The Influence Of Reperfusion In Different Time On Peripheral Nerves And Muscles After Subclavian Artery Injury

Objective:when high-level upper artery was injured, rete arteriosum of shoulder can not play a role in blood-supply,make upper limb show the signs of ischemia,eg: pallor,pain,paresthesia,paralysis,ulseless.in clinical, after high-level upper artery was injured, forearm appears ischemia ,eg:reduction of temperature, myodynamia ,endurance , necrosis and gradual fibration of partial muscle. At present, we have deeply known about the pathogenesis of ischemia reperfusion injury on and muscles ,but the pathogenesis of peripheral nerves and muscles in circumstance of partial ischemia still remain to be seen.so we designed the experiments, because upper limb`s nerve and artry of humans are as similar as rats. We make the model of partial ischemia ,rat is the experimental subject .after reperfusion in different time,we observe the influence of ischemia reperfusion injury on peripheral nerves by EMG(Electromyography); we observe the variation of sensory nerve by PWTL(Paw withdrawal thermal latency), procured specimens.we observe aerobic and anaerobic metabolism by admeasurement of enzymology .we observe the fibrillation of muscle by histomorphology,we observe the functional change of motor end plate by admeasurement of acetylcholinesterase. With the research mentioned above,by research the pathological process of high-level upper artery injury,to supply a theoretical direction how to choose method of operation,and how to evaluat prognosis of subclavian artery injury.Method:(1) Material:①Experimental animal:30 SD rats (supplyed by HEBEI Medical University Animal Lab ), clean grade, irrespective of gender, weight250-300g. ②Equipment: VIKINGQUEST myoelectricapparatus, BME-410C heat– pain stimulation instrument(light source is 12V/35W halogen light) .(2) Method:a. Management of experimental animal: Lab rats were divided into 6 groups, named ABCDEF, There were five rats in every group. A and B is control group,C,D,E and F were experimental group, A was normal, after intraperitonealanesthesia, made an incision at skin of subclavian, discovered 2nd section of subclavian artery,took the electrophsiological detection of peripheral nerve , zymologic assay and histological morphology. B was Sham operation group, discovered 2nd section of subclavian artery used same method, placed a vascular clamp between artery and nerve.30 hours later, took the electrophsiological detection of peripheral nerve , zymologic assay and histological morphology.C,D,E,F blocked the 2nd section of subclavian artery, reperfused at 6,12,18,24 hours later, took the electrophsiological detection of peripheral nerve , zymologic assay and histological morphology.b. Each group measured PWTL,befor took the electrophsiological detection of peripheral nerve , zymologic assay and histological morphology.(3) Experiment:a.Used VIKINGQUEST myoelectricapparatus(supplyed by EMG Room of 3ed affiliated hospitao of HEBEI medical university),by various current intensity, excited median nerve from subclavian to elbow about 2cm by square wave- pulse of 0.2 ms wavelength, recordde incubation and amplitude of maximum motion evoked potential.b. Used BME-410C heat– pain stimulation instrument(light source is 12V/35W halogen light), measure PWTL , repeated measure 3 times, average value was the statistical data.c. Zymologic assay: Biochemical method was used to detect Na+-k+ATPase concentration to observe the level of ischaemia of muscle and energy metabolic level . Biochemical method was used to detect acetylcholinesterase concentration to observe the change of motor end plate.d. Histological morphology:①HE staining method stain the sample of muscle below eye shot of 40 times microscope. Observe myocyte`s arrange, shape, colouration, swelling , necrosis , inflammatory cell infiltrates and so on.②MASSON staining method stain the sample of muscle eye shot of 40 times microscope. Observe fibrotic changes of muscle and fibroplasticproliferation.e. Statistical Methods: Analysis of system was SPSS11.5 software, One-way analysis of variance were carried out for incubation , amplitude of maximum motion evoked potential, PWTL, Na+-k+ATPase concentration and acetylcholinesterase concentration.Result:1. The corresponding parameters in A and B groups had no difference, influence of vascular clamp and operation was debuged.2. Result of EMG: The EMG parameters in A and C groups had no difference in incubation, The incubation parameters in D and E groups had no difference .But incubation of D and E groups is lower than A and B groups(P<0.05). incubation of F groups is lower than A B C D and E groups(P<0.05). NCV(Nerve conduction velocity) drooped,when ischemia time exceeded 12 hours .The EMG parameters in D E and F groups had no difference in amplitude of maximum motion evoked potential,but maximum motion evoked potential of D and E groups is lower than A B and C groups(P<0.05). maximum motion evoked potential reduced,when ischemia time exceeded 12 hours, then reached a plateau .3. PWTL: The PWTL parameters in A B C and D groups had no difference .PWTL was extensed at 18 hours of ischemia time, PWTL of E groups is longer than A B C and D groups (P<0.05). PWTL of F groups is longer than A B C D and E groups (P<0.05). Forearm of rat appeared dysesthesiaat E groups4. Na+-k+ATPase concentration: The parameters in A B C D and E groups had no difference in Na+-k+ATPase concentration. Na+-k+ATPase concentration of F groups decreased (P<0.05). cell of muscle appeared metabolic disorder,and it was reversible in E groups . 5. Acetylcholinesterase concentration: The parameters in all groups had no difference in acetylcholinesterase concentration. account for ,motor endplate was not injured.6. Histological morphology: Muscle was ordinary in HE staining,there was not necrosis and fibrillation. Muscle was ordinary in MASSON staining, besides fibroplasticproliferation was discovered in a part of slices of E and F groups.Conclusion:1. The time of ischemia–reperfusion have effect on NCV and maximum motion evoked potential with subclavian artery Injury , not only NCV and maximum motion evoked potential reduced at 30days age,when the time was over 12 hours,but also NCV was reduced gradually.2. The time of ischemia–reperfusion have effect on sensory nerve with subclavian artery Injury , sensory disorder recovered in 30days,when the time was not over 12 hours,.sensory disorder could ont recovere in 30days,when the time of ischemia–reperfusion was over 18 and 24 hours3. The time of ischemia–reperfusion have effect on energy metabolism of muscle with subclavian artery Injury , Na+-k+ATPase concentration could ont recovere in 30days, when the time of ischemia–reperfusion was over 24 hours.4. Function of motor end plate cuold recovere in 30days, when the time of ischemia–reperfusion was in 24 hours with subclavian artery Injury.