摘要:目的:观察针刺对卒中后痉挛大鼠海马区Na+/K+/K+-ATP酶、谷氨酸转运体(EAATs)和谷氨酸(Glu)的影响,探讨针刺抗痉挛的中枢机制。方法:SD大鼠随机分为假手术组、模型组、“阳陵泉”组和“阳陵泉”+“百会”组,每组12只。改良线栓法制作大鼠局灶性脑缺血模型。“阳陵泉”组恢刺患肢“阳陵泉”,“阳陵泉”+“百会”组恢刺患肢“阳陵泉”再平刺“百会”,均留针30 min,每日治疗1次,连续7 d。记录各组大鼠神经功能损伤(Zea Longa)评分和痉挛状态(MAS)评分,采用ELISA法检测缺血侧海马区Glu、EAAT1(GLAST)和EAAT2(GLT-1)含量,Western blot法检测缺血侧海马区Na+-ATP酶、谷氨酸转运体(EAATs)和谷氨酸(Glu)的影响,探讨针刺抗痉挛的中枢机制。方法:SD大鼠随机分为假手术组、模型组、“阳陵泉”组和“阳陵泉”+“百会”组,每组12只。改良线栓法制作大鼠局灶性脑缺血模型。“阳陵泉”组恢刺患肢“阳陵泉”,“阳陵泉”+“百会”组恢刺患肢“阳陵泉”再平刺“百会”,均留针30 min,每日治疗1次,连续7 d。记录各组大鼠神经功能损伤(Zea Longa)评分和痉挛状态(MAS)评分,采用ELISA法检测缺血侧海马区Glu、EAAT1(GLAST)和EAAT2(GLT-1)含量,Western blot法检测缺血侧海马区Na+/K+/K+-ATP酶α1(ATP1α1)的蛋白表达,实时荧光定量PCR法检测缺血侧海马区ATP1α1 mRNA表达,免疫荧光法检测缺血侧海马区GLAST、GLT-1、ATP1α1的表达。结果:与假手术组比较,模型组Zea Longa与MAS评分均升高(P<0.01);与模型组比较,两针刺组Zea Longa与MAS评分均降低(P<0.01);且“阳陵泉”+“百会”组低于“阳陵泉”组(P<0.05)。与假手术组比较,模型组缺血侧海马区Glu含量升高(P<0.01),GLAST、GLT-1含量及蛋白表达,ATP1α1蛋白和mRNA表达均降低(P<0.01);与模型组比较,两针刺组缺血侧海马区Glu含量降低(P<0.05,P<0.01),GLAST、GLT-1含量及蛋白表达,ATP1α1蛋白和mRNA表达均升高(P<0.05,P<0.01);与“阳陵泉”组比较,“阳陵泉”+“百会”组缺血侧海马区Glu含量降低(P<0.05),GLAST、GLT-1含量及蛋白表达,ATP1α1蛋白和mRNA表达均升高(P<0.05,P<0.01)。结论:针刺抗卒中后痉挛的效应可能与上调海马Na+-ATP酶α1(ATP1α1)的蛋白表达,实时荧光定量PCR法检测缺血侧海马区ATP1α1 mRNA表达,免疫荧光法检测缺血侧海马区GLAST、GLT-1、ATP1α1的表达。结果:与假手术组比较,模型组Zea Longa与MAS评分均升高(P<0.01);与模型组比较,两针刺组Zea Longa与MAS评分均降低(P<0.01);且“阳陵泉”+“百会”组低于“阳陵泉”组(P<0.05)。与假手术组比较,模型组缺血侧海马区Glu含量升高(P<0.01),GLAST、GLT-1含量及蛋白表达,ATP1α1蛋白和mRNA表达均降低(P<0.01);与模型组比较,两针刺组缺血侧海马区Glu含量降低(P<0.05,P<0.01),GLAST、GLT-1含量及蛋白表达,ATP1α1蛋白和mRNA表达均升高(P<0.05,P<0.01);与“阳陵泉”组比较,“阳陵泉”+“百会”组缺血侧海马区Glu含量降低(P<0.05),GLAST、GLT-1含量及蛋白表达,ATP1α1蛋白和mRNA表达均升高(P<0.05,P<0.01)。结论:针刺抗卒中后痉挛的效应可能与上调海马Na+/K+/K+-ATP酶和EAATs的表达有关,恢刺“阳陵泉”加平刺“百会”的抗痉挛效应优于单纯恢刺“阳陵泉”穴。Objective To observe the effect of acupuncture at “Yanglingquan”(GB34) and “Baihui”(GV20) on Na+-ATP酶和EAATs的表达有关,恢刺“阳陵泉”加平刺“百会”的抗痉挛效应优于单纯恢刺“阳陵泉”穴。
摘要:目的:观察温和灸对老年大鼠血管的抗老化作用,探讨温和灸通过调控单核细胞趋化蛋白1(MCP-1)/基质金属蛋白酶2(MMP-2)/转化生长因子β1(TGF-β1)局部促炎信号环对抗血管老化的机制。方法:50只8月龄雄性SD大鼠腹腔注射D-半乳糖300 mg·kg(-1)·d(-1)·d(-1),连续4周,制备衰老大鼠模型。24只造模成功的大鼠随机分为老年对照组、药物组和温和灸组,2月龄青年大鼠为青年对照组,每组8只。温和灸组选择“关元”及双侧“肾俞”进行温和灸治疗,1次/d, 20 min/次,治疗5 d休息2 d,持续治疗8周;药物组予丙酸睾酮注射液[7 mg·kg(-1),连续4周,制备衰老大鼠模型。24只造模成功的大鼠随机分为老年对照组、药物组和温和灸组,2月龄青年大鼠为青年对照组,每组8只。温和灸组选择“关元”及双侧“肾俞”进行温和灸治疗,1次/d, 20 min/次,治疗5 d休息2 d,持续治疗8周;药物组予丙酸睾酮注射液[7 mg·kg(-1)·(3 d)(-1)·(3 d)(-1)]腹腔注射,连续8周;老年对照组、青年对照组予0.9%氯化钠溶液腹腔注射,剂量与疗程同药物组。观察各组大鼠治疗前后游泳力竭时间;HE和Masson染色法观察各组大鼠胸主动脉形态和胶原纤维含量的变化;酶联免疫吸附测定法检测各组大鼠血清总睾酮(TT)、游离睾酮(FT)、血管紧张素2(AngⅡ)含量;免疫组织化学法检测大鼠胸主动脉中MCP-1阳性表达水平;Wes-tern blot法检测大鼠胸主动脉中MMP-2、TGF-β1、MCP-1蛋白表达量。结果:与青年对照组比较,老年对照组大鼠游泳力竭时间明显缩短(P<0.01),血清TT、FT含量明显降低(P<0.01),血清AngⅡ含量和胸主动脉胶原纤维含量、MCP-1阳性表达及MMP-2、TGF-β1、MCP-1的蛋白表达量明显升高(P<0.01)。治疗后与老年对照组比较,药物组、温和灸组游泳力竭时间明显延长(P<0.01),血清TT、FT含量明显升高(P<0.01),血清AngⅡ含量、胸主动脉胶原纤维含量、MCP-1阳性表达及MMP-2、TGF-β1、MCP-1的蛋白表达量明显降低(P<0.01,P<0.05)。与药物组比较,温和灸组血清AngⅡ含量、胸主动脉胶原纤维含量、MCP-1阳性表达均降低(P<0.05)。青年对照组大鼠主动脉内膜较薄,平滑肌细胞排列整齐;老年对照组内膜明显增厚,且平滑肌细胞排列紊乱;与老年对照组比较,药物组和温和灸组大鼠血管平滑肌细胞排列较整齐、规则,管壁变薄。结论:温和灸通过降低MCP-1/MMP-2/TGF-β1局部促炎信号环中炎性因子表达,对血管衰老起到干预作用。Objective To observe the effect of mild moxibustion on monocyte chemotaxis protein 1(MCP-1)/matrix metalloproteinase 2(MMP-2)/transforming growth factor β1(TGF-β1) pro-inflammatory signal loop in senile rats, so as to explore its mechanisms underlying improving vascular aging(VA). Methods Twenty-four male VA SD rats were randomized into senium(VA) control, medication and moxibustion groups, and other 8 young SD rats(aged 2 months) were used as the young control group. The VA model was established by intraperitoneal injection of D-galactose(300 mg·kg(-1)]腹腔注射,连续8周;老年对照组、青年对照组予0.9%氯化钠溶液腹腔注射,剂量与疗程同药物组。观察各组大鼠治疗前后游泳力竭时间;HE和Masson染色法观察各组大鼠胸主动脉形态和胶原纤维含量的变化;酶联免疫吸附测定法检测各组大鼠血清总睾酮(TT)、游离睾酮(FT)、血管紧张素2(AngⅡ)含量;免疫组织化学法检测大鼠胸主动脉中MCP-1阳性表达水平;Wes-tern blot法检测大鼠胸主动脉中MMP-2、TGF-β1、MCP-1蛋白表达量。结果:与青年对照组比较,老年对照组大鼠游泳力竭时间明显缩短(P<0.01),血清TT、FT含量明显降低(P<0.01),血清AngⅡ含量和胸主动脉胶原纤维含量、MCP-1阳性表达及MMP-2、TGF-β1、MCP-1的蛋白表达量明显升高(P<0.01)。治疗后与老年对照组比较,药物组、温和灸组游泳力竭时间明显延长(P<0.01),血清TT、FT含量明显升高(P<0.01),血清AngⅡ含量、胸主动脉胶原纤维含量、MCP-1阳性表达及MMP-2、TGF-β1、MCP-1的蛋白表达量明显降低(P<0.01,P<0.05)。与药物组比较,温和灸组血清AngⅡ含量、胸主动脉胶原纤维含量、MCP-1阳性表达均降低(P<0.05)。青年对照组大鼠主动脉内膜较薄,平滑肌细胞排列整齐;老年对照组内膜明显增厚,且平滑肌细胞排列紊乱;与老年对照组比较,药物组和温和灸组大鼠血管平滑肌细胞排列较整齐、规则,管壁变薄。结论:温和灸通过降低MCP-1/MMP-2/TGF-β1局部促炎信号环中炎性因子表达,对血管衰老起到干预作用。
摘要:目的:评价针刺联合艾灸治疗过敏性鼻炎(AR)的临床疗效及安全性。方法:选取AR患者80例,采用随机数字表法分为药物组与针刺联合艾灸组,每组40例。药物组给予氯雷他定片口服,每日1片,10 d为1个疗程,治疗3个疗程;针刺联合艾灸组以迎香(双)、印堂、合谷(双)、肾俞(双)为主穴进行针刺和艾灸,并随证加减穴位,每日治疗1次,每次30 min, 10次为1个疗程,治疗3个疗程。治疗前后分别评定临床症状评分、临床体征评分及鼻-结膜相关生活量表评分;刮取部分下鼻甲黏膜组织表面的分泌物,采用HE染色法及Sheldeny评价法进行嗜酸性粒细胞(EOS)分布程度评分;采用酶联免疫吸附法测定血清免疫球蛋白E(IgE)、维甲酸孤儿受体γt(RORγt)、叉头盒蛋白P3(Foxp3)、白细胞介素-17(IL-17)、IL-27、IL-33含量。评价两组AR患者的临床疗效,并记录治疗期间出现的所有不良反应。结果:两组患者临床症状评分、临床体征评分及鼻-结膜相关生活量表评分、EOS分布评分及血清IgE、RORγt、IL-17、IL-33含量均较本组治疗前降低(P<0.05),血清Foxp3、IL-27含量较本组治疗前升高(P<0.05)。治疗后针刺联合艾灸组临床症状评分、临床体征评分及鼻-结膜相关生活量表评分及血清IgE、RORγt、IL-33含量低于药物组(P<0.05),血清Foxp3、IL-27含量高于药物组(P<0.05)。针刺联合艾灸组总有效率为100.0%(40/40),显著高于药物组的82.5%(33/40,P<0.05)。治疗期间及治疗后,两组患者均未出现明显不良反应。结论:针刺联合艾灸治疗AR疗效显著,安全性好,其治疗机制可能与调控幼稚CD4+T细胞介导的辅助性T细胞(Th)1/Th2、Th17/调节性T细胞平衡有关。Objective To evaluate the clinical efficacy and safety of acupuncture combined with moxibustion on allergic rhinitis. Methods Using the random number table, 80 patients with allergic rhinitis were divided into a medication group and an acupuncture combined with moxibustion(acu-mox) group, 40 cases in each one. In the medication group, ioratadine tables were prescribed for oral administration, one tablet daily for 10 days as 1 session, 3 sessions of treatment were required. In the acupuncture combined with moxibustion group, bilateral Yingxiang(LI20), Yintang(EX-HN3), bilateral Hegu(LI4) and bilateral Shenshu(BL23) were selected as the main points and stimulated with acupuncture and moxibustion; and the acupoint prescription was modified according to symptoms. This combined treatment was given once every day, stimulating for 30 min each time, and 10 treatments made 1 course, for 3 courses of treatment totally. Before and after treatment, the scores for symptoms and physical signs, as well as the score of rhino-conjunctivitis related quality of life scale(R-QOL) were evaluated separately. The sample of the inferior turbinate mucosa tissue was collected and the distribution of eosinophil(EOS) was scored using HE staining and Sheldeny evaluation. Using enzyme-linked immunosorbent assay(ELISA), the contents of serum immunoglobulin E(IgE), retinoic-acid-receptor-related orphan nuclear receptor γt(RORγt), forkhead box protein P3(Foxp3), interleukin-17(IL-17), IL-27 and IL-33 were determined. The clinical efficacy was evaluated in the patients with allergic rhinitis of two groups and all the adverse reactions were recorded during treatment. Results The scores of symptoms and physical signs as well as the score of R-QOL, and EOS distribution score and the contents of serum IgE, RORγt, IL-17 and IL-33 were all reduced as compared with those before treatment in each group(P<0.05), and the contents of serum Foxp3 and IL-27 were increased as compared with those before treatment in each group(P<0.05). After treatment, the scores of symptoms and physical signs as well as the score of R-QOL, and the contents of serum IgE, RORγt and IL-33 in the acu-mox group were lower than those in the medication group(P<0.05), and the contents of serum Foxp3 and IL-27 were higher than those of the medication group(P<0.05). The total effective rate of the acu-mox group was 100.0%(40/40), significantly higher than 82.5%(33/40) in the medication group(P<0.05). No ob-vious adverse reaction was found in either group during and after treatment. Conclusion Acupuncture combined with moxibustion is significantly effective and safe in treatment of allergic rhinitis. Its effect mechanism may be related to the balance modulation of Th1/Th2 and Th17/Treg cells mediated by naive CD4+T细胞介导的辅助性T细胞(Th)1/Th2、Th17/调节性T细胞平衡有关。