Study On The Quality Standard And Its Effect And Mechanism On Osteoarthritis In Rats Of Lianweianaqi Granules

Objective:Study on the quality standard of the LWG and evaluation on arthritis in rats bone effect and mechanism.Methods:?1?the identification of the seven drugs in the LWG preparation by tlc;?2?using InertSustain C18 column?4.6mm,250mm,5um?,1%CH₃CH₂COOH methanol acetonitrile?12:11:77?as the mobile phase.The flow rate is 1.0ml/min,the detection wavelength is 334nm,the column temperature is 30°,the injection volume was 20uL,Establishment of High Performance Liquid Chromatography for Simultaneous Determination of LWG echinacoside and verbascoside;?3?SD male rats were randomly divided into normal control group,model group,positive control group,decoction group and LWG low and medium dose group;Except the normal control group,the test began in the 1d,3d and 7d respectively after rats left knee joint cavity injection of papain and cysteine solution model,in addition to the normal control and model group,other groups were administrated for 4weeks.Rats were recordedduring the whole experiment;After 4 weeks,blood was collected from abdominal aorta and the contents of COX₂ and NO were determined.The content of PGE2 in the knee joint lotion was measured.The articular cartilage and synovial membrane were extracted at the same anatomical site to make pathological sections,and the surface of synovial membrane and cartilage were observed.The experimental data were expressed by mean±standard deviation,and SPSS17.0 was used for statistical analysis,The data of multiple groups were measuredby one-way ANOVA and P<0.05 was statistically significant.Results:?1?TLC was identified seven herbs on thin layer chromatography,and no interference was found;?2?The peak area of echinacoside in the range of 37.5ug/mL-51ug/mL?r=0.999,n=6?,verbascoside in the range of 8.5ug/mL-51ug/mL?r=1,n=6?had a good linear relationship with the injection,The average recoveries of echinacoside were 101.26%and the average recoveries of cali glycosides were 99.89%;The chromatographic method used,without interference from other components,precision,recovery,good stability,to meet the needs of the determination;?3?After modeling,the model group compared with the normal group,the diameter of rat knee joint swelling significantly,the left limb toes,reduced locomotor activity;five weeks after modeling,the model group compared with normal control group,osteophyte formation;the behavior of drug treatment and model group rats than there is a certain degree of improvement;After 4 weeks of administration,from the point of view of body weight,compared to LWG in the high dose group and model group,the body weight of rats increased significantly in high dose group compared with the low dose group,body weight of rats increased significantly in a dose effect relationship;From the knee bending interaction degree,compared with the model group,LWG low,medium and high dose group rats knee activity improved significantly,LWG dose group and decoction group,positive medicine group than in the rat knee activity had no obvious improvement,the high dose group and low dose group compared rat,knee activity improved significantly and there is a dose effect relationship;From the elimination of COX₂ and NO content in serum of rats,the middle and high doses of LWG can reduce the content of NO,COX₂,LWG in each dose group and decoction group,positive drug group compared to COX₂,decreased the content of NO is not obvious,the high dose group decreased the content of COX₂ was compared with the low dose group,medium and high dose group compared with the low dose group and significantly decreased the content of NO in a dose effect relationship;Considering the elimination of PGE₂ content in joint fluid of OA rats,LWG middle dose group and high dose group significantly decreased the content of PGE₂,LWG in each dose group and decoction group,positive medicine group decreased the content of PGE₂ is not obvious,the high dose group compared with the low dose group and significantly decreased the content of PGE₂ in a dose effect relationship.Under the light microscope,the articular cartilage matrix was dyed pink,the cartilage surface was smooth,and the synovial membrane was not hyperplastic;In the model group,the articular cartilage surface was missing and the structure was not complete;In the LWG high dose group,there was no significant change in the articular cartilage layer,and the proliferation of the synovial membrane;Middle dose of LWG group,part of synovial cell swelling layer increased,from flat to round;Decoction group,articular cartilage cell swelling,skin smooth,line nest disappeared;the positive group,hyperplasia of synovial thickening and intimal cell proliferation LWG low dose group,synovial membrane cell proliferation,swelling,nuclear deep staining.Conclusion:The quality standard of TLC and content determination of LWG established in this study,a controllable method with strong specificity and good reproducibility,and simple operation;the pharmacodynamics experiment verified that LWG can effectively prevent and treat osteoarthritis in rats,its effect is equivalent to the decoction group,the mechanism may be related to the decrease in the activity.