骨癌性疼痛ラットにおけるモルヒネ寛容性に対する骨近鍼と電気鍼を組み合わせた早期介入と後根神経節におけるHDACとMORの発現に及ぼす影響

[Early intervention of bone-nearby acupuncture combined with electroacupuncture on morphine tolerance in bone cancer pain rats and its effect on the expression of HDAC and MOR in dorsal root ganglia].

• Xue-Mei Zhong
• Jun-Fan Fang
• Bin Jiang
• Jie Zhou
• Yang-Qian Cai
• Yi Liang
• Jian-Qiao Fang
• Feng Chen
• Jun-Ying Du

抄録

目的:

骨癌性疼痛モルヒネ耐性（BCP-MT）ラットの後根神経節（DRG）におけるヒストン脱アセチラーゼ1（HDAC1）、ヒストン脱アセチラーゼ2（HDAC2）、μ-オピオイド受容体（MOR）の発現に及ぼす骨近接鍼（BNA）と電気鍼（EA）の早期介入の効果を観察し、その可能性のあるメカニズムを探る。

OBJECTIVE: To observe the effect of early intervention of bone-nearby acupuncture (BNA) combined with electroacupuncture (EA) on the expression of histone deacetylase1(HDAC1), histone deacetylase 2 (HDAC2) andμ-opioid recepter (MOR) in dorsal root ganglia (DRG) of bone cancer pain-morphine tolerance (BCP-MT) rats, and to explore its possible mechanism.

方法:

SDラット35匹を、偽BCP群(=6)、BCP群(=7)、MT群(=7)、BNA+EA群(=8)、偽BNA群(=7)に無作為に割り付けた。偽BCP群を除き、他の4群では左脛骨に癌細胞接種手術を行い、骨癌疼痛モデルを確立した。MT群、BNA+EA群、偽BNA群では、モルヒネ塩酸塩の腹腔内注射を行い、モルヒネ耐性モデルを確立した。術後、BNA+EA群では「瑞山里」(ST 36)と「崑崙」(BL 60)で骨近鍼と電気鍼を併用し、拡張波、周波数2Hz/100Hz、強度0.5～1.5mAで施術した。羞恥BNA群での介入は、BNA+EA群と同じ時間とツボを適用し、電気的刺激を与えずに針を皮膚に突き刺した。針の保持は、ＢＮＡ＋ＥＡ群、羞恥ＢＮＡ群ともに、１日１回、３０分間、連続７日間行った。術前と術後10、11、15、22日後に、5群で左足離脱閾値（PWT）を測定した。DRG中のHDAC1、HDAC2、MORのレベルをウエスタンブロットで検出した。

METHODS: A total of 35 SD rats were randomized into a sham BCP group (=6), a BCP group (=7), a MT group (=7), a BNA+EA group (=8) and a shame BNA group (=7). Except of the sham BCP group, cancer cell inoculation operation at left tibia was given in the other 4 groups to establish the bone cancer pain model. In the MT group, the BNA+EA group and the shame BNA group, intraperitoneal injection of morphine hydrochloride was given to establish the morphine tolerance model. After the operation, bone-nearby acupuncture combined with electroacupuncture was applied at "Zusanli" (ST 36) and "Kunlun" (BL 60) in the BNA+EA group, with dilatational wave, 2 Hz/100 Hz in frequency, 0.5 to 1.5 mA in intensity. Intervention in the shame BNA group was applied at the same time and acupoints as those in the BNA+EA group, the needles were pierced the skin without any electrical stimulation. The needles were retained for 30 min, once a day for continuous 7 days in both BNA+EA and shame BNA groups. Before and 10, 11, 15, 22 days after the operation, the left paw withdrawal threshold (PWT) was measured in the 5 groups. The levels of HDAC1, HDAC2 and MOR in DRG were detected by Western blot.

結果:

がん細胞接種手術から10日後、BCP、MT、BNA+EA、偽BNA群のPWTは偽BCP群に比べて低下した（<0.01）。術後11日目、MT、BNA+EA、偽BNA群のPWTは偽BCP群に比べて増加した(<0.01)。術後22日目には、BCP群とMT群の間に有意差はなく(>0.05)、BNA+EA群のPWTはMT群と偽BNA群に比べて増加した(<0.01)。BCP群では、HDAC1とHDCA2のDRGレベルが上昇したが、MORレベルは偽BCP群に比べて低下した（<0.05、<0.01）。MT群では、BCP群に比べてHDAC1のDRGレベルが上昇した(<0.05)。BNA+EA群では、HDAC1のDRGレベルはMT群および偽BNA群に比べて低下したが（<0.01、<0.05）、MORレベルは上昇した（<0.01）。

RESULTS: Ten days after the cancer cell inoculation operation, the PWT of the BCP, MT, BNA+EA and sham BNA groups was decreased compared with the sham BCP group (<0.01). Eleven days after the operation, the PWT of the MT, BNA+EA and sham BNA groups was increased compared with the BCP group (<0.01). Twenty-two days after the operation, the difference was no significant between the BCP group and MT group (>0.05); the PWT of the BNA+EA group was increased compared with the MT and sham BNA group (<0.01). In the BCP group, the DRG levels of HDAC1 and HDCA2 were increased, while the level of MOR was decreased compared with the sham BCP group (<0.05, <0.01). In the MT group, the DRG level of HDAC1 was increased compared with the BCP group (<0.05). In the BNA+EA group, the DRG level of HDAC1 was decreased compared with the MT group and the sham BNA group (<0.01, <0.05), while the level of MOR was increased (<0.01).

結論:

電気鍼治療と組み合わせた骨近辺の鍼治療の早期介入は、骨癌疼痛ラットのモルヒネ耐性を和らげることができます、それはHDAC1の発現をダウンレギュレートし、背側根神経節におけるMORの発現をアップレギュレートすることに関連しているかもしれません。

CONCLUSION: Early intervention of bone-nearby acupuncture combined with electroacupuncture can relieve the morphine tolerance in bone cancer pain rats, it may relate to down-regulating the expression of HDAC1 and up-regulating the expression of MOR in the dorsal root ganglia.