Study Of Methylprednisolone,electro-acupuncture Of Hua Tuo Jiaji Acupoint And Rat Amniotic Epithelial Cells Transplantation In Therapy Of Spinal Cord Injury

Spinal cord injury (SCI) is one common central nervous system(CNS) damage disease caused by traffic,movement or mining accidents,which is also the disease that renders the most paralysis,especially when it causes lots of motor and sensory dysfunctions below the SCI section , and consequently,both family and society suffer great losses and stress. Judging from experiment results and clinical observations since 1950s,the key to SCI recovery is that nerve fibers can grow in the spinal cord,and by its growth,it can penetrate the injured cicatrix,thus repairing neural pathway in the spinal cord,eventually obtaining the recovery of the motor and sensory functions. About 30 years ago , a disastrous earthquake in Tangshan,China,took away hundreds of thousands of lives,and a great many of the survivors were SCI patients,losing the ability to take care of themselves. WenChuan earthquake which occurred on May 12th,2008,also made about 2% of more than 370,000 survivors suffer from SCI. So far,the major treatment for SCI is still operation and hormone therapy. According to statistics,the average treatment fee for SCI patients is 750,000 dollars,and America spends over 6 billion on the treatment of SCI patients. Thus an economical and efficient treatment for SCI is badly needed.Combining the treatment methods of traditional Chinese medicine and Western medicine,the present study was aimed at giving a further research on the comprehensive effect of methylprednisolone (MP),electro- acupuncture and rat amniotic epithelial cells (AECs) transplantation to recovery for SCI. In the experiment,90 adult female Wistar rats were randomly divided into five groups, with 18 in the individual group,namely-SCI control group: Improved Allen s bump equipment was employed in the present experiment,no treatment after SCI was created. MP group:MP pulse treatment after SCI. Intravenous injection MP 30mg/kg immediately after SCI,repeated the same injection 4h later,following up with two times every day,injection of MP 30mg/kg,3 days in all. MP and Huatuo spine points electro-acupuncture group: electro-acupuncture treatment on Hua Tuo jiaji acupoint 4h after SCI,based on MP group. Two paires of acupoints both above and below the SCI section,applying the left and right acupoints alternatively. The acupuncture needles were 25mm long and 0.35 mm in diameter. Therapeutic instrument (6805-II, produced in Shanghai) was set,including anode and cathode,dilatational wave, frequency(1～2Hz)and intensity(0.3～1.0 mA),kept the muscle trembled slightly still for 15min,then one time every day,with 6 days in a treatment course,after 2 days interval,the second course started,3 courses in all. MP,electro-acupuncture and AECs group:on the basis of MP and Hua Tuo jiaji acupoint electro-acupuncture , AECs 5μl (intensity=1 107/μl) were transplanted with microinjector 7d after SCI. Sham group:neural scute was opened and spinal cord was exposed without SCI. On the 30th day after the surgery,a series of tests were performed including histological testing,fluorescein red (FR) retrograde tagging , neurofilament ( NF ),5-hydroxytryptamine(5-HT),calcitonin gene related peptide(CGRP)and glial fibrillary acidic protein ( GFAP ) immunofluorescence histochemistry observation,neuro-electro-physiological tests and BBB scores (every 6d).In MP,electro-acupuncture and AECs group GFAP was discovered to have an inadequate expression compared with other SCI groups in SCI region ,a magnificent amount of FR positive nerve fibers were spotted and can be transported through SCI region,and the amount was remarkably more than other discovered in other SCI groups. The fibers were regularly arranged,almost paralleled with each other and the FR coloring appeared to be more noticeable.FR positive cells were discovered in corresponding cerebrum cortical motor area and posterior horn of gray nucleus.Meanwhile,in SCI region appeared well-grown evenly distributed transplanted AECs,labeled by Hoechst33342 (blue).In MP,electro-acupuncture and AECs group,the SCI region held numerous regularly-arranged NF,5-HT and CGRP positive nerve fibers,and the 5-HT positive nerve fibers took up 61.5% of those in sham operation group,a notable advantage over CGRP,which took up no more than 7.6%. Neuro-electrophysiological tests on both Somatosensory evoked potential (SEP) and Moto evoked potential (MEP) demonstrated that peak-to-peak value increased sharply and latency was shortened noticeably in MP ,electro-acupuncture and AECs group,and difference between this and other SCI groups is statistically significant (P<0.01). MEP recovery is better than SEP. To conclude , a combination of methylprednisolone ,electro-acupuncture Hua Tuo jiaji acupoint and AECs transplantation can restrain hyperplasy of horizontal cell and recover the pathway of nerve conduction and promote nerve fibers regeneration and hind limb function recovery for SCI rats;furthermore,motor function was prior to sensory function in the recovery. The current study bring a new idea which the combination of MP,electro-acupuncture and AECs in rat SCI treatment. It brings a new train of thought for combined therapy of SCI,and a more simple and efficient way. Meanwhile it provides a new experimental basis for clinical treatment of SCI.