[Effect of moxibustion on inflammatory pain and N-methyl-D aspartic acid receptor-nitric oxide-cyclic GMP pathway in spinal cord of adjuvant arthritis rats].

OBJECTIVE: To observe the effect of moxibustion on pain and N-methyl-D aspartic acid receptor/nitric oxide/cyclic guanosine monophosphate (NMDA-NO-cGMP) signaling pathway in the spinal cord of rats with adjuvant arthritis (AA), so as to explore its underlying mechanisms in relieving inflammatory pain of rheumatoid arthritis (RA). METHODS: SD rats were randomly divided into normal, model, moxibustion (Moxi), Moxi +NMDA receptor antagonist AP-5 (Moxi+AP-5) and Moxi +NMDA receptor agonist (NMDA) groups, with 20 rats in each group. The AA model was established by placing the rats in a wind, cold and damp environment for 12 h, once daily for 20 days and by injection of complete Freund's adjuvant into the right hind paw. Rats of the three Moxi groups received ignited moxa-stick stimulation of "Zusanli"(ST36) and "Shenshu"(BL23) alternately for 20 min, once a day for 15 days. The Moxi + AP-5 group and Moxi +NMDA group received intraperitoneal injection of AP-5 (0.7 mg·kg-1·d-1) and NMDA (5 mg·kg-1·d-1), respectively, once a day, for a total of 15 days. Mechanical pain thres-hold (MPT) was measured before and after modeling and interventions. The spinal cord tissue was sampled for detecting the expression of iNOS mRNA and protein, content of cGMP and NO, and the activity of NOS by using fluorescence quantitative PCR, Western blot, ELISA,nitrate reductase method and colorimetric method, respectively. RESULTS: Before modeling, there was no significant difference in MPT among all the 5 groups (P>0.05). After modeling, the MPT was remarkably decreased (P<0.01), the expression levels of iNOS mRNA and protein,the contents of cGMP and NO, the activity of NOS were significantly increased in the model group relevant to the normal group (P<0.01). After the interventions, the MPT was obviously increased (P<0.01), while the expression levels of iNOS mRNA and protein, the contents of cGMP and NO, the activity of NOS were significantly down-regulated in the Moxi, Moxi-AP-5 and Moxi+NMDA groups (P<0.05, P<0.01). The effect of Moxi+AP-5 group was significantly superior to that of Moxi group in raising MPT and down-regulating the expression levels of iNOS mRNA and protein, and the content of NO (P<0.05, P<0.01). The effect of Moxi+NMDA group was obviously inferior to that of Moxi group in up-regulating MPT and down-regulating the levels of iNOS mRNA and protein, and the contents of cGMP and NO, and the activity of NOS (P<0.01), suggesting a reduction of the therapeutic effects in raising MPT and down-regulating expression of iNOS mRNA and protein after administration of AP-5. CONCLUSION: Moxibustion can relieve RA inflammatory pain in AA rats, which may be related to its function in down-regulating the NMDA/NO/cGMP signaling pathway in the spinal cord.

[Effects of moxibustion on p53， SLC7A11， and GPX4 expression in synovial tissues of rats with adjuvant arthritis].

OBJECTIVE: To observe the effects of moxibustion on p53, cystine/glutamate antiporter solute carrier family 7 member 11 (SLC7A11), and glutathione peroxidase 4 (GPX4) in synovial tissues of rats with adjuvant arthritis (AA), so as to explore the mechanism of moxibustion in alleviating rheumatoid arthritis. METHODS: Eighty rats were randomly divided into normal, model, moxibustion, and medication groups (n=20 in each group). The AA model was established by exposure to wind, cold, and damp environmental factors combined with injection of complete Freund's adjuvant. Rats in the moxibustion group received suspended moxibustion at "Shenshu" (BL23) and "Zusanli" (ST36) alternately, while those in the medication group were treated with tripterygium glycoside tablet suspension (8 mg/kg) by gavage, once a day, for 15 successive days. The pathological change in synovial tissue of rat right knee joint was observed by HE staining. The protein and mRNA expression levels of p53, SLC7A11, and GPX4 in the synovial tissue were detected by Western blot and quantitative real-time PCR, respectively. The se-rum glutathione (GSH) and reactive oxygen species (ROS) contents were measured by colorimetry and fluorescence probe me-thod. RESULTS: Compared with the normal group, the model group exhibited synovial hyperplasia of the right knee joint, massive inflammatory cell infiltration, up-regulated mRNA and protein expression of p53 in synovial tissue, elevated serum ROS content (P<0.01), down-regulated mRNA and protein expression of SLC7A11 and GPX4 in synovial tissue, and lowered serum GSH content (P<0.01). Comparison with the model group showed that the synovial injuries in the moxibustion and medication groups were obviously alleviated. The mRNA and protein expression levels of p53 in the synovial tissues and the serum ROS content declined significantly (P<0.01), while the mRNA and protein expression of SLC7A11 and GPX4 in the synovial tissues and the se-rum GSH content increased (P<0.01). There was no significant difference in histopathological change of synovial tissue between the moxibustion group and medication group. However, the p53 protein expression in the synovial tissue and the level of serum ROS were significantly higher in the medication group than in the moxibustion group (P<0.05), while the GPX4 protein expression and serum GSH content were down-regulated (P<0.05). CONCLUSION: Moxibustion improves the inflammatory response in synovial tissue of AA model rats, which may be closely related to its regulation of the expression of ferroptosis-related factors.

[Implication and implementation path of the ideological and political elements in acupuncture- moxibustion courses].

To explore the implementation path of the ideological and political education according to the characteristics of teaching sections in acupuncture-moxibustion courses. Excavating the traditional Chinese culture and medical ethics contained in acupuncture-moxibustion courses helps strengthening the ideological and political quality of medical students and noble medical ethics, strengthening self-confidence in both professions and culture, and also helps students establishing a correct outlook on life, world and value. The moral education integrated with the professional teaching will helps explore ideological and political education path in acupuncture-moxibustion courses, so as to solidify them into each teaching sections and improve the teaching effect.

[Toll-like Receptor 4/Nuclear Factor-κB Signaling in Synovial Tissue Is Involved in the Anti-inflammatory Effect of Moxibustion in Rats with Rheumatoid Arthritis].

OBJECTIVE: To observe the effect of moxibustion on Toll-like receptor 4/nuclear factor-κB (TLR 4/NF-κB) signaling in the synovial tissue of the ankle joint in rats with rheumatoid arthritis (RA), so as to analyze its biological mechanism underlying improvement of RA. METHODS: Fifty male SD rats were randomly divided into normal control, model, moxibustion, moxibustion ＋ TLR 4 agonist, and moxibustion ＋ TLR 4 antagonist groups (n＝10 rats in each). The RA model was established by subcutaneous injection of Freund's complete adjuvant (FCA, 0.1 mL/rat) at the right hind-paw and by being raised in a wind (air fan blowing), cold (about 10 ℃) and wet (purling) environment for 20 days. After 3 days of modeling, mild moxibustion was applied to bilateral "Shenshu" (BL 23) and "Zusanli" (ST 36) for 20 minutes, once daily for successive 10 days. The TLR 4 agonist (lipopolysaccharide) or TLR 4 antagonist (TAK-242) (1 mg/mL) was separately administered via the tail vein 30 min before performing moxibustion every time in the agonist group and the antagonist group. The expression of NF-κB inhibitory factor ɑ (IκBɑ), IκB kinase complex ß(IκKß), myeloid differentiation factor 88 (MyD 88), TLR 4, and NF-κB p 65 proteins in the synovial tissue of the ankle joint was detected by using Western blot. RESULTS: Following modeling, the girth of the swollen ankle joint was obviously bigger (P<0.01), and the expression levels of IκBɑ， IκKß， TLR 4, MyD 88, and NF-κB p 65 proteins in the synovial tissue were considerably increased in the model group relevant to the normal control group (P<0.01). After moxibustion intervention, the girth of the swollen ankle joint and the expression levels of IκBɑ， IκKß， TLR 4, MyD 88, and NF-κB p 65 proteins were significantly down-regulated in the moxibustion, moxibustion ＋TLR 4 agonist, moxibustion＋TLR 4 antagonist groups compared with the model group (P<0.01, P<0.05). Comparison among the 3 moxibustion groups showed that the lowered levels of ankle-joint girth, and IκBɑ， IκKß， TLR 4, MyD 88, and NF-κB p 65 expression were significantly smaller in the moxibustion＋TLR 4 agonist group than in the simple moxibustion and moxibustion＋TLR 4 antagonist groups (P<0.05，P<0.01). No significant differences were found between the moxibustion and moxibustion＋TLR 4 antagonist groups in the decreased ankle joint girth and IκBɑ， IκKß， TLR 4, MyD 88, and NF-κB p 65 expression levels (P>0.05), suggesting that activation of TLR 4 reduced the anti-inflammatory effect of moxibustion intervention. CONCLUSION: Moxibustion can reduce the ankle joint swelling in RA rats, which may be closely associated with its effect in down-regulating the expression of IκBɑ， IκKß， TLR 4, MyD 88, and NF-κB p 65 proteins and in inhibiting TLR 4/NF-κB signaling in the synovial tissue of the ankle joint.

[Effect of Electroacupuncture at "Shenmen" (HT 7)- "Tongli" (HT 5) of Heart Meridian on Neuronal Activities in Paraventricular Nucleus of Hypothalamus in Myocardial Ischemia Rats].

OBJECTIVE: To observe the effect of electroacupuncture (EA) stimulation of "Shenmen" (HT 7) - "Tongli" (HT 5) segment of the Heart Meridian on neuronal electrical activities of hypothalamic paraventricular nucleus (PVN) in rats with myocardial ischemia (MI), so as to investigate its possible mechanism underlying improvement of MI. METHODS: Thirty-two SD rats were randomly divided into sham control, model, HT 7-HT 5 and "Taiyuan" (LU 9)- "Lieque" (LU 7) groups (n=8 in each group). EA preconditioning (2 Hz, 1 V, 20 min) was applied to bilateral HT 7-HT 5 and bilateral LU 9-LU 7, respectively, once everyday for 7 days. The electrical activities of the right PVN region were recorded by the implanted microelectrode array(2×4)and Plexon multi-channel acquisition system. Cluster analysis of neuronal signals was carried out by Offline Sorter software. The discharge waveforms, autocorrelation and cross-correlation of neuronal activities were analyzed by using Neuro Explorer software. RESULTS: Cluster analysis of neuronal signals showed that 2, 2, 1 and 1 interneuron in the sham, model, HT 7-HT 5, and LU 9-LU 7 groups, and 3 pyramidal neurons in the HT 7-HT 5 were acquired. Cross correlation analysis showed that the SPK 02 a and SPK 02 b neurons of the HT 7-HT 5 group had an inhibitory relationship. The total discharge frequency was significantly increased in the model group relevant to the sham group (P<0.01), and was markedly lower in the HT 7-HT 5 group than in the model group and LU 9-LU 7 group (P<0.01). Real-time spectrum analysis showed that the local field potential spectrum energy of the HT 7-HT 5 group was significantly lower than that of the model group and the LU 9-LU 7 group. CONCLUSION: EA of HT 7-HT 5 segment of the Heart Meridian can inhibit the electrical activity of interneuron and activate the electrical activity of pyramidal neuron in PVN region, and an inhibitory relationship exists between the interneuron and pyramidal neuron in MI rats, which may be a mechanism of EA in regulating activities of the ischemic heart.

[Effects of electroacupuncture on the expression of oxidized low-density lipoprotein and its receptors in rats with coronary atherosclerotic heart disease].

OBJECTIVE: To explore action mechanism of electroacupuncture for coronary atherosclerotic heart disease (CHD) in order to provide experimental support for clinical acupoint selection. METHODS: Among sixty clean-grade healthy male Wistar rats, twenty-four cases were randomly selected as a normal control group and an electroacupuncture (EA) preconditioning group, 12 cases in each one. Then rats in the EA preconditioning group and the rest 36 rats were fed with high fat diet for 12 weeks to duplicate the CHD model. When the models were successfully established, the rats were randomly divided into a model control group, an EA group and a medication group, 12 cases in each one. EA was applied with Hwa-to SDZ-IV apparatus in the EA preconditioning group at "Neiguan" (PC 6) and "Xinshu" (BL 15), 1 mA in current intensity, 2 Hz in frequency, 30 min per times, once every other day for 14 weeks. When model was established, the same acupoint and method was used in the EA group for 2 weeks while intragastric administration of atorvastatin mixed suspension, 0.25 mg/kg, once a day, was applied in the medication group for 2 weeks. The content of oxidized low-density lipoprotein (oxLDL) in the serum was tested by double antibody enzyme-linked immunosorbent assay (ELISA) while content of lectin-like oxidized low density lipoprotein receptor-1 (LOX-1) in coronary arterial tissue was test by western blot method. Expression of LOX-1 mRNA was tested by fluorogenic quantitative polymerase chain reaction (PCR). RESULTS: After model was duplicated successfully, the content of oxLDL in the serum and the expression of LOX-1 and its mRNA in coronary arterial tissue in the model control group were increased significantly compared with those in the normal control group (all P < 0.01). Compared with the model control group, the content of oxLDL in the serum and the expression of LOX-1 and its mRNA in coronary arterial tissue in the EA preconditioning group, EA group and medication group were significantly reduced (all P < 0.01). CONCLUSION: The electroacupuncture at "Neiguan" (PC 6) and "Xinshu" (BL 15) could effectively reduce the content of oxLDL in the serum and expression of LOX-1 and its mRAN in coronary arterial tissue in CHD rats. The oxidative modificatory low-density lipoprotein and its specific receptor system could be one of the ways to prevention and treatment of acupuncture for CHD.

[Effects of electroacupuncture intervention on blood lipid levels and expression of CD 40 L and MMP-9 in the coronary artery tissue in coronary heart disease rats].

OBJECTIVE: To observe the effect of electroacupuncture (EA) of "Neiguan" (PC 6) and "Xinshu" (BL 15) on levels of serum total cholesterol (TC), triglyceride (TG), high density lipoprotein (HDL) and low-density lipoprotein (LDL) and expression of T cell CD 40 L and matrix metalloproteinases-9 (MMP-9) in the coronary artery in coronary heart disease (CHD) rats, so as to reveal its mechanism underlying improvement of coronary atherosclerotic heart disease. METHODS: A total of 60 Wistar rats were randomly divided into normal control, model, EA-pretreatment, routine EA and medication groups (n = 12). Coronary atherosclerotic heart disease model was established by feeding the animals with high fat forage for 3 months and intraperitoneal injection of VD3 (2 mL/kg, 60 000 IU/100 g, once daily for 3 days). EA was applied to unilateral "Neiguan"(PC 6) and "Xinshu"(BL 15) once every other day for rats of the EA-pretreatment group from the 1st day of modeling on for 12 weeks, and once daily for rats of the routine EA group from successful modeling on for 2 weeks. Rats of the medication group were treated by intragastric administration of Atorvastatin 0. 25 mg . kg-1 . d-1 for 2 weeks. Serum TC, TG, HDL and LDL contents were detected by ELISA and the expression levels of CD 40 L and MMP-9 of the coronary artery tissue assayed by Western blot. RESULTS: Serum contents of TC, TG and LDL and expression levels of coronary arterial CD 40 L and MMP-9 in the model group were significantly higher than those of the normal control group( P<0.01), while serum HDL content was decreased considerably in the mo- del group (P<0. 01). In comparison with the model group, the contents of serum TC, TG, HDL and the expression levels of arterial CD 40 L and MMP-9 in both EA-pretreatment group and routine EA group were remarkably down-regulated (P<0. 01), and serum LDL content was obviously up-regulated (P<0. 01). The effects of the EA-pretreatment group were significantly superior to those of the medication group in down-regulating the contents of serum TC, TG, HDL and decreasing the expression of coronary arterial CD 40 L and MMP-9 proteins (P<0. 01, P<0. 05). No significant differences were found between the EA-pretreatment and routine EA groups in all the 6 indexes (P>0. 05). CONCLUSION: Both EA-pretreatment and routine EA of PC 6 and BL 15 can down-regulate blood TC, TG and HDL contents, raise serum HDL level and suppress the increased expression of CD 40 L and MMP-9 proteins of coronary artery tissue in rats with coronary atherosclerotic heart disease, which may contribute to their effect in improving CHD.