Gubi Mixture Effect On Mediated The Subchondral Bone Metabolism Research Of The Knee Osteoarthritis Model In Rat

Objective:Copy a with the human knee osteoarthritis pathogenesis process similar to the double forelegs vertical joint Hulth method in the rat model of knee osteoarthritis.And from the level of histopathology,protein function and gene level,the mechanism of bone gubi mixture on bone metabolism in the cartilage of this model was observed.Methods:The pregnant healthy SD rats 12 of 15 days in pregnancy were selected and placed in a different cage,and the young mouse was born.On the 10 th day after birth,80 healthy and non-deformed mice were selected and randomly divided into four groups according to their weight: the blank control group(the KOA model group of the two front legs),the physiological saline group,the aminosaccharide group,and the gubi mixture group,each group of 20.All the four groups were removed from the operation of the two anterior limbs under the abdominal anesthesia.After the operation,they were kept in the original mother rat cage,and the right knee joint was performed after four months(4 months of age).After the operation,they were kept in the original cage for one month.At 5 months of age,the gubi mixture group were given drug intervention for a month,and the group was given 5ml/Kg/d at 8 and 18 points.The aminosugar mein group gives the aminosaccharide to 20 mg/Kg/d,which is placed in the same volume of distilled water until it's melted and shaken,and given to the stomach at 8 and 18;The physiological saline group was treated with normal saline by 5ml/Kg/d at 8 and 18.Continuous intervention for 4 weeks;The blank control group ate only normal diet without any intervention.Rats were anesthetized and executed at 6 months of age,take a right knee line-80? cryopreservation.Some specimens were analyzed by means of fixation,decalcification,dehydration,embedding,slicing,roasting,HE staining and subchondral bone tissue morphometry.Some specimens using western blot method,testing the protein expression of COL-?,OPG and RANK in the subchondral bone of the rats.Some specimens using RT-PCR method,to examine the m RNA expression of COL-?,IL-1?,IL-6,ALP,OPG and RANK in the subchondral bone of the knee joint of the rats in each group.Result:1.The right knee HE staining of the subchondral bone tissue morphology metrology statistic results show that ammonia sugar beautiful sheen and involving the bone mixture group the subchondral bone trabecular bone volume compared with blank control group and saline group obviously higher P<0.05,the difference was statistically significant;Blank control group the subchondral bone of the bone trabecular volume compared with normal saline group slightly lower P>0.05,there was no statistically significant difference;Ammonia sugar the symplectic group of subchondral bone trabecular bone volume compared with involving the gubi mixture group slightly lower P>0.05,there was no statistically significant difference;2.Western blot method was used to detect the results of subchondral subchondral bone in each group: the expression of OPG protein in the four groups was the highest,and the content of the group was the highest in the group,which was statistically significant compared with the saline group.The content of RANK protein expression in the four groups was the lowest in the group,and it was statistically significant compared with the normal saline group and aminosaccharide group.Four groups of COL-? protein expression levels,involving the bone mixture group the highest levels and with physiological saline group,blank control group and ammonia sugar beautiful sheen comparative differences are statistically significant;3.The Real-Time PCR method to detect each group rats under the knee joint cartilage bone genes results showed that four groups of OPG m RNA expression of relative quantity,involving the bone mixture group the highest levels and with physiological saline group,blank control group and ammonia sugar beautiful sheen comparative differences are statistically significant;The relative expression of ALP m RNA in the four groups,the high physiological saline group and the blank control group were lower than that in the group,and the difference between the group and thephysiological saline group was statistically significant.Four groups of RANK m RNA expression of relative quantity,involving the bone mixture content is lower than the normal saline group and blank control group was higher than ammonia sugar beautiful sheen and involving the bone mixture group and normal saline group,blank control group,the difference had statistical significance;Four groups of COL-? m RNA expression of relative quantity,the highest levels and involving the bone mixture group compared with normal saline group and blank control group differences were statistically significant,there was no statistical difference compared with ammonia sugar the symplectic group;Four groups of IL-6 m RNA expression quantity relatively,involving the bone mixture content is lower than the normal saline group and blank control group was higher than ammonia sugar beautiful sheen and involving the bone mixture group and normal saline group,the differences between the blank control group were statistically significant,there was no statistical difference compared with ammonia sugar the symplectic group;Four groups of IL-1? m RNA expression quantity relatively,involving the bone mixture content is lower than the normal saline group and blank control group was higher than ammonia sugar beautiful sheen and involving the bone mixture group and normal saline group,the differences between the blank control group were statistically significant,there was no statistical difference compared with ammonia sugar the symplectic group.Conclusion:Involving the gubi mixture can reduce the subchondral bone IL-1?m RNA,IL-6 m RNA expression,reduce inflammation,cut the subchondral bone in RANK m RNA,and increased OPG m RNA,ALP m RNA,COL-? m RNA expression,adjust the bone metabolism and promote the subchondral bone repair,may be involving the bone mixture as part of the mechanism and play to delay the SD rat knee osteoarthritis subchondral bone degeneration process.