| Study objective: By observing the effects of Yaotuitong Capsule on lumbar disc herniation of lower extremity mechanical stimulation pain threshold in a rat model,and expression level of tumor necrosis factor-? in dorsal root ganglion and interleukin-1? and interleukin-6 and its regulation,this study explores the action target of Yaotuitong Capsule in treating lumbar disc herniation,and provide experimental basis for clinical treatment of lumbar disc herniation.Methods:Sixty SPF male Sprague-Dawley rats were randomly divided into blank group(K),model group(M),sham-operated group(J),high dose group(G),medium dose group(Z)and low dose group(D)of Yaotuitong Capsule(10 rats in each group).According to the previous study,the rat model of epidural lumbar disc herniation with autologous nucleus pulposus transplantation was established,the sham-operated group only exposed dorsal root ganglion,and the dose group was given the suspension solution of Yaotuitong Capsule and purified water daily after making the model.According to the previous study,the doses were 1.728g/kg/day/rat for G group,0.864g/kg/day/rat for Z group,0.432g/kg/day/rat for D group(dissolved in 2ml pure water),2ml pure water was given to model group and sham-operated group,and no intervention was made in blank group.Index test and record:1.The rats in each group were measured 1 day before modeling and on the 3rd,7th and 14 th day after modeling,the quality of life of the rats in each group was observed;2.The rats in each group were measured 1 day before modeling and on the 3rd,7th and 14 th day after modeling of the the withdrawal latency of mechanical stimulation of the right hind paw,and calculated the changes of mechanical stimulation pain threshold in rats;3.The rats were killed after 14 days' administration,using the ELISA method to detect the the content of tumor necrosis factor-?,interleukin-1? and interleukin-6 in dorsal root ganglion of the remaining large rat;4.two rats were randomly selected from each group,dorsal root ganglion was abstracted and made paraffin sections,with hematoxylin eosin(HE)staining,the neuronal cell morphological changes was observed under the light microscope.Research result:1.Quality of life observation: one day before the model,the rats in each group were in a good mental state,free movement,normal diet and defecation and urination.One day after the establishment of the model,the rats in each group,except group K,were mentally poor,their activities slowed down,and the right hind limb appeared limping and even dragging phenomenon.There was no incontinence,but only slow down action was appeared in J group,and no limp of right hind limb was found in J group.After 3 to 7 days of modeling,except K group and J group,the rats in each group were slightly better in spirit,easily irritated,limping and dragging of right hind limb,diet was fine.Different degrees of diarrhea appeared in Z and G group.After 8to 14 days of model making,the rats in each group,except G group,had general fine spirit and slow response,and the lower extremity activity of the intervention group was better than before,and the rats in G and Z group had different degrees of diarrhea still.The state of M group was basically the same as before.2.Measurement of mechanical stimulation pain threshold of right hind limb: 1day before modeling,there was no significant difference between the right hind limb mechanical stimulation pain threshold among the groups.The difference was not statistically significant(P>0.05),and the right hind limb mechanical pain threshold in group K was compared with each other at different observation time points.The difference was not statistically significant(P>0.05)and could be used as a reference.Compared with the intervention time point,the mechanical pain threshold of all the groups except K group was decreased.The difference was statistically significant(P<0.05);after 14 days of modeling,comparing that of 3 days,the pain threshold of D,Z and G group increased.The difference was statistically significant(P<0.05),which indicated that the treatment was effective and could raise the pain threshold.Compared from the intervention group,the pain threshold of M group and K group decreased at each time point after modeling,the difference was statistically significant(P<0.05),there was no significant difference among M,D,Z and G group in 3 days after model making(P<0.05),and between 3 days and 14 days after modeling,comparing with M group,mechanical stimulation pain threshold in D,Z,G group didnot continue to decrease,and the pain threshold was increased on the 14 th day of the modeling comparing with that of the 7th day.The difference was statistically significant(P<0.05),and it was the most obvious in G group(P<0.05).3.14 days after modeling,rats were detected with the content of tumor necrosis factor-?,interleukin-1?,interleukin-6 in dorsal root ganglion and comparison was made.M group was significantly higher than that in K group,there was a statistically significant difference(P<0.05);G group,Z group and D group were lower compared with M group,there is a statistically significant difference(P<0.05);the three groups D,Z and G decreased in sequence;J group,compared with K group had no obvious change,the difference was not statistically significant(P>0.05).4.14 days after modeling,rats were detected with the content of tumor necrosis factor-?,interleukin-1?,interleukin-6 in serum and comparison was made.M group was significantly higher than that in K group,there was a statistically significant difference(P<0.05);G group,Z group and D group were lower compared with M group,there is a statistically significant difference(P<0.05);the three groups D,Z and G decreased in sequence;J group,compared with K group had no obvious change,the difference was not statistically significant(P>0.05).5.Morphological observation of rat dorsal root ganglion HE staining under light microscope: In the blank group,the nucleus of the rat was located in the center of the cell,and the density was uniform,the nucleolus was relatively clear,there were evenly distributed granules(Nissl bodies)in the cytoplasm.The neurons in the dorsal root ganglion of the model group were obviously swollen,some of the cells were apoptotic,the nucleus was irregularly changed,the nucleus was not clear,the cytoplasm was vacuolated and the arrangement of Nissl bodies in the cytoplasm was uneven change.A small number of nuclear shifts were observed in neurons in the dorsal root ganglion in the high,middle and low groups of Yaotuitong capsules,the edge of the nucleus was clearer than the model group,the density was more uniform,the nucleolus was clearer,and the cytoplasm was smaller.The body distribution was relatively uniform,and a few vacuoles in the cytoplasm of neurons were observed.In the sham-operated group,no changes in the nucleus and nucleolus morphology anddensity were found in the neurons of the dorsal root ganglion,the Nissl bodies were dense in the cells,there was no cytoplasmic vacuole-like change and no swelling of the cells.Conclusion: The dose in Yaotuitong Capsule high,medium and low three groups could raise the mechanical stimulation pain threshold of right hind limb of rats,which may be related to the decreasing in dorsal root ganglion/serum of TNF-?,IL-1? and IL-6.Reducing the content of Inflammatory Factors in the dorsal root ganglion/serum is one of the possible effect mechanism of Yaotuitong Capsule in the treatment of lumbar disc herniation pain. |
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