Expression of human inducible nitric oxide synthase in fibroblast cell line V79 and effects of H4Bon iNOS activity

AIM: To explore the feasibility of human iNOS transfected into V₇₉ cells by gene transfer and investigate the effects of H₄B on iNOS activity. METHODS: Human iNOS was transfected into V₇₉ cells with the karyocyte expressive vector. The cloned cells were selected by G₄₁₈. The expression of iNOS mRNA was quantified by RT-PCR and iNOS expression was observed by immunofluorescence. NO product in cells was determined by measuring nitrite (NO^-₂) release using the Griess reaction. RESULTS: V₇₉ cells infected human iNOS was proved to have iNOS mRNA at 462 bp by RT-PCR, and iNOS protein in the cytochylema by immunofluorescence. When the cells were incubated without H₄B, the content of NO in pcDNA₃ cells was minimal, with NO^-₂ production (82.32±13.08) just above the normal group (74 38±9 80, P>0.05, n=6) There was no significant difference between pcDNA₃ cells incubated with or without H₄B, (P>0.05, n=6) NO^-₂ production by pcDNA₃-iNOS cells without H₄B was higher (105 58±13 33) (n=6, P<0.01vs the normal cells or pcDNA cells). However, in pcDNA₃-iNOS cells incubated with H₄B, NO^-₂ production was much higher (236 57±3183) (n=6, P<0.01vs the all former groups). CONCLUSION: iNOS activity was increased by adding H₄B in pcDNA₃-iNOS cells, and the fibroblast can be a target cell of iNOS gene transfer.     

Effects of nitric oxide on mitochondrial damage caused by exogenous calcium

AIM: To study the effects of nitric oxide (NO) on mitochondrial damage caused by exogenous calcium. METHODS: Normal myocardial mitochondria were divided into three groups; L-arginine control group (CG), Ca^2＋-damaged group (DG) and L-NAME-preserved group (PG). Mitochondria of all groups were incubated at 30 ℃ with reaction medium containing 20 μmol/L EDTA, 100 μmol/L CaCl₂ and 1 μmol/L L-NAME with 100 μmol/L CaCl₂ respectively. Then the NO₂^-/NO₃^- contents, mitochondrial viability and membrane potential were investigated. RESULTS: The NO₂^-/NO₃^- contents of DG was obviously higher than that of CG and PG, meanwhile, there was no obvious difference between CG and PG. Mitochondrial viability and membrane potential of DG were significantly lower than that of CG and PG, and negatively related to NO^-₂/NO^-₃ contents (r=-0.5297, P＜0.01; r=-0.6041, P＜0.01). But, the mitochondrial viability and membrane potential of PG were still lower than that of CG. CONCLUSION: Exogenous calcium could activate mitochondrial nitric oxide synthase resulting in NO production and the latter play an important role in decreasing mitochondrial viability and membrane potential.     

The kinetic alteration of nitric oxide formation in the lungs in the development of pulmonary fibrosis of rats

AIM: To observe the kinetic alteration of nitric oxide formation in the lungs in the development of pulmonary fibrosis in the rat. METHODS: The contents of hydroxyproline in the lungs, NO₂^-/NO₃^- (nitrite/nitrate) in out-flowing and in-flowing pulmonary blood (OPB, IPB) were assayed on the day 7, 14, 21, 30 and 70 after intratracheal administration of bleomycin A₅ . The content of NO₂^-/NO₃^- in supernatants of culture of the alveolar macrophages (AMs) and the amount of iNOS positive stain cells in lung tissue section were also observed in the rat on 14th day after-bleomycin A₅ administration. RESULTS: The content of lung hydroxyproline had no change on the 7th day, increased on the 14th day (P＜0.05), increased significantly on the 21th day, 30th day and 70th day post-bleomycin A₅ compared with control rats. On the 7th day and 14th day, the content of NO^- ₂ /NO₃^- increased in OPB and decreased in IPB (P＜0.01). On the 21th day, the content of NO₂^-/NO₃^- abated in OPB (P＞0.05) but still decreased in IPB (P＜0.01). On the 30th day and the 70th day, the NO₂^-/NO₃^- level recovered both in OPB and IPB. AMs from rats on the 14th day post-bleomycin A₅ showed significant elevation (P＜0.01) in NO₂^-/NO₃^- level. The amount of iNOS positive stain cells increased in rats on the 14th day post-bleomycin A₅. CONCLUSION: The amount of NO in the lungs was high in the initial phase of fibroproliferative reaction induced by bleomycin A₅ ,and these might be associated with the enhanced ability of AMs to release NO and the increased amount of iNOS.     

Effect of nitric oxide on proteinuria excretion in rats suffering from renal ischemia-reperfusion

AIM:To investigate the urinary protein (UP) excretion in rats suffering from renal ischemia-reperfusion (I-R) and effect of nitric oxide on it. METHODS:SD rats were used to establish the renal I-R model. Sodium nitroprusside (SNP), N^ω-nitro-L-arginine (L-NNA) and aminoguanidine (AG) were used to determine the effect of nitric oxide on UP excretion under renal I-R. Quantitative analysis of UP was made by chromatometry. UP species were separated by SDS-PAGE. RESULTS:Renal I-R caused significant increase in UP (P＜0.01). Compared with sham control, urinary proteins larger than albumin (66 kD) was found in I-R group. The increase in NO production after renal I-R was also observed and was closely correlated with the change of UP (r=0.704, P＜0.01). SNP increased UP excretion, AG and L-NNA decreased it in I-R rats. CONCLUSION:Renal I-R led to the increase in UP excretion. NO seems to play an important role in the UP excretion on rats suffering from renal I-R.     

Analysis of glomerular complement C5b-9 deposits and synthesis of NO,TNFα in the model of the rats with anti-thymocyte serum nephritis

AIM: To explore the localization and semi-quantification of the glomerular complement C5b-9 complexes and synthesis of some inflammatory mediators or cytokines such as nitric oxide (NO) and tumor necrosis factor α(TNFα) in the rats with anti-thymocyte serum nephritis(ATSN). METHODS: The animal model of rat ATSN was reproduced by a single intravenous injection of anti-thymocyte serum (ATS). Then, the deposits of glomerular C5b-9 complexes were localized and quantified by immunohistochemical staining and microscopic image scanning separately. And the glomerular mesangial cells (MC) surrounded by C5b-9 complexes were counted under microscope. In addition, the expression of glomerular MC inducible NO synthase(iNOS) mRNA and excretion of urinary NO metabolite (NO^-₂／NO^-₃) and TNF α in the rats with ATSN were detected. RESULTS: The MC in the rats with ATSN emerged necrosis followed by a rapid proliferation. In the early time of MC injury, the C5b-9 complexes were mainly seen in glomerular mesangium and MC surface. But with the progression of ATSN, the MC enclosed by C5b-9 appeared gradual decrease. Moreover, the expression of MC iNOS mRNA in early stage of ATSN obviously increased and the excretion of urinary NO^-₂／NO^-₃ and TNF α also significantly increased. However, the changes of parameters mentioned above in ATSN proliferative stage (after 7 days) alleviated gradually. CONCLUSION: The secondary lysis of MC has relation to the deposition of C5b-9 complexes and synthesis and release of NO and TNF α in rats with ATSN.     

The regulation of nitric-oxide synthase/nitric-oxide system by endogenous carbon monoxide in rats with pulmonary hypertension

AIM:To study the role and the mechanism of heme oxygenas/endogenous carbon monoxide on nitric oxide synthase/nitric oxide system in rats with pulmonary hypertension induced by hypoxic hypercapnia.METHODS:Sprague-Dawley rats were randomly divided into three groups: control group (A group), hypoxic hypercapnic group (B group), hypoxic hypercapnia+hemin group (C group). Blood CO concentration (COHb%), NO concentration, HO-1 activity, iNOS, cNOS in blood serum and lung homogenate were measured, respectively. RESULTS:① mPAP and RV/(LV+S) of B group were significantly higher than those of A and C group(P＜0.01).② Blood CO concentration, activity of HO-1in blood serum and lung homogenate in rats of B group were significantly higher than those of A group, but were significantly lower than those of C group (P＜0.01). ③ NO concentration in blood serum and lung homogenate in rats of B group were significantly lower than those of A group, those of C group were significantly higher than those of B group (P＜0.01).④The activity of iNOS in blood serum and lung homogenate in rats of B group were significantly higher than those of A group, but were significantly lower than those of C group (P＜0.01). Activity of cNOS in blood serum and lung homogenate of B group were significantly lower than those of A group (P＜0.01), and there was no significant difference between cNOS in B and C group.CONCLUSION:Endogenous carbon monoxide upregulated iNOS/NO system in rats with chronic pulmonary hypertension induced by hypoxic hypercapnia.     

The renal L-arginine/nitric oxide pathway and erythrocyte L-arginine transport in spontaneously hypertensive rats

AIM: To investigate the changes of the renal L-arginine /nitric oxide pathway and the relationship of L-arginine transport between kidney and erythrocytes in spontaneously hypertensive rat (SHR). METHODS: Sixteen week old SHR, 16 week old SHR with captopril (CAP) treated for four weeks and 16 week old WKY rats were used in the experiment. L-arginine transport, NO synthase(NOS) activity, nitrite and cyclic GMP (cGMP) content were measured in renal tissue or erythrocytes. RESULTS: In the renal tissue, compared with that of WKY group, the Vmax of high-or low-affinity L-arginine transporter, NOS activity, NO₂^- and cGMP content of SHR group were significantly decreased (P＜0.01 or P＜0.05). The Vmax of high-affinity L-arginine transporter and NOS activity of CAP group were significantly enhanced as compared with SHR group (+90%, P＜0.01; +58.6%, P＜0.05). The NOS activity had significant positive correlation with the Vmax of high-affinity L-arginine transporter (r=0.585, P＜0.05). The changes of erythrocyte L-arginine transport were the same as that of kidney. The Vmax of SHR group was lower than that of WKY group (-30%, P＜0.01), and the Vmax of CAP group was higher than that of SHR group (+26.5%, P＜0.01). Km was not significantly changed. There is a positive correlation between the Vmax of L-arginine transport in erythrocyte and the Vmax of high- or low-affinity L-arginine transporter in renal tissue, (r=0.8434, P＜0.01, high-affinity; r=0.5255, P＜0.05, low-affinity). CONCLUSION: There existed a functional inhibition in L-arginine/nitric oxide pathway in the kidney of SHR. It can be recovered obviously by captopril treatment. The changes of L-arginine transport in kidney coincide with that in erythrocyte.     

Effect of chronic hypoxia on L-Arginine/nitric oxide pathway in rat pulmonary artery

AIM: To study the effect of chronic hypoxia on L-Arginine/NO pathway in rat pulmonary artery. METHODS: Changes in pulmonary artery L-Arginine(L-Arg) transport, nitric oxide synthase (NOS) activity, plasma nitrite level and L-Arg level in HPH rats were investigated. RESULTS: (1) The mean pulmonary arterial pressure (mPAP) and weight ratio of right ventricle to left ventricle and septum (RV/LV+S) of HPH group were higher than those in control group (P＜0.01). (2) Plasma L-Arg level in HPH group was not significantly changed. (3) At low (0.2 mmol/L)or high(5.0 mmol/L)concentration of L-Arg, the velocity of L-Arg transport in HPH group was lower than that in control group (P＜0.05 or P＜0.01). (4) The activity of pulmonary artery tNOS, iNOS and cNOS in HPH group were increased by 38.0%, 32.8% and 53.0%, respectively (P＜0.01), compared with control group. (5) Plasma NO level of HPH group was decreased, which was negative correlation to mPAP and RV/LV+S (P＜0.01). CONCLUSION: The decrease of nitric oxide generation might result from L-Arg transport injury, while pulmonary artery tNOS, iNOS and cNOS activity were enhanced during chronic hypoxia.     

Role of renal cortex endothelin and NO in the development of proteinuria

AIM: To test the hypothesis that locally produced renal endothelin(ET) and nitric oxide(NO) play more important role in initiating proteinuria than circulating ET and NO. METHODS: (1) Rat nephrotic syndrome model was made by single injection of adriamycin, and the urinary protein excretion per 24 hours for every two days was measured. (2) Rat plasma and renal cortex were collected respectively 4 days, 8 days, 32 days and 56 days after the injection, and renal cortex homogenate was further prepared; ET of both plasma and renal cortex homogenate were determined by radioimmunoassay; NO of both plasma and renal homogenate were measured by the method of Griess. (3) The correlation between urinary protein and ET or NO concentration was analyzed respectively. RESULTS: (1) The urinary protein excretion of normal rat was 4-7 mg/d, that in nephrotic rat at the 4 time points was 5, 10, 241 and 201 mg/d, there was a significant increase of that at day 8, peaked at day 32, and less decrease at day 56. (2) The plasma ET in nephrotic rats of the 4 time points was 134, 150, 538 and 445 μg/L, respectively, which of day 8, day 32 and day 56 shown significant higher than that of normal control (126-129 μg/L). (3) The ET in renal cortex homogenate of nephrotic rat of the 4 time points was 364.6, 652.3, 1 526.8, 1 393.6 pg/g, all of that was much more higher than that of normal rat . (4) The plasma NO in nephrotic rat of the 4 time points was 40, 36, 8 and 11(μmol/L), there was considerable difference through day 8 to day 56 than control (42-46μmol/L, P＜0.05). (5) The NO in renal cortex homogenate of nephrotic rat was 80, 69, 8, 25 μmol/L that of normal rat was (114-124) μmol/L, there was significant difference between nephrotic rat and control from day 4 through day 56( P＜0.05). (6) The correlation coefficient of plasma ET and NO with separately urinary protein excretion shown statistic significance on only day 8 and day 56, while all of that of ET and NO in renal homogenate closely related with urinary protein from day 4 through to day 56. CONCLUSION: The ET and NO produced locally in renal play more important role in the development of proteinuria.     

Propofol inhibits activation of p38 MAPK in rat brain tissues with LPS-induced brain injury

AIM: To investigate the effects of propofol on lipopolysaccharide (LPS)-induced activation of p38 mitogen-activated protein kinase (p38 MAPK) and inducible nitric oxide synthase (iNOS) in brain tissues of rats. METHODS: Sprague-Dawley rats of both sexes were randomly divided into 3 groups (n=24 each): control group, LPS group and LPS+propofol group. The models of LPS-induced brain injury were established by injecting LPS (1 mg/kg) via left internal carotid artery in LPS group. Propofol (100 mg/kg) was given intraperitoneally immediately after the LPS was given in LPS+propofol group. The same volume of normal saline was given to the rats in control group. The rats were decapitated 6 h, 24 h, 48 h and 72 h after administration. The brains were immediately isolated to detect the water content, activation of p38 MAPK and the exepression of iNOS protein. Meanwhile, the pathological changes were observed under light microscope. RESULTS: The water content of the brain was higher in LPS group than that in control group. The protein levels of phosphorylated p38 MAPK(p-p38 MAPK) and iNOS in LPS group increased 6 h after LPS administration, reached the peak at 24 h, and still higher than those in control group at 48 h and 72 h (P<0.05). The levels of those indexes were all lower in LPS+propofol group at various time points than those in corresponding LPS group (P<0.05). The pathological changes were slighter than those in LPS group. The water content of the brain was positively correlated with the levels of p-p38 MAPK and iNOS (r=0.603, r=0.727,P<0.05). CONCLUSION: Propofol attenuates LPS-induced brain injury by inhibiting the activation of p38 MAPK and down-regulating iNOS expression.     

Immunohistochemical observation and imaging analysis of iNOS and vasoactive intestinal peptide in experimental diabetic rat lung

AIM: The pathological changes in inducible nitric oxide synthase (iNOS) and vasoactive intestinal peptide (VIP) in diabetic rat lung were investigated.METHODS: Using immunohistochemical method and imaging analysis, the changes in iNOS and VIP were observed in normal and diabetic rat lung. RESULTS: In diabetic rats, the iNOS positive or weak positive staining was localized in epithelial cells of bronchi, epithelial cells of alveolar ducts, alveolar sacs, and arteries, but negative in endothelial cells of vein and capillary. Image analysis showed the area, the integral absorbance(IA) and relative contents of iNOS were significant lower than that of control (P＜0.01, P＜0.01 and P＜0.01). In diabetic rats, the ciliated cells of bronchi epithelium showed VIP positive reaction, while the bronchial smooth muscle, surounding tracheobronchial submucosal glands, the tunica adventitia of pulmonary arteries, and alveolar septum were stained weak positive or negative for VIP. Image analysis showed the area, the IA and relative contents of VIP were significantly lower than those of control (P＜0.01, P＜0.01 and P＜0.01).CONCLUSION: The pathological changes in diabetic rat lungs are not only involving autonomic neuropathy but also NANC nerves. Nitric oxide and VIP, which are NANC neurotransmitter, may play an important role in pathogenesis of pathological change in diabetic lung.     

Influence of irbesarten on renal hypertrophy in streptozotocin-induced diabetic rats LIU Bi-cheng,LUO Dong-dong,ZHANG Xiao-liang.

AIM: To investigate the influence of irbesartan (Irb), a new angiotensin II receptor 1 antagonist, on renal hypertrophy in streptozotocin (STZ)-induced diabetic rats. METHODS: Sprague-Dawley (SD) rats were randomly divided into three groups: normal group (Group N, n=7), diabetic group (Group DN, n=6) and irbesartan treated group (Group DNI, n=7). In the experimental group, after the rats subjected to uninephrectomy, STZ was given by peritoneally injection at bolus dose of 50 mg/kg to induce diabetes. Blood glucose (BG), body weight (BW), urinary albumin excretion (Ualb), 24 hour proteinuria (24 h Upro) were measured at week 4, 8, 12, respectively. By the end of experiment at week 12, creatinine clearance (Ccr), kidney weight (KW), indicator of renal hypertrophy (KW/BW), renal total protein content (RTP), glomerular area (AG) and glomerular volume (VG) as well as glomerular basement membrane (GBM) were determined by semi-quantitative pathology technique. RESULTS: It was showed that there was no significant difference in BG between group DN and DNI, while Irb significantly reduced the increasing of Ualb, 24 h Upro in diabetic rats compared to control group (P＜0.01, respectively). Furthermore, Irb markedly inhibited the increasing of KW, KW/BW, RTP, A_G, V_G in diabetic rats (P＜0.05, P＜0.01, respectively). Of interest, Irb significantly prevented the increasing of GBM in diabetic rats. CONCLUSION: Irb exerts its early renal protective action by reducing proteinuria and inhibiting renal hypertrophy as well as the thickening of GBM.     

Inducible nitric oxide synthase and arterial pressure regulation

AIM and METHODS:To clarify the role of inducible nitric oxide synthase (iNOS) in the regulation of blood pressure,in the present study, we examined the effect of aminoguanidine (AG), a selective inhibitor of iNOS on the hemodinamical response of Dahl salt-sensitive (DS) and Dahl salt-resistant (DR) rats to low (0.3%) or high (8%) sodium chloride (NaCl) infusion by chronical in vivo hemodynamic experiment, and the effect of NaCl or NaCl plus AG infusion on urinary nitrate (NO₃)/nitrite (NO₂), the end product of nitric oxide (NO), excretion by Greiss Reaction. Furthermore, NOS activity assay was also carried out to probe the effect of NaCl and AG on calcium-dependent or independent NOS activity in renal tissue. RESULTS:1. High or low NaCl-infused DR rats and low NaCl-infused DS rats have no hemodinamical response to AG, however, the hypertensive effect of high NaCl (8%) infusion on DS rats were greatly amplified by co-infusion of AG. 2. Administration of high NaCl significantly elevated the iNOS activity of renal tissue, and greatly increased urinary NO₃/NO₂ excretion. CONCLUSION:Inducible NOS is an important modulator of arterial pressure, especially in case of higher blood pressure.     

Effect of amiodarone and metoprolol on platelet activation,fibrinolytic activity and vasoactive mediators in acute myocardial infarction in rabbits

AIM:To explore the significance of platelet activation, fibrinolytic activity and the changes of vasoactive mediators in acute myocardial infarction in rabbits and the intervention of amiodarone and metoprolol.METHODS:Fifty New Zealand white rabbits were randomly assigned to five groups, ten for each. Group Ⅰ: sham group, group Ⅱ: acute myocardial infarction(AMI) group, group Ⅲ: AMI and lidocaine group, group Ⅳ: AMI and amiodarone group, group Ⅴ: AMI and metoprolol group.The middle point of left ventricular coronary artery was ligated (groupⅡ,Ⅲ, Ⅳ and Ⅴ ) or a sham ligation(group Ⅰ). Four hours later, blood was collected for measuring plasma concentration of TXB₂, 6-Keto-PGF_1α, ET, NO, plasma activity of t-Pa and PAI.After that, the heart was taken out to evaluate the infarction size(IS).RESULTS:Plasma concentration of TXB₂, ET, NO and plasma activity of PAI were significantly higher in groupⅡ,Ⅲ, Ⅳ and Ⅴ than those in group Ⅰ(P＜0.01), but the plasma concentration of 6-Keto-PGF_1α and plasma activity of t-Pa were remarkably lower in groupⅡ,Ⅲ, Ⅳ and Ⅴ than those in group Ⅰ(P＜0.01). There were no difference in plasma concentration of TXB₂, 6-Keto-PGF_1α, t-Pa activity and infarction size in group Ⅱ,Ⅲ and Ⅳ(P＞0.05).Compared to group Ⅱ, plasma concentration of ET, NO and PAI activity were significantly decresed (P＜0.01)in group Ⅳ. Plasma concentration of TXB₂, ET, NO and plasma activity of PAI were significantly lower in groupⅤ than those in group Ⅱ(P＜0.01). Conversely, plasma concentration of 6-Keto-PGF1 and plasma activity of t-Pa were remarkably higher in group Ⅴ than those in group Ⅱ(P＜0.01). The infarction size was remarkly decrease(P＜0.01)in group Ⅴ.CONCLUSIONS:Amiodarone inhibited PAI avtivity, decreased release of ET and NO in AMI in rabbits. Metoprolol inhibited platelet activation, improved fibrinolytic, decreased release of ET and NO, and reduced myocardial infarction size in AMI in rabbits; Lidocaine had no effect above.     

Changes in serum TGF β1 in type 2 diabetic patients

AIM: To explore the changes in serum TGF β₁ in type 2 diabetes mellitus. METHODS: Forty-five cases type 2 diabetes mellitus patients were divided into three groups according to urine albumim excretion rate(UAER): normoalbuminuria(NA)group and microalbuminuria(MA) group and macroalbuminuria group (Overt DN). Serum TGF β₁, fasting blood glucose(FBG), HbA_1c,BUN,Cr,Ccr,lipidemia were detected in all cases. RESULTS: Serum TGF β₁ in NA, MA and ODN groups was higher than that in control. Serum TGF β₁ was positive correlation with Cr(r=0.390,P＜0.05), LDL(r=0.503,P＜0.01), HbA_1c (r=0.676,P＜0.01), and UARE(r=0.777,P＜0.01). CONCLUSION: Type 2 diabetes mellitus have a higher serum TGF β₁ than controls, serum TGF β₁ was positive correlation with HbA_1c and injury of renal function.     

Effect of PKC activition on cardiac myocyte apoptosis and Bcl- 2 expression during myocardial ischemia/reperfusion in rats

AIM: To explore the effect of PKC activition on cardiac myocyte apoptosis and expression of bcl- 2 during myocardial ischemia/reperfusion(I/R) in rats. METHODS: TUNEL,immunohistochemistry and in situ hybridization were used. RESULTS: The TUNEL data showed that the numbers of positive cardiac myocyte nucleus and the percentage of positive cardiac myocyte nucleus in PMA+IR_{3 h} group decreased significantly(P＜0.05,P＜0.01), compared to those in IR_3h group. The number of Bcl-2 protein positive cardiomyocytes and the percentage of Bcl-2 protein positive cardiomyocytes in PMA+IR_3h group were higher than those in IR_3h group (P＜0.01) bcl- 2 mRNA expression showed the same changes in PMA+IR_0h group compared to IR_1h group.CONCL USIONS:Activation of PKC decreased cardiomyocyte death during I/R.Upregulation of bcl-2 gene expression in cardiomyocytes during I/R may be one of the mechanisms of decreasing cardiomyocyte death by PCK activating during I/R.     

Role of endogenous nitric oxide in apoptosis of alveolar epithelial cells in the development of pulmonary fibrosis in rats

AIM: To study the role of high level of endogenous nitric oxide (NO) in apoptosis of alveolar epithelial cells in the development of pulmonary fibrosis in rats. METHODS: The content of nitrite/nitrate (NO₂^-/NO₃^-) in out-flowing pulmonary blood (OPB) was assayed by nitric acid reduction method. The apoptosis of alveolar epithelial cells was observed by TdT-mediated dUTP nick-end labeling (TUNEL) and electron microscopy, respectively. The above indices were observed on the day 14 and the day 30 after intratracheal administration of BLMA₅ alone or along with blockade of iNOS by aminoguanidine (AG) in rats. RESULTS: (1) Both the content of NO₂^-/NO₃^- in OPB and the number of apoptotic alveolar epithelial cells in lung were increased in BLMA₅ 14 d group, compared with normal control group and BLMA₅ 30 d group, respectively (P<0.05). The high level of NO₂^-/NO₃^- in OPB and the apoptosis of alveolar epithelial cells were ameliorated by AG. CONCLUSION: The apoptosis of alveolar epithelial cell is induced by high level of endogenous NO in the development of pulmonary fibrosis.     

Effect of mycobacterium bovis bacillus calmette-guerin vaccination on arterial pressure in hypertensive rats

AIM: The present study was designed to examine the effect of Bacillus Calmette-Guerin (BCG) vaccination on blood pressure, nitric oxide (NO) production and iNOS expression in hypertensive rats. METHODS: Renal hypertension (RH) were made by renal artery stenosis in SD rats and the sodium induced hypertensive (SH) rats were made by feeding the rats with high sodium chloride diet (5 g NaCl/100 g food). After four weeks, the hypertensive animals were subjected to the experiment. All the rats were vaccinated with BCG (0.1 mL, i.d) and blood pressure were examined every week. Greiss reaction was used to measure the urinary NO excretion and Western blot was applied to probe the iNOS protein expression in aortic tissue. RESULTS: It was shown that one week after BCG vaccination, the blood pressure decreased significantly in hypertensive rats induced by NaCl-overloading and renal artery stenosis, but not in normotensive control rats. Furthermore, the hypotensive effect of BCG vaccination was enhanced by co-administration of L-arginine. A significant increase in NO production was observed in hypertensive rats. Also, Western blot showed BCG vaccination led to an obvious increase in iNOS expression in the aortic tissue of hypertensive, but not of normal control rats. CONCLUSION:BCG vaccination could lower the blood pressure of hypertensive rats through activation of iNOS/NO pathway.     

Effects of Aconiti tuber polysaccharides on hypercholesterolemia and hepatic cholesterol 7α-hydroxylase expression in rats

AIM: To investigate the effects of polysaccharides isolated from Aconiti tuber (Fuzi polysaccharides,FPS) on the prevention of hypercholesterolemia induced by high-cholesterol diet and the expression of hepatic cholesterol 7α-hydroxylase (cytochrome P450 7α-1, CYP7α-1) in rats.METHODS: Fifty male Wistar rats were randomly divided into 3 groups. The rats were fed with normal diet (control group), high-cholesterol diet (HC group) or high-cholesterol diet plus FPS (224, 448 or 896 mg·kg^-1·d^-1, FPS group) for 2 weeks. The serum lipid level, body weight, food-intake and fecal amount were measured at week 2. The pathological changes of the liver were observed with HE staining. The mRNA expression of hydroxy methylglutaryl coenzyme A (HMG-CoA) reductase and CYP7α-1, the protein level of CYP7α-1, and fecal bile acid were also detected at week 2.RESULTS: FPS significantly inhibited high-cholesterol diet-induced elevation of serum total cholesterol (TC) and low-density lipoprotein cholesterol (LDL-C) (P<0.05). HE staining showed that FPS attenuated fatty degeneration in liver. Real-time PCR analysis showed that FPS significantly up-regulated the mRNA expression of CYP7α-1, but down-regulated the mRNA expression of HMG-CoA reductase (P<0.01). The protein level of CYP7α-1 was higher in FPS group than that in HC group (P<0.01). The level of fecal bile acid in HC-treated rats was higher than that in the control rats, and FPS stimulated the excretion of fecal bile acid (P<0.05).CONCLUSION: FPS significantly reduces serum cholesterol levels, which is associated with the up-regulation of hepatic CYP7α-1 expression and down-regulation of hepatic HMG-CoA reductase expression.