| Objective and significanceCervical disc disease is the neck,bone,cartilage,ligaments became degeneration, the secondary stimulation or around the organizational structure of oppression such as the spinal cord and nerve roots,blood vessels,soft tissue,sympathetic and so on, caused a group of sick syndrome eventually.Epidemiological investigations revealed that the adults in our country in the rate of cervical syndrome approximately 3.8%to 17.6%.Conservative projections based on the current population,there are at least 50 to 100 million people are afflicted by cervical syndrome.Cervical spondylotic radiculopathy is the highest incidence of the cervical syndrome,about the total incidence of 60%to 70%.CSR is the nerve root injury by various factors which led invasion and its dominant regional sensory,motor and reflex obstacles for the pain caused to the main clinical manifestations of a disease.In the medical field on cervical syndrome understanding of the concept of the earliest,most from cervical spondylotic radiculopathy.CSR's main symptoms of nerve root pain.At present,the type of cervical nerve root pain of the illness the exact mechanism is not yet entirely clear,but more and more people think that oppression mechanical and biochemical factors in the pathogenesis of factors play an important role.Disc degeneration caused Osteophyte proliferation,proliferation of osteophyte nerve root compression caused nerve membrane without the protection of the nerve root to produce nerve edema and ischemia-based pathological changes.Lots of documents indicate that the disc degeneration of the nerve root injury and multiple inflammatory neurotransmitters,such as cytokines,leukotrienes,and tumor necrosis factor,immunoglobulin and interleukin and etc.However,machinery of oppression and stimulate synergies inflammatory nerve root caused more serious damage.Therefore,the prevention and treatment of cervical nerve root pain of the illness is the most effective way to protect and repair damaged nerve root.At present, there is no effective drug treatment of cervical nerve root pain of the illness;doctors use the combination therapy treatments such as traction,acupuncture,massage manipulation,physical therapy,etc.The severity of the patients may choose surgical treatment,but indications are the restrictions,and surgery risky,costly,and there still exist the possibility of recurrence.How to play in medical treatment of cervical nerve root disease on before us has become the subject.According to the nerve root of the characteristics of cervical disease,we developed a "JingZhuiShu" clinical use for many years to achieve a satisfactory effect.Animal experiments and clinical studies suggest that "JingZhuiShu " to improve microcirculation and reduce platelet aggregation,lower blood viscosity, increased flexibility vessel wall,expanding blood vessels,can significantly improve the nerve root in patients with cervical pain,numbness,stiffness,cold,The inability of the symptoms of muscular dystrophy.Puerarin the main contain isoflavones, triterpene saponins and alkaloids,etc.Of these components,puerarin(Pur) as its main active ingredient,the chemical name is 8 -β-D-glucopyranosyl-4',7 - dihydroxy Isoflavones.Studies have shown that puerarin regulate bone metabolism and improve the body blood circulation,reducing myocardial oxygen consumption,lower blood sugar,prevent hypertension and arteriosclerosis,protect nerve cells,anti-liver drug, anti-inflammatory,expectorant,antipyretic and improve body immunity,antibacterial, antiviral and other pharmacological effect.In order to study the puerarin treatment of cervical nerve root pain Binggen of the possible mechanisms of puerarin on the cervical nerve root damage to repair,we learn from the behavior of nerve morphological,physiological and neurological perspective.Our aim is not only to explore both the puerarin cervical nerve root of the root of pain and damage repair of nerve root,but also to clear Chinese herbal compound,"JingZhuiShu" of the effect of mechanisms to provide experimental basis.Method and content1.Puerarin Lobata(Willd.) Ohwi Identification.TLC Identification According to TLC(AppendixⅥB) demands of a test solution of the lessons of these two 10μl,respectively,at the same point in sodium carboxymethyl cellulose for the silicone adhesive H plate,into strips,chloroform -methanol - water(7:2.5:0.25) for the launch of start,removed,dried,home UV light (365 nm) under review.In the chromatogram of the corresponding position for the test and reference substance two chromatograms of the same color have shown that the fluorescence spots.According to the 2005 edition of "Chinese Pharmacopoeia (1)",Puerarin in dry Puerarin Lobata(Willd.) Ohwi content of not less than 2.4%.2.Puerarin optimum extraction of the establishment and the establishment of standards for quality control.Puerarin to target for inspection,the proportion of raw materials solution (14:1,16:1,18:1),the extraction time(1,1.5,2) h,extraction times(1,2,3) for the inspection,by L9(34) orthogonal table orthogonal test,compare water extraction and ethanol extraction method to determine the best Puerarin extraction technology and quality control standards.Extraction Process in accordance with the best verification test done three batches obtained RSD value,calculating the average concentration.3.Model and intervention.All rats in the left-SLSEP preoperative detection,detection of the normal access. According to the literature Oxfam spinal surgery modeling interpolation line,slightly improved:to 0.8 cm in length,has a diameter of 0.55 mm nylon fishing line interpolation of a line,C7 laminectomy fenestration,were inserted along the longitudinal axis to the spinal cord C7-T1 and C5-C7 nerve roots,some overlapping, firmly embedded in the bone of the spinal canal.A total of seven rats in surgery or died after surgery.More than 53 rats in the first five days after receiving evaluation, selection meet the following conditions:①model around the side of E'-C6 Lat for the growth of 0.2-0.4 ms;②gait score of 2-3 points,no symptoms of spinal cord compression,a total of 47 rats into the group.Random numbers,use excel balanced generate five treatment groups,each with eight:saline group,puerarin extract of low, medium and high-dose group,control group.Control group with "Jingfukang" of particle density of 1:1 made solution,all treatment groups will be concentrated liquid capacity to 1 ml,by gavages' method of administration.4.The evaluation of the extraction of Puerarin lobata(Willd.) Ohwi to restoration of the injured nerve behavior observed in rats.Measurement of body weight in the experimental rats at the beginning and end; 9-10.30 everyday observed in rats placed in the box observation after 30 min:See whether there are rats autophagy,licking bite,scream and recorded within 5 min of the affected limbs from the cumulative time,the calculation of the average of three times;Use the method of Dubuisson and Kawakami and evaluate pain from the contraction of gait disturbance:1 = normal gait,no foot deformities;= 2 damage feet touch the glass surface,walking limp;foot or huddling malformation;3 = damaged foot lift,walking cannot touch the table,foot deformity or crouch obviously.Median nerve short-latency somatosensory evoked potentials(SLSEP) Determination.Use acupuncture needles stimulate and record the reference electrode placed 1 cm subcutaneous rats and stimulating the median nerve.Recorded and calculated operated side Erb's point C6 laminectomy,and the latency point margin(E'-C6 Lat), the C6 laminectomy point amplitude(C6 Amp),and C6 Erb's point distance measurement points laminectomy,somatosensory evoked potentials calculated conduction velocity(MCV).Detection of indicators after the completion of large doses of narcotic death, pressure line quickly removed spinal cord and nerve roots,fixed,paraffin-embedded, HE staining,was observed under light microscope axonal,and the myelin sheath of the neurons and neural patterns and endometrial stromal vascular changes,whether there are abnormal substances deposition and the infiltration of inflammatory cells, such as to the nature of specific diseases;each group respectively statistics:nerve cells vacuolar changes,necrotic foci showed loss of myelin axons distorted fish bone-shaped lesions,such as the number of cases positive.5.The statistical analysis of experimental data. Extraction of optimizing the use of orthogonal design,SPSS 11.5 single factor to the use of repeated measurement data analysis of variance(Repeated Measures), Mauchly spherical test results if P＞0.05 meets spherical assumption,without correction;if P≤0.05,spherical not meet the assumptions used s correction coefficient of Greenhouse-Geisser correction results.Measurement data with mean±standard deviation((?)±s) described before and after treatment using paired t test, each group compared with one-way analysis of variance(One-Way ANOVA).ResultThe rats had no significant weight loss,wound healing well,no self-licking macrophages and bite,no Nongxing secretions.1.The EP on the treatment of pain in rats.After 5 days,the evaluations of each group were spontaneous acts of pain and abnormal gait score difference was not significant(P＞0.05).The end of the experiment,the rats saline group still bending close upper chest,to avoid conflict with its foreleg,the touchdown site for the elbow,five minutes from the time percentile forelimb no significant changes in gait stumble,body tilt;EP high,medium and low dose treatment group pain-related behavior in rats reduced significantly limbs tend to support equal weight,or claw back contravention of the wrist,a slightly uneven gait;2.The EP on rat nerve root injury treatment.Saline group of rats observed histologically visible morphological neuronal death,false unipolar nerve cell structure disappeared,a satellite cells surrounded by empty;axons after nerve cell membrane structure and unclear,Schwann cell accumulation shaft a sudden rupture,the surrounding myelin disappeared,a hollow; spinal cord white matter after the cable,a cable lateral vacuolar degeneration of spinal cord gray matter,white matter gliosis were.Puerarin extract high,medium and low dose group and neck nerve injury rehabilitation lesser extent,there ganglion cell degeneration and necrosis and myelin fish bone changes were fewer,suggesting that the extract of Pueraria damage in rats nerve restoration and protection.3.The EP on the nerve root injury Electroneurophysiological therapeutic effect. On the group on rat(C6 Amp) impact.Score results among different groups had no significant difference(F = 6.878,P＜0.001),and score the results before and after treatment were significantly different(F = 1394.719,P＜0.001) between different groups,in addition to the normal saline group(t = 0.890,P = 0.403) with no significant difference in the rest of the group,paired t test,the results were significantly different,t = 28.714,17.540,12.529 and 21.616,respectively,P＜0.001. From different groups,the treatment did not statistically significant difference(F = 0.822,P = 0.520),post-treatment score difference in the results were significantly different(F = 14.432,P＜0.001),in which the carotid rehabilitation of the most significant group.Groups with the interaction between before and after treatment effect(F = 93.059,P＜0.001),by the table,we can see that after treatment the maximum amplitude in the neck rehabilitation group,the minimum in the saline group after treatment;On the group on rat(E'-C6Lat) impact.Score results among different groups had no significant difference(F = 20.025,P＜0.001),and score the results before and after treatment were significantly different(F = 384.062,P＜0.001) between different groups,in addition to the normal saline group(t = 0.314,P = 0.763) with no significant difference in the rest of the group,paired t test,the results were significantly different,t = 11.081,9.178,16.024 and 10.299,respectively,P＜0.001. From different groups,the treatment did not statistically significant difference(F = 0.368,P = 0.830),post-treatment score difference in the results were significantly different(F = 39.430,P＜0.001),in which the carotid rehabilitation of the most significant group.Groups with the interaction between before and after treatment effect(F = 27.133,P＜0.001),by the table,we can see that the maximum incubation period after treatment in the saline group,the minimum rehabilitation in the treatment of persistent group;On the group on rat(MCV) impact.The results among different groups score There was a significant difference(F = 3.183,P = 0.025),the score results before and after treatment were significantly different(F = 187.437,P＜0.001) between different groups,and normal saline group(t = 0.708,P = 0.502) with no significant difference between the high dose group(t = 9.642,P＜0.001) in the dose group(t = 5.170,P = 0.001) and low dose group(t = 14.803,P＜0.001) results there was significant difference.From different groups,the treatment did not statistically significant difference(F=0.394,P = 0.812),post-treatment score difference in the results were significantly different(F = 9.327,P＜0.001),with the high dose group of the most notable.Groups with the interaction between before and after treatment effect(F = 16.372,P＜0.001),by the table,we can see that the maximum rate after treatment in the neck rehabilitation group,the minimum in the saline group after treatment;Conclusions1.Extraction of Puerarin lobata(Willd.) Ohwi have better analgesic effect on nerve root type of cervical nerve root pain.When the Puerarin of Puerarin lobata(Willd.) Ohwi in the concentration of 266.1,133.0,66.5 mg ~ kg"l ~ dl,can significantly reduce pain in rats,and there are certain dose-effect relationship;2.EP each dose group can significantly improve the structure of nerve damage, reduced ganglion cell degeneration and necrosis,and the myelin sheath of fish bone changes;3.In the impact of electrophysiology study,EP-dose groups can increase the volatility,reduce the incubation period and accelerate somatosensory evoked potential conduction velocity,and prompts a certain dose-effect relationship.
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