ABSTRACT
Objective The purpose of this study was to determine the changes of expression of angiogenic factor mRNA in the spinal cord of rats with protruded intervertebral disc and the effects of electroacupuncture , and to analyze the role of electroacupuncture in the spinal microvascular angiogenesis .Method Eighteen healthy male Sprague-Dawley rats were divided randomly into 3 groups:the electroacupuncture group , model group and sham-operated group .The protruded intervertebral disc models were generated by the left lateral and ventral spinal cord compression at T 13 using a self-made silicon sheet.Only the rats in the first group were treated by electroacupuncture once a day for consecutive 7 days.Spinal cord tissue samples were taken from the compression site at 7 days after operation .The mRNA levels of Ang-1, Tie-1, Ang-2, Tie-2, VEGF, Flt-1, caspase-3 and Tsp-1 were determined by RT-PCR, and the pathological changes of the spinal cord was examined using HE staining .Results The rats in the electroacupuncture treated group showed significantly im-proved hind leg function , a relatively complete spinal cord structure , and a clear boundary between grey and white matters . The expression levels of Ang-1, Ang-2, Tie-1, Tie-2, capase-3 and Tsp-1 in the spinal cord tissues of the model group were significantly higher than those of sham-operated group ( P0.05).The expression levels of Ang-2, Tie-1, Tie-2, caspase-3 and Tsp-1 of the electroacupuncture group were significantly lower than those in the model group (P0.05).All the indexes be-tween electroacupuncture and sham-operated groups showed no significant difference ( P>0.05 ) .Conclusions Our re-sults demonstrate that mRNA expression of relevant angiogenic factors were abnormal after spinal cord compression , while electroacupuncture can down-regulate the expression of Ang-2, Tie-2,Tsp-1 and caspase-3, and modulate the promoting and inhibiting factors of angiogenesis to return towards normal , therefore , to create beneficial conditions for the repair of spinal cord injuries .
ABSTRACT
Objective To establish a canine model of intervertebral disc extrusion by surgery and observe the histological and microcirculatory changes of the spinal cord , in order to accumulate data for studies on the pathology and mechanism of treatment for intervertebral disc extrusion .Methods Normal healthy adult dogs were divided randomly into two groups:normal control group and model group .To simulate the intervertebral disc extrusion caused by spinal cord compression, 6Fr double lumen catheter with ballon was inserted into the spinal cord T 12-T13 and filled with about 5 mL Iohexol after the exposure of spinal cord L 1 by hemilaminectomy .The spinal cord blood flow ( SCBF) at the L1 level before and after compression was measured by laser-Doppler flowmetry .Morphological changes of the compressed spinal cord at 14 days after compression was examined by histopathology .Results The ( Texas spinal cord injury score ) ( TSCIS) scores of the motor function of bilateral hind limbs were highly significantly decreased (P<0.01).The blood flow of spinal cord at the L1 level was significantly decreased (P<0.05) after compression than that before .Compared with the normal control group, the model group showed abnormal vacuolization in the white matter and the number of normal neurons in the ventral horn of gray matter was significantly lower ( P<0.01 ) .Conclusions Our findings demonstrate that canine models of intervertebral disc extrusion can be successfully established by balloon catheter compression , showing local impairment of microcirculation and histological changes in the spinal cord .This canine model may provide a useful model for evaluation of therapeutic effects of acupuncture and for mechanism studies .
ABSTRACT
Objective To study the main subtypes messenger ribonucleic acid(mRNA) and the basal enzyme activity of phosphodiesterase (PDE) in rat pulmonary microvascular endothelial cells (PMVECs) through the examination mRNA expression and activity of PDE in vitro. The data were offered to reveal the relationship between PDE distributions, activity change and to accumulate data for the possibility of drug regulation of its functional alteration. Methods The cells were cultured with tissue-sticking method;the gene expression of PDEs was detected by reverse transcript polymerase chain reaction (RT-PCR), and the activity of PDEs was calculated by cyclic nucleotides content change examined with high performance liquid chromatogram (HPLC) before and after the PDE reaction( n ＝3). Results The PMVECs identified by cell immunofluorescence with polyclonal antibody of CD31 were dissociated and cultured, mRNAs of PDE1A, 1C, 2A,3A, 3B, 4A, 4D, 5A, 7A, 7B, 8A, 8B, 9A, 10A,11A were expressed in PMVECs, but there was no mRNA of PDE1B expressed in PMVECs. cAMP/cGMP-PDE in the extent of 5-20μl had a good linear correlation with its activity. Conclusion There are 17 kinds of PDE gene expression existing in PMVECs which contain of the basic enzyme with a higher activity.