Effect of electroacupuncture of "Jiaji" (EX-B2) on NOD-like receptor protein 3 mediated pyroptosis in spinal cord tissue of rats with acute spinal cord injury. / å¤¹è„Šç”µé’ˆå¯¹æ€¥æ€§è„Šé«“æŸä¼¤å¤§é¼ è„Šé«“ç»„ç»‡NODæ ·å—ä½“çƒ­è›‹ç™½ç»“æž„åŸŸç›¸å ³è›‹ç™½3介导的细胞焦亡的影响.

OBJECTIVES: To observe the effect of electroacupuncture (EA) stimulation of "Jiaji"(EX-B2) on motor function, histomorphology, and expression of NOD-like receptor protein 3 (NLRP3) and N-terminal domain of gasdermin D (GSDMD-N) in the spinal cord tissue of rats with spinal cord injury (SCI), so as to explore its mechanism underlying improvement of SCI. METHODS: Forty eight female SD rats were randomly divided into sham surgery (sham), SCI model (model), EA, and NLRP3 agonist (monosodium urate, MSU) combined with Jiaji EA (MSU+EA) groups, with 12 rats in each group which were further divided into 3 d and 7 d subgroups, with 6 rats at each time point. Two EA groups received EA stimulation of EX-B2 with a frequency of 100 Hz, electrical current of 1-2 mA for 30 min, once a day for 3 or 7 days. After 5 min, 6 h, and 24 h of modeling, rats of the MSU+EA group received intraperitoneal injection of MSU (200 µg/kg, 200 µg/mL) . The motor function was evaluated using Basso-Beattie-Bresnahan (BBB) scale, the morphological structure of rat spinal cord tissue was observed by H.E. staining. The expression of pyroptosis related factors NLRP3, cleaved Caspase-1 and GSDMD-N of the spinal cord was observed by using immunohistochemistry and Western blot separately, the expression and localization of Iba-1 and GSDMD-N in the spinal cord tissue were observed using immunofluorescence double staining method. RESULTS: Compared with the sham group, the BBB scores after modeling and on day 3 and 7 were decreased (P<0.05), while the average OD values (immunoactivity) and expression levels of NLRP3, cleaved Caspase-1 and GSDMD-N proteins, and the immunofluorescence intensity of Iba-1/GSDMD-N (co-expression) of the spinal cord tissues on day 3 and 7 were significantly increased in the model group (P<0.05). In comparison with the model group, the BBB scores on day 3 and 7 were obviously increased (P<0.05), while the immunoactivity and expression levels of NLRP3, cleaved Caspase-1 and GSDMD proteins, and the immunofluorescence intensity of Iba-1/GSDMD-N on day 3 and 7 significantly down-regulated in the EA group (P<0.05) but not in the MSU+EA group (P>0.05), suggesting an elimination of the effects of EA after administration of NLRP3 agonist (MSU). H.E. staining showed obvious bleeding area in the spinal cord tissue, loose tissue and inflammatory cell infiltration on day 3 after modeling, and basic absorption of the bleeding, loose tissue, obvious vacuolar changes of the white matter area, loss and contraction of neurons with infiltration of a large number of inflammatory cells, which was milder in the EA group but not in the MSU+EA group. CONCLUSIONS: EA of EX-B2 can improve the motor function of SCI rats, which may be related to its functions in inhibiting pyroptosis of microglia mediated by NLRP3/Caspase-1 signaling pathway.

[Research progress of acupuncture-moxibustion in treatment of spinal cord injury through mediating inflammatory reaction].

Inflammatory reaction after spinal cord injury (SCI) is the main obstacle to the recovery of neural function. In the occurrence and development of SCI, the complex regulatory mechanisms are involved in inflammatory reaction, including the activation of inflammatory cells (e.g.macrophages and microglia), and the release of cytokines (e.g. tumor necrosis factor, interleukin and chemokines). Acupuncture-moxibustion is significantly effective in clinical treatment of SCI, and its mechanism is related to adjusting the function of inflammatory cells after SCI, modulating the expression of cytokines and the activation of NLRP3 inflammatory bodies, as well as the expression of high mobility group protein B1 and calcitonin gene related peptide. This article summarizes the research progress of mechanism of inflammatory reaction after SCI and the effects of acupuncture-moxibustion intervention in recent years so that the new ideas can be provided to clarify the pathogenesis of SCI and the effect of acupuncture-moxibustion intervention.

[Professor LI Xiao-ning's experience for post-stroke dysphagia treated with penetrating-needling and swallowing technique of acupuncture].

Professor LI Xiaoning's experience for post-stroke dysphagia treated with penetrating-needling and swallowing technique of acupuncture is summarized in this paper. The key pathogenesis of this disease is dysfunction of zangfu organs, impairment of the house of mental activity and dysfunction of meridian sinew. Innovatively, the anatomic structure related to swallowing is divided into three zones. Based on the distributions of heart, kidney and stomach meridians on the neck, three lines are determined. Hence, the theory of "three-zone and three-line" as well as the penetrating-needling and swallowing technique of acupuncture are put forward. In this paper, the theoretic evidences of penetrating-needling and swallowing technique of acupuncture, point selection and manipulation in treatment of post-stroke dysphagia are introduced.

[Electroacupuncture Improves Limb Locomotor Function Possibly by Suppressing Rho-ROCK â ¡ Pathway Related Factors in Anterior Horns of Spinal Cord in Rats with Acute Spinal Cord Injury].

OBJECTIVE: To observe the effect of electroacupuncture (EA) of "Jiaji" (EX-B 2) on limb locomotor function and expression of Ras homolog gene family member A (RhoA), Rho-associated kinase Ⅱ (ROCK Ⅱ) and myosin light chain (MLC) proteins in the anterior horn of spinal cord in acute spinal cord injury (ASCI) rats, so as to explore its mechanisms under-lying improvement of SCI-induced limb locomotor dysfunction. METHODS: Forty-eight female Wistar rats were randomly divided into sham operation (sham), ASCI model (model), EA EX-B 2 (EA) and ROCK inhibitor (Fasudil) groups which were further divided into 14 d and 28 d subgroups (n=6 in each). The ASCI model was made by using weight drop striking method. Three hours after modeling, EA (100 Hz, 0.4, 0.6 mA) was applied to EX-B 2 (T 9, T 11) for 30 min, once daily for 14 d and 28 d, respectively. The ROCK inhibitor (hydrochloride Fasudil, 10 mg/kg) was administrated by intraperitoneal injection immediately after modeling, once a day, continuously for 14 d or 28 d. The expression of RhoA, ROCK Ⅱ and MLC proteins in the spinal cord anterior horn tissue (T 10) was detected by immunohistochemistry. The rats' hindlimb locomotor function was assessed according to Basso, Beattie and Bresnahan (BBB) locomotor rating scale (21-points). RESULTS: After ASCI, the BBB scores were significantly lower in the model group than in the sham group on day 14 and 28 (P<0.05), and obviously higher in the EA and inhibitor groups than in the model group (P<0.05), suggesting an improvement of the hindlimb locomotor function after EA intervention or suppression of ROCK. Immunohistochemical results indicated that the numbers of RhoA, ROCK Ⅱ and MLC immune-reaction positive cells in the anterior horn of spinal cord were significantly more in the model group than in the sham group (P<0.05), and remarkably decreased in both EA and inhibitor groups on day 14 and 28 relevant to the model group (P<0.05). The therapeutic effects of EA were markedly weaker than those of inhibitor Fasudil in up-regulating BBB score and down-regulating the number of RhoA, ROCK Ⅱ and MLC positive cells (P<0.05). CONCLUSION: EA of EX-B 2 can improve the hindlimb locomotor function in ASCI rats, which may be associated with its effect in down-regulating the expression of RhoA, ROCK Ⅱ and MLC proteins (i.e., inhibiting the RhoA/ROCK signaling pathway) in the anterior horn of spinal cord.

[Effect of Electroacupuncture Intervention at Different Time-points on Locomotor Function and Apoptosis in Rats with Acute Spinal Cord Injury].

OBJECTIVE: To observe the effect of electroacupuncture (EA) intervention at different treatment time-points on locomotor function and neuronal apoptosis in rats with acute spinal cord injury (ASCI), so as to explore its mechanism under-lying improvement of ASCI. METHODS: Seventy-two female Wistar rats were randomly divided into sham operation, ASCI model and EA groups which were further divided into 4 time-points:1, 3, 7 and 14 d (n=6 at each). The ASCI model was established by weight-drop striking the exposed spinal cord (T10) using a NYU impactor device. EA (0.4-0.6 mA, 100 Hz) was applied to bilateral "Jiaji" (EX-B 2) for 30 min, once a day for 1, 3, 7 and 14 days, respectively in the EA group. The Basso-Beattie-Bresnahan (BBB) grading score was used to assess the animals' locomotor function, and H.E. staining was used to display the pathological changes of spinal cord injury at different time points. The TUNEL staining was used to detect the nerve cell apoptosis of spinal cord tissue. RESULTS: After ASCI, the BBB scores were significantly decreased and the numbers of the apoptotic nerve cells considerably increased on day 1, 3, 7 and 14 in the model group compared with the sham operation group (P<0.05). In addition, H.E. staining showed severe damage of the spinal cord tissue, infiltration of inflammatory cells, increase of dead nerve neurons and glia cells in the number from day 3 to 7, and additional increase of vacuoles on day 14. Following EA intervention, the decrease of BBB scores and the increase of apoptotic nerve cell numbers were markedly suppressed on day 3, 7 and 14 (P<0.05). The TUNEL staining showed a decrease of the apoptotic nerve cells from day 3 on and an improvement of the injured tissue structure from day 7 to 14 in the EA group. CONCLUSIONS: EA stimulation of EX-B 2 can significantly improve locomotor function, and lighten the spinal cord injury in ASCI rats, presenting a time-dependant tendency, which may be associated with its effect in reducing neuronal apoptosis in the spinal cord.