Effect of apigenin on expression of VEGF in cerebral ischemia and reperfusion rats

AIM: To investigate the effect of apigenin on the expression of vascular endothelial growth factor (VEGF) in the rats under the condition of cerebral ischemia and reperfusion. METHODS: Ninety-one male SD rats were randomly divided into 13 groups: sham operation group (S), model groups (group M6 h, group M24 h, group M72 h, group M7 d), apigenin treatment groups (group A6 h, group A24 h, group A72 h, group A7 d) and dexamethasone treatment groups (group D6 h, group D24 h, group D72 h, group D7 d). The acute transient focal cerebral ischemia reperfusion model was established by modified method of inserting the nylon thread into middle cerebral artery, staying for 2 h and then withdrawing from the artery. In the experiment groups, the TTC staining of brain slices were performed and the neurological behavior scores were determined. The expression of VEGF by immunohistochemistry (ICH) was semi-quantitatively analyzed by measuring the integrated absorbance(IA). RESULTS: Abnormal neurological behaviors were observed in the animals of M groups, A groups and D groups, but the neurological behaviors of the rats in A7 d group were better than that in the other groups (P<0.05). Typical cortical infarct lesions in M groups, A groups and D groups were found by TTC staining, mainly in cerebral cortex and striatum. The immunnohistochemical results showed that the expression of VEGF was significantly higher in M, A and D groups than that in S group (P<0.05). Moreover,the expression of VEGF in A groups(A24 h and A72 h)was higher than that in M groups (M 24 h and M72 h,respectively)(P<0.01).The expression of VEGF in D72 h group was higher than that in M72 h group (P<0.05), and that in A7 d group was obviously higher than that in D7 d group (P<0.01).CONCLUSION: Apigenin promotes the expression of VEGF in the model of acute transient focal cerebral ischemia-reperfusion injury, improves the process of brain injury and recovers the brain functions in rats.     

Expression of aquaporin-4 in the rat contralateral hemisphere parieto-temporal cortex after intracerebral hemorrhage

AIM: To study the expression of aquaporin-4 (AQP-4) in the rat contralateral hemisphere parieto-temporal cortex after intracerebral hemorrhage. METHODS: The intracerebral hemorrhage (ICH) was established by infusing autoblood into rat caudate nucleus. MRI T2 weight and pathological observation were used to confirm edema. RT-PCR and Western blotting were used to evaluate AQP-4 mRNA and protein expression, respectively. Electron microscope was used to explore aquaporin-4 changes in contralateral hemisphere. RESULTS: As compared with the sham group, significant increase in AQP-4 mRNA and protein expression appeared on the double hemishere in rats following ICH. At 24 h, the expression of AQP-4 mRNA and protein increased in double hemishere. The expression of AQP-4 mRNA and protein reached its maximum at 72 h and decreased 3 d and 7 d after ICH, the expression of AQP-4 mRNA and protein in intracerebral hemorrhagic group were still higher than that in the sham group. Compared to ipsilateral hemisphere, espression of AQP-4 in contralateral hemisphere was low. At the same time, edema in contralateral hemisphere was not found. The result of electron microscopeic observation indicated that the neuron water degeneration occurred in ipsilateral hemisphere. Hypoxia happened in contralateral hemisphere. CONCLUSION: Expression of AQP-4 in contralateral hemisphere increases without significant edema formation. Increase in AQP-4 expression in contralateral hemisphere may be associated with hypoxia.     

Effect of Astragalus injection on expression of calmodulin after hypoxia/hypoglycemia and reoxygenation in rat hippocampal neurons

AIM: To investigate the effect of Astragalus injection on the expression of calmodulin(CaM) after hypoxia/ hypoglycemia and reoxygenation in rat hippocampal neurons.METHODS: The hippocampal neurons were cultured for 8 days and divided into 4 groups: normal control group (normal control), hypoxia/hypoglycemia and reoxygenation group (model), Astragalus injection solution group (solution control) and Astragalus injection group ( Astragalus ).The cells in all groups were treated with reoxygenation and normal medium after deprived of oxygen and glucose for 30 min except normal control group.The method of immunohistochemistry was used to measure the number of caspase-3 positive neurons.The expression of CaM at mRNA and protein levels was measured at time points of 0 h, 0.5 h, 2 h, 6 h, 24 h, 48 h, 72 h and 120 h after hypoxia/hypoglycemia and reoxygenation by RT-PCR and Western blotting, respectively.RESULTS: No difference of the parameters at all time points between model group and solution control group was found.Compared with normal control group, the numbers and the percentages of caspase-3 positive cells at all time points obviously increased in model group except at 0 h and 0.5 h (P<0.05).Compared with model group, the numbers and the percentages of caspase-3 positive cells were decreased in Astragalus injection group except at 0 h and 0.5 h (P<0.05).Compared with normal control group, the protein expression of CaM in rat hippocampal neurons at all time points obviously increased in model group (P<0.05).However, the protein expression of CaM in rat hippocampal neurons at all time points obviously decreased in Astragalus injection group as compared with model group (P<0.05).Compared with normal control group, the mRNA expression of CaM in rat hippocampal neurons at all time points obviously decreased in model group (P<0.05).The mRNA expression of CaM in rat hippocampal neurons at all time points obviously increased in Astragalus injection group as compared with model group (P<0.05).CONCLUSION: Astragalus injection inhibits the protein expression of CaM, the calcium overload and the expression of caspase-3 after hypoxia/hypoglycemia and reoxygenation, thus inhibiting hippocampal neuronal apoptosis.     

Astragalus injection inhibits expression of Apaf-1 in rat hippocampus after cerebral ischemia and reperfusion

AIM: To investigate the effect of Astragalus injection on the expression of apoptotic protease-activating factor 1 (Apaf-1) in the hippocampus of global cerebral ische-mia-reperfusion rats. METHODS: Male SD rats were randomly divided into 4 groups with 30 each: sham operation group, cerebral ischemia-reperfusion group, cerebral ischemia-reperfusion+Astragalus injection group, and cerebral ischemia-reperfusion+vehicle group. The global cerebral ischemia-reperfusion model of the rats was established by 4-vessel occlusion. The rats in cerebral ischemia-reperfusion group, cerebral ischemia-reperfusion+Astragalus injection group and cerebral ischemia-reperfusion+vehicle group were further divided into 7 subsets, according to the reperfusion time of 0 h, 0.5 h, 2 h, 6 h, 24 h, 72 h and 120 h. After reperfusion, the brains were removed at the corresponding time points. The protein expression of Apaf-1 in hippocampal neurons was detected by immunohistochemistry and Western blotting. The mRNA expression of Apaf-1 was observed by RT-PCR. RESULTS: Compared with sham operation group, the expression of Apaf-1 at mRNA and protein levels at all time points except 0 h and 120 h increased obviously in cerebral ischemia-reperfusion group (P<0.05). Compared with cerebral ischemia-reperfusion group, the expression of Apaf-1 at mRNA and protein levels at all time points except 0 h and 120 h decreased obviously in cerebral ischemia-reperfusion+Astragalus injection group (P<0.05). However, those in cerebral ischemia-reperfusion+vehicle group had no obvious change (P>0.05). CONCLUSION: Astragalus injection inhibits the expression of Apaf-1 at mRNA and protein levels in hippocampus of global cerebral ischemia-reperfusion rats, thus inhibiting the apoptosis of hippocampal neurons.     

Effects of inhibition of Cripto gene siRNA on vascular endothelial growth factor of colon cancer cell line LS-174T

AIM:To study the effects of Cripto gene on vascular endothelial growth factor (VEGF) of colon carcinoma cells.METHODS:Cripto siRNA was designed and constructed. Colon cancer LS-174T cells were divided into 4 groups: control group and different dose (3.125, 6.25 and 12.5 nmol/L) of siRNA groups. After transfected for 24, 48 and 72 h, colon cancer cells were harvested to carry on the next tests. Expression of Cripto mRNA was determined with real-time PCR, and immunofluorescence isothiocyanate (FITC) labeling assay and Northern blotting were performed to examine the expression of protein and mRNA of VEGF, respectively. The cells in control group and cells transfected with 12.5 nmol/L siRNA were inoculated into nude mice respectively. 30 days after inoculated, the mice of two groups were executed, and immunohistochemical (ICH) assay was used to evaluate the VEGF protein of mice tumor. RESULTS:siRNA down-regulated the Cripto mRNA in a dose and time dependent manner. Protein and mRNA of VEGF in transfected cells reduced in a dose and time dependent manner. Compared to control, the expression of VEGF protein from ICH assay was lowered significantly (P<0.05).CONCLUSION:Cripto gene might contribute to the regulation of angiogenesis of colon carcinoma. The down-regulation of Cripto gene by siRNA can suppress angiogenesis of human colon cancer.     

Effects of endothelial progenitor cell-conditioned medium on proliferation and differentiation of type Ⅱ alveolar epithelial cells exposed to hyperoxia

AIM: To investigate the effect of hyperoxia exposure on the paracrine function of endothelial progenitor cells (EPCs), and to explore the effects of paracrine factors of EPCs on the proliferation and differentiation of type Ⅱ alveolar epithelial cells (AECⅡ) exposed to hyperoxia. METHODS: Bone marrow-derived mononuclear cells were isolated and cultured in EGM-2MV medium for 7~10 d to obtain and identify EPCs. EPCs were cultured in room air (RA) or 60% O₂. The normoxia EPC-conditioned medium (E-CM-RA) and hyperoxia EPC-conditioned medium (E-CM-O₂) were collected. The levels of VEGF, FGF10, PDGF-BB and EGF in E-CM-RA and E-CM-O₂ were detected by ELISA. AECⅡ from adult rats were isolated, purified and cultured for 2 d, then divided into RA group, O₂ group, O₂+E-CM-RA group and O₂+E-CM-O₂ group. The proliferation of AECⅡ was detected by MTT assay and cell counting. The mRNA expression of SP-C and AQP5 was quantified by real-time PCR. RESULTS: The expression of VEGF and FGF10 in E-CM-O₂ group decreased significantly compared with E-CM-RA group (P<0.01). There were significant differences in AECⅡ viability and number among the 4 groups at 12 h, 24 h, 2 d and 3 d (P<0.01). Compared with RA group, AECⅡ viability and number in O₂ group decreased significantly at 12 h, 24 h, 2 d and 3 d (P<0.05). The AECⅡ viability and number in O₂+E-CM-RA group were significantly higher than those in O₂ group at 12 h, 24 h, 2 d and 3 d (P<0.05). However, no significant difference in AECⅡ viability and number between O₂+E-CM-O₂ group and O₂ group at 12 h, 2 d and 3 d was observed. There were significant differences in the mRNA expression of SP-C and AQP5 in the 4 groups at 24 h, 2 d and 3 d (P<0.01). Compared with RA group, the mRNA expression of SP-C in O₂ group was significantly inhibited (P<0.01), but the mRNA expression of AQP5 was promoted (P<0.01) at 24 h, 2 d and 3 d. Compared with O₂ group, the mRNA level of SP-C in O₂+E-CM-RA group and O₂+E-CM-O₂ group (P<0.05) at 24 h, 2 d and 3 d was increased, and the mRNA expression of AQP5 (P<0.01) at 2 d and 3 d was inhibited.CONCLUSION: EPCs secrete VEGF and FGF10, and hyperoxia impairs this paracrine function. Hyperoxia exposure inhibits AECⅡ proliferation and the mRNA expression of SP-C, but promotes the mRNA expression of AQP5. EPC-conditioned medium improves the proliferation of hyperoxia-exposed AECⅡ, and inhibits the transformation of AECⅡ. Hyperoxia exposure impairs the paracrine function of EPCs, and weakened the effects of E-CM-O₂ on AECⅡ.     

Effect of interlukin-22 antibody on diabetic nephropathy via inhibition on Snail1 expression

AIM:To investigate the effect of interlukin-22 (IL-22) on diabetic nephropathy (DN) and its possible mechanism. METHODS:C57BL/6 mice were randomized to normal control (NC) group,DN group, DN+recombinant IL-22 (rIL-22) group and DN+IL-22 antibody (anti-IL-22) group. After successful establishment of diabetes model for 8 weeks, the mice in DN+rIL-22 group and DN+anti-IL-22 group were intraperitoneally injected with rIL-22 (200 μg/kg) and anti-IL-22 (200 μg/kg), respectively, and the mice in NC group and DN group were intraperitoneally injected with 0.1% bovine serum albumin, twice a week for 4 weeks. After the intervention, blood glucose, kidney function, 24 h urine microalbumin (m-Alb) and 24 h urine creatinine (UCr) were measured. The pathological changes of renal tissues were observed under light microscope. The mRNA expression of Snail1 was detected by qPCR. The protein levels of fibronetin (FN) and E-cadherin were determined by Western blot. RESULTS:After the intervention, the ratio of 24 h m-Alb/UCr increased significantly in other model groups compared with NC group (P<0.05). The levels of 24 h m-Alb and 24 h UCr increased significantly in DN+rIL-22 group compared with DN group (P<0.05). However, in DN+anti-IL-22 group, the levels of 24 h m-Alb, 24 h UCr and 24 h m-Alb/UCr ratio were significantly lower than those in DN group and DN+rIL-22 group (P<0.05). The tubular epithelial cell vacuolar degeneration, protein cast formation and glomerular mesangial expansion in the renal tissues from diabetic mice were observed under light microscope. The lesions were more severe in DN+rIL-22 group, but attenuated in DN+anti-IL-22 group. The mRNA expression of Snail1 increased significantly in diabetic mice (P<0.05), but decreased significantly after a 4-week intervention by anti-IL-22 (P<0.05). The expression of FN, an extracellular matrix protein, increased significantly in DN+rIL-22 group (P<0.05). The expression of E-cadherin, an epithelial-mesenchymal transition marker, decreased significantly in DN+rIL-22 group as well (P<0.05). CONCLUSION:IL-22 neutralizing antibody may attenuate microalbuminuria and delay the progression of DN via inhibition of Snail1 expression in the renal tubular epithelial cells.     

Effect of Kang Xianling decoction on TGF-β1-Smad pathway in a unilateral ureteral obstruction rat model

AIM: To study the effect of Kang Xianling decoction,comprised of dahuang,danshen,taoren,niuxi and danggui,on TGF-β₁-Smad pathway in unilateral ureteral obstruction rat model.METHODS: Eighteen male SD rats were divided into 3 groups,sham group,model group and model group treated with Kang Xianling decoction randomly.Renal interstitial fibrosis model was established in rats by unilateral ureteral obstruction (UUO).After treatment for additional 14 d,parameters of hydroxyproline in obstructed kidney from 3 groups were analyzed.Rats were sacrificed and the pathological statuses of their kidneys were checked by HE staining and electron microscopy.Transforming growth factor-β₁ (TGF-β₁) mRNA in kidney tissue was determined by RT-PCR.TGF-β₁ receptor Ⅰ (TβRⅠ),TGF-β₁ receptorⅡ (TβRⅡ),phosphorylated Smad2 and Smad2 protein were determined by Western blotting.RESULTS: Parameters of hydroxyproline in animals of model group were significantly increased than those in sham operation group (P<0.05).The mRNA expression of TGF-β₁ and the protein expression of TβRⅠ,TβRⅡ,phosphorylated Smad2 and Smad2 in kidney tissue of animals in model group were significantly up-regulated.After intervention with Kang Xianling decoction,the above-mentioned up-regulated parameters,except TGF-β₁,were all significantly inhibited.Compared to model group,the pathological changes in renal tissues in treatment group were remarkable improved.CONCLUSION: Kang Xianling decoction inhibits the TGF-β₁-Smad pathway and the protein expression of TβRⅠ,TβRⅡ,phosphorylated Smad2 and Smad2,so as to decrease the level of collagen in obstructed kidney and to alleviate the renal interstitial fibrosis in UUO rats.     

CCK-8 up-regulats signal pathway of LPS-induced HO-1 expression in rat lungs

AIM: To study the signal pathway involved in up-regulation of LPS-induced HO-1 expression by CCK-8. METHODS: Forty-two SD rats were divided into 7 groups (six rats each) randomly as follows: control group, LPS group, LPS+SP600125 (JNK-specific inhibitor) group, CCK-8+LPS group, CCK-8+LPS+SP600125 group, CCK-8 group and CCK-8 +SP600125 group. Lungs from the rats in these 7 groups were excised 6 h after the agents were administered. HO-1 mRNA expression was examined by RT-PCR. The protein expression of HO-1 was detected by Western blotting and immunofluorescence flow cytometry (FCM). RESULTS: There were significant positive expression of HO-1 mRNA in LPS group compared to control group. CCK-8 enhanced LPS-induced HO-1 mRNA expression and CCK-8 alone induced HO-1 mRNA expression as well. The mRNA expressions of HO-1 in LPS group, CCK-8+LPS group and CCK-8 group were 3.01 (P<0.01), 5.88 (P<0.01) and 3.45 (P<0.01) times as many as that in control group, respectively. SP600125 inhibited the mRNA expression of HO-1 induced by CCK-8 and (or) LPS. The change of HO-1 protein expression was in accordance with that of HO-1 mRNA expression by Western blotting and immunofluorescence FCM. CONCLUSION: These results suggest that JNK/c-Jun signal pathway plays an important role in the up-regulation of LPS-induced HO-1 expression by CCK-8.     

Inflammatory response in rat kidney with contrast-induced nephropathy

AIM：To observe the expression of tumor necrosis factor α (TNF-α) and nuclear factor κB (NF-κB) in the renal tissue of the rats with contrast-induced nephropathy (CIN). METHODS：Male Sprague-Dawley rats (n=96) were randomly divided into control group (n=48) and CIN group (n=48). The model rats in CIN group were intravenously injected with iodinated contrast media (76% compound diatrizoate injection,10 mL/kg), while the rats in control group were injected with the same volume of saline. Six rats in each group were sacrificed at 6 h, 12 h, 24 h, 48 h, 72 h, 5 d, 10 d and 15 d after intravenous injection, respectively, and the blood and kidney samples of the rats were obtained. The renal tubular injury was assessed by histological examination (HE staining). The expression of kidney injury molecule-1 (KIM-1), TNF-α and NF-κB at mRNA and protein levels in the renal tissues were semiquantitatively measured by the methods of RT-PCR and immunohistochemistry, respectively. The correlations between the expression of TNF-α, NF-κB and tubular injury score, KIM-1 expression in renal tissue of CIN group were analyzed. RESULTS：The levels of serum creatinine (SCr) and blood urea nitrogen (BUN) in control group were not changed between different time points (P>0.05). The levels of SCr and BUN in CIN group displayed significant increases at different time points (except 15 d) compared with control group (P<0.05). The renal tubular injury score in CIN group was significantly higher at all time points than that in control group (P<0.05). The expression of KIM-1, TNF-α and NF-κB at mRNA and protein levels up-regulated significantly at 6 h and the peaking of KIM-1 expression was at 24 h, while the peaking of TNF-α and NF-κB expression was at 48 h in CIN group. The expression of KIM-1,TNF-α and NF-κB was significantly increased in CIN group compared with control group except at 15 d (P<0.05). The expression of TNF-α and NF-κB at mRNA and protein levels showed close correlations with renal tubular injury score (r=0.843, 0.758, 0.743 and 0.707, P<0.05). The expression of TNF-α and NF-κB at mRNA and protein levels was also positively correlated with KIM-1 expression (r=0.863, 0.807, 0.839 and 0.855, P<0.05). CONCLUSION：The expression of TNF-α and NF-κB at mRNA and protein levels in the renal tissues of CIN group is up-regulated and is closely related with renal tubular injury, indicating that the inflammatory response is involved in the pathogenesis of CIN.     

Macrophage migration-inhibitory factor induces myofibroblastic phenotype transformation of human embryonic lung fibroblasts via Rho-kinase in vitro

AIM: To investigate the influence and the mechanism of recombinant macrophage migration-inhibitory factor (rMIF) on fibroblasts. METHODS: Cultured human embryonic lung fibroblasts (MRC-5 cells) were divided into 2 groups: the cells in treatment group were treated with rMIF (25~100 μg/L) for 24 h or 48 h, and the control cells were without rMIF treatment. The mRNA expression of α-SMA was examined by RT-PCR. The protein level of α-SMA induced by rMIF was quantified by Western blotting. Half an hour before 100 μg/L rMIF challenge, Y27632 was added to the cells of 2 groups. After challenged for 48 h, the total RNA and protein were extracted,and the expression of α-SMA at mRNA and protein levels was determined by means of RT-PCR and Western blotting. After challenged by 100 μg/L rMIF for 6 h, 12 h, 24 h and 48 h, the cell total protein was extracted, and the protein level of α-SMA induced by rMIF was quantified by Western blotting. RESULTS: After stimulation for 24 h, rMIF did not increase the α-SMA mRNA synthesis compared with control. After stimulation for 48 h, rMIF significantly increased the expression of α-SMA mRNA and protein in a dose-dependent manner compared with control (r=0.697 and r=0.957, both P<0.01). Y27632 pretreatment prevented the increase (P<0.01). The amount of phosphorylated MYPT1 increased at 6 h (P<0.01), reached the maximum at 12 h (P<0.01), and decreased but still higher than the control at 24 h and 48 h (P<0.01). CONCLUSION: rMIF increases the α-SMA synthesis in MRC-5 cells and Rho-kinase regulates this process, suggesting that MIF may play important roles in the pathogenesis of airway remodeling.     

Quercetin inhibits endothelin-1-induced T-type Ca2+ channel expression in human umbilical arterial smooth muscle cells

AIM: To investigate the effect of quercetin on endothelin-1-induced T-type calcium channel(TCC) expression in primary cultured human umbilical arterial smooth muscle cells for exploring the protective role of quercetin in cardiovascular system. METHODS: Human umbilical arterial smooth muscle cells were verified by immunocytochemistry. The cells in 2-3 passages were used and randomly divided into control group, quercetin alone group, model group and experimental group. The cells in control group were cultured without any drugs for 24 h. The cells in quercetin alone group were cultured with 80 μmol/L quercetin for 24 h. The cells in model group were cultured with ET-1 at the concentration of 100 nmol/L for 24 h. The cells in experimental groups were pretreated with quercetin for 1 h, then coincubated with 100 nmol/L ET-1 for 24 h. The concentrations of quercetin used in this study were 20, 40and 80 μmol/L, respectively. The expression of α_1G, a TCC major subunit, was assayed at mRNA and protein levels by RT-PCR and Western blotting, respectively. The TCC currents(I_caT) were detected by the technique of whole-cell patch-clamp. RESULTS: Compared with control and experimental group, I_CaT density (P<0.01) and the expression of α_1G at mRNA (P<0.05) and protein (P<0.01) levels in model group were significantly increased. No significant difference in the results of quercetin alone group and control group was observed. CONCLUSION: The protective roles of quercetin in cardiovascular functions are related to the depressive effects of quercetin on ET-1-induced increase in both I_CaT density and the expression of α_1G at mRNA and protein levels in cultured human vascular smooth muscle cells.     

Effect of oxidized LDL on expression of connexin43 in cultured human umbilical vein endothelial cells

AIM: To investigate the effect of oxidized LDL (ox-LDL) on the expression of gap junction protein connexin43 in cultured human umbilical vein endothelial cells (HUVECs) in vitro. METHODS: Human umbilical vein endothelial cells cultured in normal condition were divided into blank control group, 50 mg/L,100 mg/L and 200 mg/L ox-LDL intervention groups. The mRNA expression of connexin43 in cultured HUVECs was detected with RT-PCR method; while the protein level of connexin43 was determined by the method of immunocytochemistry in the control and 100 mg/L ox-LDL intervention groups 24 h after ox-LDL was given. RESULTS: Different concentrations (50 mg/L, 100 mg/L, 200 mg/L) of ox-LDL up-regulated mRNA expression of connexin43 in cultured HUVECs after 24 h intervention (P<0.01). The protein level of connexin43 in cultured HUVECs intervened with 100 mg/L Ox-LDL for 24 h was up-regulated as compared to the control cells (P<0.01).CONCLUSION: Ox-LDL may up-regulate the expression of connexin43 at mRNA and protein levels in cultured human umbilical vein endothelial cells within short time, indicating that connexin43 plays an important role in the pro-atherosclerotic effect of Ox-LDL.     

Effects of cytokines on renovation of acute renal failure in mouse with bone marrow derived mesenchymal stem cell transplantation

AIM: To observe the effects of cytokines on renovation of acute renal failure (ARF) in mouse with bone marrow derived mesenchymal stem cell (MSC) transplantation. METHODS: ARF animal model was induced in mouse by subcutaneous injection of cisplatin. Mice were randomly assigned into 3 groups: normal control group, ARF group and MSC group. After 24 h cisplatin injection, animals were injected intravenously with MSC in MSC group. Animals were sacrificed at 1 d, 4 d, 7 d, 14 d and 28 d after cisplatin injection. The blood urea nitrogen (BUN) and serum creatinine (Scr) were measured. The renal morphologic changes were scored with Paller’s criterion on hematoxylin and eosin (HE) stained sections. The mRNA and protein expressions of HGF, BMP-7, TNF-α and IL-10 were detected by RT-PCR and immunohistochemistry method. RESULTS: After 4 d of cisplatin injection, the BUN and Scr values in MSC group were significantly lower than those in ARF group (P<0.01). After 7 d and 14 d, the values of BUN and Scr in MSC group were still lower than those in ARF group (P<0.01, P<0.05). The renal morphologic scores of MSC group were also lower than those of ARF group. After 7 d, the expressions of HGF, BMP-7 and IL-10 were higher in MSC group than those in ARF group, the expression of TNF-α in MSC group was lower than that in ARF group. CONCLUSION: MSC promotes the recovery of acute renal failure induced by cisplatin. The mechanism may partly depend on paracrine of growth factor and amelioration of inflammatory.     

Angiotensin-(1-7) increases expression of ABCA1 and decreases content of cholesterol in THP-1-derived foam cells via AC-cAMP pathway

AIM: To establish the THP-1-derived foam cell formation and to evaluate the effects of angiotensin-(1-7) and MDL (an inhibitor of adenylate cyclase) on the expression of ATP-binding cassete transporter A1(ABCA1) and the content of cholesterol. METHODS: THP-1-derived macrophages were treated with oxidized low-density lipoprotein(ox-LDL) to develop into foam cells. The foam cells were divided into 4 groups: control group, MDL group, Ang-(1-7) group and MDL+Ang-(1-7) group. At 24 h after treatment, the content of cAMP was measured by ELISA. The mRNA and protein levels of ABCA1 were determined by real-time RT-PCR and Western blotting, respectively. The content of cholesterol was detected by high performance liquid chromatography. RESULTS: The cAMP, the mRNA and protein levels of ABCA1 in Ang-(1-7) group were significantly higher, and the content of cholesterol was significantly lower than those in control group (P<0.05). On the contrary, the cAMP, the mRNA and protein levels of ABCA1 in MDL group were significantly lower and the content of cholesterol was significantly higher than those in control group (P<0.05). The results in MDL+Ang-(1-7) group were between Ang-(1-7) group and control group. CONCLUSION: Ang-(1-7) inhibits the formation of foam cells by promoting the expression of ABCA1 and decreasing the content of cholesterol. MDL partly antagonizes the effect of Ang-(1-7) by inhibiting the adenylate cyclase and decreasing the content of cAMP.     

Effects of vinpocetine on inflammation of brain in rats with intracerebral hemorrhage

AIM: To investigate the effects of vinpocetine on inflammation of brain in intracerebral hemorrhage (ICH) rats and to explore the underlying mechanisms. METHODS: All rats were randomly divided into sham group, ICH group, ICH with low dose of vinpocetine treatment group, ICH with medium dose of vinpocetine treatment group, and ICH with high dose of vinpocetine treatment group. Except sham group, the rats in other groups were injected with type VⅡ collagenase to establish ICH model, and then the rats in vinpocetine treatment groups were received vinpocetine at 0.5, 1.0 or 1.5 mg/kg by intraperitoneal injection once a day for 7 days. After corresponding treatment, the impairment of neurological function in the rats was scored and the water content of the brain tissues was measured. Moreover, the activity of myeloperoxidase (MPO) was determined by ELISA, and the protein expression of Toll-like receptors 4 (TLR4), nuclear factor-κB(NF-κB), intercellular cell adhesion molecule-1 (ICAM-1) and vascular cell adhesion molcule-1 (VCAM-1) in the brain tissues was determined by Western blot. RESULTS: Compared with ICH group, vinpocetine treatment significantly decreased the scores of the impairment of neurological function and the water content of the brain tissues. Moreover, the activity of MPO and the protein expression of TLR4, NF-κB, ICAM-1 and VCAM-1 were also reduced after vinpocetine treatment (P<0.05). CONCLUSION: Vinpocetine improves neurological function in ICH rats via suppression of inflammation by inhibiting NF-κB signaling and expression of ICAM-1 and VCAM-1.