隔药灸对肿瘤坏死因子-α介导克罗恩病肠上皮细胞凋亡途径的影响

目的:观察隔药灸对肿瘤坏死因子-α(TNF-α)-肿瘤坏死因子受体相关死亡结构域(TRADD)-Fas相关的死亡结构域(FADD)途径介导克罗恩病(Crohn's disease,CD)肠上皮细胞凋亡的影响,探讨其作用机制。方法:将48只SD雄性大鼠随机分成正常组、模型组、隔药灸组和西药组,每组12只。采用2,4,6-三硝基苯磺酸(TNBS)制备CD模型,造模成功后隔药灸组采用隔药饼灸"天枢""气海"穴,每日1次,每次每穴隔药灸2壮,共灸10d。西药组采用美沙拉嗪灌胃,每日2次,每次2mL,共灌胃10d。治疗结束后取各组大鼠结肠上皮组织,分离、纯化和培养结肠上皮细胞建立体外肠上皮细胞屏障模型,将100ng/mL TNF-α分别与各组肠上皮细胞孵育培养24h,应用荧光黄透过率检测各组肠上皮细胞屏障通透性,Western blot法检测各组肠上皮细胞凋亡途径相关蛋白肿瘤坏死因子受体1(TNFR1)、TRADD、受体作用蛋白1(RIP1)、FADD、锌指蛋白A20(A20)表达以及流式细胞术观察各组肠上皮细胞凋亡情况。结果:与正常组比较,模型组的荧光黄透过率显著升高(P0.001),结肠上皮细胞TNFR1、TRADD、RIP1表达量明显升高(P0.01),A20表达量明显降低(P0.01),细胞凋亡率显著增高(P0.001)。与模型组比较,隔药灸组和西药组的荧光黄透过率显著降低(P0.001),结肠上皮细胞TRADD、RIP1、FADD表达量明显降低(P0.01),A20表达量明显升高(P0.01),结肠上皮细胞凋亡率显著降低(P0.001)。结论:隔药灸可能是通过调控TNF-α介导CD肠上皮细胞凋亡途径的异常,达到保护或修复CD肠上皮屏障损伤之效应。     

隔药灸对溃疡性结肠炎大鼠结肠NF-κB 和STAT3 活性的免疫调节机制

目的：观察隔药灸改善UC大鼠结肠炎症的效应,通过NF-κB通路和STAT3磷酸化参与免疫调节的作用机制。方法：将大鼠随机分成正常组、UC组和隔药灸组,每组8只。采用4% DSS诱导UC的大鼠模型,造模成功后采用隔药灸大鼠双侧天枢穴,每穴灸2壮艾炷,每天1次,共7次。通过比较组织病理学、血清学促炎症因子的蛋白浓度观察隔药灸干预UC大鼠的效应。采用Western Blot检测结肠组织NF-κB通路及STAT3活性探讨隔药灸改善UC大鼠结肠炎症反应的潜在机制。结果：UC组大鼠结肠组织病理学提示黏膜上皮表面溃疡形成,炎症反应明显,隔药灸能修复结肠上皮损伤,减轻UC大鼠结肠组织的炎症反应。与正常组比较,UC组大鼠血清IL-1β和IL-6蛋白浓度显著升高（P < 0.05,P < 0.01）,p-STAT3磷酸化水平显著增加,pIκB-α和NF-κB p65蛋白表达均显著增加,IκB-α蛋白表达显著降低;与UC组比较,隔药灸组大鼠血清IL-1β和IL-6蛋白浓度显著降低（P < 0.05）,结肠组织NF-κB p65蛋白表达和pIκB-α磷酸化水平均降低,IκB-α蛋白表达增加。结论：隔药灸可能通过抑制NF-κB通路和STAT3磷酸化,减少了促炎症因子IL-1β和IL-6的释放,从而减轻了UC大鼠的结肠炎症反应。     

Influence of moxa smoke on mitochondrial transmembrane potential and Bax/Bcl-2 in alveolar type II epithelial A549 cells

Objective

To investigate the influence of moxibustion products on mitochondrial transmembrane potential (MTP) and mRNA expression of Bax/Bcl-2 in alveolar type II epithelial A549 cells, and to further explore influence of moxibustion products on the oxidative damage of A549 cells.

Methods

Smoke and particles generated by moxibustion were collected using the filter box for gas sampling. The moxa smoke extract (MSE) was diluted sequentially to the final concentrations of 0.05 mg/mL, 0.1 mg/mL, 0.2 mg/mL, 0.3 mg/mL and 0.4 mg/mL using the cell culture medium, and A549 cells were then intervened by the above MSE solution. Cell MTP was detected by JC-1 staining. Fluorescence quantitative polymerase chain reaction (PCR) was used to detect Bax/Bcl-2 mRNA expression of A549 cells.

Results

Compared with cells in the normal control group, MTP was significantly decreased in cells of 0.3 mg/mL and 0.4 mg/mL MSE intervention groups (P<0.01); while MTP showed no significant changes in cells of 0.05 mg/mL, 0.1 mg/mL and 0.2 mg/mL MSE intervention groups (P>0.05); compared with cells in 0.05 mg/mL MSE intervention group, MTP was decreased significantly in cells of 0.1 mg/mL, 0.2 mg/mL, 0.3 mg/mL and 0.4 mg/mL MSE intervention groups (P<0.05 ); compared with cells in 0.1 mg/mL MSE intervention group, MTP was decreased significantly in cells of 0.4 mg/mL MSE intervention group (P<0.01). Bax mRNA expression of cells in each concentration of MSE intervention group all showed no significant difference compared to that in the normal control group; Bcl-2 mRNA expression of cells was reduced with the increase of MSE intervention concentration. Wherein, Bcl-2 mRNA expressions of cells in 0.4 mg/mL and 0.3 mg/mL MSE intervention groups were significantly reduced compared with that of cells in the normal control group (P<0.05); Bcl-2 mRNA expression of cells in 0.4 mg/mL MSE intervention group was significantly reduced compared to that in 0.05 mg/mL MSE intervention group (P<0.05).

Conclusion

Certain higher concentration of moxa smoke could reduce MTP and mRNA expression of the anti-apoptosis gene Bcl-2 in alveolar type II epithelial A549 cells. Oxidative damage may be the important mechanism of apoptosis caused by the high concentration of moxa smoke solution, and further studies are necessary on the specific mechanisms.

     

Comparative study of electroacupuncture and moxibustion in influencing Tianshu (ST 25) regions mast cells in visceral hyperalgesia rats

Objective

To evaluate and compare electroacupunctures (EA) with different parameters and moxibustion at different temperatures influencing the activation of mast cells (MC) in Tianshu (ST 25) regions of visceral hyperalgesia model rats.

Methods

Rats (except for model group) respectively accepted 1 mA or 3 mA EA or moxibustion at 43 °C or 4 °C to stimulate Tianshu (ST 25) points after randomization of the fifty visceral hyperalgesia model rats, and then were compared with that in model and normal groups. Number, degranulation numbers, degranulation rates in Tianshu (ST 25) regions MC of rats in each group were observed using toluidine blue staining. Abdominal withdrawl reflex (AWR) score was used to evaluate the rat visceral hyperalgesia reactions.

Results

Compared with the normal group and the model group, MC numbers (P<0.05, P<0.01, P<0.01, P<0.01), degranulation numbers and degranulation rates (P<0.01, P<0.01, P<0.05, P<0.01) of Tianshu (ST 25) MC in regions tissues in 43 °C and 4 °C moxibustion groups, and 1 mA and 3 mA EA groups all increased significantly. Compared with the model group, AWR scores were significantly lower in 43 °C and 4 °C moxibustion groups, and 1 mA and 3 mA EA groups under the stimulation of 20 mmHg, 40 mmHg, 0 mmHg or 80 mmHg colorectal distension (CRD) (P<0.05 in 1 mA and 3 mA EA groups under the stimulation of 20 mmHg, P<0.01 in the other groups). AWR scores in 43 °C and 4 °C oxibustion groups under the stimulation of 20 mmHg, 40 mmHg, 0 mmHg or 80 mmHg CRD were not significantly different from those in the normal group (all P>0.05); AWR scores in 1 mA EA group under the stimulation of 0 mmHg or 80 mmHg were significantly higher than that in the normal group (P<0.01); AWR score in 3 mA EA group under the stimulation of 0 mmHg was significantly higher than that in the normal group (P<0.01), and AWR scores in 3 mA EA group under the stimulation of 20 mmHg or 80 mmHg were also higher than that in the normal group (P<0.05). AWR scores were higher in 1 mA EA group under the stimulation of 40 mmHg or 80 mmHg than that in 4 °C moxibustion group (P<0.05); AWR score was higher in 3 mA EA group under the stimulation of 40 mmHg than that in 4 °C moxibustion group (P<0.05).

Conclusion

There are differences among EA of different parameters and moxibustion of different temperatures in activating on Tianshu (ST 25) regions MC of visceral hyperalgesia model rats, as well as in improving the visceral hyperalgesia reaction. The effect of 4 °C moxibustion is the most significant.

     

灸法对肠易激综合征慢性内脏痛镇痛效应的研究与展望

背景：慢性内脏痛是肠易激综合征患者最主要的临床表现之一,患者的生活质量受到严重影响。近年研究表明,灸法对肠易激综合征患者慢性内脏痛的治疗取得了一定的进展。 目的：全面了解灸法对肠易激综合征患者慢性内脏痛镇痛效应的临床疗效,并对今后灸法镇痛在此领域的应用与研究发展趋势进行思考和展望,以期为今后灸法的进一步推广和应用提供一定的理论依据。 方法：以“灸法,肠易激综合征患者,内脏痛,腹痛”为中文检索词,以“moxibustion,irritable bowel syndrome,visceral pain,abdominal pain”为英文检索词,应用计算机在中国知网、维普、万方和PubMed数据库检索1990年1月至2014年10月的临床研究文献,并进行筛选、归纳与总结。 结果与结论：灸法对肠易激综合征腹泻型和便秘型患者慢性腹痛症状均有较好的镇痛效应。灸法对肠易激综合征慢性内脏痛的镇痛效应的可能是通过调节其内脏高敏感、胃肠动力障碍、脑-肠轴和神经-内分泌系统失调、免疫功能紊乱和肠道低度炎症、精神心理异常以及肠道菌群失调等产生。今后还需要更严格的随机对照试验设计同时借助现代科学定量评定内脏痛的先进设备和技术,进一步验证灸法镇痛的临床疗效并深入探讨其作用原理。 中国组织工程研究杂志出版内容重点：组织构建;骨细胞;软骨细胞;细胞培养;成纤维细胞;血管内皮细胞;骨质疏松;组织工程全文链接：     

自噬在炎症性肠病中的研究概况及展望

自噬是广泛存在于真核细胞中的生命现象,是生物在其发育、老化过程中普遍存在的净化自身多余或受损细胞器的共同机制。自噬能够在适应性免疫应答中从不同的水平维持机体对自身组织的免疫耐受,对机体起保护作用。目前,自噬在炎症性肠病发病机制中的作用越来越受到关注,自噬在肠道免疫和肠黏膜屏障方面发挥重要作用。因此,文章对近年来自噬在炎症性肠病中的调控作用研究作一综述,为炎症性肠病的发病机制研究提供参考,并试图从中医药的角度探讨自噬的作用及目前研究存在的问题。     

针药结合治疗肥胖型多囊卵巢综合征患者33例临床观察

目的 观察电针结合天癸胶囊治疗肥胖型多囊卵巢综合征(PCOS)的临床疗效.方法 将67例PCOS患者随机分为针药结合组33例和单纯电针组34例.单纯电针组取天枢、中脘、气海、三阴交、膈俞、次鼹等为主穴,采用电针治疗,每周治疗3次,1个月为1个疗程,共3个疗程后观察效果;针药结合组在此基础上同时口服天癸胶囊,每日2次,每次1.8g,连服3个月.比较两组治疗前后促卵泡刺激素(FSH)、促黄体生成素(LH)、睾酮(T)、空腹血糖(FPG)、空腹胰岛素(FINS)和胰岛素敏感指数( ISI)的变化,并观察临床疗效及3个月随访复发率.结果 针药结合组临床疗效总有效率93.9％,单纯电针组总有效率67.6％,针药结合组疗效优于单纯电针组(P＜0.05);针药结合组LH、LH/FSH、T值的降低明显优于单纯电针组(P＜0.01);针药结合组降低FINS及ISI水平也明显优于单纯电针组(P＜0.01);两组治疗后3个月随访,针药结合组复发率少于单纯电针组(P＜0.05). 结论 针药结合治疗肥胖型PCOS临床疗效肯定,可能与降低患者性激素及胰岛素水平有关.     

Observation on the efficacy of moxibustion for chronic gastritis and a clinical study of moxibustion’s effects on serum brain-gut peptides

Objective

To investigate the efficacy and mechanisms of moxibustion-based treatment of chronic gastritis (CG), and to provide an objective basis for treating CG using moxibustion.

Methods

A total of 61 CG patients were divided into an herbal cake-partitioned moxibustion group and a mild-warm moxibustion group. In both treatment groups, bilateral Tianshu (ST 25), Zhongwan (CV 12) and Qihai (CV 6) were selected for moxibustion. Before and after treatment, all the enrolled patients’ gastrointestinal disease-related traditional Chinese medicine (TCM) syndrome scores and visual analog scale (VAS) scores were measured, and the changes in the serum levels of the brain-gut peptides ghrelin, somatostatin (SS) and motilin (MTL) were observed.

Results

There was no statistically significant difference between the two groups in the clinical efficacy rate (P>0.05). After treatment, the gastrointestinal disease-related TCM syndrome scores and VAS scores were reduced to varying extents in both groups, the intra-group differences were statistically significant (all P<0.01). In both groups, the serum levels of ghrelin and MTL increased and the serum levels of SS decreased after treatment (all P<0.01). And there were no serious adverse events occurred.

Conclusion

Both herbal cake-partitioned moxibustion and mild-warm moxibustion are effective for CG; these two therapies exhibited similar therapeutic efficacy of epigastric discomfort or pain. And both the two therapies act to anti-inflammation, promote the recovery of gastric mucosa and improve the gastric motility, which is possibly their crucial action mechanism in treating CD.

     

艾灸生成物对大鼠肺形态与脏器系数的影响

目的：观察不同浓度艾灸生成物对SD大鼠肺的影响。方法40只大鼠随机分正常对照组、低剂量艾灸生成物组、中剂量艾灸生成物组、高剂量艾灸生成物组,高剂量艾灸生成物恢复组,每天在相应浓度艾灸生成物中暴露4小时,每周5天,持续2个月;高剂量艾灸生成物恢复组经2个月高剂量艾灸生成物刺激后,在空气中自然暴露21天作为恢复期。正常对照组大鼠不进行任何艾灸生成物干预,常规饲养2个月。肉眼、光学显微镜下观察各组大鼠肺的病理形态,比较各组大鼠肺脏器系数。结果肉眼下,除中、高剂量艾灸生成物组分别有1只大鼠肺部表面可见小的灰色或黑色斑点外,余各组别大鼠肺部均未见异常。光镜下,低、中、高剂量艾灸生成物组部分大鼠支气管及肺泡可见不同程度炎性细胞侵润,肺泡隔增厚,其中,高剂量艾灸生成物组较为明显。高剂量艾灸生成物恢复组大鼠支气管及肺泡炎性细胞侵润现象与肺泡隔增厚现象较高剂量艾灸生成物组大鼠明显减少。各组别大鼠肺脏器系数无统计学差异。结论艾灸生成物对大鼠肺形态的影响与艾灸生成物的浓度有一定相关性,艾灸生成物对大鼠肺形态的影响具有可逆性。