| ObjectiveTo analyze the relationship between divergent meridians classification and TCM syndrome type and syndrome elements of KOA,and to observe the clinical effect of Jingjintong decoction in the treatment of postmenopausal women with KOA divergent meridians lesions under the guidance of zang-fu differentiation,and then using the KOA model of ovariectomized rats to simulate postmenopausal women knee cartilage degeneration microenvironment,to explore the molecular mechanism of Jingjintong decoction in the treatment of ovariectomized rats with bones and muscles lesions.In order to clear Jingjintong decoction in the treatment of postmenopausal women with KOA divergent meridians lesions manifests TCM academic thought of searching for the primary cause of disease in treatment and treating both muscles and bones.MethodsSelecting the First Affiliated Hospital of Guangzhou University of Traditional Chinese Medicine outpatient postmenopausal patients with KOA,making selection criteria,patients who meet the criteria were included in the study,using the zang-fu differentiation to obtain the TCM syndrome type and syndrome elements,using divergent meridians differentiation to obtain the divergent meridians classification,building the database with the information obtained,and to observe the distribution of the divergent meridians classification in the TCM syndrome type and syndrome elements,SPSS18.0 software was used for the corresponding analysis.The selected subjects were divided into observation group and control group,the observation group was treated by Jingjintong decoction,once a day,6 times a week,2 weeks for a course of treatment,continuous taking 3 courses,the control group was treated by meridian electroacupuncture,every 2 days 1 times,3 times a week,2 weeks for a course of treatment,continuous treatment of 3 courses.After the treatment,WOMAC,JOA and Lysholm scores were used to evaluate knee function,the degree of pain relief was assessed using the VAS score,the occurrence of complications during treatment was recorded,and to observe the difference between the two groups.SPSS 18.0 software was used to statistically process the data,First,Performing homogeneity of variance,the independent sample t test was used if the variance was homogeneous,if not,Kruskal-Wallis nonparametric variance analysis was used,complications were compared using the χ2 test,the efficacy was compared using the Wilcoxon rank sum test.18 female Sprague-Dawley rats were randomly divided into OVX+KOA group,OVX group and Sham group,6 rats were in each group.The KOA model was established after 2 weeks of castration in OVX+KOA group,OVX group was treated with knee dislocation after ovariectomy,Sham group removed the adipose tissue adjacent to the ovaries and dislocated the knee,all rats run for 2 weeks at the animal test runway after modeling for 1 week,1.5 months later,serum E2 and COMP contents were measured by ELISA,the morphological changes of articular cartilage and gastrocnemius were observed by HE staining.SPSS18.0 software was used to statistically process the data,First,Performing homogeneity of variance,the independent sample t test was used if the variance was homogeneous,if not,Kruskal-Wallis nonparametric variance analysis was used.72 female Sprague-Dawley rats were randomly divided into Sham group,Model group,Jingjintong decoction high dose group,Jingjintong decoction middle dose group,Jingjintong decoction low dose group and Positive control group,12 rats were in each group.All rats run for 2 weeks at the animal test runway after modeling for 1 week,while taking drugs to intervene.The Jingjintong decoction high dose group,Jingjintong decoction middle dose group,and Jingjintong decoction low dose group were given Jingjintong decoction,the Positive control group was given celecoxib capsules.The dose of the Jingjintong decoction high dose group was 3.80g/100g/d,the Jingjintong decoction middle dose group was 1.90g/100g/d,the Jingjintong decoction low dose group was 0.95g/100g/d,the dose of the Positive control group was 2.06mg/100g/d,and Sham group and Model group were given the same volume of saline.The medication method was once a day,6 days a week,and rest 1 day,the administration time lasted for 1.5 months.After the administration,serum E2 and COMP contents were measured by ELISA,ERRα.,PGC-1 a,SOX9 and IL-1βmRNA expression in knee joint cartilage were detected by RT-PCR,the expression of type Ⅱ collagen in gastrocnemius muscle was detected by immunohistochemistry.SPSS18.0 software was used to statistically process the data,First,Performing homogeneity of variance,the independent sample t test was used if the variance was homogeneous,if not,Kruskal-Wallis nonparametric variance analysis was used.ResultsA total of 84 postmenopausal KOA patients were eligible for selection criteria.The divergent meridians lesions was mainly composed of musculature of stomach meridian,followed by musculature of bladder meridian,the three foot yin meridians and musculature of gall-bladder meridian,the proportion was 39%,23%,20%and 18%respectively.TCM syndrome type was mainly composed of blood stagnation,followed by deficiency of the liver and kidney,wind-cold-damp impediment and wind-damp-heat arthralgia,the proportion was 44.45%,30.95%,26.19%and 17.86%respectively.The analysis of correspondence between TCM syndrome type and divergent meridians classification manifested a property association,and there was a statistically significance(χ2=38.814,P=0.000).The point-to-point distance of TCM syndrome type and divergent meridians classification showed that there was closely related between musculature of stomach meridian and musculature of gall-bladder meridian and wind-cold-damp impediment and wind-damp-heat arthralgia,the three foot yin meridians and blood stagnation,musculature of bladder meridian and deficiency of the liver and kidney.Musculature of stomach meridian was mainly composed of pathogenic dampness,blood stasis and spleen deficiency,musculature of bladder meridian was mainly composed of kidney deficiency and liver deficiency,musculature of gall-bladder meridian was mainly composed of blood stasis and pathogenic dampness,the three foot yin meridians was mainly composed of blood stasis and liver deficiency.The analysis of correspondence between TCM syndrome elements and divergent meridians classification manifested there was no association and statistically significance(χ2=24.688,P=0.423).The above 84 cases of postmenopausal KOA patients were included in this study,12 cases did not meet the selection criteria to be removed,the remaining 72 patients met the selection criteria included in the study,including observation group of 38 cases,the control group of 34 cases.There were no significant differences in age,body weight,height,BMI,Menopause years,course of disease,side and Kellgren-Lawrence scores between the two groups(P>0.05).After treatment,the pain,stiffness,activity difficulty and overall score of the WOMAC score in the two groups were significantly reduced,there were significant differences comparing the pre-therapy(P<0.05),but there was no significant difference between the two groups(P>0.05),and no significant difference was found in the difference value of the two groups(P>0.05).The scores of JOA and Lysholm were significantly higher in both groups,there were significant differences comparing the pre-therapy(P<0.05),but there was no significant difference between the two groups(P>0.05),and no significant difference was found in the difference value of the two groups(P>0.05).VAS scores were significantly lower in both groups,there were significant differences comparing the pre-therapy(P<0.05),but there was no significant difference between the two groups(P>0.05),and no significant difference was found in the difference value of the two groups(P>0.05).The total effective rate was 89.47%in the observation group and 88.24%in the control group,there was no significant difference between the two groups by Wilcoxon rank sum test(Z=-0.367,P=0.714).During treatment,the two groups of patients did not complain clearly discomfort,and there were no digestive diseases and cardiovascular system symptoms,no halo needle,needle handle broken,allergic reactions and other complications were found,and other divergent meridians lesion and compound lesions were not appeared.1.5 months after modeling,three groups of body weight comparison by variance homogeneity test were P=0.527>0.05,indicating that the variance was homogeneous,the one-way ANOVA LSD method was used to find the difference was not statistically significant.The levels of serum E2 in OVX+KOA group and OVX group were significantly lower than those in Sham group(P<0.05),there was no significant difference between OVX+KOA group and OVX group(P>0.05).The COMP content of OVX group was higher than that of Sham group,but there was no significant difference between the two groups(P>0.05),the COMP content of OVX+KOA group was higher than that of OVX group and Sham group,the difference was statistically significant comparing with the Sham group(P>0.05),but there was no significant difference comparing with the OVX group(P>0.05).HE staining showed that the cartilage surface of the OVX+KOA group was rough,chapped or jagged,and fibrosis,the number of chondrocytes decreased,the nucleus was blue,the extracellular matrix was red,cartilage lacuna was vacancy,the tide mark was up shift,interrupted or multiple.The nuclei of gastrocnemius muscle were blue,distributed between the muscle fibers,and muscle fibers were polygonal.The muscle fibers of OVX+KOA group were adhesions and arranged disorder,OVX group and Sham group muscle fibers arranged neatly.Compared with Sham group,the number of nuclei of OVX+KOA group and OVX group decreased,and the muscle fiber width of OVX group was widened.Drug intervention for 1.5 months,comparison of multiple indicators among groups by the variance homogeneity test was P<0.005,that did not conform to variance homogeneity,so Kruskal-Wallis nonparametric analysis of variance was used.The serum E2 and COMP contents of Model group were statistically significant comparing with Sham group(P<0.05),the serum E2 and COMP contents of Jingjintong decoction high dose group,Jingjintong decoction middle dose group and Positive control group were statistically significant comparing with Model group(P<0.05),the serum E2 contents of Jingjintong decoction high dose group,Jingjintong decoction middle dose group were statistically significant comparing with Positive control group(P<0.05).Compared with Sham group,the expression of ERR a and SOX9 mRNA of Model group was significantly decreased(P<0.05),PGC-1α and IL-1 βmRNA expression was significantly increased(P<0.05).After Jingjintong decoction intervention,compared with Model group,the expression of ERR a and SOX9 mRNA of Jingjintong decoction high dose group was significantly increased(P<0.05),and the expression of PGC-1α and IL-1β mRNA was significantly decreased(P<0.05),the expression of ERR a mRNA of Jingjintong decoction middle dose group was significantly increased(P<0.05),and the expression of PGC-1α、IL-1β mRNA was significantly decreased(P<0.05),the expression of PGC-1α、IL-1β mRNA was significantly decreased in Jingjintong decoction low dose group(P<0.05).Compared with Positive control group,the expression of ERR a and SOX9 mRNA of Jingjintong decoction high dose group increased significantly(P<0.05),the expression of PGC-1 a and IL-1β mRNA of Jingjintong decoction middle dose group significantly increased(P<0.05),the expression of ERR a and SOX9 mRNA in Jingjintong decoction low dose group was significantly decreased(P<0.05),the expression of PGC-1α and IL-1β mRNA was significantly increased(P<0.05).Compared with Sham group,the positive score of Col2 α 1 protein in the Model group was significantly decreased(P<0.05).Compared with Model group,the positive score of Col2 α 1 protein in Jingjintong decoction high dose group,Jingjintong decoction middle dose group and Positive control group was significantly increased(P<0.05).Compared with Positive control group,the positive score of Col2α 1 protein in Jingjintong decoction high dose group and Jingjintong decoction middle dose group was significantly higher(P<0.05),Jingjintong decoction low dose group was significantly lower(P<0.05).ConclusionThere was a correspondence between KOA divergent meridians classification and TCM syndrome type,Jingjintong decoction in the treatment of postmenopausal women with KOA divergent meridians lesions under the guidance of Zang-fu differentiation could obtain good clinical efficacy.The presence of knee articular cartilage and gastrocnemius lesions in the KOA model of ovariectomized rats could better simulate the degeneration of the knee joint in postmenopausal women.Jingjintong decoction could adjust the expression of articular cartilage IL-1β/ERR a/PGC-1α/SOX9 signal pathway in the KOA model of ovariectomized rats,and increase the expression of type Ⅱ collagen in gastrocnemius,further played a role in the treatment of KOA bones and muscles lesions.Comprehensive analysis showed Jingjintong decoction in the treatment of postmenopausal women with KOA divergent meridians lesions under the guidance of zang-fu differentiation might regulate the expression of articular cartilage IL-1β/ERRα/PGC-1α/SOX9 signaling pathway,improve the expression of type Ⅱ collagen in gastrocnemius and play a therapeutic role. |
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