ABSTRACT
Objective To investigate the effect of propofol on rat′s limb ischemia reperfusion injury .Methods Sixty healthy SD rats were randomly divided into sham operate group ,ischemia-reperfusion group and propofol group (n= 20) ,each group was divided into 4 subgroups according to the different reperfusion time .To copy the right lower limb ischemia reperfusion model ,5 min before reperfusion ,use propofol injection (50 mg/kg ,intraperitoneal inject) ,various subjects in the corresponding time points (3 ,6 , 9 ,12 h) were sacrificed .TNF-α ,NF-κB of blood and MDA ,SOD of Skeletal muscle were measured ,calculate muscle wet dry weight ratio .Results Compared with ischemia reperfusion group ,propofol could significantly reduce expression of TNF-alpha ,NF-κB lev-els in serum (P< 0 .05) ,inhibit the increase of the MDA level and decrease of the SOD level in muscle (P< 0 .05) ,also reduce the extent of skeletal muscle cell edema(P< 0 .05) .Conclusion Propofol can attenuate limb ischemia reperfusion injury by inhibiting inflammation response and reducing the oxygen free radicals′ damage .
ABSTRACT
Objective To observe the proliferation and phenotype-switching of pulmonary arterial smooth muscle cell (PASMC) induced by hypoxia and interfered by Ad-PKGIα. And to investigate the potential regulative role of PKGIα gene in the molecule mechanism of hypoxia pulmonary vessel remodeling (HPVR). Methods To establish the pure PASMC cultured by tissue-sticking methods. Semi-quantitative reverse transcription and polymerase chain reaction (RT-PCR) and Western blot were used to examine the PKGIα mRNA and protein expression after PASMC were transfected by Ad-PKG. The mRNA and protein expressive change of smooth muscle α actin(SM-α-actin) determined the degree of cell phenotype-switching. The changes of PASMC proliferation were determined by flow cytometry and ~3H-TdR incorporated way. Results Ad-PKGIα could transfect into PASMC and highly express. Hypoxia down-regulated the expression of SM-α-actin protein (44. 25±5.34 in normoxia, 32. 18±4. 19 in 12 h hypoxia condition, 21.90 ±2. 44 in 24 h hypoxia condition, P < 0. 05), that could be blocked by the transfeetion of Ad-PKGIα. Hypoxia could push PASMC mitosis and proliferating(~3H-TdR incorporated way: 7570 ± 371 in normoxia,12 020± 831 in 12 h hypoxia condition,14 924 ± 1491 in 24 h hypoxia condition, P <0. 05), that could be blocked by the transfection of Ad-PKGIα, too. Conclusions The results suggested that PKGIα signaling pathway might play an important role in the molecule mechanism of HPVR. And PKGIα gene might be a target point of gene therapy.
ABSTRACT
Objective To explore the effects of hammerhead ribozyme on the expression and activity of NHE 1 and pHi in pulmonary artery smooth muscle cells (PASMCs) of rats and its role in PASMCs proliferation in vitro . Methods According to the secondary structure of NHE 1 mRNA in rats, NHE 1 specific hammerhead ribozyme was designed with the assistance of computer. The recombinant vector of retroviral plasmid pLXSN and hammerhead ribozyme, PRZ, was transfected into the cultured PASMCs. G418 resistant cell clones were screened with 100 ?g/ml G418. Then, the expression of NHE 1 mRNA was detected by RT PCR, intracellular pH(pHi) value and recovery rate of pHi after intracellular acid loading were measured by fluorescent probe BCECF. 22 Na uptake and 3H TdR incorporation were determined, respectively. Results Compared with the cells transfected with pLXSN and non transfected cells, NHE 1 mRNA, pHi value, pHi recovery rate, 22 Na uptake and 3H TdR incorporation decreased significantly in cells transfected with recombinant vector PRZ. No significance was found between the pLXSN transfected group and non transfected group. Conclusion Hammerhead ribozyme can cleave the target NHE 1 mRNA specifically, reduce the expression of NHE 1 mRNA, induce intracellular acidosis and consequently suppress the proliferation of PASMCs.
ABSTRACT
AIM:To investigate the role of bcl-2 and bax expression in apoptosis of pulmonary artery smooth muscle cells(PASMC) of rats induced by Na +/H + exchanger isoform-1(NHE-1) inhibition in vitro. METHODS:Intracellular Ca 2+ concentration ([Ca 2+ ]i) was measured using Fura-2/AM. The expression of bcl-2 and bax mRNA in cells were detected by sqRT-PCR, and the expression of Bcl-2 and Bax protein were examined immunohistochemically. RESULTS:The expression of bax mRNA and Bax protein and [Ca 2+ ]i in cells transfected with NHE-1 ribozyme gene increased significantly compared with those of cells transfected with pLXSN and nontransfected control. Meanwhile, the expression of bcl-2 mRNA and Bcl-2 protein in cells transfected with NHE-1 ribozyme gene decreased significantly. CONCLUSION:Apoptosis of pulmonary artery smooth muscle cells induced by inhibiting NHE-1 may have relevance to the increase in [Ca 2+ ]i and bax expression and the decrease in bcl-2 expression.