IMPACTS ON THE EXPRESSION OF VASCULAR ENDOTHELIAL GROWTH FACTOR IN ISCHEMIC BRAIN TISSUE TREATED WITH CLUSTER PUNC-TURE ON SCALP ACUPOINTS IN RATS

Objective To probe into the impacts on the expression of vascular endothelial growth factor （VEGF） in ischemic brain tissue treated with cluster puncture on scalp acupoints in rats, Methods 128 rats were randomized into pseudo-operation group, model group, scalp-needling group and cluster-needling group, In scalp-needling group, penetration method was used on the focal side from Bǎihuì （百会 GV20） to Qǔbīn （曲鬓 GB7）, and in cluster-needling group, penetration method was used on both sides from GV20 to GB7, and a common needling on GV 20. Needles were punctured 2 mm in depth, constantly rotated for 10min, retained for 2 h. Immunohistrochemical method was applied to determine VEGF expression and microvessel density （MVD）. Results With the intervention of cluster puncture on scalp acupoints, VEGF： expressions on every time-spot after ischemia were enhanced apparently, superior to those in scalp-needling group. On the three time-spots of the 7^th, 14^th and 21^st days, MVD was increased after cluster puncture on scale acupoints, superior to those in scalp-needling group. Conclusion Cluster puncture on scalp acupoints up-regulated VEGF expression and promoted regeneration of microvessel.

Metastasis-associated protein 1 induces VEGF-C and facilitates lymphangiogenesis in colorectal cancer

AIM:To study the correlation between high metastasisassociated protein 1(MTA1)expression and lymphangiogenesis in colorectal cancer(CRC)and its role in production of vascular endothelial growth factor-C(VEGF-C). METHODS:Impact of high MTA1 and VEGF-C expression levels on disease progression and lymphovasculardensity(LVD,D2-40-immunolabeled)in 81 cases of human CRC was evaluated by immunohistochemistry. VEGF-C mRNA and protein expressions in human LoVo and HCT116 cell lines were detected by real-time polymerase chain reaction and Western blotting,respectively,with a stable expression vector or siRNA. RESULTS:The elevated MTA1 and VEGF-C expression levels were correlated with lymph node metastasis and Dukes stages(P<0.05).Additionally,high MTA1 expression level was correlated with a large tumor size(P< 0.05).A significant correlation was found between MTA1 and VEGF-C protein expressions in tumor cells(r=0.371, P<0.05).Similar to the VEGF-C expression level,high MTA1 expression level was correlated with high LVD in CRC(P<0.05).Furthermore,over-expression of MTA1 significantly enhanced the VEGF-C mRNA and protein expression levels,whereas siRNAs-knocked down MTA1 decreased the VEGF-C expression level. CONCLUSION:MTA1,as a regulator of tumor-associated lymphangiogenesis,promotes lymphangiogenesis in CRC by mediating the VEGF-C expression.

Hypoxia-inducible factor 1 alpha and vascular endothelial growth factor overexpression in ischemic colitis

AIM: To examine the etiology and pathophysiology in human ischemic colitis from the viewpoint of ischemic factors such as hypoxia-inducible factor 1 alpha (HIF-1 alpha and vascular endothelial growth factor (VEGF). METHODS: Thirteen patients with ischemic colitis and 21 normal controls underwent colonoscopy. The follow-up colonoscopy was performed in 8 patients at 7 to 10 d after the occurrence of ischemic colitis. Biopsy samples were subjected to real-time RT-PCR and immunohistochemistry to detect the expression of HIF-1 alpha and VEGF. RESULTS: HIF-1 alpha and VEGF expression were found in the normal colon tissues by RT-PCR and immunohistochemistry. HIF-1 alpha and VEGF were overexpressed in the lesions of ischemic colitis. Overexpressed HIF-1 alpha and VEGF RNA quickly decreased to the normal level in the scar regions at 7 to 10 d after the occurrence of ischemic colitis. CONCLUSION: Constant expression of HIF-1 alpha and VEGF in normal human colon tissue suggested that HIF-1 alpha and VEGF play an important role in maintaining tissue integrity. We confirmed the ischemic crisis in ischemic colitis at the molecular level, demonstrating overexpression of HIF-1 alpha and VEGF in ischemic lesions. These ischemic factors may play an important role in the pathophysiology of ischemic colitis.

Attenuation of portal hypertension by natural taurine in rats with liver cirrhosis

AIM： To investigate the inhibitory effect of natural taurine （NTau） on portal hypertension （PHT） in rats with experimentally-induced liver cirrhosis （LC）. METHODS： Experimentally-induced LC Wistar rats （20 rats/group） were treated with either oral saline or oral NTau for 6 consecutive weeks. Evaluation parameters included portal venous pressure （PVP）, portal venous resistance （PVR）, portal venous flow （PVF）, splanchnic vascular resistance （SVR） and mean arterial pressure （NAP）. Vasoactive substance levels including nitric oxide （NO）, nitric oxide synthase （NOS） and cyclic guanosine monophosphate （cGMP） were also measured. Histological investigation of type Ⅰ and Ⅲ collagen （COL Ⅰ and Ⅲ） and transforming growth factor-β1 （TGF-β1） was also performed. RESULTS： Treatment with NTau （1） significantly decreased PVP, PVR and PVF, and increased MAP and SVP; （2） markedly increased the vascular compliance and reduced the zero-stress of the portal vein; （3） markedly decreased the amount of NO and cGMP and activity of NOS; and （4） improved the pathological status of the liver tissue and reduced the expression of COL Ⅰ, COL Ⅲ and TGF-β1. CONCLUSION： NTau inhibited the LC-induced PHT by improving hyperdynamic circulation, morphology of liver and biomechanical properties of the portal vein in experimentally-induced LC rats.

Effect of electroacupuncture on expressions of VEGF and CD31 in MCAO model rats

Objective： To investigate the effect of electroacupuncture （EA） at Ganshu （BL 18） and Shenshu （BL 23） on vascular endothelial growth factor （VEGF） and platelet endothelial cell adhesion molecule-1 （PECAM-1）/CD31 around the cerebral infarction focus in middle cerebral artery occlusion （MCAO） rats and the possible mechanism, thus to provide a new strategy for the treatment of cerebral ischemic stroke by acupuncture. Methods： A total of 180 healthy male Sprague-Dawley （SD） rats were randomly divided into a sham operation group, a model group, an acupoint group and a non-acupoint group, 45 rats in each group. MCAO model was established using the modified line-embolus method in all rats except for those in the sham operation group; rats in the acupoint group were treated with EA at Ganshu （BL 18） and Shenshu （BL 23）; rats in the non-acupoint group were treated with EA at the control points; rats in other 2 groups were only subjected to bundling without treatment. Ten rats in each group were randomly selected on the 3rd day, the 14th day and the 21st day after acupuncture stimulation to test the neurological function impairment. The expression levels of CD31 and VEGF were also detected. Results： Compared with the model group and non-acupoint group, the neurological function score of the acupoint group was decreased at each time point, and the differences were statistically significant （P〈0.05, P〈0.01）. The expressions of VEGF and CD31 in each group were the lowest on the 3rd day, reached the peak on the 14th day and still remained at high level on the 21st day. And the differences among groups were statistically significant both on the 14th day and the 21st day （P〈0.05, P〈0.01）. Compared with the model group and the non-acupoint group, the expressions of VEGF and CD31 in the acupoint group were increased, and the differences were statistically significant （all P〈0.05）. Conclusion： EA at Ganshu （BL 18） and Shenshu （BL 23） can significantly improve the neurological function score of MCAO model rats, and shows protective effect on cerebral ischemia. The protective mechanism may be related to the up-regulation of CD31 and VEGF expression around the cerebral infarction focus in the MCAO model rats and induction of angiogenesis.

An experimental study of a modified Dahuang Zhechong pill on theangiogenesis of RF/6A cells in vitro

OBJECTIVE:To investigate the effects of a modified Dahuang Zhechong Pill(MDZP) on the angiogene sis of rhesus choroid-retina endothelial(RF/6A cells and its preliminary mechanism.METHODS:A 3-(4,5-dimethylthiazol-2-yl)-5-(3-car boxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazol ium(MTS) method was used to assess the effect o a MDZP on RF/6A cell proliferation induced by vas cular endothelial growth factor(VEGF).Transwell in serts were used to assess the effect of the MDZP on RF/6A cell migration.Matrigel was used to asses the effect of the MDZP on the tube formation of RF 6A cells.Western blotting and quantitative re al-time reverse transcription polymerase chain reac tion(RT-PCR) were used to detect the protein and mRNA expression,respectively,of VEGF and matri metalloproteinase-2(MMP-2) in RF/6A cells treatedwith the MDZP.RESULTS:RF/6A cell proliferation induced by VEGF was inhibited by 0.2 mg/mL MDZP.At 0,12.5,25 and 50 mg/mL MDZP,the number of cells that migrated through Transwell membranes was 73.33± 4.51,61.33±4.04,28.67±6.66 and 17.67±4.16,respectively,and the number of tubes formed in Matrigel was 20.33±0.58,13.33±1.53,11.00±1.00 and 1.33±0.58,respectively.At 100 and 200 mg/mL MDZP,the protein and mRNA expression of VEGF and MMP-2 were inhibited in RF/6A cells.At 400 mg/mL MDZP,the expression of VEGF mRNA and MMP-2 protein were inhibited in RF/6A cells.CONCLUSIONS:MDZP inhibits the angiogenesis of RF/6A cells via the suppression of proliferation,migration and tube formation of RF/6A cells.Inhibition of the protein and mRNA expression of VEGF and MMP-2 in RF/6A cells may be an important mechanism.

Vascular endothelial growth factor promotes peripheral nerve regeneration after sciatic nerve transection in rat

Objective：To evaluate the local effect of vascular endothelial growth factor （VEGF） on transected sciatic nerve regeneration.Methods：Sixty male white Wistar rats were divided into four experimental groups randomly （n=15）.In transected group the left sciatic nerve was transected and the stump was fixed to adjacent muscle.In treatment group the defect was bridged using a silicone graft filled with 10μL VEGF.In silicone group the graft was filled with phosphate-buffered saline.In sham-operated group the sciatic nerve was exposed and manipulated.Each group was subdivided into three subgroups with five animals in each and nerve fibers were studied 4,8 and 12 weeks after operation.Results：Behavioral test,functional study of sciatic nerve,gastrocnemius muscle mass and morphometric indices confirmed a faster recovery of regenerated axons in VEGF group than in silicone group （P〈0.05）.In immunohistochemical assessment,reactions to S-100 in VEGF group were more positive than that in silicone group.Conclusion：Local administration of VEGF will improve functional recovery and morphometric indices of sciatic nerve.

Influence of Electroacupuncture on Tumor Necrosis Factor-αand Vascular Endothelial Growth Factor in Rats with Experimental Arthritis

Objective： To observe the effect of electroacupuncture （EA） on tumor necrosis factor-α （TNF-α） and vascular endothelial growth factor （VEGF） in peripheral blood and joint fluid of collagen-induced arthritis （CIA） rats. Methods： After CIA rat models were successfully established, the rats were given corresponding intervention treatments. The levels of TNF-α and VEGF in peripheral blood and joint fluid were measured. Results： The levels of TNF-α and VEGF levels in peripheral blood and joint fluid of CIA rats were higher than those of normal group （P〈0.05）; EA treatment significantly decreased the index levels, of significant difference compared with those before treatment （P〈0.05）, as well as the model group （P〈0.05）. Conclusion： EA can significantly decrease the TNF-α and VEGF levels in peripheral blood and joint fluid of CIA rats, and helps restrain further development of the disease.

Effects of electroacupuncture pretreatment on motor function and cerebral blood flow in MCAO model rats

Objective:To observe the effects of electroacupuncture(EA)pretreatment on motor function,cerebral blood flow,cerebral infarction volume,and vascular endothelial growth factor(VEGF)level in middle cerebral artery occlusion(MCAO)model rats.Methods:Twenty-four male Sprague-Dawley rats were randomly divided into a normal group,a model group,and an EA group,with eight rats in each group.The middle cerebral artery ischemia-reperfusion model was established by the suture-occluded method in the model group and the EA group,while not in the normal group.The EA group was pretreated with EA at bilateral Fengchi(GB20)before model preparation,once a day for 30 min each time for a total of 7 d.The changes in the CatWalk gait parameters,modified Bederson neurological deficit score,cerebral blood flow,cerebral infarction volume after ischemia,and VEGF level in the brain tissue of rats in each group were observed.Results:Compared with the normal group,the modified Bederson neurological deficit score in the model group and the EA group increased after modeling(P<0.05),and the CatWalk gait parameters(one-leg stance duration,gait cadence,and gait cycle)were all changed(P<0.05).Compared with the model group,the modified Bederson neurological deficit score in the EA group decreased(P<0.05),and the CatWalk gait parameters improved(P<0.05).Immediately after ischemia,the cerebral blood flow in the normal group was greater than that in the model group and the EA group(P<0.05);after reperfusion,the cerebral blood flow in the EA group was greater than that in the model group(P<0.05).Compared with the normal group,the cerebral infarction volume in the model group and the EA group increased(P<0.05).Compared with the model group,the cerebral infarction volume in the EA group decreased(P<0.05).The expression level of VEGF-positive cells in the rat brain tissue in the model group was higher than that in the normal group(P<0.05),and was higher in the EA group than in the model group(P<0.05).Conclusion:EA pretreatment improves the limb motor function in MCAO model rats,alleviates the symptoms of neurological deficits,promotes the recovery of cerebral blood flow,reduces the cerebral infarction area after MCAO modeling,and increases the VEGF expression in the brain tissue.