Effect of Shao's five-needle therapy pretreatment on airway inflammatory response in asthmatic rats based on ROS/TXNIP/NLRP3 pathway. / 基于ROS/TXNIP/NLRP3通路探讨"邵氏五针法"é¢„å¤„ç†å¯¹å“®å–˜å¤§é¼ æ°”é“ç‚Žæ€§ååº”çš„å½±å“.

OBJECTIVES: To explore the possible mechanism of Shao's five-needle therapy pretreatment on relieving airway inflammatory response in asthmatic rats. METHODS: Forty SPF-grade SD rats were randomly divided into a blank group, a model group, an acupuncture group, and a medication group, with 10 rats in each group. Except the blank group, asthma model was established by aerosol inhalation of ovalbumin in the other 3 groups. The rats in the acupuncture group were treated with acupuncture at "Dazhui" (GV 14) and bilateral "Feishu" (BL 13) and "Fengmen" (BL 12), with each session lasting for 20 min. Acupuncture was given before each motivating, once daily for 7 consecutive days. The rats in the medication group were treated with intraperitoneal injection of dexamethasone sodium phosphate solution before each motivating, once daily for 7 days. General situation of the rats was observed in each group; ELISA method was used to detect the levels of inflammatory cytokines interleukin (IL)-1ß and IL-18 in serum; immunofluorescence staining method was performed to assess the expression of reactive oxygen species (ROS) in lung tissues; Western blot method was used to measure the protein expression of thioredoxin interacting protein (TXNIP), NOD-like receptor protein 3 (NLRP3), apoptosis-associated speck-like protein containing a CARD (ASC), and Caspase-1 in lung tissues. RESULTS: The rats in the blank group exhibited normal behavior, while those in the model group showed signs of respiratory distress, ear scratching, cheek rubbing, and dysphoria. Compared with the model group, the rats in the acupuncture group and the medication group showed stable respiration and relatively agile responses. Compared with those in the blank group, the serum levels of IL-18 and IL-1ß were elevated (P<0.01), the expression intensity of ROS was increased, and the protein expressions of TXNIP, NLRP3, ASC and Caspase-1 in lung tissues were increased (P<0.01) in the model group. Compared with those in the model group, the serum levels of IL-18 and IL-1ß were reduced (P<0.01), the expression intensity of ROS was lowered, and the protein expressions of TXNIP, NLRP3, ASC and Caspase-1 in lung tissues were reduced (P<0.01) in the acupuncture group and the medication group. Compared with the medication group, the protein expression of ASC in lung tissue was reduced in the acupuncture group (P<0.05). CONCLUSIONS: Pretreatment of Shao's five-needle therapy could alleviate airway inflammatory response in asthmatic rats by reducing ROS levels and decreasing the aggregation and activation of pathway-related proteins in the ROS/TXNIP/NLRP3 pathway, ultimately leading to decreased secretion of IL-1ß and IL-18. This mechanism may contribute to the effectiveness of Shao's five-needle therapy in preventing and treating asthma.

[Professor SHAO Jing-ming's clinical experience of fire needling for surgical diseases].

Professor SHAO Jing-ming's clinical experience of fire needling for bone-joint tuberculosis, tuberculous cervical lymphadenitis, ganglion cyst and thyrophyma is summarized. Professor SHAO used fire needling to treat bone-joint tuberculosis. The acupoints included ashi points and nearby acupoints, particularly local opposite acupoints (Neixiyan [EX-LE 4] and Dubi [ST 35], Yinlingquan [SP 9] and Yanglingquan [GB 34], Xuehai [SP 10] and Liangqiu [ST 34]), and for the patients with severe yin-cold syndrome, Yanghe decoction was additionally used. For tuberculous cervical lymphadenitis, fire needling was used at different stages. In the early stage, the nucleus was punctured with fire needling; in the middle stage, the pustule was punctured with fire needling combined with cupping; in the late stage, the fire needling was inserted into the fistula or sinus tract, and the surrounding granulation tissue was treated with horizontal penetrating needling. For ganglion cyst, fire needling combined with centro-square needling was applied. For thyrophyma, the surrounding needling with filiform was used; for simple thyroid mass and thyroid nodule, the surrounding needling with fire needling was used.

[Effect of acupuncture preconditioning combined with PI3K blocker LY294002 on airway inflammation in asthmatic rats].

OBJECTIVE: To observe the effect of acupuncture preconditioning combined with PI3K blocker LY294002 on the expression of PI3K and Akt proteins and genes in the lung tissue and the contents of serum IL-12 and IL-13 in asthmatic rats, so as to explore its preprotective mechanism underlying improving asthma. METHODS: Sixty male Wistar rats were randomly divided into blank control, model, acupuncture pretreatment + blank, acupuncture pretreatment, acupuncture pretreatment + LY294002 and LY294002 groups (n=10 in each group). The asthma model was established by intraperitoneal injection of mixture solution of OVA and Al(OH)3 and followed inhalation of 1%OVA for 30 min, once daily for 7 days. Rats of the blocker groups received inhalation of atomized LY294002 solution for 30 min before inhalation of 1% OVA, and acupuncture was applied to "Feishu"(BL13), "Dazhui"(GV14) and "Fengmen"(BL12) for 20 min, once daily for 7 days before modeling. H.E. staining was used to assess histopathological changes of the lung tissue, and ELISA was used to detect the contents of serum IL-12 and IL-13. The immunoactivity of PI3K and Akt and expression of Akt mRNA of the lung tissue were detected by using immunohistochemistry and fluorescence quantitative real-time PCR, separately. RESULTS: Compared with the blank control group, the content of serum IL-12 was significantly decreased (P<0.01), and the content of serum IL-13, the expression levels of PI3K, Akt protein and Akt mRNA were remarkably increased (P<0.01) in the model group. In comparison with the model group, the content of serum IL-12 in the pretreatment, pretreatment + LY294002 and LY294002 groups was significantly increased (P<0.01, P<0.05), while the content of IL-13 and the expression levels of PI3K, Akt protein and Akt mRNA were considerably decreased (P<0.01, P<0.05) in the acupuncture pretreatment, acupuncture pretreatment+LY294002 and LY294002 groups. The therapeutic effect of acupuncture pretreatment+LY294002 was obviously superior to that of simple acupuncture pretreatment and LY294002 (except PI3K and Akt in the LY294002 group) in up-regulating serum IL-12 level, and in down-regulating serum IL-13, and PI3K and Akt protein levels in the lung tissue (P<0.01). H.E. staining showed severe inflammatory factor infiltration in the bronchus and pulmonary interstitium, and obvious bronchial lumen narrowing with increased exudate in rats of the model group, which was relatively milder in rats of the acupuncture pretreatment, acupuncture pretreatment+LY294002 and LY294002 groups. There were no significant diffe-rences between blank control and pretreatment+blank groups in all of the above indicators （P>0.05）. CONCLUSION: Acupuncture preconditioning can inhibit airway inflammation in asthmatic rats, which may be associated with its functions in down-regulating the levels of pulmonary PI3K and Akt and serum IL-13 and up-regulating the content of serum IL-12. Acupuncture preconditioning combined with LY294002 has the best effect.

[Effect of acupuncture and moxibustion at "Feishu""Dazhui"and "Fengmen"on the expression of CC chemokine ligand 1 and CC chemokine receptor 8 in lung tissue of rats with asthma].

OBJECTIVE: To investigate the protein expression of CC chemokine ligand 1 (CCL1) and CC chemokine receptor 8 (CCR8) in the lung tissue of rats and the mechanism of acupuncture and moxibustion at "Feishu"(BL13), "Dazhui" (GV14) and "Fengmen"(BL12) in the treatment of asthma. METHODS: Sprague-Dawley rats were randomly divided into blank， model, acupuncture and moxibustion groups，n=10 in each group. Ovalbumin sensitization via intraperitoneal injection was performed to establish a model of asthma. The rats in the acupuncture group and the moxibustion group were given acupuncture for 20 min or circling moxibustion for 10 min at BL13, GV14 and BL12, once a day for 7 days. H.E. staining was used to observe the morphological changes of lung tissue. Real-time fluorescence quantitative PCR was used to measure the mRNA expression of signal transducer and activator of transcription 6 (STAT6) in lung tissue and immunohistochemistry was used to measure the protein expression of CCL1 and CCR8 in lung tissue. RESULTS: H.E. staining showed that the rats in the blank group had regular bronchial lumens and alveolar arrangement, with no inflammatory cell infiltration and aggregation around the bronchi; the rats in the model group had the infiltration and aggregation of a large number of inflammatory cells around the bronchi, stenosis of bronchial lumens, wall thickening, and alveolar structural disorder; compared with the model group, the acupuncture group and the moxibustion group had lower degrees of inflammatory cell infiltration and aggregation around the bronchi, stenosis of bronchial lumens, and wall thickening, as well as regular alveolar arrangement. The model group had significantly higher protein expression of CCL1 and CCR8 and mRNA expression of STAT6 than the blank group (P<0.05), and the acupuncture group and the moxibustion group had significantly lower protein expression of CCL1 and CCR8 and mRNA expression of STAT6 (P<0.05). CONCLUSION: Acupuncture and moxibustion can intervene against airway inflammation by inhibiting the protein expression of CCL1 and CCR8 and STAT6 signal transduction in lung tissue, which may be one of the mechanisms of acupuncture and moxibustion in the treatment of asthma.

["SHAO's five-needle method" as the main treatment for allergic rhinitis and asthma syndrome: a multi-center randomized controlled trial].

OBJECTIVE: To compare the clinical effect differences between "SHAO's five-needle method" and routine acupoint selection on allergic rhinitis and asthma syndrome. METHODS: A total of 210 patients with allergic rhinitis and asthma syndrome were randomly divided into an observation group (105 cases, 4 cases dropped off) and a control group (105 cases, 4 cases dropped off). The patients in the observation group were treated with "SHAO's five-needling method", and the acupoints of Feishu (BL 13), Dazhui (GV 14), Fengmen (BL 12), Yintang (GV 29), Shangyingxiang (EX-HN 8) and Hegu (LI 4), etc. were selected; the patients in the control group was treated with routine acupuncture, and the acupoints of Feishu (BL 13), Zhongfu (LU 1), Taiyuan (LU 9), Dingchuan (EX-B 1), Danzhong (CV 17), Yintang (GV 29), Fengmen (BL 12) and Zusanli (ST 36), etc. were selected. The treatment in the two groups was given once a day, 6 times a week, for 4 weeks. The score of symptoms and signs was observed before and after treatment as well as 1 month, 2 months and 3 months after treatment. The forced expiratory volume in 1 second (FEV1), peak expiratory flow (PEF) and eosinophils in peripheral blood were measured before and after treatment in the two groups. After treatment, the clinical therapeutic effect was compared between the two groups. RESULTS: The total effective rate was 98.0% (99/101) in the observation group, which was superior to 94.1% (95/101) in the control group (P<0.01). Compared before treatment, the total score of symptoms and signs in the two groups was significantly decreased at 1, 2, 3 and 4 weeks of treatment (P<0.01); after treatment and at each time point of follow-up, the total score of symptoms and signs in the observation group was lower than that in the control group (P<0.01). Compared with 4 weeks of treatment, the total score of symptoms and signs at each time point of follow-up was not statistically different in the observation group (P>0.05), and the total score of symptoms and signs in the third month of follow-up in the control group was significantly increased (P<0.05). After treatment, FEV1 and PEF in the two groups were increased (P<0.01), eosinophil count in peripheral blood was decreased (P<0.01), and the improvement in the observation group was greater than that in the control group (P<0.01, P<0.05). CONCLUSION: "SHAO's five-needle method" can improve the clinical symptoms and pulmonary function, reduce the count of eosinophils in peripheral blood in patients with allergic rhinitis and asthma syndrome, and the curative effect is better than routine acupuncture.

[Effect of acupuncture intervention on airway remodeling and transforming growth factor-beta1 expression in asthma rats].

OBJECTIVE: To observe the effect of acupuncture stimulation of "Dazhui" (GV 14), bilateral "Fengmen" (BL 12) and "Feishu" (BL 13) on the thickness of the bronchial tubal wall and smooth muscle and expression of transforming growth factor-beta1 (TGF-beta1) in asthma rats so as to reveal its mechanism underlying improvement of asthma. METHODS: Forty SD rats were randomly assigned to control, model, acupuncture and medication groups. The asthma model was induced by subcutaneous injection of 1% egg albumin solution and forced inhalation of atomized ovalbumin. Rats of the acupuncture group were treated by acupuncture stimulation of GV 14, bilateral BL 12 and BL 13 (once daily for 10 days), and rats of the medication group treated by intraperitoneal injection of Aminophylline Injection (100 mg/kg, once daily for 10 days). The thickness of the bronchial tubal wall and airway smooth muscle of the lung tissue were measured by image analyzer for assessing the degree of airway remodeling. The expression of TGF-beta1 in the small airway was detected by immunohistochemistry. RESULTS: Compared to the control group, the thickness of the airway wall and smooth muscle, and the expression level of TGF-beta1 were remarkably increased in the model group (P < 0.01). After acupuncture intervention and medication treatment, the thickness of the airway wall and smooth muscle, and the expression level of TGF-beta1 were remarkably down-regulated in both groups in comparison with those of the model group (P < 0.05, P < 0.01), and the expression of TGF-beta1 in the acupuncture group was obviously lower than that of the medication group (P < 0.05). CONCLUSION: Acupuncture stimulation of GV 14, bilateral BL 12 and BL 13 can down-regulate bronchial asthma-induced increase of TGF-[, expression in the lung tissue in asthma rats, which may contribute to its effect in improving airway remodeling.

[Talking about "moxibustion treatment for before sick" from "ischemic preconditioning"].

"Treatment for before sick" is a theory of TCM, reflecting preventing thought of "prevention being better than cure" and "nipping in the bud", while "moxibustion treatment for before sick" is highly praised by doctors of past ages. Moxibustion can activate human vital-qi and increase immunologic function of the organism, playing a preventive role for before sick, which is similar to the essence of "ischemic preconditioning" raised in recent years. Because of convenient manipulation, no adverse effect, it has very important position in the field of preventive medicine.

Effects of moxibustion pretreating on SOD and MDA in the rat of global brain ischemia.

OBJECTIVE: To probe into the mechanism of moxibustion preconditioning in preventive brain-protecting effect. METHODS: The global brain ischemia rat model was developed by blocking 4 arteries. Seventy-eight Wistar male rats were randomly divided into 5 groups: a normal control group, a sham-operation group, a brain ischemia group, a brain ischemia preconditioning group, a moxibustion pretreating group. The brains in the 5 groups were taken at 24 h, 48 h, and 72 h after operation respectively. Superoxide dismulase (SOD) activity was determined with xanthine oxidase method and malondialdehyde (MDA) content with thiobarbituric acid method. RESULTS: After the operation, in the moxibustion preconditioning group, SOD activity significantly increased, especially 24 h after moxibustion preconditioning; and MDA content decreased, with a very significant difference as compared with that of the cerebral ischemia group (P<0.01). CONCLUSION: Moxibustion preconditioning protects the ischemic and anoxic brain tissue by increasing the activity of endogenous antioxidase.

[Effects of moxibustion preconditioning on SOD and MDA in rats with global brain ischemia].

OBJECTIVE: To reveal the mechanism of moxibustion preconditioning in preventive brain-protecting action. METHODS: The rat model of global brain ischemia was made with 4-artery ligation method. 78 Wistar rats were randomly divided into 5 groups: normal control group, sham-operation group, brain ischemia group, brain ischemia preconditioning group and moxibustion preconditioning group. The brain was taken 24 h, 48 h and 72 h after operation in the all groups, respectively, for determination of SOD activities by xanthine oxidase method and MDA content by thibabituric acid method. RESULTS: The SOD activity significantly increased, especially 24 h after the moxibustion preconditioning, and the MDA content decreased significantly with a very significant difference as compared with the ischemia group(P<0.01). CONCLUSION: Moxibustion preconditioning exerts the protective action on the brain tissue of ischemia and anoxia through increasing the endogenous anti-oxidase activity.