| ObjectiveWith the increase of the spinal-epidural anesthesia in our country this years, about 50%of the call cases(＞10,000,000 per year),more and more complications of the anesthesia and pain medicine increased in the number of reports.So,the neurotoxicity of the local anesthesia and whether drugs can inhibit the apoptosis of neuron becomes one of the hot studies on Anesthesiology and pain medicine filed. The severity of histopathology and sensor motor function impairment is directly related to the local anesthetic potency,position,and rate of administration.Follow-up of patients who received uncomplicated spinal anesthesia revealed that some of them developed pain in the lower extremities after an initial full recovery.This painful condition that occurs in the immediate postoperative period was named "transient neurological symptoms"(TNS).the etiology,and occurred mechanism were not clear by now,clinical symptoms always appears in 24h of the anesthesia,a few case lasts for 2 or 5d even 10d.Chiral local anesthetics,such as ropivacaine,have been introduced into clinical anesthesia because of their potential advantage over racemic mixtures in terms of reduced toxic effects,especially to convulsions and cardiac dysrhythmias.Peripheral neurotoxicity of the ropivacaine,the duration,reversibility,etiology,occurred mechanism becomes the hot study.P38 mitogen-activated protein kinas(p38) are a member of MAP kinas family, which includes at least ERK1/2,JNK,ERK5,and P38.Its wide-spectrum roles in the control of energy metabolism have been indicated in numerous studies.Recent investigations demonstrate that LA induces apoptosis in neuron cultures in vitro.P38 MAPK inhibitor is effective in attenuating LA-induced neurotoxicity.Polydatin(PD) can take pharmacologic action through hybridization of resveratrol(Res).Some researchers have found that Res can attenuate spinal cord injury to promote nerve regeneration and functional restoration.Whether Res can be used to prevent the neurotoxicity induced by LA or not,it is unknown and need to be proved through further research.So we want to do some tests to investigate effects of intravenous(i.v.) PD on the neurotoxicity induced by ropivacaine injected intrathecally(i.t.) in rats.MethodsPart 1 the effect of polydatin(PD) on sensor motor function of rats after Continuous Spinal Anesthesia With 2%RopivacaineObjective To study the effect of polydatin(PD) on sensor motor function after Continuous Spinal Anesthesia With 2%RopivacaineMethods Eighteen healthy,adult,male Spraque-Dawley(SD) rats were randomly divided in to sham-operated group,ropivacaine group,and PD-treated group according to completely random design.Each group has six rats.We established intrathecal models.After the surgery,the rats were fed in the cage alone for 2 days.We checked the performance of the rats every day.The rats which had behavior disorder were excluded.Group sham-operated received 40μl artificial cerebrospinal fluid i.t 3 times at 1.5 h interval and served as control;group R received 2%ropivacaine 40μl i.t 3 times at 1.5 h interval;group PD received polydatin i.v. 10mg/kg once before the inthrathecal and one time/each day afar,this group received 2%ropivacaine 40μl i.t also 3 times at 1.5 h interval.Spontaneous locomotion in an open field,foot splays in hind limb landing,nervous sensory to pain and heat in hind limbs were assessed one day before and daily after the first intrathecal administration. Using SPSS 13.0 software for statistical treatment,the results were expressed as mean±standard deviation.Comparisons among groups were made by Repeated Measures.Differences were considered significant at P＜0.05.Results Group of rats given 2%ropivacaine developed significant increase in the threshold of sensory nerve to heat and pain,but it had no significant change in sham-operated group.The hind limb landing foot splays was increased more in rats given 2%bupivacaine also.PD-treated group makes less neurotoxicity than 2% ropivacaine.Conclusion PD can decrease the sensor motor function change induced by continuous spinal anesthesia with 2%ropivacaine in rats.Part 2 the effect of polydatin(PD) on the number of apoptotie cells and the expression of P-P38 protein of rats after Continuous Spinal Anesthesia With 2% Ropivaeaine.Objective To observe HE dyeing and measure the number of apoptotic cells and p-p38 protein positive cells in nerve root with TUNEL or immunohistochemistry after continuous spinal anesthesia with 2%ropivacaine.To make sure if PD execute protection on the nerve injury by inhibiting apoptosis.Methods Thirty-six healthy,adult,male Spraque-Dawley(SD) rats were divided into control group(n=6),sham-operated group(n=6),R group(n=6), SB203580-treated group(n=6) and PD-treated group(n=6).To establish models and enroll rats as the same as part 1.There was done nothing to control group.Group sham-operated received 40μl artificial cerebrospinal fluid i.t 3 times at 1.5 h interval and served as control;group R received 2%ropivacaine 40μl i.t 3 times at 1.5 h interval;group PD received polydatin i.v.10mg/kg once before the inthrathecal and one time/each day afar,this group received 2%ropivacaine 40μl i.t also 3 times at 1.5 h interval.SB203580-treated group intrathecal SB203580 5ug 2h before 2% ropivacaine 40μl i.t.Observe indexes:1.path biology:After 3 days animals were anesthetized with chloral hydrate(30ml/kg) and fixed with normal saline 250ml and 4%Para formaldehyde 300ml through apex of heart.The nerve roots were removed from the spinal.The nerve roots were cut and used for HE staining and TUNEL for apoptosis and immunohistochemical analysis for p-p38 protein.Using SPSS 13.0 software for statistical treatment,the results were expressed as mean±standard deviation.Comparisons among groups were made by analysis of variance(ANOVA). Differences were considered significant at P＜0.05.Results 1,HE stain:Compared with the R group,SB203580-treated group and PD-treated group decreased neuronal degeneration,ischemia,cellular necrosis and interstitial edema.2.Compare SB203580-treated group and PD-treat group with R group,the count of TUNEL and Bcl-2 positive cells increased,and p-p38 positive cells decreased.Conclusion PD executed protection on the nerve injury induced by continuous spinal anesthesia with 2%ropivacaine.It decreased the number of apoptotic cells in the initial three days of ropivacaine injury by and down-regulating p-p38.Part 3 the effect of Extract from Rabbit Skin Inflamed by Vaccinia Virus for Injection on sensor motor function of rats after Continuous Spinal Anesthesia with 2%RopivacaineObjective To study the effect of Extract from Rabbit Skin Inflamed by Vaccinia Virus for Injection(AGC) on sensor motor function after continuous spinal anesthesia with 2%ropivacaineMethods Thirty-six healthy,adult,male Spraque-Dawley(SD) rats were randomly divided in to sham-operated group,ropivacaine group,and AG-treated group according to completely random design.Each group has 12 rats.We established intrathecal models.After the surgery,the rats were fed in the cage alone for 2 days.We checked the performance of the rats every day.The rats which had behavior disorder were excluded.Group sham-operated received 40μl artificial cerebrospinal fluid i.t 3 times at 1.5 h interval and served as control;group R received 2%ropivacaine 40μl i.t 3 times at 1.5 h interval;group AGC received Ext ract from Rabbit Skin Inflamed by Vaccinia Virus for Injection i.v.10u/kg/d once before the inthrathecal and one time/each day after,this group received 2%ropivacaine 40μl i.t also 3 times at 1.5 h interval.Spontaneous locomotion in an open field,foot splays in hind limb landing,nervous sensory to pain and heat in hind limbs were assessed one day before and daily after the first intrathecal administration.Using SPSS 13.0 software for statistical treatment,the results were expressed as mean±standard deviation.Comparisons among groups were made by Repeated Measures. Differences were considered significant at P＜0.05.Results Group of rats given 2%ropivacaine developed significant increase in the threshold of sensory nerve to heat and pain,but it had no significant change in sham-operated group.The hind limb landing foot splays was increased more in rats given 2%bupivacaine also.AGC-treated group makes less neurotoxicity than 2% ropivacaine.Conclusion AGC can decrease the sensor motor function change induced by continuous spinal anesthesia with 2%ropivacaine in rats. |
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