Effect Of Liuwei Dihuang Pill On Extracellular Matrix Components Of Intervertebral Disc In Rabbit Intervertebral Disc Degeneration Model

Objective:By observing the influence of extracellular matrix components of liuwei dihuang pills on the rabbit models of intervertebral disc degeneration, to study the efficacy of prevention and treatment of intervertebral disc degeneration of liuwei dihuang pills.Method:80clean6months’ age New Zealand rabbits were randomly divided into only liuwei dihuang pills group(group A), model group(group B), the control group(group C), and blank control group(group D), each group of20.The model and liuwei dihuang pills groups to establish animal model of intervertebral disc degeneration. Sterile drapes were whisked after general anesthesiain. Via the anterolateral of left abdomen skin incision in turn, cut the external oblique muscle, internal oblique muscle and the transverse abdominal muscle. Along the psoas major muscle gap opened the retroperitoneum and large blood vessels, exposure the L4/5and L5/6intervertebral discs, with10μl trace syringe injected1ul TNF alpha solution of10ng/μl into randomly chosen disc, mark the experimental animals and vertebral bodies after the injection. Flushed with physiological saline flushing. In accordance with theand put them into the cages.Control group of the same approaches into the lumbar spine, then hierarchy suture.Complexing iodic inunction disinfection to the incision.The4millions units of penicillin intramuscular injection for the postoperatively infection,1time a day, for consecutive3days. After liuwei dihuang capsules dissolved in physiological saline, the liuwei dihuang pill group was given10mg/kg lavage, liuwei dihuang capsules solution1time a day.The model group, the control group and the blank control group were filled in the amount of normal saline,1time a day, and natural feeding.Last4hour after lavage in2weeks,4weeks,6weeks,8weeks in batches to be put to death animals, kept the intervertebral disc specimens.Determination the intervertebral disc endplate total sugar, amino polysaccharide (glycosaminoglycan, GAG), chondroitin sulfate, chondroitin sulfate, CS) and keratin sulfate (keratan sulfate, KS) ratio with enzyme-linked immunoassay,ria method for the determination of hyaluronic acid (hyaluronic acid, HA) content. Rt-PCR detection proteoglycan and collagen type I, II mRNA content of collagen type, Western Blot detection proteoglycan and collagen type I, the expression of collagen type II.Results:New Zealand rabbits of control group compared with model group, model group protein polysaccharide and mRNA expression, chondroitin sulfate, sugar, amino polysaccharide content and the ratio of keratin sulfate, hyaluronic acid content, the expression of collagen type II and mRNA levels were significantly lower than control group, and mRNA expression of type I collagen content were significantly higher than the control group, P＜0.05, the difference was statistically significant, the rabbit models of intervertebral disc were built successfully.Model group compared with the group of liuwei dihuang pills, model group’s protein polysaccharide and mRNA expression in rabbit, chondroitin sulfate, sugar, amino polysaccharide content and the ratio of keratin, hyaluronic acid, sulfate content, the expression of collagen type Ⅱ and mRNA levels were significantly lower than that of liuwei dihuang pill group, and the concentration of type Ⅰ collagen mRNA expression significantly higher than that of liuwei dihuang pill group, P<0.05, the difference was statistically significant.Indicating after suspension liquid lavage of liuwei dihuang pills,it has a certain therapeutic effect.In liuwei dihuang pills group,2weeks and4weeks after surgery,4weeks and6weeks,6weeks compared with8weeks, the change of protein expression, polysaccharide and mRNA content, sugar, amino polysaccharide content, the ratio of chondroitin sulfate and sulfuric acid keratin, hyaluronic acid content, the expression of type I collagen and mRNA content, the expression of collagen type II and mRNA content is not big, and stable, P>0.05, there was no statistically significant difference. Indicating that liuwei dihuang pills can effectively delay the degeneration of intervertebral disc;In the model group,2weeks and4weeks,4weeks and6weeks,6weeks compared with8weeks, the expression of protein, polysaccharide and mRNA content, sugar, amino polysaccharide content, the ratio of chondroitin sulfate and sulfuric acid keratin, hyaluronic acid content, the expression of collagen type Ⅱ and mRNA levels dropped, type I collagen content and mRNA expression rose,P<0.05, the difference was statistically significant, and intervertebral disc degeneration model successfully established;Conclusion:Liuwei dihuang pill can increase sugar amino polysaccharide content, chondroitin sulfate/acid ratio of keratin, the hyaluronic acid content in intervertebral disc degeneration model, and modestly lower the expression of type I collagen in intervertebral disc degeneration model, increase the expression of type II collagen, stabilized the proteoglycan content.So to a certain extent,delay the degeneration of the intervertebral disc.