独活寄生汤含药血清对退变软骨细胞PERK/Bip通路关键调控蛋白的影响
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摘要
目的观察独活寄生汤含药血清对大鼠退变软骨细胞PERK/Bip信号通路关键调控蛋白的影响。方法采用IL-1β诱导建立体外退变软骨细胞模型,分别采用空白血清和独活寄生汤含药血清干预24 h、48 h、72h后,收集软骨细胞,采用Western Blot法检测退变软骨细胞中PERK、Bip、eIF-2α、ATF-4、GADD153、Caspase-9和Caspase-3蛋白表达变化。结果随着干预时间的延长,空白血清组和含药血清组PERK、Bip、eIF-2α、ATF-4、GADD153、Caspase-9、Caspase-3蛋白表达均逐渐降低,含药血清组降低更明显,差异均有统计学意义(P<0.05)。结论独活寄生汤可能是通过调控PERK/Bip信号通路,抑制软骨细胞凋亡,从而起到治疗膝骨关节炎的作用。
Objective: To observe the effect of serum medicated with Duhuo Jisheng Decoction(DHJSD) on the key regulation proteins of PERK/Bip signaling pathway in rat's degenerated chondrocytes. Methods: The in vitro degenerated chondrocytes models were established, which were intervened by blank serum and serum medicated with Duhuo Jisheng Decoction for 24 h, 48 h and 72 h,the chondrocytes were then collected for Western Blot to detect the protein expression of PERK, Bip, eIF-2α, ATF-4, GADD153, Caspase-9 and Caspase-3 in degenerated chondrocytes after serum intervention. Results: With prolonging of intervention, the protein expression of PERK, Bip, eIF-2α, ATF-4, GADD153, Caspase-9 and Caspase-3 in both groups were gradually decreased, and those of medicated serum group showed more significant. The difference between each time point was statistically significant(P<0.05). Conclusion: DHJSD might inhibit the apoptosis of chondrocytes by regulating the PERK/Bip signaling pathway, which may, in part, explain its clinical efficacy in the treatment of knee OA.
Objective: To observe the effect of serum medicated with Duhuo Jisheng Decoction(DHJSD) on the key regulation proteins of PERK/Bip signaling pathway in rat's degenerated chondrocytes. Methods: The in vitro degenerated chondrocytes models were established, which were intervened by blank serum and serum medicated with Duhuo Jisheng Decoction for 24 h, 48 h and 72 h,the chondrocytes were then collected for Western Blot to detect the protein expression of PERK, Bip, eIF-2α, ATF-4, GADD153, Caspase-9 and Caspase-3 in degenerated chondrocytes after serum intervention. Results: With prolonging of intervention, the protein expression of PERK, Bip, eIF-2α, ATF-4, GADD153, Caspase-9 and Caspase-3 in both groups were gradually decreased, and those of medicated serum group showed more significant. The difference between each time point was statistically significant(P<0.05). Conclusion: DHJSD might inhibit the apoptosis of chondrocytes by regulating the PERK/Bip signaling pathway, which may, in part, explain its clinical efficacy in the treatment of knee OA.
引文
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[19]熊飞,韦宜山.内质网应激与软骨细胞凋亡[J].中国组织工程研究,2017,21(36):5886-5891.
[20]ZHANG P,MCGRATH B,LI S,et al.The PERK eukaryotic initiation factor 2 alpha kinase is required for the development of the skeletal system,postnatal growth,and the function and viability of the pancreas[J].Mol Cell Biol,2002,22(11):3864-3874.
[21]WANG F M,GALSON D L,ROODMAN G D,et al.Resveratrol triggers the pro-apoptotic endoplasmic reticulum stress response and represses pro-survival XBP1 signaling in human multiple myeloma cells[J].Exp Hematol,2011,39(10):999-1006.
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[23]GRISHKO V,XU M,WILSON G,et al.Apoptosis and mitochondrial dysfunction in human chondrocytes following exposure to lidocaine,bupivacaine,and ropivacaine[J].J Bone Joint Surg Am,2010,92(3):609-618.
[24]SEOL J W,LEE H B,LEE Y J,et al.Hypoxic resistance to articular chondrocyte apoptosis-a possible mechanism of maintaining homeostasis of normal articular cartilage[J].FEBS J,2009,276(24):7375-7385.
[25]潘彩彬,冯阳,刘献祥.独活寄生汤的内涵与外延探讨[J].风湿病与关节炎,2013,2(6):39-41.
[26]顾程泷,朱艳伟,孙玉明.独活寄生汤在骨伤科的运用[J].河南中医,2013,33(4):605-607.
[2]SHAKOOR M A,RAHMAN M S,AZAD A K,et al.Effects of isometric quadriceps muscle strengthening exercise on chronic osteoarthritis of the knee[J].Bangladesh Med Res Counc Bull,2010,36(1):20-22.
[3]李宁华,薛庆云,张毅,等.中国六城市中老年人群X线膝骨关节炎流行病学分析[J].实用医学杂志,2008,24(16):2887-2888.
[4]周腊梅,王亚克.独活寄生汤治疗膝骨关节炎的临床疗效观察[J].成都中医药大学学报,2014,37(3):46-48,78.
[5]邝高艳,严可,柴爽,等.独活寄生汤加减治疗膝骨关节炎疗效及安全性的Meta分析[J].中医药导报,2016,22(17):59-62.
[6]万美珍,邓紫玉,代龙金.独活寄生汤治疗肝肾不足型膝骨关节炎的临床疗效观察[J].中国社区医师,2017,33(14):92-93.
[7]刘云,张敏.独活寄生汤联合运动疗法治疗膝骨关节炎临床观察[J].风湿病与关节炎,2017,6(2):29-32.
[8]陈俊,吴广文,黄云梅,等.独活寄生汤对膝骨关节炎大鼠关节软骨基质的影响[J].福建中医药,2017,48(6):24-26.
[9]郑文伟,翁霞萍,黄绥心,等.独活寄生汤对骨关节炎软骨退变的影响及其作用机制[J].中医正骨,2017,29(7):5-11.
[10]吴广文,王武炼,潘彩彬,等.独活寄生汤含药血清对大鼠退变软骨细胞线粒体凋亡通路的影响[J].风湿病与关节炎,2014,3(10):5-9.
[11]黄继汉,黄晓晖,陈志扬,等.药理试验中动物间和动物与人体间的等效剂量换算[J].中国临床药理学与治疗学,2004,9(9):1069-1072.
[12]叶锦霞,付长龙,林洁,等.透骨消痛胶囊对毒胡萝卜素诱导的内质网应激(PEKR信号通路)介导的大鼠体外培养关节软骨细胞凋亡的影响[J].中医正骨,2017,29(6):1-7.
[13]PARK K,HOFFMEISTER B,HAN D K.Therapeutic ultrasound effects on interleukin-1beta stimulated cartilage construct in vitro[J].Ultrasound Med Biol,2007,33(2):286-295.
[14]吴广文.独活寄生汤对骨性关节炎软骨细胞增殖与凋亡影响的实验研究[D].福州:福建中医药大学,2014:39-40.
[15]ROUSH J K,CROSS A R,RENBERG W C,et al.Evaluation of the effects of dietary supplementation with fish oil omega-3 fatty acids on weight bearing in dogs with osteoarthritis[J].J Am Vet Med Assoc,2010,236(1):67-73.
[16]李西海,梁文娜.软骨下骨骨重塑与骨关节炎的关系[J].国际骨科学杂志,2007,28(1):35-37.
[17]MALEMUD C J.Protein kinases in chondrocyte signaling and osteoarthritis[J].Clin Orthop Relat Res,2004(427 Suppl):S145-151.
[18]MISTRY D,OUE Y,CHAMBERS M G,et al.Chondrocyte death during murine osteoarthritis[J].Osteoarthritis Cartilage,2004,12(2):131-141.
[19]熊飞,韦宜山.内质网应激与软骨细胞凋亡[J].中国组织工程研究,2017,21(36):5886-5891.
[20]ZHANG P,MCGRATH B,LI S,et al.The PERK eukaryotic initiation factor 2 alpha kinase is required for the development of the skeletal system,postnatal growth,and the function and viability of the pancreas[J].Mol Cell Biol,2002,22(11):3864-3874.
[21]WANG F M,GALSON D L,ROODMAN G D,et al.Resveratrol triggers the pro-apoptotic endoplasmic reticulum stress response and represses pro-survival XBP1 signaling in human multiple myeloma cells[J].Exp Hematol,2011,39(10):999-1006.
[22]RASHEVA V I,DOMINGOS P M.Cellular responses to endoplasmic reticulum stress and apoptosis[J].Apoptosis,2009,14(8):996-1007.
[23]GRISHKO V,XU M,WILSON G,et al.Apoptosis and mitochondrial dysfunction in human chondrocytes following exposure to lidocaine,bupivacaine,and ropivacaine[J].J Bone Joint Surg Am,2010,92(3):609-618.
[24]SEOL J W,LEE H B,LEE Y J,et al.Hypoxic resistance to articular chondrocyte apoptosis-a possible mechanism of maintaining homeostasis of normal articular cartilage[J].FEBS J,2009,276(24):7375-7385.
[25]潘彩彬,冯阳,刘献祥.独活寄生汤的内涵与外延探讨[J].风湿病与关节炎,2013,2(6):39-41.
[26]顾程泷,朱艳伟,孙玉明.独活寄生汤在骨伤科的运用[J].河南中医,2013,33(4):605-607.