当归四逆汤对奥沙利铂神经毒性大鼠背根神经节TRPs通道的影响
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摘要
目的探讨当归四逆汤对奥沙利铂神经毒性大鼠背根神经节TRPs通道的影响。方法使用奥沙利铂腹腔注射造成慢性神经毒性大鼠模型,60只大鼠分空白组、模型组、及当归四逆汤高、中、低剂量组。通过大鼠疼痛行为学、机械疼痛阈值测定检测当归四逆汤对奥沙利铂神经毒性的缓解作用,并进一步通过RT-PCR、Western blot观察大鼠背根神经节TRPs(TRPV1、TRPA1、TRPM8)mRNA及蛋白表达水平变化。结果大鼠行为学测定显示,与空白组比较,模型组、当归四逆汤低、中剂量组机械疼痛阈值下降(P<0.05),而高剂量组未出现明显的下降(P>0.05)。与空白组比较,其余各组大鼠背根神经节TRPA1、TRPM8、TRPV1蛋白表达量显著增加(P<0.01)。与模型组比较,当归四逆汤低剂量组可降低TRPM8蛋白表达(P<0.05),中剂量组可降低TRPA1(P<0.01)、TRPM8(P<0.01)、TRPV1(P<0.05)蛋白表达,高剂量组明显降低三者蛋白表达量(P<0.01)。与空白组比较,其余各组大鼠背根神经节TRPV1、TRPA1、TRPM8mRNA表达显著增加(P<0.01),与模型组比较,当归四逆汤各组均可显著下调三者表达(P<0.01)。结论当归四逆汤可以预防奥沙利铂慢性神经毒性,减缓大鼠机械疼痛阈值下降程度,其机制可能与下调TRPs通道蛋白表达有关。
OBJECTIVE To investigate the effect of Danggui Sini Decoction on TRPs channels in the dorsal root ganglia of the oxaliplatin-induced neurotoxic rats.METHODS A rat model of chronic neurotoxicity was induced by intraperitoneal injection of oxaliplatin.60 rats were divided into blank group,model group,and Danggui Sini Decoction high,medium and low dose groups.The neurotoxicity of oxaliplatin was detected by rat pain behavior and mechanical pain threshold.The mRNA and protein expression levels change of TRPs(TRPV1,TRPA1,TRPM8)in rat dorsal root ganglion were further observed by RTPCR and Western blot.RESULTS Compared with the blank group,the model group,the low-and medium-dose group of Danggui Sini Decoction showed a significant decrease in the mechanical pain threshold(P<0.05),while the high-dose group did not show a significant decrease.Compared with the blank group,the expression of TRPA1,TRPM8 and TRPV1 protein in dorsal root ganglion of each group increased with oxaliplatin(P<0.01).Compared with the model group,the low dose group of Danggui Sini Decoction reduced TRPM8 protein expression(P<0.05),and the middle dose group decreased TRPA1(P<0.01),TRPM8(P<0.01),TRPV1(P<0.05)while high dose group can significantly reduce the expression of TRPs(TRPV1,TRPA1,TRPM8)(P<0.01).Compared with the blank group,the expression of TRPV1,TRPA1 and TRPM8 mRNA in the dorsal root ganglion of the rats in each group were significantly increased after application of oxaliplatin(P<0.01).Compared with the model group,each group could be significantly down-regulated after applying the Danggui decoction.Expression(P<0.01).CONCLUSION Danggui Sini Decoction can prevent the chronic neurotoxicity of oxaliplatin and slow down the decline of mechanical pain threshold in rats.The mechanism may be related to the down-regulation of TRP channel protein expression.
OBJECTIVE To investigate the effect of Danggui Sini Decoction on TRPs channels in the dorsal root ganglia of the oxaliplatin-induced neurotoxic rats.METHODS A rat model of chronic neurotoxicity was induced by intraperitoneal injection of oxaliplatin.60 rats were divided into blank group,model group,and Danggui Sini Decoction high,medium and low dose groups.The neurotoxicity of oxaliplatin was detected by rat pain behavior and mechanical pain threshold.The mRNA and protein expression levels change of TRPs(TRPV1,TRPA1,TRPM8)in rat dorsal root ganglion were further observed by RTPCR and Western blot.RESULTS Compared with the blank group,the model group,the low-and medium-dose group of Danggui Sini Decoction showed a significant decrease in the mechanical pain threshold(P<0.05),while the high-dose group did not show a significant decrease.Compared with the blank group,the expression of TRPA1,TRPM8 and TRPV1 protein in dorsal root ganglion of each group increased with oxaliplatin(P<0.01).Compared with the model group,the low dose group of Danggui Sini Decoction reduced TRPM8 protein expression(P<0.05),and the middle dose group decreased TRPA1(P<0.01),TRPM8(P<0.01),TRPV1(P<0.05)while high dose group can significantly reduce the expression of TRPs(TRPV1,TRPA1,TRPM8)(P<0.01).Compared with the blank group,the expression of TRPV1,TRPA1 and TRPM8 mRNA in the dorsal root ganglion of the rats in each group were significantly increased after application of oxaliplatin(P<0.01).Compared with the model group,each group could be significantly down-regulated after applying the Danggui decoction.Expression(P<0.01).CONCLUSION Danggui Sini Decoction can prevent the chronic neurotoxicity of oxaliplatin and slow down the decline of mechanical pain threshold in rats.The mechanism may be related to the down-regulation of TRP channel protein expression.
引文
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[12]STEPHAN W,HOHMANN CA,SORIN T,et al.The G2Areceptor(GPR132)contributes to oxaliplatin-induced mechanical pain hypersensitivity[J].Sci Rep,2017,7:446.
[13]TAKAHITO M,SAKI N,ZHAO M,et al.Distinct mechanism of cysteine Oxidation-dependent activation and cold sensitization of human transient receptor potential ankyrin 1channel by high and low oxaliplatin[J].Front Physiol,2017,8:878.
[14]TSUGUNOBU A,SHIZUKA M,YASUSHI K.Shakuyakukanzoto attenuates oxaliplatin-induced cold dysesthesia by inhibiting the expression of transient receptor potential melastatin 8in mice.[J].Tradit Complement Med,2017,7(1):30-33.
[15]MIZUNO K,KONO T,SUZUKI Y,et al.Goshajinkigan,a traditional Japanese medicine,prevents oxaliplatin-induced acute peripheral neuropathy by suppressing functional alteration of TRP channels in rat[J].J Pharmacol Sci,2014,125(1):91-98.
[16]PAN YW,ZHAO GP,CAI ZJ,et al.Synergistic effect of ferulic acid and z-ligustilide,major components of A.sinensis,on regulating cold-sensing protein TRPM8and TPRA1In Vitro[J].Evid Based Complement Alternat Med,2016:3160247.
[17]唐宗湘.温度敏感的TRP通道与中药药性四气理论[J].南京中医药大学学报,2015,31(4):301-304.
[2]HAMA A,NATSUME T,OGAWA S,et al.Gaps in understanding mechanism and lack of treatments:potential use of a nonhuman primate model of Oxaliplatin-induced neuropathic pain[J].Pain Res Manage,2018,2018:1630709.
[3]曹鹏,李松林,霍介格.奥沙利铂相关性周围神经病变以血痹论治析[J].中国中医基础医学杂志,2013,19(4):430-431.
[4]丁蓉,霍介格,汪悦.当归四逆汤防治奥沙利铂周围神经毒性的临床观察[J].南京中医药大学学报,2014,30(5):432-433.
[5]胡莹,霍介格,曹鹏,等.当归四逆汤防治奥沙利铂致慢性周围神经病变[J].中国实验方剂学杂志,2013,19(20):255-258.
[6]MIHARA Y,EGASHIRA N,SADA H,et al.Involvement of spinal NR2B-containing NMDA receptors in oxaliplatin-induced mechanical allodynia in rats[J].Mol Pain,2011,20(7):8.
[7]KAWASHIRI T,EGASHIRA N,WATANABE H,et al.Prevention of oxaliplatin-induced mechanical allodynia and neurodegeneration by neurotropin in rat model[J].Eur J Pain,2011,15(4):344-350.
[8]SPROWL JA,CIARIMBOLI G,LANCASTER CS,et al.Oxaliplatin-induced neurotoxicity is dependent on the organic cation transporter OCT2[J].Proc Natl Acad Sci USA,2013,110(27):11199-11204.
[9]BAKOGEORGOS M,GEORGOULIAS V.Risk-reduction and treatment of chemotherapy-induced peripheral neuropathy[J].Expert Rev Anticancer Ther,2017,17(11):1045-1060.
[10]PARK SB,LIN CSY,KRISHNAN AV,et al.Dose effects of oxaliplatin on persistent and transient Na+conductances and the development of Neurotoxicity[J].PLoS ONE,2011,6(4):e18469.
[11]ARGYRIOU AA.Updates on Oxaliplatin-induced peripheral neurotoxicity(OXAIPN)[J].Toxics,2015,3(2):187-197.
[12]STEPHAN W,HOHMANN CA,SORIN T,et al.The G2Areceptor(GPR132)contributes to oxaliplatin-induced mechanical pain hypersensitivity[J].Sci Rep,2017,7:446.
[13]TAKAHITO M,SAKI N,ZHAO M,et al.Distinct mechanism of cysteine Oxidation-dependent activation and cold sensitization of human transient receptor potential ankyrin 1channel by high and low oxaliplatin[J].Front Physiol,2017,8:878.
[14]TSUGUNOBU A,SHIZUKA M,YASUSHI K.Shakuyakukanzoto attenuates oxaliplatin-induced cold dysesthesia by inhibiting the expression of transient receptor potential melastatin 8in mice.[J].Tradit Complement Med,2017,7(1):30-33.
[15]MIZUNO K,KONO T,SUZUKI Y,et al.Goshajinkigan,a traditional Japanese medicine,prevents oxaliplatin-induced acute peripheral neuropathy by suppressing functional alteration of TRP channels in rat[J].J Pharmacol Sci,2014,125(1):91-98.
[16]PAN YW,ZHAO GP,CAI ZJ,et al.Synergistic effect of ferulic acid and z-ligustilide,major components of A.sinensis,on regulating cold-sensing protein TRPM8and TPRA1In Vitro[J].Evid Based Complement Alternat Med,2016:3160247.
[17]唐宗湘.温度敏感的TRP通道与中药药性四气理论[J].南京中医药大学学报,2015,31(4):301-304.