| Objective1.Clinical study:to evaluate the clinical efficacy and safety of acupressure on the pain,dysfunction,soft tissue tension of the waist and hips,and tenderness threshold in patients with lumbar disc herniation.2.Animal experiment:to investigate the effects of acupressure on the effects of acupressure on erector spinal muscles and lumbar intervertebral discs,as well as the effects on the Wnt/β-catenin signal pathway in the rats with lumbar intervertebral disc degeneration.Part Ⅰ Clinical StudyMethodsA randomized control clinical trial was performed.Eighty-six patients,diagnosed with lumbar intervertebral disc herniation,were equally randomized into 2 groups,which were the test group and the control group.Acupressure was administered in the test group,while tuina was applied in the control group.Both treatments were conducted twice a week,for 4 weeks.The Oswestry Disability Index(ODI)and visual analogue scale(VAS)were observed at 4 time points,which were before treatment,at 2 weeks,at the end of the course of treatment(4 weeks),and at 1-month follow up.Soft tissue tension and tenderness threshold were evaluated at 3 time points,which were before treatment,at 2 weeks,and at the end of the course of treatment(4 weeks).The demographic characteristics of the 2 groups were analyzed by baseline,and then the effectiveness and safety indicators of the two groups were compared.Normally distributed measurement data is described by the mean ±standard deviation(x±s),skewed distribution measurement data is described by the median(interquartile range)[M(Q)],and the number of count data use cases and percentage description.SPSS 22.0 statistical software was used for data analysis.The baseline comparison of measurement data uses analysis of variance or rank-sum test.The baseline comparison of count data uses chi-square test or Fisher test.The repeated measures analysis of variance was used to analyze the data at different observation time-points.The pairwise comparisons between groups at each time point adopted multivariate analysis of variance,and the repeated measures of binary data were statistically analyzed by generalized estimating equations.P<0.05 will be considered as statistically significant.The descriptive statistics is used for safety analysis,including the incidence of adverse events and specific descriptions of adverse events.ResultsDuring the course of treatment,2 cases fell off.The data of 86 cases were analyzed.There were 43 patients in each group.1.There was no statistically significant difference in the demographic characteristics and baseline conditions of the disease,and the pre-treatment level of each evaluation index between the test group and the control group(P>0.05).2.At the two-week time-point,the ODI and VAS scores of the two groups were significantly lower than pre-treatment,and the differences were statistically significant(P<0.05).At the four-week time-point,the ODI and VAS continued to decrease,and the difference was statistically significant(P<0.05).In the one-month follow-up,the downward trend of ODI and VAS scores stopped,and the difference was not statistically significant,compared with that at the four-week time-point.However,the ODI and VAS scores at the 1-month follow-up were still lower than those of pre-treatment,and the difference was statistically significant.3.As to the displacement values of the erector spinae and gluteus maximus under a load of 0.5 kg,upward trends were shown in the displacement values of the two muscles.In each group,the values of soft tissue displacement at 2-and 4-week time-point were significantly higher than that of pre-treatment(P<0.05).Within each group,the value of soft tissue displacement measured at 4-weeks was also higher than that of 2-weeks,and the difference was statistically significant(P<0.05).4.As to the tenderness threshold of erector spinae and gluteus maximus,upward trend was observed in the tenderness threshold within each group.The tenderness threshold measured at 2-and 4-week were both higher than that of pre-treatment,and the difference was statistically significant(P<0.05).The tenderness threshold at 4-weeks was also higher than that at 2-weeks,and the difference was still statistically significant(P<0.05).5.There were no significant differences in the ODI and VAS scores between the test group and the control group at the time-point of 2-,4-weeks,and 1 month of follow-up.At 2-and 4-week time-point,there was no statistically significant difference in the soft tissue tension of the erector spinae and gluteus maximus between two groups.At 2-week time-point,the tenderness threshold of erector spinae and gluteus maximus in the test group was larger than that of the control group,and the difference was statistically significant(P>0.05).However,there was no significant difference in the tenderness threshold between the two groups at 4 weeks of treatment.The results of the comparison of effects between subjects further indicated that there was no significant difference in the improvement of tenderness threshold between the two groups,suggesting that the two groups had a similar effect in improving tenderness threshold.6.In the test group,2 cases were cured,11 cases were markedly improved,27 cases were moderately improved,and 3 cases were ineffectively relieved.The total effective rate was 93%.In the control group,3 cases were cured and 8 cases were markedly relieved.Twenty-eight cases were moderately relieved but 4 cases were ineffectively alleviated.The total effective rate was 90.6%,and the difference was not statistically significant(P>0.05).ConclusionAcupressure could relieve the pain and disability symptoms caused by lumbar intervertebral disc herniation.And the soft-tissue tension and tenderness threshold were found alleviated after acupressure treatment.Part Ⅱ Animal ExperimentMethods1.AnimalsFifty-four Sprague Dawley(SD)rats participated in the experiment.Ten rats were randomized into the control group(CG).Fore limbs amputation surgeries were performed on the other 44 rats for modelling the bipedal rats.2.Experiments ProcedureWe established the lumbar intervertebral disc degeneration(IVDD)model by inducing bipedal rats to maintain upright posture for 5 months.In the last week of the modelling procedure,the lumbar intervertebral discs of 4 random rats were harvested to evaluate whether IVDD modelling was successful.The results showed that IVDD modelling had caused significant degenerative changes in the lumbar IVDs.As IVDD modelling finished,the 40 IVDD rats were equally randomized into 4 groups(n=10):acupressure group(AG),sham acupressure group(SAG),DKK-1 group(DG),and model group(MG),and interventions were given respectively once a day for 30 consecutive days.In AG,acupressure was applied with cotton swabs on the selected lumbar acupoints of rats,while sham acupressure was applied to the the acupoints of rats in SAG without pressure.The rats in DG received intraperitoneal injections of DKK-1(0.1μg/100g),which is the Wnt/β-catenin pathway antagonist.The rats in MG and CG received no intervention.The erector spinae were stained with hematoxylin and eosin(HE),while en bloc L4/5 intervertebral discs(IVDs)were stained with HE and safranin 0-fast green staining.The histological changes of IVDs,cartilage endplates(CEP),and erector spinae were observed.The histological changes of IVDs and CEPs were further evaluated by the Boos grading scale.Immunochemistry staining,western blot,and Real-Time Quantitative Polymerase Chain Reaction(RT-qPCR)were applied to detect the expression of Wnt3a,β-catenin,GSK-3β,and MMP-13 in IVD tissues.ResultsHistological evaluation showed that both acupressure and DKK-1 offered significant protective effects on erector spinae,IVDs and CEPs.As demonstrated by immunochemistry staining,WB,and RT-qPCR,Wnt3a expression was downregulated significantly in AG and DG,when compared with MG and SAG.Such downregulation led to upregulation of GSK-3β and promoted proteasomal degradation of β-catenin,consequently inhibiting the expression of MMP-13.ConclusionAcupressure could protect IVDs against degeneration by suppressing the Wnt/β-catenin signal pathway.This may be one of the mechanisms of acupressure treatment on lumbar intervertebral disc herniation. |
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