Electroacupuncture for myocardial ischemia injury in rats via AMPK-HDAC5-HIF-1α signaling / 中国针灸

OBJECTIVE@#To explore the impact of electroacupuncture (EA) on the AMPKα-HDAC5-HIF-1α signaling in the heart of the rats with myocardial ischemia (MI) via detecting the expressions of AMP-activated protein kinase α (AMPKα), histone deacetylase 5 (HDAC5), hypoxia inducible factor (HIF-1α) and vascular endothelial growth factor (VEGF).@*METHODS@#Thirty-six healthy male SD rats were randomized into a sham operation group (6 rats), a sham + EA group (6 rats), a model group (12 rats) and an EA group (12 rats). We ligated the left anterior descending artery (LAD) for MI model, and exposed the heart of rats after opening the chest without ligation for the rats in the sham operation gorup and the sham + EA group. On the 2nd day after LAD ligation, EA was applied at "Neiguan" (PC 6) with 2 Hz/15 Hz and 1.5-2 mA for 30 min in the EA group and sham+EA group, once a day for 4 days. The same fixation was used in the sham operation group and the model group, without EA. Myocardial infarction area was observed by TTC staining and serum cardiac troponin T (cTnT) was detected by radioimmunoassay. The expression of VEGF mRNA was detected by real time PCR. The protein expressions of AMPKα, HDAC5, HIF-1α and VEGF were detected by western blot.@*RESULTS@#Compared with the sham operation group, 4 days after LAD ligation, the myocardial infarction was obvious and the expression of serum cTnT increased in the model group (0.05). After EA for 4 days, the myocardial infarction area and cTnT expression decreased in the EA group (both <0.01); the VEGF mRNA and protein expressions and AMPKα, HDAC5, HIF-1α protein expressions increased (<0.05, <0.01).@*CONCLUSION@#EA could regulate the AMPKα-HDAC5-HIF-1α signaling in myocardial tissue, which may activate VEGF expression for angiogenesis signaling, reduce myocardial infarction area so as to achieve cardioprotective effect.

Effects of electroacupuncture on inflammatory response of cardiac muscle tissue in mice with acute myocardial ischemia / 中国针灸

<p><b>OBJECTIVE</b>To observe the effects of electroacupuncture (EA) on inflammatory reaction of acute myocardial ischemia (MI) in mice, and to explore its action mechanism.</p><p><b>METHODS</b>Forty adult male C57BL/6 mice were randomly divided into a control group, a sham operation group, a model group and an EA group, 10 mice in each one. The model was established in the model group and EA group by ligating the left anterior descending branch of coronary artery. The mice in the EA group were treated with EA at "Neiguan" (PC 6) with 2 mA of intensity and 2 Hz /100 Hz of frequency; EA was given 30 min per treatment, once a day for totally 5 days. The mice in the control group and model group were treated with immobilization and no EA was given. The mice in the sham operation group were not treated with ligating at the left anterior descending branch of coronary artery, but the remaining procedure was identical to the model group. The electrocardiogram was recorded and △ST was calculated to evaluate the model. TTC and HE staining methods were applied to evaluate the infarct size and pathologic change of myocardial tissue, respectively. Western blot method was applied to test the protein expression levels of tumor necrosis factor-α (TNF-α), nuclear factor-κB p65 (NF-κB p65), interleukin-1β (IL-1β) and interleukin-8 (IL-8).</p><p><b>RESULTS</b>Compared with the sham operation group, the S-T segments in the model group and EA group were increased obviously after modeling (both <0.01), indicating the MI model was established successfully. The TTC and HE staining results indicated, compared with the sham operation group, the model group had larger infarction size (<0.01), more myocardial fibers injury and inflammatory infiltration; compared with the model group, the infarction size of the EA group was significantly reduced (<0.01), and the myocardial fibers injury and inflammatory infiltration were improved. Compared with the control group, the protein expression levels in the sham operation group were similar (all >0.05); compared with the sham operation group, the expression levels of TNF-α, NF-κB p65, IL-1β and IL-8 were significantly increased in the model group (<0.01, <0.05); compared with the model group, the expression levels of TNF-α, NF-κB p65, IL-1β and IL-8 were significantly reduced in the EA group (all <0.05).</p><p><b>CONCLUSION</b>EA might reduce the protein expression levels of TNF-α, NF-κB p65, IL-1β and IL-8 in cardiac muscle tissue to inhibit inflammatory reaction and achieve myocardial protective effect in mice with acute myocardial ischemia.</p>

Effects of electroacupuncture on the expression of adenosine receptors in the heart tissue of myocardial ischemia rats / 中国针灸

<p><b>OBJECTIVE</b>To explore the impact of electroacupuncture (EA) on the protein expression of adenosine receptors in the heart of the rats with myocardial ischemia (MI).</p><p><b>METHODS</b>Thirty healthy male SD rats were divided randomly into a control group (=6), a model group (=12) and an EA group (=12). We ligated the left anterior descending artery (LAD) for MI model in the model group and EA group, and exposed the heart after opening the chest without ligation in the control group. EA, 2 Hz /15 Hz and 1.5-2 mA, was applied at bilateral"Neiguan"(PC 6) in the EA group for 20 min, once a day for continuous 5 days. No intervention except grabbing and fixation was used in the control group and model group. We applied 2% TTC staining to observe the infarct size of myocardium, colorimetry to analyze serum lactic dehydrogenase (LDH), creatine kinase (CK), creatine kinase isoenzyme (CK-MB), radio-immunity assessment to detect cardiac troponin T (cTnT), Western blot to evaluate the adenosine A1 receptor (A1AR), A2aAR, A2bAR and A3AR.</p><p><b>RESULTS</b>After treatment, myocardial infarction of (27.56±3.24)% was obvious in the model group; the myocardial infarction in the EA group was (21.04±3.61)%, with statistical significance (<0.05). The expressions of serum LDH, CK, CK-MB and cTnT levels in the model group increased compared with those in the control group (all<0.01), and the expressions of LDH, CK, CK-MB and cTnT levels in the EA group decreased compared with those in the model group (<0.05，<0.01). The A1AR expression in the model group was not different from that in the control group (>0.05), and A2aAR、A2bAR、A3AR expressions decreased (<0.05,<0.01). A2aAR and A2bAR expressions in the EA group increased compared with those in the model group (both<0.01), and there was no statistical significance between A1AR and A3AR expressions (both>0.05). .</p><p><b>CONCLUSION</b>EA may achieve cardioprotective effect by regulating the expressions of A2aAR and A2bAR in myocardial tissue, which induce the corresponding signal cascade for reducing myocardial infarction area.</p>

Application of contrast-enhanced ultrasonography in the evaluation of the stability of carotid atherosclerotic plaque / 中国基层医药

Objective To study the application of contrast-enhanced ultrasonography (CEUS) in the evaluation of the stability of carotid atherosclerotic plaque (CAP).Methods 162 patients with CAP were selected as study group.Meanwhile,34 patients with carotid artery strong echo plaques were selected as control group.The color doppler ultrasound was used to observe the CAP.Results The proportions of lipid type,fiber type,calcification and ulcer plaque in the study group were 21.60%,33.33%,34.57% and 10.37%,which were higher than those of the control group (5.88%,2.94%,2.94% and 2.94%),the differences were statistically significant (χ2=4.537,12.859,13.629 and 3.855,all P0.05).The plaque diameter (4.13±0.75)mm diagnosed by CEUS was significantly larger than that of conventional ultrasound [(3.62±1.14)mm],the difference was statistically significant (t=4.757,P=0.000).Conclusion The CEUS can qualitatively detect the atherosclerotic plaque angiogenesis,can quantitatively assess plaque,evaluate the stability of the plaques,and the sensitivity is high.

Protective Effect of Catalpol on Vascular Endothelial Cell Injured by High Glucose / 世界科学技术-中医药现代化

This study was aimed to investigate the protective effect of catalpol against high glucose induced injury on vascular endothelial cells (ECV-304). ECV-304 cells were culturedin vitro. MTT method was applied to detect cell viability. NO and LDH level were measured by spectrophotometer method. The effect of catalpol against high glucose induced injury on ECV-304 was observed. The results showed that there were no obvious changes in cell viabilities of ECV-304 cultured in normal concentration of glucose medium among groups with different concentrations of catalpol. Compared with the control group, cell viability was significantly decreased (P 0.05); while the LDH level was reduced (P< 0.01). Comparison between the high dose catalpol (100 ng·μL-1 catalpol + 30 mmol·L-1 glucose) group and the high glucose model group showed that the cell viability was obviously increased (P< 0.01); NO secretion was increased (P< 0.05); and the LDH level was obviously reduced (P< 0.01). It was concluded that catalpol had protective effect against high glucose induced ECV-304 injury through the regulation of NO and LDH level. The effect of high dose was obvious.

Analysis on the academic cultivation of young TCM scholars based on Professor LIU Zhicheng's academic track / 中国针灸

In view of the public academic papers of Professor LIU Zhicheng, his academic track is analyzed, including validating effect, exploring mechanism, improving efficacy, expanding disease variety, deepening mechanism, optimizing program and seeking breakthrough. On this basis, combined with the self-character of TCM, the potential path for the academic cultivation of young scholars is refined: focusing on clinical practice, confirming clinical efficacy, deepening mechanism exploration, improving clinical therapeutic effect, optimizing clinical program, extending clinical disease spectrum, inheriting, seeking, breaking through, innovating, expecting and topping over. It is believed that during the academic cultivation, the importance is to form the stable research direction and continuity, organize research team, absorb actively new knowledge and thoughts, and adopt multidisciplinary cross and coordination so as to achieve the greater success ultimately.

Wnt signaling pathway is involved in catalpol-induced proliferation of rat bone marrow mesenchymal stem cells / 中国病理生理杂志

AIM:To investigate the changes of Wnt signaling pathway in catalpol-induced proliferation of rat bone marrow mesenchymal stem cells (BMSCs).METHODS:The BMSCs were isolated from SD rats , purified by differ-ential time adherent method and divided into control group and catalpol (1.0 mg/L) group.Flow cytometry was used to de-tect the proliferation index of BMSCs .The mRNA levels of Wnt3a, Wnt5a, Wnt11 and β-catenin was evaluated by real-time PCR.In addition, the protein expression level of β-catenin was determined by Western blotting .RESULTS:Prolife-ration index was increased from 8.90%±0.46% to 17.93%±1.68% after treatment with catalpol (P<0.01).Com-pared with control group , the mRNA expression of Wnt5a, Wnt11 andβ-catenin was all increased with catalpol treatment . No difference of Wnt3a mRNA expression between control group and catalpol group was observed .Meanwhile, the protein expression of β-catenin was increased in catalpol group compared with control group .CONCLUSION:Catalpol promotes BMSCs going into the cell cycle .Classical and non-classical Wnt signaling pathways are activated in this process .

The study of the correlation between serum C-reactive proteln,tumor necrosis factor-alpha and atherosclerosis in patients with subclinnical hypothyroidism / 中国医师进修杂志

ObjectiveTo investigate the correlation between serum C-reactive protein(CRP),tumor necrosis factor-alpha(TNF-α) and atherosclerosis in patients with subclinical hypothyroidism.Methods Seventy-five patients with subclinical hypothyroidism were divided into two groups according to the level of TSH:mild group (42 cases) with TSH between 5.5 and 10.0 mU/L and severe group (33 cases) with TSH ＞10.0 mU/L.Thirty healthy physical examinees or volunteers were selected as control group.TG,LDL-C,HDL-C,TSH,FT4,FT3,CRP,TNF-α and carotid intima-media thickness(CIMT) of all cases were detected and measured.ResultsCIMT of mild,severe and control groups were(0.88 ± 0.20),( 1.12 ±0.21 ),(0.62 ±0.21 ) mm,respectively.CIMT of mild and severe groups were both significantly higher than that of control group,CIMT of severe group was significantly higher than that of mild group (P ＜ 0.01 ).There were no significant differences of CRP and TNF- α among three groups (P ＞ 0.05 ).LDL-C of mild and severe groups were significantly higher than that of control group [ (3.22 ± 0.37),(3.49 ± 0.38 ) mmol/L vs.(2.48 ±0.41 )mmol/L,P ＜0.01 ].HDL-C and TG between mild and control groups had no statistical significance (P ＞0.05).Compared with control and mild groups,HDL-C of severe group decreased [(0.92 ±0.10)mmol/L vs.( 1.21 ± 0.14),( 1.17 ± 0.11 ) mmol/L] and TG increased [ ( 1.50 ± 0.49) mmol/L vs.( 1.11 ±0.53),(1.27 ±0.47) mmol/L](P＜ 0.01 or ＜ 0.05).The levels of TG,LDL-C,TSH,CRP and TNF-α had positive correlation with CIMT (r =0.52,0.37,0.48,0.39,0.45;P ＜0.05 or ＜0.01 ).FT4 was negatively correlated with CIMT (r=-0.24,P＜0.05).HDL-C had no relation with CIMT (r =0.06,P ＞0.05).ConclusionsThere are many abnormalities such as metabolism disorder of blood lipid in patients with subclinical hypothyroidism.The risk of atherosclerosis increases in these patients and inflammatory reaction may not be an important factor of subclinical hypothyroidism complicated with atherosclerosis.

Effect of TNF-? and Yigu capsule drug-containing sera on osteoblasts apoptosis in coculture system / 中国病理生理杂志

AIM:To observe the effect of TNF-? and Yigu capsule drug-containing sera on osteoblasts apoptosis in osteoblasts-osteoclasts coculture system.METHODS:(1)Twenty female Sprague-Dawley rats,10 months old,were randomly assigned into 2 groups,NS and Yigu capsule groups,to prepare blank sera and drug-containing sera.(2)The DNA gel electrophoresis method was used to detect apoptosis of osteoblasts treated with different concentration of TNF-? in order to determine the best dosage in the co-culture system.(3)The cells were divided into four groups,TNF-? group,normal group,TNF-? + blank serum group and TNF-? + drug-containing serum group.DNA gel electrophoresis,acridine orange staining and flow cytometry were used to observe osteoblast apoptosis in these groups.RESULTS:(1)After induced by TNF-? for 72 h at a concentration of 60 ?g/L,relatively typical DNA ladder appeared in TNF-? group.(2)Only two DNA brands appeared and most of cells were well-proportioned stained in TNF-? + drug-containing serum group,the rate of osteoblasts apoptosis in TNF-? + drug-containing serum group(9.60%?0.26%)was obviously lower than that in the TNF-? group(26.90%?0.06%)and TNF-? + blank serum group(18.10%?0.06%).CONCLUSION:TNF-? induces osteoblast apoptosis in the co-culture system,and Yigu capsule drug-containing serum prevents osteoblast apoptosis induced by TNF-?.

Effect of yigu capsule drug-containing serum on proliferation, ALP activity and expression of interleukin-11 mRNA in rat osteoblasts in the co-culture / 中国病理生理杂志

AIM: To investigate the effect of yigu capsule drug-containing serum on proliferation, ALP activity and expression of interleukin-11 mRNA in Sprague-Dawley rat osteobalsts in the co-culture system. METHODS: (1) Osteoblasts and osteoclasts were isolated from 1 and 5-day-old Sprague-Dawley rats, respectively. The osteoblasts-osteoclasts co-culture system was built to prevent the two kinds of cells from contact and allow the media to exchange. The experiment included two groups, drug-containing sera group and control group. (2) The osteoblasts proliferation, ALP activity and expression of interleukin-11 mRNA were detected by the MTT, 4-aminoantipyrine spectrometric methods and FQ-PCR,respectively. RESULTS: In drug-containing sera group, the osteoblasts proliferation, ALP activity, expression of IL-11 mRNA were higher than those of control group (P

Effect of BBC-containing serum on the activity and apoptosis of osteoclasts in osteoblasts and osteoclasts co-culture system / 中国病理生理杂志

AIM: To study the effect of Bone-benefiting capsule(BBC)-containing serum on activity and apoptosis of osteoclasts(OC) in osteoblasts(OB) and osteoclasts co-culture system.METHODS:(1) OB were separated from the skull of SD rats aged 1 d and OC from the thighbone and shinbone of SD rats aged 5 d,the plane OB and OC co-culture system was established,in which the cells in each culture system did not intermix,but the culture medium communicated,and BBC-containing serum group and control group were concluded in the experiment.(2) Female SD rats aged 10 months were divided into BBC group and control group to prepare BBC-containing serum and control serum.(3) The TRAP secreted by OC was measured by the diazol method and the absorption of lacuna formed by OC was observed by microscope in OB and OC co-culture system to determine the activity of OC with the effect of BBC-containing serum.(4) The effect of BBC-containing serum on OC apoptosis in OB and OC co-culture system was examined by optical microscopes and fluorescence microscopes.RESULTS: In OB and OC co-culture system,the activity of TRAP secreted by OC of BBC-containing serum group were obviously lower than that of control group in 48 h,72 h and 96 h,the OC surviving numbers were obviously lower than that of control group and the rate of OC apoptosis was markedly higher than that of control group which showed an obvious time-efficiency relationship;the amount of the absorption of lacuna formed by OC in BBC-containing serum group was evidently lower than that of control group(P