Study on Xuefu Zhuyu Decoction on Changes of Serum Cytokines MMP-13 and TNF-α in Rats with Knee Osteoarthritis

Purpose: The purpose of this study is to study the effect of Xuefu Zhuyu Decoction on the serum levels of MMP-13 and TNF-α in experimental rats with knee osteoarthritis through animal experiments and observe the morphological changes of cartilage injury. After establishing the rat knee osteoarthritis model, through 4 weeks of experimental intervention, the knee cartilage of each group of experimental rats was taken out for histomorphological observation and comparison, and immunohistochemical methods were used to detect the MMP- in the serum of each group of rats. 13 and TNF-α content, the data is analyzed and processed using statistical methods to explore the possible mechanism of action of Xuefu Zhuyu Decoction on the prevention and treatment of knee osteoarthritis. Arthritis provides scientific experimental basis. Methods: The 60 SD male rats needed for the experiment, fasting weights of about 200 to 250 g, were fed with ordinary pellets, and the rats were divided into 4 groups according to the random number method, 15 rats in each group, respectively sham operation group (A): Only open the joint cavity, do not damage the ligaments and meniscus, suture the skin, and wrap the bandage aseptically; model group (B): All rat osteoarthritis models use surgical knee medial tibial incision, carefully dissect the medial collateral ligament Cut off and remove the internal molars of approximately 0.5cm, and then use 80,000U Qingdamer sulfate injection to instill the surgical field to prevent joint cavity infection. After aseptic bandaging, suture layer by layer; Western medicine group (C): mixed with ibuprofen Suspension was administered to the stomach; Chinese medicine group (D): Xuefu Zhuyu Decoction was administered to the stomach. One week after the operation, all the molded rats were alive. The surgical wound was red, the wound was basically healed, and the excretion was normal. The rats were forced to move once a day for 30 minutes each time for 1 week; after 2 weeks Imaging and histopathological observations revealed that the articular surface of the lower end of the femur and tibial plateau of the knee joint was flattened, the joint deformed and the articular cartilage defect, the chondrocyte arrangement was disordered, the trabecular area of the femur and tibia was significantly reduced, and the bone marrow cavity cells proliferated. Successfully established a rat knee osteoarthritis model. On the first day after successful modeling, 5 rats from each group were taken from each group and 5ml of abdominal aortic blood was taken after anesthesia, and 1ml of liquid was taken after centrifugation. The serum levels of MMP-13 and TNF-α in each group of rats were detected by ELISA And start to give drug intervention to the rest of the rats at the same time. The frequency of administration in each group is once a day. The dosage is: the sham operation group and the model group are respectively given normal saline (1ml/100g), and the western group is given Ibuprofen suspension was given by intragastric administration (1ml/100g), and the traditional Chinese medicine group was given Xuefu Zhuyu Decoction (1ml/100g) by intragastric administration; after 4 weeks of drug intervention, the rats in each group were anesthetized and placed on a clean table in the laboratory On the upper rat plate, take 5ml of abdominal aortic blood according to the V-shaped incision, take 1ml of liquid and store it in the refrigerator after centrifugation, again use ELISA method to detect the content of MMP-13 and TNF-α in the serum of each group of rats; then use All the rats were killed by air embolization, the opening and closing cavities were cut, the knee cartilage of each group was taken out and fixed in neutral polyoxymethylene fixative, and the morphological and tissue changes of the knee cartilage of each group were observed after staining with HE staining method. Results: 1 Observation of tissue morphology(1) The surface of the knee cartilage of the rats in group A is smooth without degeneration, no defect area, no obvious thickening of the joint capsule, and normal shape of the joint synovium without obvious hyperplasia;(2) The surface of the knee cartilage of the rats in group B is severely degenerated, with ulcers formed on the surface, some areas are defective, the joint capsule is thickened, the joint synovium is obviously proliferated, and the color is dim;(3) The knee cartilage of rats in group C has obvious degeneration, the surface is slightly rough, there are cracks, the joint capsule is slightly proliferated, and there is no ulcer formation;(4) The knee cartilage of the rats in group D was slightly degenerated, the surface was smoother, there was no obvious cartilage defect, the ulcer formation was small, and the synovial membrane was not hyperplasia; 2 Changes in rat serum mmp-13There was no significant change in serum mmp-13 of rats in group A; mmp-13 was expressed in a large amount in the serum of rats in group B; the levels of mmp-13 in serum of rats in group C and D were both decreased compared with group B, but in group D serum The decrease was more obvious than that in group C, and the difference in serum mmp-13 content between the two groups of rats was statistically significant (P <0.05).3 Changes in serum TNF-α content in ratsThe serum TNF-α content of rats in group A did not change significantly; the serum TNF-α content of rats in group B increased significantly, and the serum TNF-α content of rats in group C and D decreased compared with group B, but group D was larger The level of serum TNF-α in rats decreased more significantly than that in group C, and the difference in serum TNF-α levels between the two groups of rats was statistically significant (P <0.05). Conclusions: 1 The rat medial collateral ligament (about 0.5cm in length) is surgically removed and removed, and the meniscus is removed, to replicate the model of rat knee osteoarthritis;2 Xuefu Zhuyu Decoction can significantly improve the abnormal tissue morphology of the knee cartilage in experimental rats;3 The serum MMP-13/TNF-α content of rats in group C and D increased significantly before treatment, suggesting that the content of serum MMP-13/TNF-α may be related to the onset of knee osteoarthritis;4 Xuefu Zhuyu Decoction can significantly reduce the levels of mMP-13 and TNF-α in the serum of rats with knee osteoarthritis, and the effect is better than the western medicine group.Tabled 1Changes of serum MMP-13 levels in rats in each groupGroupPrior treatmentPost treatmentB Group210.684±38.23248.746±32.39C Group210.421±39.96177.805±26.41*D Group209.883±12.73160.951±12.43* Open table in a new tab Tabled 1Changes of serum TNF-α levels in rats in each groupGroupPrior treatmentPost treatmentB Group89.57±38.8288.34±36.18C Group89.44±40.7947.61±15.03*D Group90.13±37.1831.28±9.23* Open table in a new tab