| Objective: Objective to study the therapeutic effect of halofuginone on the early osteoarthritis model in ALCT mice.Methods : Healthy male C57BL6 J 90 mice were randomly divided into a test group and two months in experimental group,and then divided into sham operation group(sham group)and placebo group(ALCT+Vehicle group)and treatment group(ALCT+Halofuginone group),15 mice in each group,sham operation group only open the right knee joint capsule in mice no damage to other parts of the body,after treatment;the other two groups were made for osteoarthritis model in mice to cut off the anterior cruciate ligament,treatment group for one month: placebo third days Postoperatively with distilled water to 30 days;the treatment group after third days to start irrigation by 0.5mg/kg weight the stomach to 30 days.According to the volume of distilled water and halofuginone 0.2ml daily by intragastric administration once continuous intragastric administration were killed after 30 days;two months of treatment group: placebo for third days postoperatively with distilled water to 30 days;the treatment group after third days by intragastric 0.5mg/kg to 30 days.According to the volume of distilled water and halofuginone 0.2ml daily by intragastric administration once continuous intragastric administration were killed after 60 days;and sacrificed mice under the right knee,HE staining and safranin fast green staining was used to observe the morphology and structure of the joint tissue;the expression of immunohistochemical detection of TGF-1 in articular cartilage of beta 1,MMP-13,COLX protein;for immunohistochemical detection of subchondral bone in the Psmad protein expression and immunohistochemical detection of subchondral bone in osteoblast numbers markers Osterix and TRAP cells.Results: 6 groups of animals survived to the end of the experiment,the wound healing well.Histological observation,one month group: sham group articular cartilage normal structure,clear lines,cells arranged neatly,no cartilage surface wear traces;ALCT + Vehicle group articular cartilage layer was thinning,and even partial damage to the tide line,cartilage surface The chondrocytes of ALCT + Halofuginone group were more clear and arranged neatly,the lines were clear and the cell morphology was more regular;two months: sham group articular cartilage structure is normal,the tide line is clear,the cells are arranged neatly,cartilage surface is slightly rough,slightly wear marks;ALCT + Vehicle group articular cartilage structure has deformed,articular cartilage layer thinning,calcification area The majority of the damage is very serious,even the damage to the tide line,cartilage surface wear is very serious and fragmentation phenomenon,the cell arrangement is extremely disorder;ALCT + Halofuginone group chondrocytes stratified more clearly and arranged neatly,clear lines,cells Shape more rules,a small part of the surface of the cartilage wear;(HC / CC)were:(1.47 ± 0.20),(0.77 ± 0.13)and(1.2 ± 0.09)in the three groups of sham,ALCT + Vehicle,ALCT + Halofuginone in three groups.(P <0.05).The number of clear cartilage thickness / calcified cartilage(HC / CC)in sham,ALCT + Vehicle and ALCT + Halofuginone were:(1.24 ± 0.25),(0.58 ± 0.15)and(1.04 ± 0.11).There was significant difference between the three groups(P <0.05).The scores of sham,ALCT + Vehicle,ALCT + Halofuginone group were(0.53 ± 0.52),(9.27 ± 2.60)and(4.53 ± 1.30),respectively.There was significant difference between the three groups(P <0.05)(1.53 ± 0.55),(12.27 ± 2.81)and(6.85 ± 1.52)points in the sham,ALCT + Vehicle and ALCT + Halofuginone groups,respectively.There was significant difference between the three groups(P < 0.05).(P <0.05).The expression of TGF-?1 positive cells / chondrocytes in sham,ALCT + Vehicle and ALCT + Halofuginone were(0.054 ± 0.013),(0.239 ± 0.032),(0.157 ± 0.013)(P <0.05).At 2 months,the expression of TGF-?1 positive cells / chondrocytes in sham,ALCT + Vehicle and ALCT + Halofuginone were significantly higher than those in the control group(P <0.05),and the difference was statistically significant(0.069 ± 0.018),(0.331 ± 0.041)and(0.213 ± 0.026),respectively.There was significant difference between the three groups(P <0.05).(18.33 ± 1.43),(109 ± 11.16)and(35 ± 2.83),respectively.The expression of TRAP positive cells in the subchondral cartilage of sham,ALCT + Vehicle and ALCT + Halofuginone(P <0.05).The number of Osterix positive cells / cells in the subchondral bone of sham,ALCT + Vehicle and ALCT + Halofuginone were(14.33 ± 0.95),(206 ± 23.46),and the difference was statistically significant),(85 ± 17.89)points;the difference between the three groups was statistically significant(P <0.05).Conclusion: Bushen ketone 0.5mg /(kg · d)can partially prevent the early osteoarthritis of the articular cartilage in mice.The mechanism may be to reduce the expression of TGF-?1,which affects the Psmad pathway and reduce osteoclast Resulting in delaying the progression of osteoarthritis. |
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