shock were established by maintaining MBP at that level for 30 -60min, then those rats were treated with saline or benthiactzine (i.m). RESULTS: In circulatory failure rats inducedby DDV, SBP, DBP, MBP, HR, LVDP, IP, + dp/dtmax , -dp/d tmax, Vpm and +dp/d tmax IP dramatically decreased (P <0.01) compared with pre-poisoning,and some kinds of arrhythmia were observed. Benthiactzine affected each parameter in 30 s to 3 min. CONCLUSION: Treated with benthiactzine, DDV poisoning rats can get away from circulatory failure in a short time. Benthiactzine can be a favourable medicine against circulatory failure induced by organophosphate insecticides to save time for other clinic treatments.

Operational guidance :information needed to support clinical trials of herbal products

WANG Xiu-qin, Revised by XIONG Ning-ning, BO Qin-yan, JU Wen-zheng

2007, 12(5): 582-585.

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