| Purpose:Through the use of our hospital's traditional empirical prescription "Tongbi Zhitong Liniment" for the treatment of limb arthritis,the osteoarthritis rat model was treated,and the anti-inflammatory and repairing effects of the drugs on the lesions were observed from multiple levels of indicators.The mechanism of action of Tongbi Zhitong Liniment will provide the preliminary experimental basis for the further application of in-hospital preparations,new drugs and other clinical applications.Material and method:84 wistar rats were randomly divided into 7 groups,namely the blank group,the model group,the Tongbi Zhitong Liniment high concentration group,the Tongbi Zhitong Liniment medium concentration group,the Tongbi Zhitong Liniment low concentration group,and the western medicine positive control group.Group and Chinese patent medicine positive control group.Except for the blank group,the modified Hulth method was used to make the model.After one week,each group was given different intervention measures: the blank group did not do any intervention,the model group was rubbed with normal saline,and the three groups of Tongbi Zhitong liniment were rubbed with different concentrations of experimental drugs.(High 0.15g/kg,medium 0.075g/kg,low0.0375g/kg),the positive control group of western medicine was rubbed with Futarin ointment,the positive control group of Chinese patent medicine was rubbed with Guyouling liniment,the above groups were treated twice a day Times for four weeks.Observe and record the mental state,active diet,etc.of each group of rats every day.Measure the swelling of the knee joints of each group 24 hours after the last administration.Take samples and test the following indicators:(1)Observe roughly the degree of damage to the knee joint cartilage in each group,and then HE staining was used to observe the histopathology of the lesions.(2)ELISA method was used to detect the serum levels of IL-1?,TNF-? and PGE2 in each group of rats to observe the inhibitory effect of each group's intervention measures on the inflammation of rat articular cartilage.(3)The Real Time PCR method was used to detect the expression levels of MMP-13 m RNA and type II collagen m RNA in the cartilage of each group of rats,and the regulatory effects of each intervention on the main indicators of cartilage matrix degradation and synthesis were observed from the gene level.(4)The immunohistochemical technique was used to detect the distribution and optical density of MMP-13 and type II collagen in the cartilage of each group to observe the influence of each group's intervention measures on the expression of the two.Results:1.General situation: the model group's fur is dull and loses its luster,hind limbs are unwilling to touch the ground,obviously limp and activity restriction is accompanied by reduced diet;the mental state,active diet,etc.of the rats in each drug intervention group are better than the model group;blank The rats in the group had no abnormal behavior changes.The knee joint diameter of rats in the model group was significantly thicker than that in the blank group(P<0.01),and the swelling of the knee joint in each drug intervention group was improved.2.Gross knee joint observation and HE staining results: Except for the blank group,the other six groups of joints were swollen and congested to varying degrees,and the joint surface was rough and dark in color.Except for the blank group by HE staining,the cartilage surface morphology of each group was abnormal and accompanied by different degrees of defects.The model group had the most serious lesions.The joint surface smoothness of the high-concentration group of Tongbi Zhitong Liniment and the western medicine control group was better than that of the other groups;the cartilage layer of the middle and low-concentration group and the Chinese patent medicine control group tended to become thinner than the previous two groups.3.ELISA test results: Compared with the model group,the levels of inflammatory factors IL-1?,TNF-? and PGE2 in each drug intervention group were significantly reduced(P<0.01).Among them,the index level of Tongbi Zhitong Liniment high concentration group and western medicine positive control group was not significantly different(P>0.05),and it was lower than that of Chinese patent medicine positive control group(P<0.05).The levels of inflammatory indexes in the high,medium and low groups of Tongbi Zhitong Liniment gradually increased.4.Real Time PCR test results: MMP-13 m RNA: Compared with the model group,the content of each drug intervention group was significantly reduced(P<0.01),and there was no significant difference between the high concentration group and the western medicine positive control group in index levels(P>0.05)),and was significantly lower than the positive control group of proprietary Chinese medicines(P<0.01).The content of cartilage in the high,medium and low groups of Tongbi Zhitong Liniment gradually increased.Type II collagen m RNA: the content of Tongbi Zhitong Liniment in the high and middle concentration groups and the western medicine control group was significantly increased(P<0.01);while the content in the low concentration group was higher than that in the model group,but the difference was not statistically significant.(P>0.05);There was no significant difference in the index level between the high concentration group and the western medicine positive control group(P>0.05),and the content was higher than that of the Chinese patent medicine positive control group(P<0.05).The content of cartilage in the high,medium and low groups of Tongbi Zhitong Liniment gradually decreased.5.Immunohistochemical test results: Compared with the model group,the positive expression intensity of MMP-13 staining in the tissues of each drug intervention group was reduced,and the expression intensity of Tongbi Zhitong Liniment was high,medium concentration and western medicine group was low.In the low-concentration and Chinese patent medicine groups;the positive expression intensity of type II collagen staining in the tissues of each drug intervention group increased.Among them,the expression intensity of Tongbi Zhitong Liniment high,medium concentration group and western medicine group was significantly stronger than the low concentration group and medium concentration group.Medicine group.The expression of MMP-13 and type II collagen is negatively correlated.Conclusion:Tongbi Zhitong Liniment can effectively alleviate joint inflammation by reducing the levels of IL-1?,TNF-? and PGE2 inflammatory factors in the serum of model rats,and by regulating the genes of MMP-13 and type II collagen in cartilage tissue Expression,inhibit the secretion of MMP-13 and promote the expression of type II collagen,reduce and repair cartilage damage caused by the imbalance between the degradation and synthesis of cartilage extracellular matrix.Among them,the high-concentration group of Tongbi Zhitong Liniment has the same level of indicators as the western medicine group,and is better than the Chinese patent medicine group.As a Chinese medicine external liniment,it has the advantages of simple and cheap test and small side effects,and it is worthy of clinical promotion and application. |
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