| Objectives1.Assess the efficacy of acupuncture at Gongsun,Neiguan,Xin Yu and Ju Que points in the treatment of angina pectoris of the Qi-deficiency-and-blood-stasis type;2.Study the mechanism of acupuncture treatment at Xin Yu,Ju Que,Gong Sun and Neiguan in patients with angina pectoris after PCI by measuring angina pectoris symptoms,TCM symptom score,quality of life,anxiety score,inflammatory factors and endothelial function;3.Study the efficacy of acupuncture at Xin Yu,Ju Que,Gong Sun and Neiguan in myocardial ischemia by an ischemia-reperfusion rat model,and investigate a possible mechanism of acupuncture-induced improvements through a SPHK1/TIMP1 signaling pathway.MethodsSystem assessment:The relevant literature from PubMed,MEDLINE,Cochrane Library,CNKI,Wanfang Database and VIP Chinese Journal Database were selected by using"angina pectoris","chest pain","acupuncture or acupuncture and moxibustion" and "qi deficiency and blood stasis" as the main search words,and the search date was on July 31,2022.A randomized controlled trial was conducted to compare the efficacy of acupuncture and other methods in the treatment of angina pectoris in patients with angina pectoris.The quality of the literature included in this study was evaluated using the Review Manager 5.3 software,and statistical analyses were conducted by using R4.1.2.Clinical study:A total of 120 patients with UAP who continued experiencing chest tightness and chest pain after PCI were randomly divided into an acupuncture point group(Group A),an acupuncture non-meridian and non-acupoint group(Group B)and a control group at a ratio of 1:1:1.The control group was treated according to the guidelines of the treatments of coronary heart disease.Group A was treated with 30-minute-long-acupuncture at Xin Yu,Ju Que,Gong Sun and Neiguan once every other day for 4 weeks.Group B was treated with 30-minute-long acupuncture at sites that were 2 centimeter from Xin Qiu,Ju Que,Gong Sun and Neiguan(non-menstrual non-acupuncture points)once every other day for 4 weeks.Baseline data were analyzed and compared with the Seattle Angina Scale scores,angina pectoris efficacy,TCM symptom score,number of 24h ischemic episodes,total ischemic time,longest ischemic time,ECG efficacy,changes in left ventricular diameter based on echocardiogram and EF values,inflammatory indexes Hs-CRP,TNF-α,IL-6,IL-8,endothelial damage related P-selectin,EMPs,endothelial particles,moesin antibodies,positive changes and negative conversion rates,and changes in HAMA anxiety scores.Experimental study:75 SD rats(200g-250g)during ad libitum consumption of food and water were used for this study.10 were randomly selected as the sham-operated group(control)and the remaining 65 were treated with ligation of the anterior descending branch for 45 min,followed by a removal of ligature.Rats were closely monitored after surgery.There were four types of treatments assigned randomly to the rats after a successful surgery:a model group(no effective treatment),an acupuncture group(acupuncture at Gong Sun,Neiguan,Ju Que,Xin Yu),an SPHK1 activator group,and an SPHK1 activator+acupuncture group.For the acupuncture group,an acupuncture treatment started on the 1st day after being assigned.A 20-minute-long acupuncture at Gong Sun,Neiguan,Ju Que and Xin Yu was given once every other day,resulting in a total of 14 acupuncture sessions in 4 weeks.The SPHK1 activator group received intravenous injection of SPHK1 activator(MHP).The SPHK1 activator+acupuncture group received both intravenous injection of SPHK1 activator(MHP)and the acupuncture treatments Next,the model group received nonmenstrual non-acupuncture subcutaneous sham acupuncture near Gong Sun,Neiguan,Ju Que and Xin Yu for 20 minutes once every other day for a total of 14 acupuncture sessions in 4 weeks.Finally,the sham-operated control group received non-transverse nonacupuncture subcutaneous sham needling near Gong Sun,Neiguan,Ju Que and Xin Yu for 20 minutes once every other day for a total of 14 needling sessions in 4 weeks.Rats were euthanized after the last intervention.The myocardial infarction area was measured by TTC.Morphological changes were measured by HE staining.Blood levels of creatine kinase(CK)and lactate dehydrogenase(LDL-C)were measured by immunoturbidimetric method using a fully automated biochemical analyzer.The expression levels of SPHK1 and TIMP-1 were measured by Western Blot.ResultsSystem assessment:A total of 13 articles were chosen,including 994 subjects.Results showed that acupuncture,moxibustion/acupuncture and moxibustion in conjunction with medicine for angina pectoris of the Qi-deficiency-and-blood-stasis type are more effective than the use of medicine alone,as manifested by a lowered TCM syndrome score.Clinical study:There were no statistical differences among three groups at baseline(P>0.05).suggesting a good quality baseline in all groups.Detailed comparisons among three groups are shown below.1.In physical activity(PL),angina stable state(AS),angina attack,treatment satisfaction(TS)and disease perception(DP)sections of the Seattle Angina Scale(Group A:PL 57.33±11.82、AS 87.62±8.23、AF 86.03± 7.58、TS 88.97±4.37、DP 78.10±9.94;the control group:PL 47.17 ± 12.95、AS 69.97 ± 14.62、AF 65.38 ± 12.90、TS 66.55 ± 8.18、DP 65.98 ± 9.30;GroupB:PL 48.42± 10.74、AS 71.37 ± 12.48、AF 65.11 ± 10.83、TS 66.21± 12.94、DP 66.03 ± 9.68),Group A outperformed both the control group and Group B(P<0.05).2.In the total effective rate of improving angina pectoris,Group A(94.87%)staticially significantly outperformed the control group(67.50%)and Group B(71.05%).3.In the total effective rate of TCM symptom scores,Group A(94.87%)statistically significantly outperformed the control group(80.0%)and Group B(84.21%).4.In inflammatory factors Hs-CRP,TNF-α,IL-6 and IL-8,Group A showed a statistically significant reduction compared to the control group and Group B(P<0.05),HsCRP(mg/ml)、TNF-α(pg/ml)、IL-6(pg/ml)and IL-8(pg/ml)were showed:Group A:2.60±0.28、42.31 ± 11.24、0.48±0.17、0.52±0.17,Group B:2.80±0.32、61.91 ± 15.62、0.75±0.18、0.76±0.19,Control group:2.76± 0.29、66.52± 19.33、0.79±0.20、0.77±0.21.5.In the comparison of endothelial function,acupuncture group A was able to reduce the levels of endothelial damage indicators E selectin and EMPs endothelial cell particles(E selectin and EMPs:Group A:26.99± 3.84、831.08± 65.84,Group B:31.35± 5.06、1134.94 ± 100.08,Control group:32.42± 4.10、1221.09±90.81),and was better than the control group and acupuncture group B.The difference was statistically significant(P<0.05).6.In the positive detection rate of moesin antibody,three groups had the same level at baseline.After treatments,Group A(61.54%)showed a lowered level compared to the control(77.50%)and Group B(73.68%)but this result did not reach statistical significance(P>0.05).7.In the HAMA scale score,Group A(7.85 ± 4.59)statistically significantly outperformed the control group(12.47± 5.45)and Group B(12.00 ± 6.11)(P<0.05).8.To further explore the individual effects of different treatment groups on each indicator in the Seattle scale,the baseline information of the study subjects were included together in a generalised linear model for regression analysis.Three models were developed for each outcome indicator.Model 1 represented a model without adjustments for baseline indicators.Model 2 represented a model with adjustments for two baseline indicators(age and gender).And Model 3 was a fully adjusted model with adjustments for all baseline indicators.The results of the regression analyses were as follows.(1)Seattle Angina Scale regression analysis.All three models suggested an independent effect(P<0.05)of Group A on five aspects of the Seattle Scale:the physical activity limitation(PL),angina stable state(AS),angina attack,treatment satisfaction(TS),and disease perception(DP),with stable effect values.In Model 3,using the control group as a reference,acupuncture increased a score by 9.51.17.19,20.08,23.00 and 11.72 points in the physical activity(PL),angina stable state(AS),angina attack,treatment satisfaction(TS)and disease perception(DP),respectively.(2)Regression analysis of angina pectoris efficacy and TCM efficacy.All three models suggested an independent effect of acupuncture on angina pectoris efficacy and TCM efficacy(P<0.05)with stable effect values.In model 3,using the control group as a reference,acupuncture reduced the likelihood of an ineffective angina pectoris outcome by 73%and reduced the likelihood of an ineffective TCM syndrome score outcome by 78%.(3)Regression analysis of inflammatory indexes.All three models suggested an independent effect of acupuncture on HS-CRP,TNF,IL-6 and IL-8(P<0.05)with stable effect values.In model 3,using the control group as a reference,acupuncture caused a decrease of 0.18 mg/ml in HS-CRP,23.86 pg/ml in TNF,0.33 pg/ml in IL-6 and 0.28 pg/ml in IL-8.(4)Regression analysis of endothelial injury indicators.All three models suggested an independent effect of acupuncture on both E selectin,and EMPs(P<0.05)with stable effect values.In model 3,using the control group as a reference,acupuncture decreased E selectin by 5.62 pg/ml and EMPs by 400.95/ul.(5)HAMA scale regression analysis.All three models suggested an independent effect of acupuncture on HAMA scale score(P<0.05)with stable effect values.In model 3,using the control group as a reference,acupuncture resulted in a 3.80-point decrease in the HAMA scale score,indicating that the intervention of the acupuncture alleviated the patients’anxiety.Experimental study:1.An ischemic infarct was present in a white and pale color as opposed to red or pink as healthy heart tissue using TTC staining.The presence of infarct in various sizes in all except for the sham-operated control suggested successful ligation-induced myocardial ischemia.The infarction size of the heart tissue of the acupuncture group was smaller than the model group.The infarction size of the SPHK1 activator group was greater than both the model group,and the SPHK1 activator+acupuncture group.2.In the HE sections,under light microscopic,local necrosis was shown in the heart tissue of the model group,with pathologies such as myocardial fiber breakage,atrophy,necrosis and disorganization,hemorrhage and inflammatory cell infiltration.The morphological results suggested that the MIRI method of coronary ligation and perfusion caused necrosis to the myocardium and that the MIRI modeling was successful.While no pathological changes were seen in the control group,the SPHK1 activator group had the heaviest relative degree of cardiac histopathological changes,followed by the model group,the SPHK1 activator+acupuncture group with relatively mild cardiac histopathological changes,and the acupuncture group with the least relative degree of cardiac histopathological changes.3.To assess myocardial injury,the blood levels of CK-MB(U/L),cTnI(ng/ml),CK(U/L)and LDH(U/L)were showed:acupuncture group:26.37±0.92、5.24 ± 0.19、626.83 ±6.20、760.10 ± 1.95,control group:23.78 ± 0.96、4.87 ± 0.20、354.60± 8.34、509.83± 6.71,model group:31.05 ± 0.37、5.95 ± 0.07、779.40 ± 2.99、898.57 ± 2.92,SPHK1 activator group33.39±0.19、6.27 ± 0.16、999.57 ± 31.81、910.23±1.21,SPHK1 activator+acupuncture group:28.64 ± 0.44、5.57 ± 0.32、659.83± 3.41、870.53 ± 5.02. the levels of CK-MB,cTnI,CK and LDH increased in the model group compared with the control group(P<0.01),while the acupuncture group showed a statistically significant reduction in the levels of CK-MB,cTnl,CK and LDH compared with the model group(P<0.01,P<0.05,P<0.01,P<0.01).The activator group showed increased levels of CK-MB,cTnI,CK,LDH compared to the model group(P<0.01,P>0.05,P<0.01,P<0.05),and the SPHK1 activator+acupuncture group showed an increased CK-MB,cTnI,CK,LDH levels compared to the model group(P<0.01,P>0.05,P<0.01,P<0.05),while the SPHK1 activator+acupuncture group showed an decreased levels of CK-MB,cTnI,CK,LDH compared with the SPHK1 activator group(P<0.01,P<0.05,P<0.01,P<0.01).4.To assess inflammation improvement,the levels of inflammatory factors TNF-α,IL6 and IL-1β in the blood of the rats(acupuncture group:54.23±1.80、12.05 ± 0.59、57.97± 0.82,control group:47.85 ± 2.13、10.90±0.49、51.45 ± 1.68,model group:61.50 ±2.06、14.35 ± 0.37、59.64 ± 0.93,SPHK1 activator group:65.85±3.06、16.26 ± 0.58、65.93 ± 0.54,SPHK1 activator+acupuncture group:57.10 ± 1.84、13.65 ± 0.17、55.80 ±1.22)in the ischemia-reperfusion model group were increased compared with those in the control group(P<0.01),suggesting that ischemia-reperfusion caused an inflammatory response.The acupuncture group showed a reduced levels of TNF-α and IL-6 and a marginal level of reduction in IL-1β(P<0.05,P<0.01.P>0.05),suggesting that acupuncture reduced inflammation.The SPHK1 activator group showed increased IL-6 and IL-1β levels and a marginal increase in TNF-α level compared to the model group(P<0.01,P<0.01,P>0.05).,The SPHK1 activator group showed increased TNF-α and IL-6,levels and a marginal increase in IL-1 β(P>0.05,P<0.01,P<0.01)compared to the model group,while the SPHK1 activator+acupuncture group showed decreased TNF-α,IL-6,IL-1β levels compared with the SPHK1 activator group(P<0.01).5.To speculate the SPHK1/TIMP1 signaling pathway,we found that the expression of SPHK1 protein and TIMP1 protein in myocardial tissues of rats(acupuncture group:1.23 ±0.05、1.08 ± 0.10、9.70 ± 0.60,control group:1.00 ± 0.09、1.00 ± 0.13、7.54 ± 1.17,model group:1.56± 0.04、1.61± 0.08、12.21±0.77,SPHK1 activator group:1.77±0.04、1.90±0.02、13.42± 0.63,SPHK1 activator+acupuncture group:1.31±0.08、1.38± 0.05、10.98 ± 0.54)with ischemia/reperfusion was increased(P<0.01),and the level of SPHK1 in blood was increased(P<0.01),suggesting that the SPHK1/TIMP1 signaling pathway was involved in the process of ischemia/reperfusion and was activated in this process.while the acupuncture The SPHK1 activator group showed decreased expression of SPHK1 protein and TIMP1 protein compared to the model group(P<0.01,P<0.05),and increased blood levels of SPHK1(P<0.01,P<0.05),suggesting that acupuncture has an inhibitory effect on SPHK1/TIMP1 signaling.and the trend of elevated blood SPHK1 levels(P>0.05),while the SPHK1 activator+acupuncture group was able to reduce the expression levels of SPHK1 protein and TIMP1 protein(P<0.01)and the trend of blood SPHK1 levels(P>0.05)compared with the SPHK1 activator group,suggesting that the SPHK1/TIMP1 signaling pathway The SPHK1/TIMP1 signaling pathway exacerbated the process of ischemiareperfusion injury,while acupuncture could inhibit SPHK1/TIMP1 signaling and thus attenuate the effect of ischemia-reperfusion injury.6.The percentage of apoptotic cells in the heart of rats in the model group was statistically significantly increased compared with the control group(P<0.01).The percentage of apoptotic cells in the heart of rats in the SPHK1 activator group was decreased compared with the model group,but not statistically significant(P>0.05).The percentage of apoptotic cells in the heart of rats in the acupuncture group was statistically significantly decreased compared with the model group(P<0.01).The percentage of apoptotic cells in the hearts of rats in the SPHK1 activator+acupuncture group decreased,with extremely significant statistical significance(P<0.01);compared with the SPHK1 activator group,the percentage of apoptotic cells in the hearts of rats in the SPHK1 activator+acupuncture group decreased,with extremely significant statistical significance(P<0.01);compared with the SPHK1 activator group,the percentage of apoptotic cells in the hearts of rats in the SPHK1 activator+acupuncture group decreased,with extremely The percentage of apoptotic cells in the heart of the SPHK1 activator+acupuncture group was reduced compared with that of the SPHK1 activator group,which was statistically significant(P<0.01).7.In the assay of Bax,Bcl-2 and Caspase-3 content,compared with the control group,the Bax content was increased,the Bcl-2 content was decreased and the Caspase-3 content was increased in the hearts of rats in the model group,with extremely significant statistical significance(P<0.01);compared with the model group,the Bax content was decreased,the Bcl-2 content was increased and the Caspase-3 content was increased in the hearts of rats in the acupuncture group,with extremely significant statistical significance(P<0.01).Bax content was decreased,Bcl-2 content was increased and Caspase-3 content was decreased in the hearts of rats in the SPHK1 activator+acupuncture group,with extremely significant statistical significance(P<0.01);compared with the SPHK1 activator group,Bax content in the hearts of rats in the SPHK1 activator+acupuncture group was Compared with the SPHK1 activator group,the Bax content in the hearts of rats in the SPHK1 activator+acupuncture group decreased,and the Bcl-2 content increased and the Caspase-3 content decreased,all of which were statistically significant(P<0.05).ConclusionThe addition of acupuncture to conventional treatments can improve the symptoms of angina pectoris in patients with coronary heart disease and reduce the TCM syndrome score.However,due to limited literature,small sample sized studies,and inconsistent intervention time among studies,future research is needed to establish the best treatment strategy with acupuncture for patients with angina pectoris.Results suggested that acupuncture at Xin Yu,Ju Que,Gongsun and Neiguan improved scores of the Seattle Angina Scale,symptoms of angina pectoris and the scores of TCM syndromes as well as myocardial ischemia,HAMA anxiety score,inflammation,and vascular endothelial functions.Acupuncture at Gong Sun,Neiguan,Ju Que and Xin Yu reduced local necrosis,myocardial fiber breakage,atrophy and disorder of arrangement.It reduced pathological changes such as myocardial cell hemorrhage,inflammatory cell infiltration,myocardial infarction,the levels of CK-MB,cTnI,CK,LDH enzymatic and injury markers in MIRI rats.It also reduced the levels of TNF-α,IL-6,IL-1βinflammatory factors in AMI rats.The mechanism of acupuncture-induced effects may be related to the inhibition of SPHK1/TIMP1 signaling pathway to exert its anti-inflammatory,anti-myocardial necrosis and apoptosis inhibitory and protective effects. |
PDF Full Text Request