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OBJECTIVE: To observe the effect of "Neiguan" (PC6)-electroacupuncture (EA) or moxibustion (Moxi) pretreatment on myocardial apoptosis and expression of autophagy related proteins light chain (LC) 3-Ⅰ and LC3-Ⅱ in rats with myocardial ischemia/reperfusion injury (MIRI), so as to explore their mechanisms underlying improvement of MIRI. METHODS: Forty SD rats (half male and half female) were randomly divided into sham operation, model, ischemic preconditioning (IP), EA and Moxi groups (n＝8 in each group). EA (10 Hz/50 Hz, 1 mA) or Moxi (ignited moxa stick) was respectively applied to bilateral "Neiguan" (PC6) for 20 min, once daily for 7 days. The MIRI model was established by occlusion of the anterior descending branch of the left coronary artery for 40 min, followed by reperfusion for 60 min. The ultrastructural changes and autophagy of myocardial cells were observed by electron microscopy (EM), and the myocardial cellular apoptosis [apoptotic index ＝ (number of apoptotic cells/total number of cardiomyocytes)×100%] was detected by the terminal deoxyribonucleotidyl transferase mediated dUTP nick end labelling (TUNEL) method. The expressions of LC3-Ⅰ and LC3-Ⅱ proteins (markers for autophagy) in myocardial tissue were detected by Western blot. RESULTS: Following MI, EM observation revealed a vague structure of cardiomyocytes and muscular horizontal grain, dissolution of myofibers, mitochondrial swelling, some autophagic vacuoles and autophagic lysosomes at different degrees and surrounded by a double membrane in the model group, these situations were apparently milder in the EA and Moxi groups. The apoptosis index, myocardial LC3-Ⅰ and LC3-Ⅱ protein expression levels, and the ratio of LC3-Ⅱ/Ⅰ were significantly increased in the model group relevant to the sham operation group (P0.05), and no significant differences were observed in the apoptosis index and levels of LC3-Ⅱ and LC3-Ⅱ/Ⅰ between the IP and Moxi groups (P>0.05)．. CONCLUSION: Both EA and moxibustion pretreatments, similar to IP, have a positive role in reducing myocardiocyte apoptosis and regulating autophagy-related protein expression in MIRI rats, which maybe contribute to their protective effects on ischemic myocardium.
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OBJECTIVE: To observe the effect of moxibustion pretreatment at different time on serum hormone levels in diminished ovarian reserve (DOR) rats, so as to explore its protective mechanisms. METHODS: Forty female SD rats were randomly divided into control, model, moxibustion-1 (moxibustion was given 4 weeks before modeling), moxibustion-2 (moxibustion was given 2 weeks before modeling and 2 weeks from the 1st day on after modeling ) and moxibustion-3 (moxibustion was given 4 weeks from the 1st day on after modeling) groups (n=8 rats in each group). The DOR model was established by gavage of Tripterygium Glycosides (75 mg/kg) once daily for 14 days. Grain-moxibustion was applied to "Shenshu" (BL23) and "Guanyuan" (CV4) for 7 cones, 5 times a week for 4 weeks. The body weight and the ovary weight were recorded for calculating the ovarian index. The levels of serum anti-mullerian hormone (AMH), follicle stimulating hormone (FSH), estradiol (E2), androgen (T) and dehydroepiandrosterone (DHEA) were detected by ELISA. RESULTS: After modeling, ovarian index and serum AMH levels were obviously decreased (P<0.05), and the levels of serum FSH, E2, T and DHEA were significantly increased in contrast with the control group (P<0.05). Following intervention and compared with the model group, the serum FSH and DHEA levels of each moxibustion group were significantly reduced (P<0.05), the AMH levels significantly increased and E2 and T contents significantly decreased in the moxibustion-2 and moxibustion-3 groups (P<0.05). The serum FSH, E2 and T contents in moxibustion-2 group were obviously lower than those of the moxibustion-1 and moxibustion-3 groups (P<0.05). CONCLUSION: Moxibustion pre-treatment can improve ovarian reserve function in DOR rats, while the effect is different with different intervention time, and the best intervention time is pre-occurrence and early stage of DOR.
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OBJECTIVE: To observe the effect of electroacupuncture (EA) and moxibustion (Moxi) pretreatment on myocardial pathological and structural changes and expression of autophagy related protein LC 3 Ⅰ/Ⅱ and Beclin 1 in rats with myocardial ischemia-reperfusion injury (MI/RI), so as to explore their mechanisms underlying improving MI/RI. METHODS: Forty SD rats were randomly divided into sham operation, model, ischemic preconditioning (IP), EA and Moxi groups (n＝8 in each group). EA (10 Hz/50 Hz，1 mA) or Moxi (ignited moxa stick) was respectively applied to bilateral "Neiguan"(PC 6) for 20 min, once daily for 7 days. The MI/RI model was established by occlusion of the anterior descending branch of the left coronary artery for 40 min, followed by reperfusion for 60 min. The left ventricular (LV) tissue samples were collected and analyzed for pathological (H.E. staining) and ultrastructural changes, for myocardial apoptosis (apoptotic index＝ number of apoptotic cells/total number of cardiomyocytes×100%) with terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) method, and for the expression of LC 3 and Beclin 1 in myocardial cells with Western blot. RESULTS: Following MI/RI, H．E. staining revealed a disorder of arrangement of cardiomyocytes with vague border, inflammatory cell infiltration, intracellular swelling with bleeding, necrosis and dissolution of partial striated muscles of the left ventricle under light microscope, and dual staining of Uranyl acetate and leadnitrate showed atrophy, arrangement disorder, dissolution, necrosis, and interstitial edema of partial myocardial fibers, mitochondrial structural disorder, vacolation, and large body of autophagosomes with bilayers, etc. in ultrastructure, which was relatively lighter in both EA and Moxi groups. The apoptosis index, expression levels of myocardial LC 3 Ⅱ and Beclin 1 and the ratio of LC 3 Ⅱ/LC 3 Ⅰ were significantly higher in the model group than those in the sham operation group (P<0.01), but the expression level of LC 3 Ⅰ was considerably down-regulated in the model group relevant to the sham operation group (P<0.01). Following the intervention and MI preconditioning, the increased apoptosis index and expression levels of LC 3Ⅱ and Beclin 1 proteins and the ratio of LC 3Ⅱ/LC 3 Ⅰ were obviously down-regulated in the IP, EA and Moxi groups relevant to the model group (P<0.01), and the decreased expression of LC 3 Ⅰ protein was up-regulated obviously in the 3 treatment groups (P<0.05，P<0.01). The effects of EA were significantly superior to those of IP and Moxi groups in down-regulating apoptosis index and expression of LC 3 Ⅱ and Beclin 1 and the ratio of LC 3 Ⅱ/LC 3 Ⅰ and in up-regulating expression of LC 3 Ⅰ (P<0.05, P<0.01). CONCLUSION: Both EA and Moxi preconditioning of PC 6 have a protective effect on ischemic myocardium in MI/RI rats, which is probably related to their effects in regulating expression of myocardial autophagy proteins as LC 3 Ⅰ/Ⅱ and Beclin 1.
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@#ObjectiveTo observe the effect of dressing and moxibustion-pretreatment on the local joint swelling and stress hormone in hypothalami of rats with adjuvant arthritis (AA) at early and secondary stages.MethodsForty Wistar rats were randomly divided into 5 groups: normal group, early and secondary model groups, early and secondary pre-dressing and moxibustion (PDM) groups. The dressing with Chinese herb and moxibustion was stuck on Dazhui point (GV14) before the AA model established. The effects of dressing and moxibustion-pretreatment on the feet swelling and corticotropin-releasing-hormone (CRH), beta-endorphin (β-EP) and neuropeptide-Y (NPY) in hypothalami were observed.ResultsThe right feet swelling rate at early and secondary stages obviously increased after modeling ( P<0.01), and it became lower in early and secondary PDM groups than in model groups at the same phases ( P<0.05～0.01). The level of hypothalamic CRH was higher after modeling ( P<0.05～0.01), compared with early model group, it had a tendency to going down in early PDM group, moreover, in secondary PDM group the level was similar with the normal group. The level of hypothalamic β-EP increased significantly in early model group ( P<0.01), and lightly changed in secondary model group, it decreased in early PDM group but increased in secondary PDM group, compared with model groups at the same stages ( P<0.05). The level of hypothalamic NPY increased significantly after modeling, and it declined in the secondary PDM group ( P<0.05).ConclusionDressing and moxibustion-pretreatment can relief feet swelling of AA rats, which may be related with its regulative effect on the level of hypothalamic CRH, β-EP and NPY.