| Lumbar disc herniation (LDH) is a common reason that leads to low back pain, and it have a negative impact on the quality of people' s daily life, the incidence of which is going up gradually. The epidemiologic dates in the world indicates that the incidence of LDH in adults is 2%, and there is no sexual difference. The primary cause leading to low back pain is that hernial disc crackdowns nerve roots directly. But clinic evidences proved that low back pain is often associated with radiculitis. Furthermore, the inflammation may induce spinal cord injury in the vicinity, which may affect the nerve fibre regeneration. There are two crucial steps to treat radiculitis, and as the following: (l)Alleviating radiculitis inflammation as soon as possible;(2)Preventing and alleviating the secondary sequential spinal cord injury.The low back pain is a severe problem for the patients and family as well as the society, but until now it has no effective remedy and therapy method. Surgery may take out hernial disc, but the pain continues to exist. It is so urgent for us to develop a novel effective agent. Mailuoning, the compounds of Chinese traditional medicine is composed of hyssop, figwort, honeysuck, shihu, and so on. It can activate blood circulation, acesodyne and relief the inflammation, since 1989, professor Zhi have treated low back pain with mailuoning mixed with 2% lidocaine and 5% baking soda(mailuoning compound, MC ) through epidural injecting. The curative effects is satisfying. The elementary experiments indicate: mailuoning compounding does no harm to ligament, arachnoid membrane, nerve root, spinal cord, and so on. And long-term curative effects is good. This project is a research funded by CMDF, and its object is to systematically study the curative effects and mechanisms of mailuoning compound on radiculitis in rats and the experiment includes two parts.1. The experimental study on therapeutic effect of MC on radiculitis by epidural injecting.1.1 Objective to study the therapeutic effect of MC on radiculitis by epidural injecting, and change histopathology and also observe the effect of MC on regulating the reactive astrogliosis and promoting the regeneration of injured nerve fiber.1. 2 Material and methods There were 30 Wistar rats involved and 24 out of them were modeled in radiculitis at L2-L3 level of the cord, and then divided into three groups randomly: MC-treated group(A, 6 rats);Hormone-treated group(H, 6 rats);sham operation group(K, 6 rats);model group(S, 6 rats). The rats in A & H groups were treated with MC and hormone respectively by epidural injecting with a dosage of 6ml/kg each time, and the total was 4 times. The rats in K group were feed with equal Saline. The other 6 rats were the control(Z). After 18 days, the rats were killed for histopathological examination. The process was that rats were first narcosised by injecting 1% soluble pentobarbital into abdominal cavity and then perfused with 4% polyformaldehyde solution from a syringe needle in the aortic through the left cardial ventricle. The phlogistic nerve root and adjacent spinal cord were removed from the rats, and made into paraffin sections. The sections were examed with some methods included the Nissl staining, Weil' s method, and immunohistochemistric staining for Glial fibrillary acidic protein (GFAP), Growth-associated protein-43(GAP-43) and Neurofilament (NF). The histopathological changes of the phlogistic nerve root and adjacent spinal cord were observed by light microscope and then the other markers were analyzed by 3-Y micro image analysis instrument including the optical density mean(OD) of normal neuron and neural fiber, and the optical density mean(OD) of GFAP> GAP-43^ NF immunopositive expressions. 1.3 results and conclusions1.3. 1 experimental study in effect of MC on nerve root and spinal cord histopathologic change after radiculitis in ratsResults: (l)In Nissl staining: OD of normal neuron in S group was decreased obviously, and the group K was as much as The group Z. after therapy, the OD of degeneration neuron became less and the normal neurons were increased in A and H group. (2) In Weil' s staining: The OD of the neural fiber in S group has evident differentiation compared with other groups, after treated, The OD of the neural fiber were increased obviously.Conclusion: (1) It is positive that injecting mailuoning compounding in epidural deals with radiculitis rats. (2) MC is provided with protective effect for the neuron in the spinal cord adjacent phlogistic nerve root and also can facilitate the recovery of degenerating neuron. (3) MC can reduce the nervous fiber demyelination induced by radiculitis.1. 3. 2 The experimental study on alleviating the reactive astrogliosis in spinal cord following radiculitis with MC in rats.Results: (1) Reactive astrogliosis in implicated spinal area was obvious in S group, and the spinal damaged focus gradually developed a large and compact scarafter radiculitis, and usually involved in the left gray in the scar. The reactive astrogliosis was lighter and the scar is smaller in A and H groups. There was no the compact scar in the area near to central gray of the spinal cord. (2) The OD of GFAP protein was obviously increased in S group. Compared with Z and K group, these two parameters were significant higher in S group (P<0. 01). (3) There was no significant differentiation between Z groups and K groups of OD. (4) In group A and group H, there was no significant differentiation (P>0. 05)of the OD, compared with Z and K groups, there was also no significant differentiation (P>0. 05).Conclusion: (1) Reactive astrogliosis and cicatrization of glia-connective tissue was significantly induced by radiculitis in the spinal cord in adult rats. (2) mailuoning compounding can obviously mitigate the reactive astrogliosis and prevent from cicatrization induced by radiculitis. It is possible to improve microenvironment in the spinal cord injured resulting in radiculitis to make for repairing and regenerating of nervous tissue, that MC is.1. 3. 3 The experimental study on promoting regeneration of nervous fiber by MC Results: The observation of GAP-43 immunohistochemitric section: ? S group: There was a plenty of GAP-43-positive nerve fiber, and the OD of GAP-43-positive immunoreactivity granules was increased obviously at the phlogistic nerve root and the adjacent spinal cord. There was much different between S group and the others (P<0. 01) . (2) A and H groups: GAP-43-positive nerve fiber and the OD of GAP-43-positive granules were obviously decreased. There was no distinct different between the two groups (P>0. 05) . however there was different between A> H groups and Z> K groups.Conclusion: (1) The regeneration nerve fiber may extend into spinal cord after radiculitis. (2) injecting mailuoning compounding in epidural can shorten the regenerative time, and facilitate the nerve fiber regeneration.2. The experimental study in mechanism of curative effect and action of MC on radiculitisThe result of first part the topic show that MC is good effect and action on radiculitis in rats. MC can protect neuron from damage, inhibit reactive astrogliosis and promote repair and regeneration of injured nerve root. So we want to lucubrate the mechanism of effect and action of MC by second aspects including the change of inflammatory media and the mediation apoptosis related genes.2.1 Objective: To observe the effect of MC on changes of inflammatory media in blood serum and plasma, and the action of MC on apoptosis related genes, and to discuss the mechanism of curative the effect and action of MC.2. 2 Material and methods2.2. 1 material and methods of inflammatory media in blood: The model was ibid, after 18 days, samples of blood were taken from left ventricle of heart, and laid them on cuvettes respectively, and then taken out to radioimmunoassay. The assay included IL-6, IL-8, TXB2^ PRA> AT-II.2. 2. 2 material and methods of apoptosis related genes bcl-2 and Bax: The sample came from the spinal cord of rats that were radiculitis. According to request of SP immunohistochemistric staining, the paraffin section was made, the OD of bcl-2- and Bax-immunoactivity positive markers were measured with 3-Y micro image analysis system, and the data were treated by One-Way ANOVA. 2.3 results and conclusions: 2. 3.1 radioimmunoassay of inflammatory mediaResults (1) S group: the inflammatory media were obviously increased. Compared with other groups, there was distinct different (P<0. 01);(2) K group: there was no distinct different between K group and Z group (P>0. 05). (3) A and H groups: With mailuoning compounding and hormone respectively, the inflammatory media decreased. Compared with Z group, there was no distinct different (P>0. 05). Conclusions: (1) the change of inflammatory media in the blood can be resulted in the injury of radiculitis. (2) MC can reduce the inflammatory media in the blood. (3) The results showed that MC can protect the injured nervous tissue by rectification of the abnormal change of microcirculation and keep with limits the injury to a certain extent.2. 3. 2 Immunohistochemistric staining of gene expression related to the apoptosis Results: (1) S group: the OD of Bax-positive was obviously increased. Compared with Z group, there was distinct different (P<0.01);A and H groups: the OD of Bax-positive was decreased, compared with Z group, there was no distinct different (P>0.05). (2) S group: the 0D of bcl-2-positive was increased appreciably, however, compared with Z and K groups, there were no distinct different (P>0. 05). by therapy, the 0D of bcl-2-posotive was increased obviously, compared with S group, there was distinct different (P<0. 05), but there was no distinct different between A> H groups and Z> K groups. (3) Bax/bcl-2 ratio: Bax-positive neuron increased obviously after radiculitis, so Bax/bcl-2 invert. Through therapy, Bax-positive neuron decreased, at the same time, bcl-2-positive neuron increased, the Bax/bcl-2 ratio balanced again.Conclusions: MC can improve neural viability in spinal inflammation injured area by modulating the expression of genes related to apoptosis of the neuron and facilitating the neural repair by protecting neuron from apoptosis process induced by inflammation injury. It can be one of the mechanisms of MC on protectingneuron.Conclusion1. MC in epidural is provided with protective effect for the neuron in the inflammatory spinal cord and also can facilitate the recovery of degeneration neuron.2. MC in epidural can reduce the nervous fiber demyelination induced by radiculitis.3. MC in epidural can obviously mitigate reactive astrogliosis and prevent from glial scar induced by radiculitis, that is availability to CNS regeneration.4. MC can promote and support repair and regeneration of nerve fiber after radiculitis.5. MC has benign adjustment to disturbance of inflammatory factor in blood induced by radiculitis.6. MC can mitigate the apoptosis of neuron induced by radiculitis.7. MC is a needle dosage that be composed of herbs with supplement the vital energy and activating blood circulation effects including honeysuckle, hyssop, figwort, and so on. So the results of this study shows that mailuoning compounding with lidocaine and baking soda has large potential for treating low back pain resulting in radiculitis and promoting the regeneration of CNS. |
PDF Full Text Request