| ObjectivesObservation of spinal massage on cerebral palsy rats brain white matter inflammatory cytokines interleukin 6(IL-6) and interleukin 10(IL-10) effects and molecular mechanism of manipulation of the spinal column reconstruction of cerebral palsy inflammation steady-state provides the basis of the preliminary study.MethodsThe 108 3 day old SD rats, were randomly divided to 3 groups: blank control group(40 rats)and model group(a total of 40) and experimental group(a total of 28), model control group and the experimental group adopted the left common carotid artery ligation combined with hypoxia 3H method to prepare the model of cerebral palsy, the control group was not given any treatment, the normal feeding. At 24 h after modeling, from the blank control group and model control group randomly removed 6 rats were sacrificed for TTC staining, then randomly removed 6 rats were sacrificed for brain wet weight ratio test; on the same day, the experimental group began with spine massage method for early intervention, 1 times / day, 15 minutes each time and blank control model group and controls the rest of rats not any intervention. Three groups maintain the same temperature, moderate living environment and feeding method, at the same time detected and recorded in young rats, the mental state, hair moist degree and weight. 4 weeks(28 days postnatal,P28) and 8 weeks(P56) two time points, each group were taken randomly 7 rats, the cerebral white matter of the pathological changes were observed by HE staining; each of the remaining only 7 immunofluorescence detection of cerebral white matter in the region of IL-6 and IL-10 protein expression levels. Balance Test Beam(BBT) was performed in the first 1 week before death in order to observe the balance ability.Results1. Experimental group 4 weeks of mental state, reaction generally in good condition, and blank compared to the control group had no significant difference; compared with the model control group, model control group rats: the response is slow, binocular unequal. Experimental group 8weeks when the hair shiny gloss, good mental state, sensitive reaction, normal development, and blank control group compared with almost no difference; in the model control group rats hair color is dim, unresponsive, growth retardation, eyes of different sizes(eyes open, after the left eye fission is small). Daily weight gain: the blank control group were significantly increased compared with the model group(P<0.05). P13, p17, P19, p21 and p23, P25, the experimental group at 4weeks, the amount of diurnal variation of body weight were compared with model control group increased significantly(P < 0.05); P7, P11, p17, P19, p21 and p23, p27, P29, P31, P33, p35, P37,P39, p41, p43 P45 and P47, p53, the experimental group 8 weeks weight, variable volume were compared with the model control group increased significantly(P < 0.05). That spinal manipulation can accelerate the growth rate of body weight were promoting cerebral palsy. With a certain degree of dependence on the massage time, along with the increase of the number of massage, weight growth faster.2. The balance beam test, the experimental group at 4 weeks, compared with the 8 week number by balance time and found that the hind limbs decline through the balance beam time had no significant difference(P>0.05), but significant differences in the number of double hindlimb decline(P<0.001); at 4 weeks, compared with the control group, there were no significant differences in each index(P>0.05), compared with the model group, the balance time was significantly reduced(P<0.05), bipedal decline was significantly reduced(P<0.05); at 8 weeks,compared with the control group, all indexes were no significant difference(P>0.05), compared with the model group through the balance beam, shorten the time(P<0.05), bipedal decline was significantly reduced(P<0.05). The results showed that the spine massage can promote the improvement of ability to balance the motions of cerebral palsy rats, and massage time length,massage for a longer period of time, improve the better.3. HE staining showed that the experimental group at 4 weeks, there are different degrees of edema, necrosis and pycnosis of cerebral white matter in part neurons. The experimental group at8 weeks and 4 weeks compared partially damaged neurons edema, necrosis and nuclear pyknosis were significantly improved. At 4 weeks, found that the cerebral white matter in some neurons exist different degree edema, necrosis and nuclear pyknosis, 4 weeks and compared with the control group, the difference was not statistically significant, compared with the model group, the neuron cell edema, necrosis and nuclear pyknosis relatively light; compared with the blank control group, almost indistinguishable, compared with the model control group, neuronal cell edema,necrosis and nuclear pyknosis relatively light. The results indicate that spinal manipulation can promote the repair and regeneration of neurons in the cerebral white matter area, and the longer the massage time, the more obvious the recovery effect.4. Immunofluorescence showed that the abnormal expression of the experimental group at 4weeks within the cerebral white matter consists of inflammatory cytokines IL-6 and IL-10 were significantly improved. The experimental group at 8 weeks compared with 4 weeks found differences in expression of IL-6, improve the degree of inflammatory factor IL-10 protein level was significant(P<0.01). At 4 weeks, compared with the blank control group, IL-6 and IL-10 protein expression were significantly increased(P < 0.05), and model control group, IL-6 protein expression decreased significantly(P < 0.05) and IL-10 protein expression increased significantly(P < 0.05); at 8 weeks, compared with the blank control group, IL-6 and IL-10 protein expression were significantly increased(P < 0.05), and model control group, IL-6 protein expression decreased significantly(P < 0.05) and IL-10 protein expression increased significantly(P < 0.05).That spinal manipulation can effectively reduce the level of IL-6 protein in the white matter of the brain, and improve the level of IL-10 protein expression. Through massage can effectively participate in the regulation of abnormal expression of inflammatory cytokines IL-6 and IL-10,through the manipulation of the spinal column can modulate Pro inflammatory cytokines IL-6 and anti-inflammatory cytokine IL-10 protein level expression, and the regulating effect and massage time, massage the longer, the better.Conclusions1. Spinal massage can effectively promote the growth and development indexes of cerebral palsy rats, such as weight, diet, mental state etc.. And related to the length of massage, massage time is longer, the better the growth and development2. Manipulation of the spinal column can effectively improve the balance ability of rats with cerebral palsy(CP), and massage time length, massage for a longer period of time, improve the better.3. Spinal manipulation can improve the pathological morphology of neurons in the cerebral white matter area, reduce the level of IL-6 protein and increase the expression of anti-inflammatory factor IL-10 protein, and the relative steady state of inflammation. The linked in a way consistent with expression of inflammatory factors in serum mentioned in the literature,suggest that spinal manipulation of brain white matter pathology and spinal manipulation of CP rats of mobility and balance ability improvement and inflammatory cytokines expression.4. Manipulation of the spinal column can be by reducing the brain white matter pro inflammatory cytokines IL-6 and improve the anti-inflammatory cytokine IL-10 protein expression to reconstruct the inflammatory state of cerebral palsy, and promote the comprehensive rehabilitation of movement and other functions, for further study on regulation mechanism of lay the preliminary foundation... |
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