TRPA1在大鼠骨癌痛机械性痛觉过敏及温度异常性疼痛中的作用
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摘要
目的采用Walker256乳腺癌细胞注入大鼠胫骨髓腔构建骨癌痛模型,探讨TRPA1是否参与骨癌痛的发生与发展。方法12只雌性SD大鼠,随机分为假手术组(胫骨接种0.9%氯化钠注射液10μl)和骨癌痛模型组(胫骨接种Walker256肿瘤细胞10μl),每组6只,分别检测术前1 d及术后3d、5 d、7 d、9 d、11 d、13 d、14d两组大鼠机械性缩足阈值及温度缩足潜伏期变化。术后第14天对各组大鼠行X线及病理切片检查,western blot法检测两组大鼠TRPA1蛋门表达情况。另10只雌性SD大鼠,随机分为骨癌痛+TRPA1拮抗剂(A967079)组10 mg/kg 10μl、骨癌痛+0.9%氯化钠注射液组10μl,每组5只,分别检测给药前1h、给药后2 h、4 h、6 h、8 h机械性缩足阈值及温度缩足潜伏期变化。结果术后14d大鼠行X线及HE病理切片检测,骨癌痛大鼠肿瘤生长,出现异肿瘤细胞,成功建立骨癌痛模型。术后14d,与假手术组大鼠相比,骨癌痛组大鼠体质量、机械性缩足反射阈值、热痛潜伏期、冷痛潜伏期显著降低(P均<0.01),TRPA1蛋白表达量显著升高(P<0.01)。模型建立第7天时应用TRPA1拮抗剂A967079对骨癌痛大鼠进行干预,2～4 h时骨癌痛大鼠机械性痛敏升高(P均<0.01);给药后4～6 h热痛潜伏期升高(P均<0.01)。结论TRPA1对骨癌痛模型机械性痛觉过敏及温度异常性疼痛可能有介导作用。
Objective To establish bone cancer pain model by injecting Walker 256 tumor cells into tibial medullary canal of rats,and investigate the role of TRPA1 in occurrence and development of bone cancer pain.Methods Twelve female SpragueDawley rats were randomly divided into sham operation group(10μl of 0.9 sodium chloride injection)and bone cancer pain model group(10μl of Walker 256 tumor cells),with 6 rats in each group.The paw mechanical withdraw threshold and the paw thermal withdraw latency in the two groups at 1 day before operation and at 3,5,7,9,11,13,14 days after operation were detected X-ray and pathological examination were performed at 14 days after operation.The expression of TRPA1 protein in the two groups was detected by western blot.Another 10 female Sprague-Dawley rats were randomly divided into bone cancer pain+TRPA1 antagonist(A967079)group(10 mg/kg 10 μl)and bone cancer pain+ saline group(10μl),with 5 rats in each group.The paw mechanical withdraw threshold and the thermal withdraw latency in each group were measured at 1 hour before administration and at 2,4,6 and 8 hours after administration.Results At 14 days after operation,X-ray imaging showed that tibial bone was destructed,and HE pathological sections showed atypia tumor cells,indicating that the bone cancer pain model was successfully induced.Compared with sham operation group,the weight,paw mechanical withdraw threshold,thermal withdraw latency-hot,thermal withdraw latencycold in rats of bone cancer pain group were significantly lower(all P<0.01),and the relative protein expression of TRPA1 in rats of bone cancer pain group was higher(P<0.01).After TRPA1 antagonist(A967079)was administrated at 7 days after operation,mechanical allodynia alleviated at 2 h and 4 h after treatment in rats with bone cancer pain; and the paw thermal withdraw latencyhot was also elevated(all P<0.01).Conclusion TRPA1 mediates mechanical hyperalgesia and temperature allodynia in bone cancer pain model.
Objective To establish bone cancer pain model by injecting Walker 256 tumor cells into tibial medullary canal of rats,and investigate the role of TRPA1 in occurrence and development of bone cancer pain.Methods Twelve female SpragueDawley rats were randomly divided into sham operation group(10μl of 0.9 sodium chloride injection)and bone cancer pain model group(10μl of Walker 256 tumor cells),with 6 rats in each group.The paw mechanical withdraw threshold and the paw thermal withdraw latency in the two groups at 1 day before operation and at 3,5,7,9,11,13,14 days after operation were detected X-ray and pathological examination were performed at 14 days after operation.The expression of TRPA1 protein in the two groups was detected by western blot.Another 10 female Sprague-Dawley rats were randomly divided into bone cancer pain+TRPA1 antagonist(A967079)group(10 mg/kg 10 μl)and bone cancer pain+ saline group(10μl),with 5 rats in each group.The paw mechanical withdraw threshold and the thermal withdraw latency in each group were measured at 1 hour before administration and at 2,4,6 and 8 hours after administration.Results At 14 days after operation,X-ray imaging showed that tibial bone was destructed,and HE pathological sections showed atypia tumor cells,indicating that the bone cancer pain model was successfully induced.Compared with sham operation group,the weight,paw mechanical withdraw threshold,thermal withdraw latency-hot,thermal withdraw latencycold in rats of bone cancer pain group were significantly lower(all P<0.01),and the relative protein expression of TRPA1 in rats of bone cancer pain group was higher(P<0.01).After TRPA1 antagonist(A967079)was administrated at 7 days after operation,mechanical allodynia alleviated at 2 h and 4 h after treatment in rats with bone cancer pain; and the paw thermal withdraw latencyhot was also elevated(all P<0.01).Conclusion TRPA1 mediates mechanical hyperalgesia and temperature allodynia in bone cancer pain model.
引文
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2 Basile A,Giuliano G,Scuderi V,et al.Cementoplasty in the management of painful extraspinal bone metastases:our experience[J].Radiol Med,2008,113(7):1018-1028.
3 Suva LJ,Washam C,Nicholas RW,et al.Bone metastasis:mechanisms and therapeutic opportunities[J].Nat Rev Endocrinol,2011,7(4):208-218.
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7 Lam DK,Dang D,Zhang J,et al.Novel animal models of acute and chronic cancer pain:a pivotal role for PAR2[J].J Neurosci,2012,32(41):14178-14183.
8 Koivisto A,Chapman H,Jalava N,et al.TRPA1:a transducer and amplifier of pain and inflammation[J].Basic Clin Pharmacol Toxicol,2014,114(1):50-55.
9孙武平,周倩,肖礼祖.TRP通道在内脏痛中的研究进展[J].中国疼痛医学杂志,2018,24(1):54-59.
10郑晓娇,万宏梅,段雨岑,等.在冷环境中大鼠下丘脑TRPA1参与体温调节的作用[J].解剖科学进展,2018,24(4):378-381.
11 Chen D,Liu N,Li M,et al.Blocking PAR2 Alleviates Bladder Pain and Hyperactivity via TRPA1 Signal[J].Transl Neurosci,2016,7(1):133-138.
12 Asgar J,Zhang Y,Saloman JL,et al.The role of TRPA1 in muscle pain and mechanical hypersensitivity under inflammatory conditions in rats[J].Neuroscience,2015,310:206-215.
13 Shenoy PA,Kuo A,Vetter I,et al.The Walker 256 Breast Cancer Cell-Induced Bone Pain Model in Rats[J].Front Pharmacol,2016,7:286.
14刘艳红,张宏,徐龙河.大鼠骨癌痛模型的制备及电压依赖性钠通道Nav1.8在其背根神经节的表达研究[J].解放军医学杂志,2007,32(4):319-322.
15邱方,刘艳红,米卫东,等.基质细胞源性因子及其受体在慢性疼痛中的作用[J].解放军医学院学报,2015,(4):396-398.
16 Chaplan SR,Bach FW,Pogrel JW,et al.Quantitative assessment of tactile allodynia in the rat paw[J].J Neurosci Methods,1994,53(1):55-63.
17 Yalcin I,Charlet A,Freund-Mercier MJ,et al.Differentiating thermal allodynia and hyperalgesia using dynamic hot and cold plate in rodents[J].J Pain,2009,10(7):767-773.
18 Jasmin L,Kohan L,Franssen M,et al.The cold plate as a test of nociceptive behaviors:description and application to the study of chronic neuropathic and inflammatory pain models[J].Pain,1998,75(2-3):367-382.
19 Erami E,Azhdari-Zarmehri H,Ghasemi-Dashkhasan E,et al.Intra-paragigantocellularis lateralis injection of orexin-A has an antinociceptive effect on hot plate and formalin tests in rat[J].Brain Res,2012,1478:16-23.
20 Medhurst SJ,Walker K,Bowes M,et al.A rat model of bone cancer pain[J].Pain,2002,96(1-2):129-140.
21 Honore P,Luger NM,Sabino MA,et al.Osteoprotegerin blocks bone cancer-induced skeletal destruction,skeletal pain and painrelated neurochemical reorganization of the spinal cord[J].Nat Med,2000,6(5):521-528.
22 Nelson JB,Carducci MA.The role of endothelin-1 and endothelin receptor antagonists in prostate cancer[J].BJU Int,2000,85Suppl 2:45-48.
23 Pacharinsak C,Beitz A.Animal models of cancer pain[J].Comp Med,2008,58(3):220-233.
24 Sabino MA,Luger NM,Mach DB,et al.Different tumors in bone each give rise to a distinct pattern of skeletal destruction,bone cancer-related pain behaviors and neurochemical changes in the central nervous system[J].Int J Cancer,2003,104(5):550-558.
25 Wacnik PW,Eikmeier LJ,Ruggles TR,et al.Functional interactions between tumor and peripheral nerve:morphology,algogen identification,and behavioral characterization of a new murine model of cancer pain[J].J Neurosci,2001,21(23):9355-9366.
26 Wacnik PW,Kehl LJ,Trempe TM,et al.Tumor implantation in mouse humerus evokes movement-related hyperalgesia exceeding that evoked by intramuscular carrageenan[J].Pain,2003,101(1-2):175-186.
27 Menendez L,Lastra A,Fresno MF,et al.Initial thermal heat hypoalgesia and delayed hyperalgesia in a murine model of bone cancer pain[J].Brain Res,2003,969(1-2):102-109.
28 Antoniazzi CTD,Nassini R,Rigo FK,et al.Transient receptor potential ankyrin 1(TRPA1)plays a critical role in a mouse model of cancer pain[J].Int J Cancer,2019,144(2):355-365.
29张春鹏,周庆辉.骨癌痛模型研究进展[J].中国疼痛医学杂志,2018,24(10):774-777.
30 Sasaki A,Yoneda T,Terakado N,et al.Experimental bone metastasis model of the oral and maxillofacial region[J].Anticancer Res,1998,18(3a):1579-1584.
31 Xia H,Zhang D,Yang S,et al.Role of ATP-sensitive potassium channels in modulating nociception in rat model of bone cancer pain[J].Brain Res,2014,1554:29-35.
32 Menendez L,Juarez L,Garcia E,et al.Analgesic effects of capsazepine and resiniferatoxin on bone cancer pain in mice[J].Neurosci Lett,2006,393(1):70-73.
33 Asai H,Ozaki N,Shinoda M,et al.Heat and mechanical hyperalgesia in mice model of cancer pain[J].Pain,2005,117(1-2):19-29.
34 Hartel M,di Mola FF,Selvaggi F,et al.Vanilloids in pancreatic cancer:potential for chemotherapy and pain management[J].Gut,2006,55(4):519-528.
35宋晓文,陈金波,吴欣彤.瞬时受体电位A1通道在偏头痛中的研究进展[J].中国疼痛医学杂志,2018,24(2):122-125.
2 Basile A,Giuliano G,Scuderi V,et al.Cementoplasty in the management of painful extraspinal bone metastases:our experience[J].Radiol Med,2008,113(7):1018-1028.
3 Suva LJ,Washam C,Nicholas RW,et al.Bone metastasis:mechanisms and therapeutic opportunities[J].Nat Rev Endocrinol,2011,7(4):208-218.
4 Ross JR,Saunders Y,Edmonds PM,et al.Systematic review of role of bisphosphonates on skeletal morbidity in metastatic cancer[J].Bmj,2003,327(7413):469.
5 Smith TJ,Staats PS,Deer T,et al.Randomized clinical trial of an implantable drug delivery system compared with comprehensive medical management for refractory cancer pain:impact on pain,drug-related toxicity,and survival[J].J Clin Oncol,2002,20(19):4040-4049.
6 Jones B,Finlay I,Ray A,et al.Is there still a role for open cordotomy in cancer pain management?[J].J Pain Symptom Manage,2003,25(2):179-184.
7 Lam DK,Dang D,Zhang J,et al.Novel animal models of acute and chronic cancer pain:a pivotal role for PAR2[J].J Neurosci,2012,32(41):14178-14183.
8 Koivisto A,Chapman H,Jalava N,et al.TRPA1:a transducer and amplifier of pain and inflammation[J].Basic Clin Pharmacol Toxicol,2014,114(1):50-55.
9孙武平,周倩,肖礼祖.TRP通道在内脏痛中的研究进展[J].中国疼痛医学杂志,2018,24(1):54-59.
10郑晓娇,万宏梅,段雨岑,等.在冷环境中大鼠下丘脑TRPA1参与体温调节的作用[J].解剖科学进展,2018,24(4):378-381.
11 Chen D,Liu N,Li M,et al.Blocking PAR2 Alleviates Bladder Pain and Hyperactivity via TRPA1 Signal[J].Transl Neurosci,2016,7(1):133-138.
12 Asgar J,Zhang Y,Saloman JL,et al.The role of TRPA1 in muscle pain and mechanical hypersensitivity under inflammatory conditions in rats[J].Neuroscience,2015,310:206-215.
13 Shenoy PA,Kuo A,Vetter I,et al.The Walker 256 Breast Cancer Cell-Induced Bone Pain Model in Rats[J].Front Pharmacol,2016,7:286.
14刘艳红,张宏,徐龙河.大鼠骨癌痛模型的制备及电压依赖性钠通道Nav1.8在其背根神经节的表达研究[J].解放军医学杂志,2007,32(4):319-322.
15邱方,刘艳红,米卫东,等.基质细胞源性因子及其受体在慢性疼痛中的作用[J].解放军医学院学报,2015,(4):396-398.
16 Chaplan SR,Bach FW,Pogrel JW,et al.Quantitative assessment of tactile allodynia in the rat paw[J].J Neurosci Methods,1994,53(1):55-63.
17 Yalcin I,Charlet A,Freund-Mercier MJ,et al.Differentiating thermal allodynia and hyperalgesia using dynamic hot and cold plate in rodents[J].J Pain,2009,10(7):767-773.
18 Jasmin L,Kohan L,Franssen M,et al.The cold plate as a test of nociceptive behaviors:description and application to the study of chronic neuropathic and inflammatory pain models[J].Pain,1998,75(2-3):367-382.
19 Erami E,Azhdari-Zarmehri H,Ghasemi-Dashkhasan E,et al.Intra-paragigantocellularis lateralis injection of orexin-A has an antinociceptive effect on hot plate and formalin tests in rat[J].Brain Res,2012,1478:16-23.
20 Medhurst SJ,Walker K,Bowes M,et al.A rat model of bone cancer pain[J].Pain,2002,96(1-2):129-140.
21 Honore P,Luger NM,Sabino MA,et al.Osteoprotegerin blocks bone cancer-induced skeletal destruction,skeletal pain and painrelated neurochemical reorganization of the spinal cord[J].Nat Med,2000,6(5):521-528.
22 Nelson JB,Carducci MA.The role of endothelin-1 and endothelin receptor antagonists in prostate cancer[J].BJU Int,2000,85Suppl 2:45-48.
23 Pacharinsak C,Beitz A.Animal models of cancer pain[J].Comp Med,2008,58(3):220-233.
24 Sabino MA,Luger NM,Mach DB,et al.Different tumors in bone each give rise to a distinct pattern of skeletal destruction,bone cancer-related pain behaviors and neurochemical changes in the central nervous system[J].Int J Cancer,2003,104(5):550-558.
25 Wacnik PW,Eikmeier LJ,Ruggles TR,et al.Functional interactions between tumor and peripheral nerve:morphology,algogen identification,and behavioral characterization of a new murine model of cancer pain[J].J Neurosci,2001,21(23):9355-9366.
26 Wacnik PW,Kehl LJ,Trempe TM,et al.Tumor implantation in mouse humerus evokes movement-related hyperalgesia exceeding that evoked by intramuscular carrageenan[J].Pain,2003,101(1-2):175-186.
27 Menendez L,Lastra A,Fresno MF,et al.Initial thermal heat hypoalgesia and delayed hyperalgesia in a murine model of bone cancer pain[J].Brain Res,2003,969(1-2):102-109.
28 Antoniazzi CTD,Nassini R,Rigo FK,et al.Transient receptor potential ankyrin 1(TRPA1)plays a critical role in a mouse model of cancer pain[J].Int J Cancer,2019,144(2):355-365.
29张春鹏,周庆辉.骨癌痛模型研究进展[J].中国疼痛医学杂志,2018,24(10):774-777.
30 Sasaki A,Yoneda T,Terakado N,et al.Experimental bone metastasis model of the oral and maxillofacial region[J].Anticancer Res,1998,18(3a):1579-1584.
31 Xia H,Zhang D,Yang S,et al.Role of ATP-sensitive potassium channels in modulating nociception in rat model of bone cancer pain[J].Brain Res,2014,1554:29-35.
32 Menendez L,Juarez L,Garcia E,et al.Analgesic effects of capsazepine and resiniferatoxin on bone cancer pain in mice[J].Neurosci Lett,2006,393(1):70-73.
33 Asai H,Ozaki N,Shinoda M,et al.Heat and mechanical hyperalgesia in mice model of cancer pain[J].Pain,2005,117(1-2):19-29.
34 Hartel M,di Mola FF,Selvaggi F,et al.Vanilloids in pancreatic cancer:potential for chemotherapy and pain management[J].Gut,2006,55(4):519-528.
35宋晓文,陈金波,吴欣彤.瞬时受体电位A1通道在偏头痛中的研究进展[J].中国疼痛医学杂志,2018,24(2):122-125.
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