| Objective:In the first experiment,the model of acute closed soft tissue injury in SD rats was established.The symptoms of limb injury in rats were observed and recorded at different time points after the administration of Rumo ointment.The contents of TNF-α,IL-1β,PGE-2and SOD in the injured tissues were detected.The changes of histomorphology and b FGF content in the tissues were observed under the microscope,and the curative effect of Rumo ointment on acute closed soft tissue injury was preliminarily discussed.In experiment 2,the anti-inflammatory effect of Rumo ointment was investigated by carrageenan-induced rat foot swelling experiment and xylene-induced mouse ear swelling experiment.The analgesic effect of Rumo ointment was studied by hot plate test(physical stimulation)and acetic acid writhing test(chemical stimulation)in mice,which provided a theoretical basis for the clinical application of Rumo ointment.Methods:In the first experiment:Sixty SD rats(6 spare)were fed for 1 week,and 54 were randomly selected for experiments.Among them,45 were modeled by heavy hit method(acute closed soft tissue injury model).After successful modeling(the first day),SD rats were randomly divided into 5 groups(n = 9),namely group B(model control group),group C(Qingpeng ointment group),group D(high-dose Rumo ointment group),group E(middle-dose Rumo ointment group),group F(low-dose Rumo ointment group);nine SD rats without modeling were group A(blank control group);the 6 groups of rats were randomly divided into 3 groups(n = 3)according to the second day,the seventh day and the fourteenth day.Rats in groups C,D,E and F were given quantitative external application at the same time every day,group B was given corresponding normal saline,and group A was not given anything.On the 2nd,7th and 14 th day after modeled,the rats were divided into groups to observe and compare the injury syndrome index score,detect the contents of TNF-α,IL-1β,PGE-2 and SOD in the damaged tissue,observe the morphological changes of the damaged tissue and the changes of b FGF content in the tissue under the microscope.In the second experiment:hot plate test(physical stimulation),acetic acid writhing test(chemical stimulation),xylene-induced ear swelling test in mice,50 Kunming mice,number grouping,high dose group of Rumo ointment,middle dose group of Rumo ointment,low dose group of Rumo ointment,Qingpeng ointment group and blank group,10 in each group,each group of animals were abdominal hair removal,the area is about 2.0 × 1.5cm2;qingpeng ointment group and Rumo ointment group were applied with corresponding drugs at the abdominal hair removal site,and fixed with breathable tape.The dressing was changed once a day for a total of 7 days.The blank group was only depilated without sticking.The mouse hot plate experiment(physical stimulation)cold and hot pain tester measured the pain threshold of each group of mice before administration and the pain threshold at 0.5 h,1.0 h,and 1.5 h after administration;acetic acid writhing test in mice(chemical stimulation)One hour after the last administration,mice in each group were intraperitoneally injected with 0.6 % acetic acid solution.The time of the first painful writhing and the number of writhing reactions within 15 minutes were recorded.Xylene-induced ear swelling in mice After the last administration,the left ears of mice were exposed to xylene cotton balls(about 50 μl of xylene)for 15 min,and the discs were weighed on the same part of the left and right ears with a 6 mm punch.The difference in weight between the two ears of each group was calculated.Carrageenan-induced rat foot swelling experiment,50 SD rats were numbered and grouped,Rumo ointment high-dose group,Rumo ointment medium-dose group,Rumo ointment low-dose group,Qingpeng ointment group and blank group,with 10 rats in each group.The drug group applied the corresponding drugs to the left hind foot of the rats,and the patch was cut into the same size during the application.The application was firm,and the external tape and bandage were wound several times to fix,and the dressing was changed once every 24 hours for a total of 7 days.The blank group did not do any treatment.After the last application,0.1 ml of 1 % carrageenan solution was injected subcutaneously into the middle of the left hind paw of the rat,and the dorsal thickness of the left hind paw before inflammation was measured.Results:In the first experiment:(1)General condition of SD ratsThe rats in group B showed lethargy and burnout,and a small number of rats had messy and dull coats.The first 7 days after modeling,the right lower limb was swollen and bruised,and the right lower limb was claudication.The symptoms and signs gradually disappeared after 7 days.The rats in each drug group were gradually improved during the treatment period,such as curled up and lethargy,hair clutter and other conditions after modeling.(2)Injury symptom index scoreOn the 2nd,7th and 14 th day after modeling,the injury symptom index scores of each group and the overall distribution of the 6 groups were statistically different(P < 0.05).On the 7th day,there was a statistical difference between group A and group B(P < 0.05),between group A and group F(P < 0.05),and between group B and group D(P < 0.05).On the 14 th day,except that there was a statistical difference between group A and group B(P <0.05),there was no statistical difference in the injury symptom index scores of each drug group(P > 0.05).(3)Results of TNF-α content in rat tissues of each group after modelingOn the 2nd,7th and 14 th day after modeling,the content of TNF-α in the 6 groups was statistically different(P < 0.05),and there was statistical difference between the other two groups(P < 0.05).On the 14 th day,there were statistical differences between group A and group B,group A and group E,group A and group F,group B and group C,group B and group D,group B and group E,group C and group E,group C and group F,group D and group E,group D and group F,group E and group F(P < 0.05).(4)The results of IL-1β content in rat tissues of each group after modelingOn the 2nd,7th and 14 th day after modeling,the content of IL-1β in the 6 groups was statistically different(P < 0.05),and there was statistical difference between the other two groups(P < 0.05).On the 14 th day,there were significant differences between group A and group B,group A and group E,group A and group F,group B and group C,group B and group D,group B and group E,group C and group E,group C and group F,group D and group E,group D and group F,group E and group F(P < 0.05).(5)Results of PGE-2 content in tissues of rats in each group after modelingOn the 2nd day,7th day and 14 th day after modeling,there was a statistical difference in the overall distribution of PGE-2 in the 6 groups(P < 0.05),and there was a statistical difference between the other two groups(P < 0.05).On the 14 th day,there were significant differences between group A and group B,group A and group E,group A and group F,group B and group C,group B and group D,group B and group E,group C and group E,group C and group F,group D and group E,group D and group F,group E and group F(P < 0.05).There was no significant difference between the other two groups(P > 0.05).(6)The results of SOD content in each group after modelingOn the 2nd,7th and 14 th day after modeling,the content of SOD in the 6 groups was statistically different(P < 0.05),and there was statistical difference between the other two groups(P < 0.05).On the 14 th day,there were significant differences between group A and group B,group A and group E,group A and group F,group B and group C,group B and group D,group B and group E,group C and group E,group C and group F,group D and group E,group D and group F,group E and group F(P < 0.05).There was no significant difference between the other two groups(P > 0.05).(7)Morphological observation results(1)On the second day after modeling : Under the microscope,most of the muscle fibers were broken,arranged distorted and disordered,and the cells were broken,dissolved and necrotic,and inflammatory cell infiltration was observed locally.No significant difference was found in each group after modeling.(2)On the 7th day after modeling,inflammatory cell infiltration was observed under the microscope in each model group,and fibrous connective tissue and granulation appeared occasionally,and new cell morphology was observed.Among them,some tissue fiber hyperplasia,thickening and mild edema were observed in each group.Among them,a small amount of fibrous tissue hyperplasia and tissue edema were observed in group B(model control group);group C(Qingpeng ointment group),group D(Rumo ointment high dose group)and group E(Rumo ointment medium dose group)were significantly improved compared with group F(Rumo ointment low dose group)and group B(model control group).(3)On the 14 th day after modeling,microscopically,the muscle tissue cells were better,the muscle fibers were neatly arranged,and the local tissue fibers were hyperplasia without obvious inflammation.Among them,the improvement of group B was worse than that of each group,and some muscle fiber fibrosis was observed.The improvement of group C(Qingpeng ointment group)and group D(Rumo ointment high dose group)was the most obvious.(8)Expression of b FGF in tissues(1)On the second day after modeling,there was no significant difference in the average optical density of b FGF protein in each group(F = 2.800,P > 0.05).(2)On the 7th day after modeling,the average optical density of b FGF protein in each group was statistically different(F = 22.738,P < 0.05).Pairwise comparison showed that there were significant differences between group A and group C,group A and group D,group A and group E,group B and group C,group B and group D,group B and group E,group C and group E,group C and group F,group D and group E,group D and group F,group E and group F(P < 0.05).(3)On the 14 th day after modeling,the average optical density of b FGF protein in each group was statistically different(F = 14.749,P < 0.05).Pairwise comparison showed that there were significant differences between group A and group B,group B and group C,group B and group D,group B and group E,group B and group F(P < 0.05).In the second experiment:(1)After 0.5 h of intervention,the pain threshold time of high dose group of Rumo ointment was significantly different from that of middle dose group of Rumo ointment,low dose group of Rumo ointment and blank control group(P < 0.05).The middle dose group of Rumo ointment was significantly different from that of Qingpeng ointment group and blank control group(P < 0.05).The low dose group of Rumo ointment was significantly different from that of Qingpeng ointment group and blank control group(P < 0.05).There was significant difference between Qingpeng ointment group and blank control group(P < 0.05).The time of pain threshold at 1.0 h after intervention was statistically different between the high dose group of Rumo ointment and the low dose group of Rumo ointment and the blank control group(P < 0.05).There was a statistical difference between the middle dose group of Rumo ointment and the blank control group(P < 0.05).There was a statistical difference between the low dose group of Rumo ointment and the Qingpeng ointment group and the blank control group(P < 0.05).There was a statistical difference between the Qingpeng ointment group and the blank control group(P < 0.05).After 1.5 h of intervention,the pain threshold time was statistically different between the high dose group of Rumo ointment and the middle dose group of Rumo ointment,the low dose group of Rumo ointment and the blank control group(P < 0.05).The middle dose group of Rumo ointment was statistically different from the Qingpeng ointment group and the blank control group(P < 0.05).The low dose group of Rumo ointment was statistically different from the Qingpeng ointment group and the blank control group(P < 0.05).The Qingpeng ointment group and the blank control group were statistically different(P < 0.05).(2)The latency time of pain after intervention was statistically different between the high-dose Rumo ointment group and the low-dose Rumo ointment group and the blank control group(P < 0.05).There was a statistical difference between the medium-dose Rumo ointment group and the blank control group(P < 0.05).There was a statistical difference between the low-dose Rumo ointment group and the Qingpeng ointment group(P < 0.05).There was a statistical difference between the Qingpeng ointment group and the blank control group(P < 0.05).The number of writhing within 15 minutes after intervention,the high dose group of Rumo ointment was statistically different from the low dose group of Rumo ointment and the blank control group(P < 0.05),the middle dose group of Rumo ointment was statistically different from the blank control group(P < 0.05),the low dose group of Rumo ointment was statistically different from the Qingpeng ointment group(P < 0.05),and the Qingpeng ointment group was statistically different from the blank control group(P <0.05).(3)After the intervention,the weight of the left ear was statistically different between the high-dose Rumo ointment group and the low-dose Rumo ointment group and the blank control group(P < 0.05).There was a statistically significant difference between the middle-dose Rumo ointment group and the low-dose Rumo ointment group,the Qingpeng ointment group and the blank control group(P < 0.05).There was a statistically significant difference between the low-dose Rumo ointment group and the Qingpeng ointment group(P< 0.05).There was a statistically significant difference between the Qingpeng ointment group and the blank control group(P < 0.05).The difference in the weight of the two ears after the intervention was statistically different between the high-dose group of Rumo ointment and the low-dose group of Rumo ointment and the blank control group(P < 0.05).There was a statistical difference between the middle-dose group of Rumo ointment and the Qingpeng ointment group and the blank control group(P < 0.05).There was a statistical difference between the low-dose group of Rumo ointment and the Qingpeng ointment group and the blank control group(P < 0.05).There was a statistical difference between the Qingpeng ointment group and the blank control group(P < 0.05).(4)The swelling degree at 0.5h after intervention was statistically different between the high-dose Rumo ointment group and the blank control group(P < 0.05).There was a statistically significant difference between the medium-dose Rumo ointment group and the Qingpeng ointment group and the blank control group(P < 0.05).There was a statistically significant difference between the low-dose Rumo ointment group and the Qingpeng ointment group and the blank control group(P < 0.05).At 1.0 h after intervention,the swelling degree of the high-dose Rumo ointment group was statistically different from that of the blank control group(P < 0.05).There was a statistically significant difference between the medium-dose Rumo ointment group and the blank control group(P < 0.05).There was a statistically significant difference between the Qingpeng ointment group and the blank control group(P < 0.05).After 2.0 hours of intervention,there was a statistically significant difference in swelling between the high-dose Rumo ointment group and the blank control group(P < 0.05).There was a statistically significant difference between the medium-dose Rumo ointment group and the low-dose Rumo ointment group and the Qingpeng ointment group(P < 0.05).There was a statistically significant difference between the low-dose Rumo ointment group and the Qingpeng ointment group(P < 0.05).There was a statistically significant difference between the Qingpeng ointment group and the blank control group(P <0.05).At 3.0 h after intervention,the swelling degree of Rumo ointment high-dose group was statistically different from that of Rumo ointment low-dose group and blank control group(P< 0.05).There was a statistically significant difference between Rumo ointment low-dose group and Qingpeng ointment group(P < 0.05).There was a statistically significant difference between Qingpeng ointment group and blank control group(P < 0.05).The swelling degree at 5.0 h after intervention was statistically different between the Qingpeng ointment group and the blank control group(P < 0.05).The swelling degree of 7.0 h after intervention,there was a statistical difference between the high dose group of Rumo ointment and the low dose group of Rumo ointment and the blank control group(P < 0.05),there was a statistical difference between the middle dose group of Rumo ointment and the blank control group(P < 0.05),there was a statistical difference between the low dose group of Rumo ointment and the Qingpeng ointment group(P < 0.05),there was a statistical difference between the Qingpeng ointment group and the blank control group(P < 0.05).Conclusion:(1)Rumo ointment can significantly improve the general state of rats with acute closed soft tissue injury and promote the recovery of limb injury.(2)Rumo ointment can inhibit the increase of TNF-α,IL-1β and PGE-2,enhance the activity of SOD and increase the expression of b FGF in the local tissue of the injured model rats,and can promote the repair of the local tissue of the injured rats.(3)Rumo ointment and Qingpeng ointment have similar efficacy in the treatment of acute closed soft tissue injury in rats.(4)Rumo ointment high dose group and Qingpeng ointment in anti-inflammatory analgesic effect. |
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