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1.
AIM: To study the expression of glycine receptor α1 subunit in neonatal rat myocardial cells and to investigate the effect of lipopolysaccharide (LPS), hypoxia/reoxygenation, isoproterenol (ISO) and high concentration of glucose (HG) on the expression of glycine receptor α1 subunit in the neonatal rat myocardial cells. METHODS: Neonatal rat myocardial cells were cultured in vitro. The expression of glycine receptor α1 subunit was detected by Western blotting. The neonatal rat myocardial cells were treated with LPS (20 mg/L), ISO (100μmol/L) or high concentration of glucose (25 mmol/L) for 24 h, or were exposed to hypoxia for 3 h followed by reoxygenation for 3 h. Subsequently, the cell viability was measured by CCK-8 assay, and the expression of glycine receptor α1 subunit was determined by Western blotting. RESULTS: The expression of glycine receptor α1 subunit in the neonatal rat myocardial cells was positively detectable by Western blotting. Compared with control group, no significant difference of the cell viability (P>0.05) in LPS group, ISO group, hypoxia/reoxygenation group and HG group was observed. The expression of glycine receptor α1 subunit was increased (P<0.01) in LPS group, ISO group and hypoxia/reoxygenatio group, but decreased (P<0.01) in HG group. CONCLUSION: Glycine receptor α1 subunit exists in the neonatal rat myocardial cells. A certain concentration of LPS or ISO, or hypoxia/reoxygenation for a certain period upregulate the expression of glycine receptor α1 subunit, but HG downregulates the expression of glycine receptor α1 subunit in cultured neonatal rat myocardial cells.  相似文献   

2.
AIM: To determine the beneficial effects of icariin on streptozotocin (STZ)-induced diabetic testopathy in rats. METHODS: The diabetic animal model was induced in male Sprague-Dawley rats by an injection of streptozotocin (40 mg/kg, iv). The rats were randomly divided into 3 groups: control group, model group and icariin (80 mg/kg, ig) group. Twelve weeks after injected with streptozotocin, all rats were anaesthetized and killed to remove the testes from scrotum. Serum concentrations of glucose and testosterone, and the levels of succinate dehydrogenase (SDH), acid phosphatase (ACP), γ-glutamyl transpeptidase (γ-GT) and lactate dehydrogenase (LDH) in testes were measured. The morphology of the testicular tissues was observed under light microscope. Immunohistochemistry was employed to determine the protein levels of TGF-β1 and type Ⅳ collagen. RESULTS: Compared with control group, the content of serum glucose increased while the serum level of testosterone and the activitiy of SDH, ACP, γ-GT and LDH in testis decreased in model group (P<0.01). The histopathological examination showed that the diameters of seminiferous tubules and various grades of spermatocytes in the testis were markedly decreased. Compared with control group, the expression of TGF-β1 and collagen Ⅳ was significantly increased in model group. These alterations were significantly attenuated in icariin group (P<0.01). CONCLUSION: Icariin evidently relieves testicular damage in rats with diabetic testopathy by improving the secretion of testosterone and reducing the expression of TGF-β1 and collagen Ⅳ at protein level.  相似文献   

3.
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.  相似文献   

4.
AIM:To investigate the effect of tenuigenin(TEN) on hyperphosphorylation of tau protein in neurons of amyloid β-peptide1-40(Aβ1-40) -induced Alzheimer disease(AD) rats. METHODS:Aβ1-40 was injected into hippocampus CA1 region of the rats to establish AD model. TEN at different doses(18.5 mg/kg, 37.0 mg/kg and 74.0 mg/kg) was intragastrically administered. The protein expression of protein kinase A(PKA),protein phosphatase 2A(PP2A), total tau and p-tau(Ser396) in the neurons was observed by the method of immunohistochemistry. The protein content of total tau and p-tau(Ser396), and the expression level of PKA and PP-2A were detected by Western blotting analysis. RESULTS:In Aβ1-40 group, the level of total tau, the phosphorylation of tau protein and the expression of PKA were significantly increased compared with those in sham operation group. Meanwhile, the expression of PP2A in Aβ1-40 group was lower than that in sham operation group. In TEN treatment group, the level of total tau, the phosphorylation of tau protein and the expression of PKA were markedly decreased, and the expression of PP2A was increased as compared with Aβ1-40 group. CONCLUSION:TEN may protect the neurons from the toxic effect of Aβ1-40 and reduce the hyperphosphorylation of tau(Ser396) in the neurons of AD rats by activating the expression of PP2A and inhibiting the expression of PKA.  相似文献   

5.
AIM:To study the possible molecular mechanism of beta-amyloid peptide1-40-induced apoptosis in rat cortical neurons.METHODS:40 mg/L beta-amyloid peptide1-40 (Aβ1-40) was used to induce apoptosis in cultured rat cortical neurons. The level of CDK4, phosphorylated pRB were detected by flow cytometry and immunoblotting; RT-PCR was used to examine the mRNA expression of E2F1 while fluorescent spectrofluorometer was used to measure caspase-3 activity. All of the above study was designed to observe whether the level of CDK4, phosphorylated pRB and E2F1 mRNA expression could be affected by Aβ1-40.RESULTS:(1)The level of CDK4, phosphorylated pRB increased markedly 2-4 hours after treatment with Aβ1-40, and caspase-3 activity elevated remarkably 12-24 hours after treatment with Aβ1-40; (2) E2F1 mRNA expression was upregulated 3 hours after incubation with Aβ1-40.CONCLUSION:Aβ1-40 may induce apoptosis in rat cortical neurons in a manner dependent on CDK4-pRB-E2F1 pathway.  相似文献   

6.
AIM: To explore the influence of angiotensin-(1-7) on angiotension II (Ang II)-induced activation and extracellular matrix secretion in rat renal interstitial fibroblasts (NRK-49F cells). METHODS: The NRK-49F cells were maintained and sub-cultured, then the cells were divided into control group, Ang II group, Ang-(1-7) group and Ang II+Ang-(1-7) group. The expression of α-smooth muscle actin(α-SMA),transforming growth factor β1 (TGF-β1) and insulin-like growth factor I(IGF-I) was detected by the method of immunocytochemistry when the cells were cultured for 72 h. The content of TGF-β1, IGF-I and collagen type I(Col I) in the cultured supernatants were measured by ELISA. RESULTS: In control group and Ang-(1-7) group, only basic expression of α-SMA and almost no expression of TGF-β1, IGF-I and Col I were observed. Compared with control group, the expression of α-SMA, TGF-β1, IGF-I and Col I was increased in Ang II group. Compared with Ang II group, the expression of α-SMA, TGF-β1, IGF-I and Col I was significantly decreased in Ang II+Ang-(1-7) group.CONCLUSION: Ang-(1-7) inhibits the activation of renal interstitial fibroblasts and decreases the Ang II induced secretion of Col I by suppressing TGF-β1 and IGF-I expression.  相似文献   

7.
AIM: To investigate the effect of platelet-derived growth factor (PDGF) on β3 integrin gene expression and the role of β3 integrin on adhesion, migration and proliferation of vascular smooth muscle cells (VSMC) induced by PDGF. METHODS: β3 integxin gene expression was detected by RT-PCr. After β3 integrin extracellular do-main was blocks, VSMC adhesio, migration and proliferation were measured by adhesion assay awound-culture model an [3H]-TSR incorporation respectively.RESULTS: After the interaction between β3 integrin and extracellular matrix was blocked, VSMC proliferation was inhibited in some degree and the rate of [3H]-TdR incorporation into VSMC decreased 39%. The cell adhesion and migration were significantly inhibited when 10 mg/L anti-β3 integrin antibody was added (P＜0.05). When VSMC were treated by PDGF for 6 hours, the expression of β3 integrin gene was 87% higher than that of control. CONCLUSION: PDGF significantly induces expression of β3 integrin gene in VSMC, and the interaction between β3 integrin and ECM protein may play an important role in VSMC adhesion and migration.  相似文献   

8.
AIM: To investigate whether chrysophanol alleviates amyloid β-protein (Aβ)-induced cognitive dysfunction and the underlying antioxidative mechanism.METHODS:Adult male Wistar rats (230~250 g) were randomly divided into control group, Aβ1-42 group, chrysophanol group, and Aβ1-42+chrysophanol (1, 10 and 100 mg/kg) groups. Aβ1-42 was delivered by intracerebroventricular injection under the guidence of a brain stereotaxic apparatus. Y-maze test, open-field test and Morris water maze test were performed 1 week after Aβ1-42 injection to evaluate the ability of rat spacial learning and memory. Chrysophanol was intraperitoneally injected once daily for 5 consecutive days. After the behavioral tests, the animals were sacrificed immediately by decapitation, and the hippocampus were collected. The malondialdehyde (MDA) content and the activity of superoxide dismutase (SOD), glutathione peroxidase (GSH-Px) and catalase (CAT) in the hippocampus were measured.RESULTS:Multiple (7 consecutive days, once daily) but not single (once a day) chrysophanol treatment at 1, 10 and 100 mg/kg effectively prevented Aβ1-42-induced cognitive function deficits in a dose-dependent manner as shown by Y-maze test and Morris water maze test. Moreover, the Aβ1-42-induced increase in MDA content and decrease in the activity of antioxidant enzymes (SOD, GSH-Px and CAT) in the hippocampus of the rats were also attenuated by multiple chrysophanol treatment.CONCLUSION:Repeated chrysophanol treatment attenuates Aβ1-42-induced cognitive deficits and synaptic plasticity dysfunction, and the mechanisms underlying the neuroprotective effects are likely due to its antioxidant activity.  相似文献   

9.
AIM:To study the effect of Bene Jones protein (BJP) from multiple myeloma(MM) patient and TGF-β1 on cultured renal proximal tubular cell(PTC) proliferation.METHODS:[H3]TdR incorporation was used to study the effect of λBJP and TGF-β1 on cultured rat NRK.52E PTC proliferation, the expression of TGF-β1 in the supernatant of PTC cultured with BJP was assessed with ELISA.RESULTS:① [H3]TdR incorporation of PTC was inhibited by BJP in a dose-dependent manner, when co-cultured with 100-800 μmol/L BJP and 2.0 μg/L TGF-β1, the [H3]TdR incorporation was lower than that of BJP alone, especially when BJP≥400 μmol/L;②The expression of TGF-β1 in the supernatant of PTC cultured with BJP was increased, especially when BJP≥400 μmol/L(P＜0.05);③ The [H3]TdR incorporation of PTC was also inhibited by exogenous TGF-β1 in a dose-dependent manner.CONCLUSION:λBJP has antiproliferative effect on rat PTC in vitro, The effect is related with stimulating the PTC to produce excessive TGF-β1, which also has antiproliferative effect on PTC in some degree.  相似文献   

10.
AIM: To investigate the effect of Thymosin α1 on the development and matutation of thymocytes. METHODS: The proportion of CD4+CD8+ thymocytes and the expression of smoothened (Smo) of the hedgehog (Hh)-signaling in CD4-CD8-thymocytes were examined to observe the effect of thymosin α1 on thymocyte development and matutation. RESULTS: Flowcytometric analysis showed that thymosin α1 showed activity at a low dose of 30 μg/kg, and 30 μg/kg thymosin α1 accelerated the replenishment and maturation of thymocytes according to the expression of Smo of the Hh-signaling in CD4-CD8-thymocytes, the potent negative regulator of proliferative responses. CONCLUSION: Thymosin α1 can accelerates thymocyte development from CD4-CD8- to CD4+CD8+.  相似文献   

11.
AIM: To investigate whether transforming growth factor-β1 (TGF-β1) participates in hypoxia-induced bronchial epithelial-mesenchymal transition (EMT) through lysyl oxidase (LOX). METHODS: Sprague-Dawley (SD) rats were exposed to hypoxia to establish the animal model and were treated with LOX inhibitor β-aminopropionitrile (β-APN). Furthermore, primary rat bronchial epithelial cells were cultured in vitro and exposed either to normoxia or to hypoxia. TGF-β1, TGF-β1 receptor inhibitor (SB431542) or β-APN was used in the cell experiments. The content of collagen was measured by colorimetric method. The expression of TGF-β1, LOX, and 2 EMT-related proteins (namely, the epithelial marker E-cadherin and the mesenchymal marker vimentin) were determined by immunohistochemistry and We-stern blot, respectively. RESULTS: The expression of TGF-β1, vimentin and LOX and cross-linking of collagen were enhanced in hypoxia-exposed rat and in hypoxia-exposed bronchial epithelial cells, but the enhancement was impaired by the treatment with β-APN. In contrast, the expression of E-cadherin was reduced in hypoxia-exposed rat, and was reversed by treatment with β-APN. In vitro experiments demonstrated that TGF-β1 and hypoxia led to the morphological phenotype characteristic of EMT in rat bronchial epithelial cells, in which the morphology of rat bronchial epithelial cells was switched from cobble-stone shape in normoxia-exposed group to spindle fibroblast-like morphology in hypoxia-or TGF-β1-exposed group (P<0.01). Additionally, both β-APN and SB431542 partially prevented TGF-β1 and hypoxia induced EMT in rat bronchial epithelial cells. TGF-β1was able to dose-dependently up-regulate LOX expression in rat bronchial epithelial cells, which was blocked by concurrent incubation with SB431542. The up-regulation of TGF-β1, vimentin, LOX and cross-linking of collagen and down-regulation of E-cadherin in hypoxia-exposed rat bronchial epithelial cells was significantly reversed by incubation with SB431542. CONCLUSION: TGF-β1 regulates hypoxia-induced EMT in bronchial epithelial cells via activation of the LOX.  相似文献   

12.
AIM: To study the correlation between phasic change of the relative quantity of major crystallins with aging in rats. METHODS: Six groups of SD rats (age 1 d， 8 d，2 weeks，8 weeks，8 months and 1.5 years) were raised routinely. Water soluble crystallins were extracted and separated by two－dimensional polyacrylamide gelelectrophoresis. After comassize blue staining，the crystallins patterns were scanned and analyzed. RESULTS: (1) Out of the eighteen water soluble major rat crystallins tested in each group, seven showed gradual phasic changes in relative quantity of crystallins, but there were no significant changes in total quantity of water soluble crystallins. (2) Phasic changes in these crystallins presented four different patterns: increasing (βB4、αB2、αA2、βA1), decreasing (β7、β8、γ2,3、γ5,6),relatively stable(βA3、βB5), and irregular. (3) The ratio of βB4 /αA2 increased gradually with the rat aging process. CONCLUSION: The gradual phasic changes in relative quantity of crystallins reflect the aging status of rat crystalline.  相似文献   

13.
AIM: To investigate the role of transforming growth factor β1 (TGF-β1)/Smads and extracellular signal-regulated kinase(ERK) expression in vascular remodeling induced by high-salt diet in Wistar rats. METHODS: Wistar rats were randomly divided into 3 groups: normal control group (n=13), high salt (8%) model group and high salt+telmisartan group (n=13). Tail-cuff arterial pressure was determined every 2 weeks. After 24 weeks, the rats in high salt model group were divided into model animals with hypertension group (MH, n=12) and model animals without hypertension group (MN, n=12). The remodeling of aorta and mesenteric artery was observed by HE and Masson staining. In addition, the techniques of immunohistochemistry and real-time PCR were applied to detect the expression of proliferating cell nuclear antigen (PCNA), TGF-β1, p-Smad2/3, p-ERK1/2 and Smad7 at both protein and mRNA levels. RESULTS: Compared with normal control group, blood pressure in MH group was much higher, and media thickness (MT) and collagen volume fraction (CVF) of arteries in MH and MN groups were higher.The mRNA expression of TGF-β1, Smad2 and Smad7 in the aorta was significantly increased, and the protein levels of PCNA, p-ERK1/2, TGF-β1 and p-Smad2/3 in the aorta and mesenteric artery media were elevated, but Smad7 decreased. After telmisartan treatment, MT and CVF were much lower,and the protein levels of PCNA, TGF-β1, p-Smad2/3 and p-ERK1/2 were significantly reduced, whereas Smad7 was increased. CONCLUSION: The abnormal expression of TGF-β1/Smads and ERK may be involved in the mechanism of remodeling of aorta and mesenteric artery induced by high-salt diet. Telmisartan prevents the vascular remodeling via regulating TGF-β1/Smads and ERK signal pathways mediated by angiotensinⅡ type 1 (AT1) receptor, at least in part.  相似文献   

14.
AIM: To observe the expression of transforming growth factor β1 (TGF-β1), MAPK1/3 and fibronectin (FN) in the development of renal tubulointerstitial disease. METHODS: Wistar male rats were randomly divided into normal control group, diabetic group of 1week, 2 weeks, 4 weeks and 8 weeks. Diabetic model was induced by peritoneal injection of streptozotocin. Immunohistochemistry was employed to detect the expression of TGF-β1, MAPK1/3 and FN in the kidney. TGF-β1 protein in the renal cortex was checked by Western blot. BG, Scr and UP were analysed by biochemical methods, and the morphological changes in renal tubulointerstitium were also examined under microscopy on sections stained with HE and PAS. RESULTS: The expression of MAPK1/3 and FN was observed, but not the expression of TGF-β1 in normal renal tissue. Positive staining of TGF-β1 was observed in the renal tubulo-interstitium in 1-week diabetic group and thereafter it increased in the course of diabetes. A continuous increase in the expression of MAPK1/3 and FN was also observed in two - week diabetic rats. Chronologically the expression of TGF-β1,MAPK1/3 and FN and the ratio of KW/BW were positively correlative with each other in diabetic animals except one -week diabetic rats. There was also a positive correlation between MAPK1/3 and FN in l -week diabetic rats. CONCLUSION: Our data suggest that TGF-β1 appears in the renal tubulointerstitium in early period of diabetes and then its signal is mediated by MAPK1/3 cascades to accelerate production of FN ,and in turn leads to renal hypertrophy and tubulointerstitial fibrosis.  相似文献   

15.
AIM: To study the inhibitory effect of metformin on alveolar epithelial-mesenchymal transition (EMT) in rats with pulmonary fibrosis and the possible mechanism. METHODS: SD rats (n=48) were used, 12 of which were set up as normal control group, and 36 of which were induced by bleomycin (5 mg/kg) by tracheal instillation to establish pulmonary fibrosis. The pulmonary fibrosis rats were randomly divided into bleomycin group, low dose (100 mg/kg) of metformin group, and high dose (300 mg/kg) of metformin group. The rats in metformin groups were given the corresponding dose of metformin daily for 4 weeks. HE staining and Masson staining were used to observe the changes of lung histopathology and collagen deposition. Real-time PCR, Western blot and innunohistochemical staining were used to detect the mRNA and protein expression of α-smooth muscle actin (α-SMA), E-cadherin, vimentin, zonula occludens-1 (ZO-1), collagen I, collagen III and transforming growth factor-β1 (TGF-β1), and the protein phosphorylation levels of Smad2/3 and extracellular signal-regulated kinase 1/2 (ERK1/2) were also determined. RESULTS: Metformin up-regulated the expression of E-cadherin and ZO-1, down-regulated the expression of α-SMA, vimentin, collagen I and collagen III, and the protein phosphorylation levels of Smad2/3 and ERK1/2 were also decreased (P<0.05). CONCLUSION: Metformin inhibits alveolar EMT in the rats with pulmonary fibrosis, and its mechanism may be related to the inhibition of TGF-β1 signal transduction pathway.  相似文献   

16.
AIM:To investigate the expression of soluble guanylate cyclase protein and its mRNA in rat pulmonary artery after exposure to hypoxia and hypercapnia.METHODS:Male Sprague-Dawley rats were randomly split into 4 group, which were hypoxic hypercapnic (HH 1 week, HH 2 weeks, HH 4 weeks) group and control group, to copy pulmonary hypertensive animal model. The expression of sGCα1 and β1 subunits protein of medial and small pulmonary artery was performed by immunohistochemistry with a polycolonal antibody. In situ hybridization was performed on the rat lung tissue using sGC oligonuclear probe to assay the expression of sGCα1subunit mRNA.RESULTS:The sGCα1 and β1 subunits protein and sGCα1 subunit mRNA were faint staining in the pulmonary small and medium artery in HH1 week, HH 2 weeks and HH 4 weeks groups compared with control group (all P＜0.01).CONCLUSION:sGC subunit mRNA and protein expression in pulmonary small and medium artery were decreased after exposure to hypoxia and hypercapnia, which took part in the development of the pulmonary hypertension.  相似文献   

17.
AIM: To investigate the effects of 1,25-dihydroxyvitamin D3 on the proliferation of passively-sensitized human airway smooth muscle cells (HASMCs), and to explore its potential role in asthmatic airway remodeling.METHODS: HASMCs were passively sensitized with 10% serum from asthmatic patients.1,25-(OH)2D3 was used as the interventor.The effect of 1,25-(OH)2D3 on the cell proliferation and its optimal concentration were determined by MTT colorimetric assay.The cell cycle analysis was performed by flow cytometry.The expression of proliferating cell nuclear antigen (PCNA) was measured by the method of immunocytochemical staining.RESULTS: 1,25-(OH)2D3 at the concentrations of 10-9-10-7 mol/L markedly inhibited the cell proliferation and the maximum effect was observed at the concentration of 10-7 mol/L.This concentration of 1,25-(OH)2D3 markedly suppressed the PCNA-positive rate and hampered the G1/S transition in HASMCs passively-sensitized by asthmatic serum.CONCLUSION: 1,25-(OH)2D3 has direct inhibitory effects on the proliferation of passively-sensitized HASMCs in vitro, which may be concerned with the beneficial role of 1,25-(OH)2D3 on the prevention and therapy of asthmatic airway remodeling.  相似文献   

18.
AIM: To study the up-regulation of inducible nitric oxide synthase (iNOS) in lung of pulmonary fibrosis and its relationship with fibrosis. METHODS: The changes of amount of iNOS positive stain cells and type Ⅰ?Ⅲ collagen were examined on the day 7, 14 and 30 after intratracheal administration of bleomycin A5. The contents of NO2-/NO3- (nitrite/nitrate) in out-flowing pulmonary blood (OPB), hydroxyproline in lung and the histological changes were detected after iNOS was blocked by aminoguanidine (AG). RESULTS: (1) The number of iNOS-positive stain cells increased significantly in BLMA5 7 d, 14 d and 30 d groups compared with that in control group (P<0.01). Furthermore, the increment of the number of iNOS-positive stain cells in BLMA5 7 d, 14 d groups was more than that in BLMA5 30 d group. There was an increment of collagen in BLMA5 14 d group and in BLMA5 30 d group , with an increase in type Ⅲ collagen in BLMA5 14 d group and an increase in type Ⅰcollagen in BLMA5 30 d group. (2) The high level of NO2-/NO3- in OPB and hydroxyproline level in lung could be reversed by AG, a selective inhibitor of iNOS. Large amount of fibroblasts and macrophages were also abated by AG. CONCLUSION: In the development of pulmonary fibrosis, the expression of iNOS is up-regulated, which induces nitric oxide (NO) production and promotes propagation of pulmonary fibrosis.  相似文献   

19.
AIM:To study the effect of human complement C5b~9 complex on nitric oxide(NO) synthesis of glomerular mesangial cells (MC). METHODS: First, the human complement C5b~9 complexes were isolated and glomerular MC of rats were cultured. Second, the MC were stimulated with C5b~9 complex and changes of metabolism products of NO(NO3- and NO2) in MC culture supernatant at 6,24 and 48 hours after C+5b~9 stimulating were detected. Moreover, cGMP levels in cultured MC were also measured. RESULTS: NO3-／NO2- contents from culture supernatant and cGMP levels in MC were increased parallelly after C5b~9 complex stimulation. Further, NO synthesis was inhibited by L-NG-nitro-arginine-methylester(L-NAME). CONCLUSION: NO synthesis of rat glomerular MC was incerased by human complement C5b~9 stimulation.  相似文献   

20.
AIM: To investigate the effects of curcumin (Cur) on the expression of High mobility group box 1 protein (HMGB1), interleukin-1β (IL-1β), tumor necrosis factor-α (TNF-α) in amyloid-β (Aβ)-induced primary rat microglial cells. METHODS: Microglia were derived from the cerebral cortices of postnatal rat brains. The cells were identified by immunocytochemistry using mouse anti rat Iba-1 monoclonal antibody. A cell model using primary rat microglial cells incubated with Aβ25-35 as an inflammation model of Alzheimer's disease (AD) was set up. The morphological characters of primary rat microglial cells were observed. The concentration of Aβ25-35 and the treatment concentration of curcumin were selected by CCK-8 assay. Cultured primary rat microglial cells were divided into 5 groups:normal cell group, Aβ25-35 group, Cur group, Aβ25-35+Cur group and Aβ25-35+DMSO group. The expression of HMGB1, NF-κB, and receptor for advanced glycation end products (RAGE) was detected by Western blot. The levels of HMGB1, IL-1β, and TNF-α in the culture supernatant were measured by ELISA. RESULTS: The purity of primary microglias determined by Iba-1 immunofluorescence was more than 95%. The protein levels of HMGB1, RAGE and NF-κB were significantly increased after Aβ25-35 stimulation. After treatment with Cur, the protein levels of HMGB1, RAGE and NF-κB were significantly decreased (P<0.05). The levels of HMGB1, IL-1β and TNF-α in the supernatant were significantly increased after Aβ25-35 stimulation. Cur significantly decreased the level of HMGB1, IL-1β and TNF-α in the supernatant. CONCLUSION: Curcumin significantly inhibits neuroinflammation stimulated by Aβ25-35 in primary rat microglial cells.  相似文献   

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