Effect of intestinal endotoxemia on ICAM-1 expression in the liver of rats

AIM: To investigate the effect of intestinal endotoxemia on intercellular adhesion molecule-1(ICAM-1) expression in the hepatic tissue.METHODS:Rat intestinal endotoxemia was induced by thioacetamide(TAA).Changes in ICAM-1 protein in the liver were detected by Western blot. RESULTS: The molecular weight of the ICAM-1 is 95 kD.Western blot analysis of hepatic tissue from control rats and rats injected with TAA within 6 hours revealed low ICMA-1 expression. ICAM-1 expression upregulation occured in rats with intestinal endotoxemia in experimental liver injury induced by TAA 12 h after TAA injection. ICAM-1 expression, plasma endotoxin level and alanine transaminase activity is of equal rank.CONCLUSION: Intestinal endotoxemia can upregulate ICAM-1 expression in the hepatic tissue and the latter is related to liver injury.     

Effects of nicotine on activation of PMNs and the ICAM-1 mRNA expression of HUVEC

AIM: To investigate the effects of nicotine on activation of PMNs, adhesion of PMNs-HUVEC and expression of ICAM-1 mRNA in HUVEC. METHODS: Activation of PMNs was measured by detecting the activity of β-glucuronidase and lysozym of PMNs. Adhesion of PMNs and HUVEC was observed. Northern blot was conducted for quantitating ICAM-1 mRNA. RESULTS: Nicotine could increase the activity of β-g [(8.76± 1.01)μg/10⁷·h vs(14.87±2.00)μg/10⁷·h,P<0.05]and Lysozym [(20.0±1.5)μg/10⁷·h vs(36.5±4.4)μg/10⁷·h,P<0.05], and also could promote adhesion of PMNs-HUVEC(38.5±9.8 vs 61.0±4.4,P＜0.05). The expression of ICAM-1 mRNA was induced by nicotine in dose-dependent fashion (10^-5-10^-3mol/L).After a 2 h treatment of HUVEC with nicontine(10^-4mol/L), the level of ICAM-1 mRNA is above the control(1.23 vs 1.63) and the highest level (2.03) is at a 12 h treatment. 764-3 can obviously counteract the above effect of nicotine. CONCLUSIONS: Nicotine could activate PMNs, enhance adhesion of PMNs-HUVEC and increase the expression of ICAM-1 mRNA in HUVEC.     

丹参开花与繁育特性研究

 通过田间观察和人工授粉等方法, 对丹参( Salvia miltiorrhiza Bge. ) 的开花生物学和繁育特性进行了初步研究。结果表明: 丹参为无限开花植物, 总状花序上通常有两性花近50朵, 单花花期5～7d, 从花前2 d到开花6 h花粉活力和柱头可授性都较大, 两者有效可遇期约为1.5 d, 可同花自交; 丹参自交结实和自然结实主要是通过同株异花传粉获得; 结合花粉/胚珠比、杂交指数以及繁育处理的结果认为,丹参不存在无融合生殖, 繁育类型为兼性异交, 自交亲和, 需要传粉者; 传粉媒介主要是蜂类。初步推断丹参为常异花授粉植物, 并对丹参育种中适宜采用的杂交方法及育种途径进行了探讨。     

丹参细胞分裂素调控因子序列特征分析

细胞分裂素通过双元信号系统感知外界环境刺激并作出应答。其中细胞分裂素响应调控因子（response regulator,RR）是该双元信号系统中的重要构成。采用生物信息学的方法,对丹参（Salvia miltiorrhiza）细胞分裂素调控因子进行序列特征和组织表达特性分析。结果发现,丹参基因组中共有15个丹参细胞分裂素调控因子SmRR,序列比对与系统发生分析表明,SmRR1 ~ SmRR7与拟南芥A类RR亲缘关系比较接近,划分为A类RR,SmRR8 ~ SmRR15与拟南芥B类RR亲缘关系近,归为B类;在A类SmRR中,REC接受域含有保守的DDK残基,因此这些基因被推测有完整的功能,B类SmRR具有高度保守Myb_SHAQKYF结构域。两类SmRR的内含子、外显子组成也具有相对的保守性,顺式调控元件的分析发现,SmRR存在很多胁迫和生长调节物质的响应元件。表达量热图显示A类和B类SmRR基因分别在叶和根中表达量较高。     

Chlamydia pneumoniae enhanced the expression of ICAM-1 in cultured human umbilical vein endothelial cells

AIM: To investigated the effects of Chlamydia pneumoniae (C.pneumoniae) infection on the expression of intercellular adhesion molecule-1 (ICAM-1) in cultured human umbilical vein endothelial cells (HUVECs). METHODS: After propagated in HEP-2 cells, C. pneumoniae organisms were infected to HUVECs. The infection was assessed by ectromicroscope and PCR. The expression of ICAM-1 on HUVECs was detected by flow cytometry before infection and 12 h, 24 h, 48 h, 72 h after infection. RT-PCR was used to detect the ICAM-1 mRNA expression. RESULTS: C.pneumoniae was able to infect cultured HUVECs. After infection, the expression of ICAM-1 on the surface of cultured HUVECs was increased,, the peak was at the time of about 24-48 h; The light quantative RT-PCR analysis demonstrated that the infection also enhanced the ICAM-1 mRNA expression. CONCLUSION: The ability of C.pneumoniae to grow in HUVECs and to stimulate the expression of ICAM-1 protein and mRNA suggests that C.pneumoniae may playa role in atheriosclerosis.     

NF-κB site of promotor mediates nicotine-induced ICAM-1 expression in human umbilical vein endothelial cells

AIM:To study the mechanisms of nicotine-induced expression of intercellular adhesion molecule-1(ICAM-1).METHODS:Related luciferase reporter gene plasmids were constructed with molecular cloning techniques;above plasmids and intracontrol plasmid pSV-β-gal were co-transfected into human umbilical vein endothelial cells(HUVECs) with eukaryotic gene transfection techniques; the relative luciferase activities were detected in the transfected HUVECs.RESULTS:Series of luciferase reporter gene containing different sequences of human ICAM-1 promotor and site-directed mutants of NF-κB and Sp-1 in promotor were successfully constructed; Nicotine could increase the expression of luciferase reporter gene plasmid containing-579 bp(pGL3E-579/+36),-230 bp(pGL3E-230/+36) and mutated Sp-1 version(pGL3E-Sp-1-MU)(P＜0.05 vs control) of ICAM-1 promotor in the transfected HUVECs, whereas deletion derivative (pGL3E-134/+36) and mutation (pGL3E- NF-κB -MU) of downstream NF-κB site of ICAM-1 promotor prevent nicotine-induced increase in expression of luciferase reporter gene plasmid.CONCLUSION:NF-κB site of promotor mediates nicotine-induced ICAM-1 expression in human umbilical vein endothelial cells.     

白花丹参组织培养技术的研究

以白花丹参叶为外植体,研究了外植体种类、激素比例、琼脂浓度、活性炭等培养条件对组培快繁的影响。结果表明:培养基MS+6-BA 2.0mg/L+NAA 1.0mg/L+琼脂7%适宜2种愈伤组织诱导;培养基MS+KT 1.5mg/L+NAA 0.5mg/L+琼脂8%+活性炭0.3%能有效预防玻璃化苗,适宜健壮芽苗增殖;培养基1/2MS+IBA 0.2mg/L+琼脂8%(或同时添加0.3%活性炭)有利于试管苗根系的生长,生根率达到100%。试管苗移栽成活率达到95%。     

丹参新品种‘丹杂1号’和‘丹杂2号’

 丹参新品种‘丹杂1号’和‘丹杂2号’是以‘冀丹2号’为母本,分别以‘D0501’和‘冀丹1号’为父本进行杂交选育而成。两品种产量高,药用成分含量高,抗根腐病能力较强。‘丹杂1号’产量为5 629.5 kg · hm^-2,比河北安国栽培丹参增加14.7%,丹参酮ⅡA含量为0.25%,丹酚酸B含量为8.10%;‘丹杂2号’产量为5 578.5 kg · hm^-2,比安国栽培丹参增加13.7%,丹参酮ⅡA含量为0.29%,丹酚酸B含量为11.7%。     

Induction of apoptosis with Salvia miltiorrhiza monomer IH764-3 via downregulating focal adhesion kinase in H2O2-stimulated hepatic stellate cells

AIM: To investigate the effect of IH764-3 on the apoptosis of H₂O₂-stimulated hepatic stellate cells(HSCs) and the alteration of focal adhesion kinase (FAK). METHODS: The proliferation of HSCs was examined by direct cell count and the apoptosis was determined by Annexin-V/PI labeling, while the morphological change was observed by light microscopy and transmission electron microscopy. In addition, FAK mRNA was detected by RT-PCR. RESULTS: H₂O₂ promoted the proliferation of HSCs. IH764-3 induced the apoptosis of HSCs in a dose-dependent manner. The HSC apoptotic rates of different groups were 6. 35%,9. 28%,15. 10%,19. 69%,respectively, after treated with different concentrations of IH764-3 for 48 h while H₂O₂ group showed 2. 30%. In 30 mg/L group, the apoptosis rates were 6. 73%、10. 34%、15. 10% for the indicated time periods(12 h, 24 h, 48 h). In the presence of IH764-3, FAK mRNA decreased. The FAK mRNA reduction began at 2 h after adding IH764-3. CONCLUSION: IH764-3 induced the apoptosis of HSCs. Down-regulating the expression of FAK mRNA may be one of its mechanisms.     

灵丹菌质双向发酵工艺及药效品质研究

采用双向固体发酵工程技术,以灵芝为发酵菌株,在含有不同比例丹参边角料的培养基中进行发酵,观察了不同培养基中菌丝的生长情况,测定了发酵产物（灵丹菌质）中多糖、蛋白质、三萜、丹参酮的含量变化,并研究了其提取物对小鼠活血化瘀功效的影响。结果表明灵丹菌质的最佳发酵组合为组合4,发酵40d,多糖含量26.48μg·g-1,蛋白质含量为397.24μg·g-1,灵芝酸三萜含量为4.92%。药效学实验显示,灵丹菌质组合4具有活血化瘀的药效,优于丹参药材。     

Expression of ICAM-1 gene in the ischemia-reperfusion liver

AIM: To investigate the expression of ICAM-1 mRNA in hepatic sinusoidal cells (HSCs) during liver ischemia-reperfusion injury and its effects. METHODS: Three groups of rabbit livers experiencing ischemia for 15, 30 and 60 minutes respectively, and reperfusion for 60 minutes were conducted. The hepatic ICAM-1 mRNA expression was observed and leukocytes in hepatic sinus were counted. RESULTS: The lower expression of ICAM-1 mRNA showed in HSCs before reperfusion. Besides, leukocyte counts could no change. However, the more expression of ICAM-1 mRNA was found at the point of 60 minutes reperfusion. Furthermore, the higher expression of ICAM-1 mRNA showed in the more prolonged ischemic liver. Meanwhile, there was significant positive correlation between ICAM-1 mRNA contents and leukocyte counts. CONCLUSION: The high ICAM-1 mRNA expression induced by hepatic ischemia in HSCs increased adherence of HSCs and sequestrated more leukocytes in liver.     

Effect of compound Salvia militorrhiza injection on LPS-induced disseminated intravascular coagulation in rabbits

AIM: To evaluate the effect of compound Salvia militorrhiza(SM) injection on lipopolysaccharide (LPS)-induced experimental disseminated intravascular coagulation (DIC) in rabbits. METHODS: The LPS-induced DIC model in rabbits was performed by continuous infusion of LPS at 100 μg·kg^-1·h^-1 for 6 h. Treatment with compound SM injection or heparin was begun simultaneously with LPS infusion through the contralateral marginal ear vein.Activated partial thromboplastin time (APTT), prothrombin time (PT), platelet count and fibrinogen concentration were determined. The plasma levels of alanine aminotransferase (ALT) and blood urine nitrogen (BUN) were detected. The activity of protein C and antithrombi III (ATIII) was also measured.RESULTS: Continuous infusion of LPS induced gradual impairment of hemostatic parameters and damages in renal and liver functions. The intravenous administration of compound SM injection attenuated the increase in plasma levels of APTT and PT, and the decrease in plasma levels of fibrinogen and platelet induced by LPS infusion. Compound SM injection decreased the levels of ALT and BUN, reduced the damages in the liver and kidney, and also significantly inhibited the decreased plasma levels of protein C and ATIII in LPS- treated animals.CONCLUSION: Compound SM injection has a protective effect against DIC in rabbits.     

Salvia miltiorrhiza antagonized the alterations of contraction and intracellular calcium of rat ventricular cardiomyocytes induced by anoxia and reoxygenation

AIM:To study the effect of salvia miltiorrhiza (SM) on cell contraction and intracellular calcium of enzymatically isolated rat ventricular myocytes during normoxia and anoxia/reoxygenation.METHODS:Contraction and intracel ular calcium were determined with video tracking system and spectrofluorometric method,and the chemical anoxic method was employed. RESULTS:The ±dL/dt_max, dL of cell contraction and the amplitude of [Ca²⁺]_i in the cardiomyocytes following SM treatment were decreased in a dose-dependent manner. During anoxia, the ±dL/dt_max, dL and amplitude of [Ca²⁺]_i were decreased, while the diastolic Ca²⁺ level was elevated compared with control group. All the contractile parameters and the diastolic Ca²⁺ level were back toward pretreatment values during reoxygenation, but could not return to control level. After the treatment with SM (3 g/L), ±dL/dt_max and dL of cell contraction and the amplitude of [Ca²⁺]_i were higher and the diastolic Ca²⁺ level was lower than those in anoxia/reoxygenation group. CONCLUSION:SM antagonized effects of anoxia and reoxygenation on cell contraction and intracellular calcium in isolated ventricular myocytes.     

氮磷钾配施对丹参生长和药用成分的影响

以"川丹参1号"为试材,采用氮、磷、钾三因素四水平正交实验设计,研究在底肥中施入不同比例氮磷钾对丹参各部位生物量和主要药用成分的影响,以期找出适宜丹参生长的最佳氮磷钾配施比例。结果表明:氮肥能增加丹参根条数以及根鲜质量,但过量的施用氮肥会抑制丹参酮类的积累;磷肥对株高存在抑制作用;钾肥对根长生长存在抑制作用。综合产量和品质因素考虑,在该试验条件下,以N₃P₂K₂效果最好,在大田应用中,尿素、过磷酸钙、硫酸钾施肥参数分别为495、1260、300 kg·hm^-2。     

Effect of homocysteine on expression of interleukin-8 mRNA and protein in THP-1 macrophages

AIM: To investigate the effect of homocysteine (Hcy) on expression of interleukin-8 (IL-8) mRNA and protein in THP-1-derived macrophages (THP-1 macrophages). METHODS: Cultured THP-1 monocytes were induced to macrophages by 0.1 μmol/L PMA treatment for 72 hours, then the differentiated THP-1 macrophages were incubated with homocysteine (0.01 mmol/L-0.20 mmol/L) for 24 hours, or with 0.10 mmol/L Hcy for various time up to 48 hours. IL-8 protein in THP-1 supernatants was measured by ELISA, and IL-8 mRNA expression was detected by semiquantitive RT-PCR. RESULTS: Compared with control, Hcy significantly increased the expression of IL-8 protein in a concentration-dependent manner. 0.05 mmol/L, 0.10 mmol/L and 0.20 mmol/L Hcy increased IL-8 production by 1.28 fold, 1.32 fold and 1.55 fold, respectively (P＜0.01). IL-8 production were elevated significantly 3 h after treatment with 0.10 mmol/L Hcy. In addition, Hcy also increased IL-8 mRNA expression in a concentration-and time-dependent manner. CONCLUSION: Hcy may contribute to atherogenesis by inducing IL-8 expression and secretion in THP-1 macrophages.     

Effect of erigeron on intercellular adhesion molecule-1 and its mRNA expression during cerebral ischemia and reperfusion in rats

AIM: To investigate the effect of erigeron on intercellular adhesion molecule-1 (ICAM-1) and mRNA expression during cerebral ischemia/reperfusion. METHODS: The rat models of middle cerebral artery (MCA) focal cerebral ischemic reperfusion were established with the suture method in the study. The ICAM-1 mRNA and protein expression were measured by RT-PCR and immunohistochemistry techniques, respectively. RESULTS: By down-regulating the expression of ICAM-1 protein and mRNA and alleviating inflammation in cerebral ischemic region, erigeron exerted a protective effect in cerebral ischemia and reperfusion. CONCLUSION: The results suggest that erigeron protects the brain against cerebral ischemia and reperfusion injury via inhibiting ICAM-1 expressino.     

Effects of oxidized HDL on the levels of MCP-1, ICAM-1 and free calcium in cultured human umbilical venous endothelial cells

AIM:To study the effects of oxidized high-density lipoprotein (oxHDL) on the expression of monocyte chemoattractant protein-1(MCP-1) and intercellular adhesion molecule-1(ICAM-1) and intracellular free calcium concentration ([Ca²⁺]i) level in cultured human umbilical venous endothelial cells(HUVECs). METHODS:The MCP-1 protein content in the medium of conditioned HUVEC was measured by ELISA, and the ICAM-1 on HUVECs was detected by indirect immunofluorescence, and [Ca²⁺]i was determined by Fluo-3/AM, the injury of cells was observed by scanning electron microscopy (SEM).RESULTS:oxHDL could induce the expression of MCP-1 and ICAM-1 in HUVECs. In oxHDL group (HUVECs were incubated with 100 mg protein/L oxHDL for 24 h), the levels of MCP-1, ICAM-1 and [Ca²⁺]i increased by 160%, 60% and 70% respectively compared with the control group (P＜0.01). When HUVECs were incubated with 300 mg protein/L oxHDL for 24 h, cells were injured obviously. CONCLUSION:By inducing the expression of ICAM-1 and MCP-1 in endothelial cells, oxHDL may promote monocyte-endothelium adhesion and monocyte migration to intima, it may promote atherosclerosis as oxidized low-density lipoprotein (oxLDL).     

Effects of emodin on activity of sphingomylinase and content of ceramide in rabbit aorta of experimental atherosclerosis

AIM: To study the changes of the sphingomylinase activity and ceramide content in rabbit aorta of experimental atherosclerosis and investigate the effects of emodin on them. METHODS: The qualified rabbits were fed with food containing 1% cholesterol and 5% lard for 10 weeks to establish the animal models. The concentration of cholesterol (TC) was assayed by a enzyme method. Trace-fast-test method was used to test the activity of superoxide dismutase (SOD) and motified- BAMuGuoFu methods was employed to assay the content of myocardial malondialdehyde (MDA). Radiolabeled -enzyme-tracing was used to detect the activity of the sphingomyelinase,and thin-layered scanning was conducted to analyze the content of the ceramide in aorta. RESULTS: The ceramide content in aorta and the sphingomyelinase activity were markedly increased in the rabbits with experimental atherosclerosis. The increase was positively correlated with the content of TC and MDA and negatively correlated with the activity of SOD in blood. Compared to the model animals, emodin at concentration of 5 mg·kg^-1, 10 mg·kg^-1 and 20 mg·kg^-1 respectively reduced the area of plague on endothelium in rabbit's aortic artery and elevated the activity of SOD (P<0.05). The activity of sphingomylinase and the content of ceramide were decreased at the same time (P<0.05). 10 mg·kg^-1 emodin proved to be more effective than 5 mg·kg^-1 and 20 mg·kg^-1 emodin (P<0.01). CONCLUSIONS: Our data suggest that atherosclerosis is related to ceramide signal transduction initiated by factors such as oxidative stress in hypercholesterolemia. The emodin prevents the development of atherosclerosis probably by interfering with the above pathway.