Electroacupuncture inhibits IL-17/IL-17R and post-receptor MAPK signaling pathways in a rat model of chronic obstructive pulmonary disease.

OBJECTIVE: Interleukin (IL)-17, as a T-helper 17 cell (Th17) cytokine, plays a key role in chronic obstructive pulmonary disease (COPD) pathophysiology including chronic inflammation and airway obstruction, which lead to decreased pulmonary function. The aim of this study was to investigate the effect of acupuncture on IL-17, its receptor (IL-17R) and the mitogen-activated protein kinase (MAPK) signaling pathway, in a rat model of COPD. METHODS: The COPD model was induced in Sprague Dawley rats by exposure to cigarette smoke for 12 weeks. The model rats were treated with electroacupuncture (EA) at BL13 and ST36. The lung function and histology of the rats were observed. IL-17, tumor necrosis factor (TNF)-α, and IL-10 were detected by enzyme-linked immunosorbent assay (ELISA) in bronchoalveolar lavage fluid (BALF) and in plasma. The leukocytes and macrophages in the BALF were counted. The expression levels of IL-17R were assayed in lung tissue by real-time polymerase chain reaction (PCR), western blotting, and immunohistochemistry. MAPK signaling pathway molecules including c-Jun N-terminal kinase (JNK), extracellular signal-regulated kinase (ERK)1/2 and p38, and their phosphorylated forms, were observed in the lung by western blotting. RESULTS: Compared with the control group rats, lung function decreased and there was a severe inflammatory infiltration of the pulmonary parenchyma in the COPD rats. EA effectively improved lung function and alleviated the inflammatory infiltration in the lungs of COPD rats. EA also reversed the elevated total leukocyte and macrophage counts, the high levels of IL-17 and TNF-α, and the low IL-10 content in COPD rats. Meanwhile, EA downregulated the increased mRNA and protein expression of IL-17R, and significantly inhibited the elevated levels of phosphorylated JNK, ERK1/2, and p38 in the lungs of COPD rats. CONCLUSION: Our results suggest that the protective effects of acupuncture therapy on the lungs of COPD rats are likely related to inhibition of IL-17/IL-17R and the post-receptor MAPK signaling pathways.

[Protective effect of electroacupuncture intervention on neurovascular unit in rats with focal cerebral ischemia-reperfusion injury].

OBJECTIVE: To observe the effect of electroacupuncture (EA) intervention on behavior changes, expression of cerebral vascular endothelial growth factor (VEGF), nerve growth associated protein-43 (GAP-43), synaptophysin (SYN), myelin basic protein (MBP), neurite outgrowth inhibitor-A (Nogo-A) in cerebral focal ischemia-reperfusion injury (CI/RI) rats, so as to study its mechanism underlying improvement of ischemic cerebral vascular disease. METHODS: Sixty male SD rats were randomly divided into sham-operation group, model group and electroacupuncture (EA) group. CI/RI model was established by occlusion of the middle cerebral artery (MCAO) and reperfusion. EA was applied to bilateral "Neiguan" (PC 6), "Sanyinjiao" (SP 6), "Shuigou" (GV 26) and "Baihui" (GV 20) for 30 min, once a day for 14 days. The neurologic deficits were evaluated by Longa 5-grade standard (the higher the score, the severer the neurologic deficit). The immunoactivity of cerebral VEGF, GAP-43, SYN, MBP (important in the process of myelination of nerves in the nervous system) and Nogo-A (inhibiting axonal regeneration) in the focal ischemic cerebral tissue was detected by immunohistochemistry. RESULTS: The Longa's score of the model group was significantly increased after MCAO in comparison with the sham-operation group (P < 0.01). In comparison with the model group, Longa's score of the EA group was evidently lower on day 14 after CI/RI (P < 0.05), suggesting an improvement of the neurological deficits after EA intervention. In comparison with the sham-operation group, the immunoactivity of cerebral VEGF, GAP-43 and Nogo-A was significantly upregulated on day 7 and 14 in the model group (P < 0.01), while that of cerebral SYN was remarkably down-regulated in the model group on day 7 and 14 after CI/RI (P < 0.05). Compared with the model group, cerebral VEGF, GAP-43, SYN and MBP expression levels were considerably upregulated on day 7 and 14 following CI/RI in the EA group (P < 0.01, P < 0.05), while that of cerebral Nogo-A was significantly decreased at the two time-points in the EA group (P < 0.01). CONCLUSION: EA intervention can effectively improve neurological function in cerebral infarction rats, which is closely related to its effects in upregulating cerebral VEGF, GAP-43, SYN and MBP expression, and down-regulating Nogo-A protein, suggesting a protective effect on neurovascular unit.

[Wuzi yanzong pills increases sperm mitochondrial membrane potential and protects its ultrastructure in oligo-asthenozoospermia model rats].

OBJECTIVE: To study the effects of Wuzi Yanzong Pills (WYP) on sperm mitochondrial membrane potential (MMP) and its ultrastructure in oligo-asthenozoospermia model rats. METHODS: Oligo-asthenozoospermia models were made in 50 male rats weighing 200 - 220 g by intragastric administration of Tripterygium Glucosides at 30 mg per kg per d for 8 weeks, and then equally allocated to a model control, a Huangjing Zanyu Capsule (HZC) control, a low-dose WYP, a medium-dose WYP, and a high-dose WYP group. Another 10 age-matched normal male rats were included as normal controls. The rats in the model and normal control groups were given intragastrically distilled water at 10 ml/kg, those in the HZC group administered HZC at 3.01 g/kg, and those in the low-, medium- and high-dose WYP groups medicated with WYP at 2.30, 4.60 and 9.20 g/kg, respectively, once daily for 30 days. At 30 minutes after the last administration, we detected the sperm MMP by JC-1 fluorescent staining and flow cytometry, and examined the sperm ultrastructure under the JEM-1230 transmission electron microscope. RESULTS: JC-1 + % and its fluorescence intensity were (33.77 +/- 6.19)% and 1 468 +/- 496 in the model control, (56.34 +/- 10.35)% and 3 277 +/- 895 in the HZC control, (40.80 +/- 10.40)% and 2 016 +/- 767 in the low-dose WYP, (59.40 +/- 6.51)% and 3 897 +/- 643 in the medium-dose WYP, and (60.71 +/- 7.81)% and 3 371 +/- 647 in the high-dose WYP group, significantly reduced in comparison with (70.80 +/- 4.92)% and 4 360 +/- 945 in the normal control group (P < 0.05), but remarkably higher in the medium- and high-dose WYP groups than in the model controls (P < 0. 05). After modeling, the sperm membrane was loose and degenerated, the mitochondria swelling, variously sized and with incomplete membrane, and the axonemal structure unclear or ruptured. After 30 days of WYP administration, compared with the model control group, the rats exhibited integrated sperm membrane and mitochondrial membrane, reduced mitochondrial swelling and basically normal axonemal and microtubular structures. CONCLUSION: Tripterygium Glucosides could decrease the sperm mitochondrial membrane potential and damage the mitochondrial structure, while WYP could significantly increase the sperm mitochondrial membrane potential and reduce the sperm mitochondrial structure damage. The protection of the integrity of sperm mitochondrial structure and function is one of the mechanisms of WYP acting on oligo-asthenozoospermia.

[PI 3 K/Akt signaling pathway contributed to the protective effect of acupuncture intervention on epileptic seizure-induced injury of hippocampal pyramidal cells in epilepsy rats].

OBJECTIVE: To observe the protective effect of acupuncture stimulation on pyramidal cells in hippocampal CA 1 and CA 3 regions and to analyze the involvement of phosphatidy linositol-3-kinase (PI 3 K)/protein kinase B(PKB or Akt) signaling pathway in the acupuncture effect in epilepsy rats. METHODS: A total of 120 SD rats were randomly divided into normal control group, model group, LY 294002 (a specific antagonist for PI 3 K/Akt signaling) group, acupuncture+ LY 294002 group and acupuncture group (n = 24 in each group, 12 for H. E. staining, and 12 for electron microscope observation). Epilepsy model was established by intraperitoneal injection of pentylenetetrazol (PTZ, 5 microL). Manual acupuncture stimulation was applied to "Baihui" (GV 20) and "Dazhui" (GV 14) once daily for 5 days. Dimethyl Sulfoxide (DMSO, 5 microL, a control solvent) was given to rats of the normal, model and acupuncture groups, and LY294002 (5 microL, dissolved in DMSO) given to rats of the LY 294002 and acupuncture+ LY 294002 groups by lateral ventricular injection. Four hours and 24 h after modeling, the hippocampus tissues were sampled for observing pathological changes of CA 1 and CA 3 regions after H. E. staining under light microscope and for checkin ultrastructural changes of the pyramidal cells under transmission electron microscope. RESULTS: In comparison with the normal control group, the numbers of pyramidal cells of hippocampal CA 3 region in the model group were decreased significantly 4 h and 24 h after epileptic seizure (P < 0.01). While compared to the model group, the pyramidal cells of hippocampal CA 3 region in the acupuncture group were increased considerably in the number at both 4 h and 24 h after seizure (P < 0.01). No significant differences were found between the LY 294002 and model groups, and between the acupuncture+ LY 294002 and model groups in the numbers of pyramidal cells at 4 h and 24 h after seizure (P > 0.05). Findings of the light microscope and electron microscope showed that the injury severity of pyramidal cells of hippocampal CA 1 and CA 3 regions was moderate 4 h after epileptic seizure and even worse 24 h after seizure in the model group, LY 294002 group and acupuncture+ LY 294002 group, but relatively lighter in the acupuncture group. These results suggested an elimination of the acupuncture effect after blocking the PI 3 K/Akt signaling pathway by lateral ventricular injection of LY 294002 in epilepsy rats. CONCLUSION: Acupuncture intervention has a protective effect on pyramidal cells of hippocampal CA 1 and CA 3 regions in epilepsy rats, which is associated with the normal function of intracellular PI 3 K/Akt signaling pathway.

[Effect of electroacupuncture on epileptic EEG and intracellular Ca2+ content in the hippocampus in epilepsy rats].

OBJECTIVE: To study the underlying mechanism of electroacupuncture (EA) in relieving epilepsy in pentylenetetrazole (PTZ)-induced epilepsy rats. METHODS: Twenty SD rats were randomly divided into normal control, model, EA, Nimodipine groups, with 5 cases in each. Epilepsy model was established by intraperitoneal injection of PTZ (32.0 mg/kg), once daily for 28 days. EA (100 Hz, 0.6 mA) was applied to "Baihui" (GV 20) and "Dazhui" (GV 14), once daily for 7 days. For Nimodipine group, the rats were given with nimodipine (0.25 mg/kg, i.p.), once daily for 7 days. Electroencephalogram (EEG) was recorded and the fluorescence intensity of Ca2+ of the hippocampus tissue sections was detected by laser scanning confocal microscope (LSCM) after incubation in artificial cerebrospinal fluid containing Flou-3/AM (10 micromol/L) and pluronic F-127 (5 microl). RESULTS: Compared with model group, the latencies of epileptic EEG seizure prolonged obviously (P < 0.05), and epileptic EEG seizure frequencies decreased significantly (P < 0.05) in EA and Nimodipine groups. The fluorescence intensity of intracellular Ca2+ in hippocampus tissue in model group was obviously higher than that in control group (P < 0.01). In comparison to model group, Ca2+ levels in EA and Nimodipine groups lowered considerably (P < 0.05, P < 0.01). No significant differences were found between EA and Nimodipine groups in the aforementioned 3 indexes (P > 0.05). CONCLUSION: EA has an obvious anti-epileptic effect, which may be closely related to its effect in downregulating the increased hippocampal Ca2+ level in PTZ-kindled epilepsy rats.