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《The Journal of Traditional Chinese Orthopedics and Traumatology》 2017-01
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Effect of Qiangjin Zhuanggu Qufeng Heji on acid- sensing ion channel 3 of dorsal root ganglia in rats with inflammation caused by nucleus pulposus

WANG Dong;PAN Hao;ZHU Hang;ZHU Li;HE Yongjiang;WANG Jian;JIA Gaoyong;Hangzhou Hospital of Traditional Chinese Medicine;  
Objective: To observe the effect of Qiangjin Zhuanggu Qufeng Heji( 强筋壮骨祛风合剂,QJZGQFHJ) on acid- sensing ion channel 3( ASIC3) of dorsal root ganglia( DRG) in rats with inflammation caused by nucleus pulposus. Methods: Forty clean adult male SD rats were selected and randomly divided into blank group,sham- operated group,model group,Amiloride group and TCD group,8 cases in each group. The surgeries were performed on rats in sham- operated group,model group,Amiloride group and TCD group to expose L5 DRG. The surgical incisions were sutured in sham- operated group right after the L5 DRG were exposed. The nucleus pulposuses in tails were fetched out and were placed on the denudate L5 DRG in model group and TCD group,and then the surgical incisions were sutured. The nucleus pulposuses in tails were fetched out and were placed on the denudate L5 DRG in Amiloride group,and Amiloride solution( 1 ml),containing hydrochloric acid( 0. 1 mg),was dropped arround the denudate L5 DRG,then the surgical incisions were sutured. Since the 1st day after modeling,the rats in TCD group were intragastric administrated with QJZGQFHJ( 5ml / kg),twice a day for 30 consecutive days. No drug interventions were performed on rats in blank group,sham- operated group and model group. The 50% paw withdrawal threshold( 50% PWT) of left posterior feet were measured by using Up- Down methods at 1 day before modeling and at 3,5,9,15,23 and 29 days after modeing respectively. After the end of drug interventions,all rats in TCD group were executed and the L5 DRGs were fetched out and were divide into 2 parts. One of them was used for calculating the ASIC3 positive areas by using immunohistochemical method,and the other potion was used for detecting the ASIC3 protein expression by using Western Blot assay. Results: Left posterior foot eversion and toe dorsal extension were found in some rats of model group,Amiloride group and TCD group within 3 days after modeling. There was statistical difference in 50% PWT between different timepoints before and after modeling,in other words,there was time effect( F = 50. 132,P = 0. 000).There was statistical difference in 50% PWT between the 5 groups in general,in other words,there was group effect( F = 288. 219,P =0. 000). Except at 1 day before modeling( F = 0. 332,P = 0. 854),there was statistical difference in 50% PWT between the 5 groups( F =288. 015,P = 0. 000; F = 308. 588,P = 0. 000; F = 374. 945,P = 0. 000; F = 560. 713,P = 0. 000; F = 343. 043,P = 0. 000; F = 459. 914,P = 0. 000). At different timepoints after modeling,the 50% PWT were lower in model group compared to blank group,sham- operated group,Amiloride group and TCD group( P = 0. 010,P = 0. 011,P = 0. 001,P = 0. 001; P = 0. 023,P = 0. 022,P = 0. 000,P = 0. 004; P =0. 013,P = 0. 013,P = 0. 003,P = 0. 001; P = 0. 012,P = 0. 002,P = 0. 002,P = 0. 002; P = 0. 004,P = 0. 000,P = 0. 000,P = 0. 000; P =0. 003,P = 0. 005,P = 0. 003,P = 0. 013). There was no statistical difference in 50% PWT between blank group and sham- operated group( P = 0. 640,P = 0. 710,P = 0. 590,P = 0. 480,P = 0. 330,P = 0. 390) and the 50% PWT were higher in Amiloride group compared to TCD group( P = 0. 021,P = 0. 011,P = 0. 013,P = 0. 002,P = 0. 001,P = 0. 005) at different timepoints after modeling. There was interaction between time factor and group factor( F = 2. 358,P = 0. 002). There was statistical difference in ASIC3 positive areas and ASIC3 protein expression in DRG between the 5 groups( F = 205. 890,P = 0. 030; F = 101. 740,P = 0. 001). ASIC3 positive areas and ASIC3 protein expression were less in blank group,sham- operated group,Amiloride group and TCD group compared to model group( P = 0. 004,P =0. 003,P = 0. 001,P = 0. 007; P = 0. 000,P = 0. 000,P = 0. 000,P = 0. 000),and were greater in Amiloride group compared to TCD group( P = 0. 001; P = 0. 000). Conclusion: QJZGQFHJ can reduce the ASIC3 level in diseased DRG and increase the pain threshold in rats with inflammation caused by nucleus pulposus,while it is inferior to Amiloride in effectiveness.
【Fund】: 浙江省中医药科技计划项目(2015ZA157);; 浙江省杭州市科技计划重点专病专科项目(20150733Q60)
【CateGory Index】: R285.5
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