| Objective: Zhengshi Renjian Powder is often used in the treatment of joint stiffness after bone and joint injury,and it has remarkable curative effect on joint blood accumulation and joint movement limitation after injury.In this study we traumatic knee joint stiffness model,by establishing the rat to the cheng soft firm scattered topical treatment,observation of zheng soft firm dispersion gels traumatic stiff knee joint function of rat,joints,joint capsule fibroblasts,pathological changes and the influence of the expression of TGF-beta 1,to study the effect of zhengshi soft firm are scattered in the treatment of joint stiffness and its mechanism,It provides theoretical basis for clinical application.Methods: Sixty-six male SD rats were selected,18 rats were randomly selected as the blank group,and the remaining 48 rats were used to establish a post-traumatic right knee flexion stiffness model according to the Hildebrand modified method,which was randomly divided into the model control group and the Ruanjian San external application group after successful modeling.The blank group and the model control group were fed routinely without intervention;the Ruanjian powder external application group was treated with Zhengshi Ruanjian powder external application,each application time was 6 hours,once a day.The range of motion of the right knee joint of rats was measured and tissues were sampled at 4 W,6 W,and 8 W phases,and the histological changes of the joint capsule were observed by HE staining,and the images were taken for pathological scoring and fibroblast counting;the joint capsule was taken and the thickness of the joint capsule was measured by a spiral micrometer;and the expression of TGF-?1 in the joint capsule was determined by immunohistochemistry.Data were analyzed using SPSS 26.0 statistical software.Results:(1)Right knee range of motion: The range of motion of each group at different time points was compared,and overall analysis revealed that there were significant differences between time points(F = 54.966,p = 0.000)and groups(F = 566.420,p =0.000),and the interaction between time points and groups was also statistically significant(F = 23.200,p = 0.000).At the three time points,the joint range of motion in the model control group and Ruanjian powder external application group was less than that in the blank group(p < 0.05);the joint range of motion in the model control group and Ruanjian powder external application group increased with time,and on the 8th day,there was a significant difference between the model control group and Ruanjian powder external application group(p < 0.05).Comparison of the improvement range of motion: The improvement of joint range of motion in the external application group of Ruanjian Powder was significantly higher than that in the model control group,and the improvement of range of motion in the external application group of Ruanjian Powder was significantly different from that in the model control group from the 6th week to the8 th week(p < 0.05).Comparison within the group: The improvement of joint range of motion in the control group was greater than that from the 6th week to the 8th week(p <0.05).(2)Knee joint capsule thickness: The joint capsule thickness of each group at different time points was compared,and the overall analysis revealed that there was no significant difference between the populations at different time points(F = 1.269,p =0.374),there were significant differences between the populations in different groups(F= 181.281,p = 0.000),and there was no significant interaction between time points and groups(F = 0.023,p = 0.999).At the 4th and 8th week,compared with the blank group,the thickness of joint capsule in the model control group and Ruanjian powder external application group was statistically different(p < 0.05).On the 6th day,there was significant difference between the blank model control group(p < 0.05)and the Ruanjian powder external application group(p > 0.05).At the three time points,there was no significant difference in capsule thickness between the model control group and the Ruanjian San external application group(p > 0.05).(3)Pathological tissue sections: The pathological scores of each group at different time points were significantly different(H = 38.745,p = 0.000).On the 4th day,compared with the blank group,there were statistically significant differences in the synovial pathological scores between the model control group(p = 0.011)and the Ruanjian powder external application group(p = 0.016),and the synovial tissue of the joints at each time point was more severely damaged in the model control group and Ruanjian powder external application,the number of synoviocytes in the synovial layer increased,the fibrous tissue in the subsynovial layer proliferated,and some new blood vessels were observed,and there was no significant difference in the pathological scores at each time point between the model control group and Ruanjian powder external application(p > 0.05).(4)Fibroblasts: The number of fibroblasts in each group was compared at different time points,and overall analysis revealed that there were significant differences between time points(F = 38.248,p = 0.000)and groups(F = 132.847,p = 0.000),and the interaction between time points and groups was also statistically significant(F =17.911,p = 0.000).At three time points,the number of fibrocytes in the model control group and Ruanjian powder external application group was greater than that in the blank group,and the difference was significant(p < 0.05).At the 6th and 8th week,the difference was significant between the model control group and Ruanjian powder external application group(p < 0.05).Comparison of the decrease range of fibroblasts:The decrease range of the number of fibrocytes in the external application of Ruanjian powder from the 4th week to the 6th week was more than that in the model control group,and the difference was statistically significant(p < 0.05).Comparison within the group: The decrease of fibroblast number in the model control group was greater than that in the model control group from the 4th to the 6th week,and the decrease of fibroblast number in the external application group was significantly different from that in the model control group(p < 0.05).(5)Joint capsule TGF-?1 expression: The proportion of positive TGF-?1expression in joint capsule of each group at different time points was compared,and the overall analysis revealed that there were significant differences between time points(F= 19.216,p = 0.002)and groups(F = 14.710,p = 0.005)overall,and the interaction between time points and groups was also statistically significant(F = 6.919,p = 0.004).On the 4th day,compared with the blank group,there were significant differences in the positive proportion of TGF-?1 expression between the model control group and the Ruanjian powder external application group(p < 0.05),on the 6th day,there were significant differences between the blank model control group(p < 0.05),and there were significant differences between the model control group and the Ruanjian powder external application group(p < 0.05).Comparison of the decrease of the positive proportion of TGF-?1 expression: In the two time periods,there was no significant difference in the decrease of the positive proportion of TGF-?1 expression between the Ruanjian San external application group and the model control group(p > 0.05).Comparison within the group: The decrease of the positive proportion of TGF-?1expression in the Ruanjian Powder external application model control group from the4 th week to the 6th week was greater than that from the 6th week to the 8th week,and the decrease of the positive proportion of TGF-?1 expression in the Ruanjian Powder external application group was significantly different from that in the model control group(p < 0.05).Conclusion:1.Zhengshi Ruanjian Powder has obvious therapeutic effect on traumatic knee joint stiffness in rats and can effectively improve knee joint function.External application of Ruanjian Powder can obtain faster and better functional recovery than natural recovery,but it cannot change the trend of pathological changes.2.Zhengshi Ruanjian Powder can effectively reduce the number of fibroblasts and inhibit the expression of TGF-?1 in the joint capsule,and the inhibition of fibroblast proliferation and the expression of TGF-?1 in the joint capsule by Zhengshi Ruanjian Powder may be the mechanism of its treatment of traumatic knee stiffness.3.The effect of Zhengshi Ruanjian Powder on traumatic knee joint stiffness is more obvious in the early stage of intervention.Early application of Zhengshi Ruanjian Powder can promote the functional recovery of traumatic knee joint stiffness and inhibit the development of knee joint capsule fibrosis. |
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