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Objective: We have demonstrated that needle knife (acupotomy) treatment can improve knee osteoarthritis (KOA) in rabbits. The present study was designed to examine its effect on expression of phosphorylated focal adhesion kinase (p-FAK), phosphinositides 3 kinase (p-PI 3 K) and Aggrecan genes and proteins in the knee-joint cartilage tissues of KOA rabbits, so as to explore its partial molecular mechanism underlying improvement of KOA., Methods: Forty-nine New Zealand male rabbits were randomly divided into normal control, model, model +inhibitor, needle knife, needle knife+ inhibitor, electroacupuncture (EA), EA +inhibitor groups ( n =7 in each). The KOA model was established by modified Videman method (left hindlimb extension immobilization and ankle dorsal flexion 60°). Acupotomy relaxing manipulation was applied to the lateral collateral ligament and patellar ligament of the left knee-joint, two times a week for 4 weeks, and EA (2 Hz/100 Hz, 3 mA) was applied to the left "Liangmen" (ST 21), "Xuehai" (SP 10), "Neixiyan " (EX-LE 4) and "Dubi" (ST 35) for 20 min, three times a week, for 4 weeks. About 2 h before every needle-knife or EA treatment or at the corresponding time-point, intra-articular cavity injection of PF-562271(a specific antagonist of FAK, 200 μmol/L, 0.5 mL)was performed in the three inhibitor groups. The expression levels of p-FAK, p-PI 3 K, Aggrecan genes and proteins in the cartilage tissues were measured with quantitative Real-time PCR and Western blot, separately., Results: After modeling, the expression levels of p-FAK and p-PI 3 K genes and proteins were significantly up-regulated ( P <0.05, P <0.01), while those of Aggrecan protein and mRNA considerably down-regulated in the model group in comparison with the normal group ( P <0.01). Following 4 weeks' needle-knife or EA treatment, the expression levels of p-FAK and p-PI 3 K and Aggrecan proteins in both EA and needle-knife groups, and Aggrecan mRNA in the needle knife group were significantly up-regulated ( P <0.05, P <0.01). After administration of p-FAK antagonist, modeling-induced upregulation of expression of p-FAK mRNA and protein and p-PI 3 K protein, as well as modeling-induced down-regulation of Aggrecan mRNA and protein were significantly suppressed in the model+inhibitor group ( P <0.05, P <0.01), and needle knife-induced and EA-induced up-regulation of expression of p-FAK, p-PI 3 K and Aggrecan mRNAs and proteins was notably suppressed respectively in comparison with the needle knife and EA groups ( P <0.05, P <0.01). The effect of needle knife was significantly superior to that of EA in up-regulating p-PI 3 K and Aggrecan mRNAs as well as p-FAK, p-PI 3 K and Aggrecan proteins ( P <0.05, P <0.01)., Conclusion: Needle knife intervention can up-regulate the expression levels of p-FAK, p-PI 3 K and Aggrecan proteins and mRNAs in the cartilage tissue of the knee-joint in KOA rabbits, suggesting an involvement of FAK-PI 3 K signaling in the needle knife-induced improvement of KOA.

Objective: To observe the effect of acupotomy lysis (AL) on hypothalamic proopiomelanocortin (POMO) mRNA and preproenkephalin (PPE) mRNA expression in rats with the third lumbar vertebrae transverse process syndrome (TLVTPS) so as to study its underlying mechanism in relieving symptoms of lumbar muscle strain., Methods: Twenty-four SD rats were randomly divided into normal control group, model group, AL group and electroacupunture (EA) group. The TLVTPS model was established by inserting a piece of gelatin sponge into the posterior of the left 3rd lumbar vertebrae transverse process. AL and EA were applied to the left "Shenshu"(BL 23) and "Yaoyangguan" (GV 3) respectively. The POMC mRNA and PPE mRNA expression levels in the hypothalamus were detected by in situ hybridization., Results: In comparison with the normal group, the integrated optical density (IOD) values of hypothalamic POMC mRNA and PPE mRNA positive cells in the model group were increased significantly (P < 0.01); while compared with the model group, those of POMC mRNA and PPE mRNA positive cells in both left and right hypothalamus were increased further considerably in both AL and EA groups (P < 0.01). No significant differences were found between AL and EA groups in POMC mRNA and PPE mRNA expression levels (P > 0.05)., Conclusion: AL and EA therapies can increase the expression of POMC mRNA and PPE mRNA in hypothalamus in rats with TLVTPS, which may contribute to its effect in relieving pain in the treatment of lumbar muscle strain.

Objective: To observe changes of SP and CCK-8 contents in the hypothalamus and spinal cord after performing acupotomy lysis so as to study its central analgesic effect in rats with transverse-process syndrome of the third lumbar vertebra., Methods: Twenty-eight SD rats were randomly divided into normal control group, model group, acupotomy lysis group and electroacupuncture (EA) group, with 7 cases in each one. The model of the third lumbar vertebra transverse process syndrome was established by inserting a piece of gelatin sponge into the deep subfascial part of the 3rd lumbar transverse process. Acupotomy lysis was conducted 1 time/week for 2 weeks. EA (2/100 Hz, 1-2 mA) was applied to the left "Shenshu" (BL 23)-"Yaoyangguan"(GV 3) for 20 min/time, 3 times/week and for 2 weeks. SP and CCK-8 contents in the hypothalamus and spinal cord were detected with enzyme linked immunosorbent assay (ELISA)., Results: In model group, SP and CCK-8 contents in the hypothalamus and spinal cord were increased significantly in comparison with normal control group (P < 0.01). Compared with model group, both SP and CCK-8 contents in hypothalamus and SP content in spinal cord in acupotomy lysis and EA groups were apparently lower (P < 0.01, 0.05). CCK-8 content in spinal cord in cupotomy lysis group was lower markedly than that of model group (P < 0.01), while no significant difference was found between model group and EA group in CCK-8 level in spinal cord (P > 0.05)., Conclusion: Acupotomy lysis can downregulate SP and CCK-8 contents in hypothalamus and spinal cord, which may contribute to its effect in relieving pain from transverse process syndrome of the third lumbar vertebra.

Objective: To observe the effect of small needle-knife lysis on plasma calcitonin gene-related peptide (CGRP), endothelin (ET), 6-keto-prostaglandin F1alpha (6-keto-PGF1alpha), thromboxane A2 (TXA2) contents in rats with experimental third lumbar vertebra transverse process syndrome (TLVTPS) so as to explore its underlying mechanism in clinical treatment., Methods: Forty SD rats were randomly divided into normal control, model, lysis and EA groups. TLVTPS model was established by embedding a piece of gelatin sponge (0.5 cm x 0.5 cm) to the transverse process of the 3rd lumbar vertebra under anesthesia. EA (2/100 Hz, 1-2 mA) was applied to left "Shenshu" (BL23) -"Yaoyangguan" (GV3) for 20 min, once every other day, 6 times altogether. For animals of lysis group, the lysis was performed by using a small needle-knife in the induration spot or cord-like region near the incision, once a week and twice altogether. Four weeks later after modeling, plasma CGRP, ET, 6-keto-PGF1alpha and TXA2 contents were detected by using radioimmunoassay and enzyme linked immunosorbent assay (ELISA)., Results: Compared with normal control group, plasma CGRP, ET, TXA2 and 6-keto-PGF1alpha increased significantly in model group (P<0.01); in comparison with model group, plasma CGRP, TXA2 and 6-keto-PGF1alpha in both EA and lysis groups decreased considerably (P<0.05, 0.01). No significant differences were found between EA and lysis groups in plasma CGRP, ET and 6-keto-PGF1alpha levels (P>0.05)., Conclusion: Both EA and lysis of acupotomology have an adjusting effect on vasoactive substances (CGRP, TXA2 and 6-keto-PGF1alpha) levels in TLVTPS rats, which may contribute to their effects in improving local blood circulation and relieving soft tissue injury in the treatment of third lumbar vertebra transverse process syndrome.