Ⅱ型胶原变构肽对胶原性关节炎软骨破坏的影响

Inhibition of cartilage destruction in collagen-induced arthritis by altered CII 263-272 peptide:experiment with rats

目的研究连续替换Ⅱ型胶原(CⅡ)268-270位T细胞受体(TCR)结合氨基酸的CⅡ变构肽sub268-270对胶原性关节炎(CIA)关节软骨损伤的抑制作用。方法使用牛CⅡ在Lewis大鼠诱发CIA。将CIA大鼠分为30μg组、5μg组、1μg组和空白对照组,每组8只,每次分别接受CⅡ变构肽sub268-27030μg、5μg、1μg及磷酸盐缓冲液静脉注射,每周2次。从关节炎症积分、病理学滑膜炎症积分、血管翳评分、关节软骨损伤评分和骨破坏积分评价变构肽疗效,用血清软骨寡聚基质蛋白(COMP)水平和关节组织切片中软骨番红花红含量来评价变构肽对关节软骨损伤的抑制作用。结果30μgCⅡ变构肽组关节炎症积分为5·6±2·63,显著低于5μg组为(9·67±5·61)、1μg组为(10·02±5·06)及空白对照组为(11·8±5·34)(P<0·01)。与5μg变构肽、1μg变构肽和PBS相比,30μg变构肽可以显著降低CIA大鼠滑膜炎积分(P<0·01)、血管翳积分(P<0·01)、软骨损伤积分(P<0·01)和骨破坏积分(P<0·01)。30μg变构肽组血清COMP水平(μg/ml)为2·21±0·76,5μg变构肽组为5·63±1·73,1μg变构肽组为6·04±1·76,空白对照组为7·00±1·46,30μg治疗组与空白对照组比P<0·01。关节组织切片中软骨蛋白聚糖含量(A值)30μg组为2·35±0·76,5μg组为1·57±0·63,1μg组为1·37±0·53,空白对照组为1·00±0·41,30μg组与空白对照组比P<0·01。结论30μgCⅡ变构肽sub268-270可以有效缓解CIA,显著抑制CIA中的关节软骨破坏。

Objective To study the inhibitory effect of altered collagen Ⅱ （ C Ⅱ ） 263-272 peptide （sub268-270） with three consecutive substitutions of TCR- contacting residues on joint inflammation and cartilage destruction in collagen-induced arthritis （CIA）. Methods Thirty-two Lewis rats were injected intracutaneously with bovine collagen type Ⅱ so as to establish models of arthritis and then were randomly divided into 4 equal groups to be injected intravenously with sub268-270 30 μg, 5 μg, or 1 μg and PBS twice a week for 3 weeks. The therapeutic effect of the altered peptide on arthritis was evaluated by arthritis score. After the treatment the rats were killed and their ankle joints were taken to undergo pathological examination to observe the existence of synovitis, pannus formation, cartilage damage, and bone erosion. Blood samples were collected to detect the serum cartilage oligomeric matrix protein （COMP）. Cartilage proteoglycan-speciflc dye safaranin O was used on the joint sections to observe the coloration of the dye in the cartilage. Results The arthritis score in rats treated by 30 μg altered C Ⅱ peptide was （5. 6 ± 2. 63 ）, significantly lower than those of the 5 μg, 1 μg, and blank control groups [ （9. 67 ± 5. 61 ）, （ 10. 02 ± 5. 06） , and （ 11.8 ±5. 34） respectively, all P 〈0. 01 ]. The synovitis score of the 30 μg group was （ 1.11 ± 0. 43） , significantly lower than those of the 5 μg, 1 μg, and blank control groups [ （ 1.87 ± 0. 78 ） , （2. 11 ±0. 83） , and （2. 25 ±0.73） respectively, all P 〈0. 01 ]. The pannus score of the 30 μg group was （ 1.11 ± 0. 43 ）, significantly lower than those of the 5 μg, 1 μg, and blank control groups [ （ 1.83 ± 0. 79 ） , （ 2. 07 ± 0. 91 ） , and （ 2. 27 ± 0. 71 ） respectively, all P 〈 0. 01 ]. The cartilage damage score of the 30 μg group was 0. 56 ±0. 23）, significantly lower than those of the 5 μg, 1 μg, and blank control groups [ （ 1.91 -±0. 83） , （2. 13 ±0.79） , and （2. 29 ±0. 69） respectively, all P 〈0. 01 ]. The bone erosion score of the 30 μg group was （0. 53 ±0. 21 ）, significantly lower than those of the 5 μg, 1 μg, and blank control groups [（1.71±0.67）, （1.88 ±0.93）, and （2.01 ±0.93） respectively, all P〈0.01]. The serum COMP of the 30 μg group was （2.21 ±0. 76）, significantly lower than those of the 5 μg, l μg, and blank control groups [（5.63±1.73）, （6.04±1.76）, and （7.00±1.46） respectively, all P〈0.011. The content of safranin O （ A value） in the joint section of the 30 μg group was （2. 35 ± 0. 76）, significantly higher than those of the 5 μg, 1 μg, and blank control groups [（1.57 ±0.63）, （1.37±0.53）, and （ 1.00 ± 0.41 ） respectively, all P 〈 0. 011. Conclusion The altered C Ⅱ peptide sub268-270 effectively ameliorates CIA and inhibits the cartilage damage in CIA, and may modify the disease course of rheumatoid arthritis.