骨痹方对大鼠膝骨关节炎PI3K/AKt信号通路及组织形态学的影响
Effects of Gubi Recipe on the PI3K/AKt Pathway and Synovial Tissue of Knee Joint in the Rats Model of Knee Osteoarthritis
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摘要: 探讨骨痹方对KOA大鼠软骨细胞PI3K/AKt信号通路及膝关节滑膜病理组织形态学的影响。方法 按照改良Hulth's法复制40只KOA大鼠模型随机分为5组:模型组、阳性药组(维固力组)、骨痹方高、中、低剂量组,每组8只,另假手术组(n=8)大鼠术中暴露前后交叉韧带后即可,正常组(n=8)大鼠无需任何处置。每日灌胃1次,共8周。8周后取各组大鼠患膝滑膜,观察其组织形态学变化;应用Western blot法检测膝关节软骨细胞PI3K/AKt信号通路AKt及其下游靶蛋白Bcl-2、Bax的表达。结果 模型组大鼠膝关节滑膜组织形态学检查评分比正常组显著增高(P<0.01);骨痹方中、高剂量组大鼠膝关节滑膜损伤评分较模型组均有所下降(P<0.05)。模型组大鼠AKt及下游靶蛋白Bcl-2表达低于正常组,骨痹方高剂量组AKt表达量显著高于正常组(P<0.01),骨痹方中剂量组AKt表达量与正常组接近;与模型组比较,骨痹方高剂量组AKt表达量及下游靶蛋白Bcl-2显著升高(P<0.01),Bax表达降低。结论 骨痹方可以有效地减少膝骨关节炎病变过程中软骨细胞的凋亡,一定程度地改善膝骨关节炎病理组织形态学,其作用机制可能与其干预KOA大鼠软骨细胞PI3K/AKt信号通路密切有关。
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关键词:
- 膝骨关节炎 /
- 骨痹方 /
- 软骨细胞凋亡 /
- PI3K/AKt信号通路
Abstract: OBJECTIVE To study the effects of Gubi recipe on the PI3K/AKt pathway and synovial tissue of nnee joint in the rats model of knee osteoarthritis. METHODS SD rats(n=40) were duplicated by Hulth's technique and randomly divided into the model group, the Glucosamine Sulfate capsules group, the low dose group of Gubi recipe, the dose group of Gubi recipe and the high dose group of Gubi recipe. Also, the rats in the sham group (n=8) were exposed to the cruciate ligament after the operation, and the normal group (n=8) did not need any disposal. All rats were given the solution by filling the stomach for 8 weeks. 8 weeks later, the the synovial membrane was taken from each rat for observing; Western blot technic of articular cartilage PI3K/AKt pathway AKt activation extent and detect the expression of Bcl-2 and Bax. RESULTS Gubi recipe can improve the knee synovial membrane injury score of rats, effectively reduce the inflammatory reaction in knee synovial membrane. The expression of AKt and downstream target protein Bcl-2 was lower than that of normal group, the expression level of AKt in the high dose group of Gubi recipe was much higher than that in the normal group(P<0.01), the expression level of AKt in the middle dose group was close to that in the normal group; compared with the model group, the expression of AKt and the downstream target protein Bcl-2 in the high dose group were significantly higher (P<0.01), and the expression of Bax was decreased. CONCLUSION Gubi recipe can effectively reduce the chondrocyte apoptosis of osteoarthritis of the knee lesions in the process, to some extent improve the pathology of knee osteoarthritis, the mechanism may be related to the intervention of PI3K/AKt pathway of knee osteoarthritis cartilage tissue of rat. -
[1] FRANSEN M, BRIDGETT L, MARCH L, et al. The epidemiology of osteoarthritis in Asia[J]. Int J Rheum Dis, 2011, 14(2): 113-121.〖ZK)〗 [2] ULICI V, HOENSELAAR KD, GILLESPIE JR, et al. The PI3K pathway regulates endochondral bone growth through control of hypertrophic chondrocyte differentiation[J]. BMC Dev Biol, 2008, 8(8): 40.〖ZK)〗 [3] TONG KM, SHIEH DC, CHEN CP, et al. Leptin induces IL-8 expression via leptin receptor, IRS-1, PI3K, Akt cascade and promotion of NF-kappaB/p300 binding in human synovial fibroblasts[J]. Cell Signal, 2008, 20(8): 1478-1488.〖ZK)〗 [4] HSIEH YS, YANG SF, LUE KH, et al. Upregulation of urokinase-type plasminogen activator and inhibitor and gelatinase expression via 3 mito-gen-activated protein kinases and PI3K pathways during the early develop-ment of osteoarthritis[J]. J Rheumatol, 2007, 34(4): 785-793.〖ZK)〗 [5] ROGART JN, BARRACH HJ, CHICHESTER CO. Articular collagen degradation in the Hulth-Telhag model of osteoarthritis[J]. Osteoarthritis Cartilage, 1999, 7(6):539-547.〖ZK)〗 [6] 李文雄,孙赫,沈玮,等.SD大鼠骨关节炎造模方法的筛选及优化研究[J].牧畜与兽医,2014,46(12):71-73.〖ZK)〗 [7] CARPTEN JD, FABER AL, HORN C, et al. A transforming mutation in the pleckstrin homology domain of AKT1 in cancer[J]. Nature, 2007, 448(7152): 439-444.〖ZK)〗 [8] GAROFALO RS, ORENA SJ, RAFIDI K, et al. Severe diabetes, age-dependent loss of adipose tissue, and mild growth deficiency in mice lacking Akt2/PKB beta[J]. J Clin Invest, 2003, 112(2): 197-208.〖ZK)〗 [9] YANG ZZ, TSCHOPP O, BAUDRY A, et al. Physiological functions of proteinkinase B/A Kt[J]. Biochem Soc Trans, 2004, 32(Pt2): 350-354.〖ZK)〗 [10] TONG KM, SHIEH DC, CHEN CP, et al. Leptin induces IL-8 expression via leptin receptor, IRS-1, PI3K, Akt cascade and promotion of NF-kappaB/p300 binding in human synovial fibroblasts[J]. Cell Signal, 2008, 20(8): 1478-1488.〖ZK)〗 [11] HOFLER A, NICHOLS T, GRANT S, et al. Study of the PDK1/AKT signaling pathway using selective PDK1 inhibitors, HCS, and enhanced biochemical assays[J]. Anal Biochem, 2011, 414(2): 179-186.〖ZK)〗 [12] HUSSAIN AR, UDDIN S, AHMED M, et al. Prognostic significance of XIAP expression in DLBCL and effect of its inhibition on AKT signalling[J]. J Pathol, 2010, 222(2): 180-190.〖ZK)〗 [13] MALLA R, GOPINATH S, ALAPATI K, et al. Downregulation of uPAR and cathepsin B induces apoptosis via regulation of Bcl-2 and Bax and inhibition of the PI3K/Akt pathway in gliomas[J]. PLoS One, 2010, 5(10): e13731.〖ZK)〗 [14] GU S, PAPADOPOULOU N, NASIR O, et al. Activation of membrane androgen receptors in colon cancer inhibits the prosurvival signals Akt/bad in vitro and in vivo and blocks migration via vinculin/actin signaling[J]. Mol Med, 2011, 17(1/2): 48-58.〖ZK)〗 [15] KLOSOWSKA K, VOLIN MV, HUYNH N, et al. Fractalkine functions as a chemoattractant for osteoarthritis synovial fibroblasts and stimulates phosphorylation of mitogen-activated protein kinases and Akt[J]. Clin Exp Immunol, 2009, 156(2): 312-319.〖ZK)〗 [16] WONG ML, KAYE AH, HOVENS CM. Targeting malignant glioma survival signalling to improve clinical outcomes[J]. J Clin Neurosci, 2007, 14(4): 301-308.〖ZK)〗
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