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The aim of this study is to explore the tissue distribution of PEGylated puerarin in acute myocardial ischemia model rats. Healthy male SD rats were randomly divided into two groups (30 each). Both were given PEGylated puerarin at a dose of 488 mg x kg(-1). After 5 min of medication, one group was normal rats, another group with acute myocardial ischemia was established by peritoneal injection of 50 mg x kg(-1) isoprenaline. After administration, the animals were executed at 30, 60, 90, 120, 150 and 180 min, then heart, liver, spleen, lung, kidney were extracted. The content of puerarin in organ tissue was determined by HPLC. The results showed that the AUC of tissue distribution of PEGylated puerarin in normal rats was liver > kidney > heart ≈ spleen > lung > brain. While the AUC of tissue distribution of PEGylated puerarin in acute myocardial ischemia model rats was liver ≈ heart > kidney > lung ≈ spleen > brain. AUC(heart) of PEGylated puerarin in acute myocardial ischemia model rats was 1.7 times than that of the normal rats, and there was significant difference (P < 0.05). Thus, PEGylated puerarin had a good heart-targeting property in early myocardial infarction area, drugs could accumulate in the ischemic myocardium. It provided important information for further study and clinic use of PEGylated puerarin.
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<p><b>OBJECTIVE</b>To study the effects of focal adhesion kinase (FAK) phosphorylation on smooth muscle cells (SMCs) adhesion and migration stimulated by fibronectin.</p><p><b>METHODS</b>Adhesion and migration of cultured SMCs were stimulated by different concentrations of fibronectin (FN), FAK and its phosphorylation were detected by immunoprecipitation and Western blot. FAK antisense oligodeoxynucleotides (ODNs) were transfected into SMCs by cationic lipid to investigate its modulatory effects on tyrosine phosphorylation. SMCs adhesion and migration were also measured by morphological enumeration and modified Boyden Chambers, respectively.</p><p><b>RESULTS</b>FAK were expressed when SMCs adhesion and migration were successfully simulated by different concentrations of FN. FAK phosphorylation were detected only at 20 microg/ml FN or more. FAK antisense ODNs were transfected efficiently by cationic lipid and FAK phosphorylation was inhibited substantially. The SMCs migration rate in the 5 - 60 microg/ml FN groups was reduced by 17.89% - 27.67%. Cell migration stimulated by FN at 10, 20, 40 and 60 microg/ml were reduced by 23.26%, 21.63%, 19.31% and 17.88%, respectively (P < 0.05).</p><p><b>CONCLUSIONS</b>FAK phosphorylation and FAK-mediated signal transduction play important roles in SMCs adhesion and migration stimulated by ECM. The process can be inhibited effectively by FAK antisense ODNs.</p>
Subject(s)
Animals , Rats , Cell Adhesion , Cell Movement , Cells, Cultured , DNA, Antisense , Genetics , Physiology , Dose-Response Relationship, Drug , Enzyme Activation , Fibronectins , Pharmacology , Focal Adhesion Kinase 1 , Focal Adhesion Protein-Tyrosine Kinases , Muscle, Smooth, Vascular , Cell Biology , Phosphorylation , Protein-Tyrosine Kinases , Genetics , Metabolism , TransfectionABSTRACT
AIM To investigate effect of urotensin Ⅱ (UⅡ)on proliferation of aort a smooth muscle cells （ASMC）of rat and study the signal transduction pathway o f it. METHODS In cultured ASMC of rat, UⅡ was used to stimulate proliferation of these cells and levels of ［3H］-TdR incorporation were used to evaluate the speed of DNA synthesis, and different inhibitors were used to s tudy the action of different signal transduction pathway of mitogenic effect of UⅡ on VSMC. RESULTS 1×10-9～1×10-7 mol*L- 1 UⅡ caused marked concentration-dependent increasing of ［3H］-TdR i ncor poration of ASMC. ［3H］-TdR incorporation of 1×10-9,1×10-8 and 1×10-7 mol*L-1 UⅡ were 22%（P<0.05）, 57%（P<0.01）and 65%（P<0.01）higher than control. Nicardipine, H7, W7 and PD98059 , which are inhibitors of calcium channel, PKC, CaM-PK and MAPK respectively, inhibited the effects of UⅡ obviously, with the inhibitory rate by 55%（P< 0.01）, 27%（P<0.01）,18%（P<0.05）and 16%（P<0.05）respectively . CONCLUSION UⅡ is a strong mitogen for VSMC and the mitogenic e ffect of UⅡ is probably mediated by Ca2+, PKC, CaM-PK and MAPK signal tr ansduction pathway.
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AIM: To observe the alteration of urotensin II (UII) receptors and contractile response to UII in rat aorta after balloon angioplasty injury. METHODS: The plasma membrane isolated from balloon injured aorta was used to study the binding of [ 125 I]-UII to the membrane and the contractile potency of UII on rat aorta was assayed. RESULTS: In contrast to the normal aorta, the contractile potency to UII enhanced in balloon injury artery and the calculated maximal number of specific binding sites (Bmax) was increased about 44% and 36% respectively in rat artery after balloon injury 3 and 21 days ( P
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Objective To observe the expression and distribution of IL 10 in local vessel after balloon injury in rat and study its significance in the response of artery injury Methods RT PCR and Western Blot as well as immunohistochemical method were used to assay the changes of IL 10′s mRNA as well as expression and distribution of protein Results There is no expression of IL 10 in vascular wall of normal rat After Balloon injury, the level of IL 10 mRNA and product of IL 10 was up regulated, and immunohistochemiscal staining showed immunoreactive IL 10 mainly in smooth muscle cells Conclusion IL 10 is expressed in balloon injured aorta and may contribute to the modulation of the local inflammatory response
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AIM: To study the effect of focal adhesion kinase (FAK) phosphorylation on the adhesion and migration of smooth muscle cells (SMCs) stimulated by fibronectin (FN). METHODS: Cultured SMCs were stimulated by different concentrations of FN.FAK expression and phosphorylation were detected by immunoprecipitation and Western blot. To investigate the modulating effect of FAK on tyrosine phosphorylation and SMCs adhesion and migration, FAK antisense oligodeoxynucleotides (ODNs) were transfected into SMCs by actionic lipid method. RESULTS: FAK expression increased when SMCs adhesion and migration were induced by FN at different concentrations used in the experiment. However, FAK phosphorylation was only observed by FN stimulation at concentration of 20 mg/L. FAK antisense ODNs inhibited FAK phosphorylation magnificently. The migration rates of SMCs were reduced by 17.89%-27.67% when FN was used at concentration from 5 mg/L to 60 mg/L. The decreased migrating cell numbers were showed the same patterns. The apoptotic SMCs were 33.57% higher than that control ( P
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AIM To investigate effect of urotensin Ⅱ (U Ⅱ )on proliferation of aorta smooth muscle cells (ASMC) of rat and study the signal transduction pathway of it. MEfHODS in cultured ASMC of rat, U Ⅱ was used to stimulate proliferation of these cells and levels of [3H]-TdR incorporation were used to evaluate the sped of DNA synthesis, and different inhibitors were ed to study the action of different signal transduction pathway of mitogenic effect of UⅡ on VSMC. RESULTS. 1 ? 10-9~l ? 10-7 mol. L-l U Ⅱ caused marked concentration-de pendent increasing of [3H]-TdR incorporation of ASMC [3H]-TdR incorporation of 1 ? 10-9, 1 ? 10-8 and １ ? 10-7 mol. L－l U Ⅱ were 22%'(P
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AIM: To observe the effect of adrenomedullin(ADM)on proliferation of vascular smooth muscle cells(VSMC) induced by urotensin Ⅱ(UⅡ). METHODS: DNA synthesis of cultured rat aortic VSMC was measured by -TdR incorporation. The activities of mitogen activated protein kinase(MAPK) were determined by isotope tagged with [?- 32 P]-ATP. RESULTS: UⅡ(10 -8 mol/L) significantly increased -TdR incorporation of VSMC and MAPK activities by 38%( P0.05 ), 32%( P0.05 ), 32%( P
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AIM:To investigate the effect of batroxobin on thromboxane A 2(TXA 2)level in canine heart ischemia/reperfusion (I/R) injury and the cardioprotective mechanism of batroxobin against I/R injury. METHODS: Canine heart ischemia was induced by ligation of the left anterior descending coronary artery for 30 min followed by 90 min reperfusion.Batroxobin was intravenously administered before heart ischemia and 15 min before reperfusion.The level of plasma thromboxane (TXA 2) ,creatine phosphokinase (CK), lactic dehydrogenase (LDH ) and the concentration of myocardial TXA 2 were measured. The pathological changes in I/R myocardium were observed. RESULTS: In I/R group, TXA 2 levels of both plasma and myocardium increased significantly,and myocardium was injured obviously. Myocyte cells of central zone of I/R heart showed intracellular edema, swollen and damaged mitochondria, and fragmentation of cristae and concentrated nucleus. Plasma CK and LDH level was also elevated. Both ways of batroxobin administration reduced TXA 2 level apparently and plasma CK?LDH were also reduced significantly. LVEDP lowered while +dp/dt max elevated greatly,the mortality of I/R canine reduced obviously. CONCLUSION: Batroxobin decreased TXA 2 levels of both plasma and myocardium significantly, and could afford protection from I/R injury.