灸疗抗炎免疫作用的实验研究

应用Ｆｒｅｕｎｄ＇ｓ完全佐剂致炎，观察艾灸“肾俞”穴区对佐剂性关节炎大鼠的炎症和免疫功能的影响。结果，灸治组大鼠的活动、体重等一般情况均好于致炎组，局部炎症肿胀明显好转，跖围减小（Ｐ＜０．００１），并能一定程度地预防或减轻其迟发性变态反应──多发性关节炎。淋巴细胞功能测定表明，灸治组大鼠对ＣｏｎＡ与ＩＬ－２诱导的脾细胞增殖反应明显高于正常组和致炎组（Ｐ＜０．００１），表明了机体免疫应答的增强和免疫能力的提高。以上提示，灸疗具有较好的抗炎免疫作用。     

艾灸抗炎免疫作用的实验观察与分析

本实验建立大鼠佐剂性关节炎动物模型,观察艾克“肾俞”穴区的抗炎免疫作用。结果显示：灸疗能减轻局部炎症反应,消退足跖肿胀,使跖围减小;能预防或减轻多发性关节炎,维持体重,缩短病程。免疫功能观察表明,艾灸能恢复和促进ConA诱导的脾淋巴细胞增殖反应,促进IL－2的产生,降低IL－1的含量。以上指标与对照组比较均有非常显著差异。实验表明,艾灸有直接的抗炎消肿作用,并通过提高免疫应答水平,催化、激活和调整免疫功能,增强机体的抗炎免疫能力,起到抗炎免疫和抗变态反应的作用。     

艾灸对佐剂性关节炎大鼠滑膜细胞凋亡及病理组织的观察

目的:探讨艾灸治疗类风湿性关节炎(RA)的作用机理.方法:复制佐剂性关节炎(AA)大鼠模型,应用放免、细胞凋亡和病理学等方法,观察艾灸"肾俞"穴的抗炎免疫作用和对关节滑膜组织形态学的影响.结果:(1)灸治能减轻关节的肿胀和多发性关节炎,减少血清中肿瘤坏死因子(TNF)、白细胞介素I(IL-1)含量,提高IL-2和胸腺指数,与模型组比较差异显著(P＜0.05),表明艾灸具有抗炎免疫的作用;(2)艾灸能诱导AA大鼠关节滑膜细胞的调亡,能提高凋亡指数,减轻炎症反应;(3)病理学观察证实,灸治能减轻关节滑膜的充血水肿、炎细胞浸润、滑膜细胞增生、组织增厚等渗出性变.结论:艾灸主要通过抑制滑膜炎起到治疗关节炎的作用.抑制炎症细胞因子的产生、释放,诱导滑膜细胞凋亡、是艾灸治疗RA抗炎免疫作用的主要机制.     

艾灸抗炎免疫作用及对神经递质影响的实验研究

目的 :探讨灸法治疗 RA抗炎免疫作用的中枢神经递质调控机理。方法 :建立 AA大鼠模型 ,观察艾灸肾俞穴对炎症肿胀、细胞因子 IL- 6、IL- 2以及下丘脑内神经递质 NE、5- HT和 NO的影响。结果 :1 .灸治 AA大鼠能减轻炎症肿胀 ,降低 IL- 6,提高 IL- 2 ,与对照组比较差异显著 ( P<0 .0 1 ) ,表明灸疗能抗炎消肿 ,抑制炎症因子 ,提高机体免疫功能 ;2 .灸疗能上调 NE、5- HT的水平 ,降低 NO的含量 ,提示艾灸能激活或调整 HPA轴抗炎免疫的功能活动 ,有利于 AA的缓解与转归。结论 :艾灸有抗炎免疫的作用 ,下丘脑神经递质参与对该作用的中枢调节。     

大鼠肾上腺摘除对艾灸抗炎免疫作用的影响

为探讨艾灸抗炎免疫作用机理，本实验观察了ＡＡ大鼠肾上朱摘除对灸疗作用的影响。结果表明，在肾上腺完整的情况下，艾灸能抗炎消肿，增强免疫功能，其跖转减小、胸脾指数改善、ＩＬ－６、ＩＬ－２含量趋于正常，与对照组比较差异显著（Ｐ〈０．０１）。肾上腺摘除后，灸疗的抗炎免疫作用被部分阻断或削弱，上述指标与肾上腺摘除对照组比较差异不显著（Ｐ〉０．０５）。提示灸疗可能通过激活ＨＰＡ系统对皮质激素的调控参与并影响其     

针灸治疗大鼠溃疡性结肠炎的机理研究

采用免疫学方法,制备大鼠溃疡性结肠炎模型,并观察隔药灸、电针天枢和气海穴对溃疡性结肠炎大鼠结肠粘膜β－ＥＰ、ＶＩＰ含量的影响。研究结果表明,模型组大鼠结肠粘膜β－ＥＰ、ＶＩＰ含量显著高于正常对照组（Ｐ＜０．０５,Ｐ＜０．０１）,而隔药灸组与电针组均明显下降,表明隔药灸与电针具有调节溃疡性结肠炎大鼠结肠粘膜β－ＥＰ、ＶＩＰ的作用。     

艾灸抗炎免疫作用中松果腺褪黑素高位调节的研究

目的研究松果腺褪黑素(MT)对炎症与免疫的调节,探讨艾灸抗炎免疫作用的机理.方法建立佐剂性关节炎大鼠模型,分别损毁其海马内NA神经和摘除肾上腺,观察艾灸对MT及细胞因子IL-6、IL-2的影响.结果艾灸"肾俞"穴能恢复和促进大鼠MT的分泌,使IL-6降低、IL-2升高,与对照组比较差异显著(P＜0.05,P＜0.01).失去肾上腺和海马交感神经的支配,松果腺MT增多,IL-6、IL-2含量上升,灸疗的抗炎免疫作用被削弱或部分阻断.结论提示松果腺MT参与艾灸抗炎免疫作用的高位调节.     

迷迭香酸抗炎作用研究

目的：研究迷迭香酸的抗炎作用。方法：采用鼠耳肿胀法、腹腔毛细血管通透性法等急性炎症模型、慢性肉芽肿模型及大鼠佐剂性关节炎模型观察迷送香酸对急、慢性炎症和免疫性炎症的抗炎作用。结果：迷迭香酸能明显抑制二甲苯所致小鼠耳肿胀、醋酸所致小鼠毛细血管通透性增高及大鼠棉球肉芽肿性炎症，对Freund＇s完全佐剂介导的大鼠佐剂性关节炎足肿胀的抑制作用强弱与剂量大小呈较好的量效关系。另外，迷迭香酸能降低炎症模型动物血清中炎症细胞因子TNF-α、IL-β、CRP的含量。结论：迷迭香酸有明显的抗炎和免疫抑制作用，降低炎性细胞因子TNF-α、IL-1β、CRP的产生可能是迷迭香酸减轻炎症反应机制之一。     

灸法对脾虚大鼠血清D—木糖含量及红细胞免疫功能的影响

目的观察灸法对实验性脾虚大鼠血清Ｄ－木糖含量及细胞免疫功能的影响。方法采用复合造模法复制出实验性脾虚大鼠,用麦粒灸灸“脾俞”、“胃俞”、“足三里”穴。结果灸治组血清Ｄ－木糖值较模型组显著升高（Ｐ＜０．００１）;红细胞Ｃ３ｂ受体环率显著上升（Ｐ＜０．００１）,ＩＣ花环率显著下降（Ｐ＜０．００１）,旦与中药对照组比有显著性差异（Ｐ＜０．０５）。结论灸法能提高细胞免疫功能     

槌果藤对佐剂性关节炎大鼠模型外周血中细胞因子影响的研究

目的观察槌果藤对佐剂性关节炎大鼠模型外周血中细胞因子TNF-α及sICAM-1的影响。方法建立佐剂性关节炎大鼠模型,观察槌果藤对AA大鼠足肿胀、踝关节组织病理变化及血清中促炎细胞因子TNF-α及sICAM-1水平的影响。结果槌果藤能有效抑制关节肿胀及减轻炎症的发展,降低佐剂性关节炎大鼠血清中促炎细胞因子TNF-α及sI-CAM-1的水平。结论槌果藤抗RA作用与抑制促炎细胞因子TNF-α及sICAM-1的水平有关。     

双清口服液解热抗炎作用研究

目的:观察双清口服液的药理作用。方法:对双清口服液的解热、抗炎及减少炎症介质等作用进行了实验研究。结果:双清口服液对伤寒副伤寒四联疫苗引起的大鼠及大白兔的发热反应有明显解热作用(P<0.01),对醋酸引起的小鼠腹膜炎症及巴豆油引起的耳廓炎症反应均有抑制作用(P<0.05),可减少炎症介质(前列腺素E、5-羟色胺)在炎症组织中产生(P<0.05),并能明显提高单核细胞吞噬功能。结论:双清口服液有明显的解热抗炎作用。     

麦粒灸对实验性RA大鼠局部组织炎症介质的影响

目的　观察艾灸对实验性 RA大鼠炎症局部炎症介质的影响。方法　麦粒灸佐剂性关节炎大鼠“肾俞”、“足三里”,采用荧光免疫、放射免疫方法检测炎症局部 5 -羟色胺 (5 - HT)、组胺 (HA )和前列腺素 E2 (PGE2 )含量。结果　造模后 ,5 - HT、HA、PGE2 含量均显著升高 (P<0 .0 1) ;艾灸治疗后 ,三者含量又显著降低 (P<0 .0 1) ;摘除肾上腺后艾灸 ,HA含量有显著性下降(P<0 .0 5 ) ,而 5 - HT、PGE2 含量下降不明显 (P>0 .0 5 )。结论　艾灸治疗可明显清除炎症局部组织中 5 - HT、HA、PGE2 ,从而减轻局部炎症反应、水肿、疼痛等症状。肾上腺在艾灸清除炎症介质过程中有明显作用。     

Potential anti-inflammatory effects of Melaleuca alternifolia essential oil on human peripheral blood leukocytes

The fungicidal and bactericidal actions of the essential oil (EO) of Melaleuca alternifolia seem well established, but their anti-inflammatory and antioxidative effects remain unclear. This study investigated in vitro the possible role of whole Melaleuca alternifolia EO as a modulator of the inflammatory/non-specific immune response by exploring the chemotaxis and kinetic radical oxygen species (ROS) production of leukocytes and cytokine secretion in peripheral blood mononuclear cells (PBMCs) in humans. The influence of Melaleuca alternifolia EO on the chemotaxis under agarose of isolated neutrophils (PMNs) was evaluated. The kinetics of ROS production by stimulated total circulating leukocytes was followed over 2 h by recording the fluorescence intensity of oxidized dihydrorhodamine 123. The effects of this EO on pro-(interleukin IL-2) and anti-(IL-4 and IL10) inflammatory cytokine secretions were determined by ELISA following incubation of PBMCs with the EO for 24 h. Melaleuca alternifolia EO was inefficient on the chemotaxis of PMNs. It exerted an antioxidant effect, reducing ROS production throughout the kinetic study. Melaleuca alternifolia EO inhibited PBMC proliferation, as revealed by a reduction in IL-2 secretion by stimulated lymphocytes. This EO at 0.1% directly increased the secretion of the anti-inflammatory cytokine IL-4 compared with IL-4 secretion without EO (18.5 +/- 10.0 vs 3.3 +/- 1, p < 0.05), and also increased IL-10 secretion at 0.01% (94.9 +/- 38.7 vs 44.1 +/- 18, ns). Melaleuca alternifolia EO may not only act as an anti-inflammatory mediator through its antioxidant activity but may also efficiently protect the organism by reducing the proliferation of inflammatory cells without affecting their capacity to secrete anti-inflammatory cytokines.     

艾灸对类风湿性关节炎家兔滑膜细胞JAK-STAT通路负反馈调节家族细胞因子信号抑制因子的影响

目的:观察艾灸对实验性类风湿性关节炎(RA)家兔的抗炎消肿作用,探索艾灸抗炎效应的滑膜细胞JAK-STAT信号通路负反馈调节机制。方法:日本大耳白兔随机分为对照组、模型组和艾灸组,各14只。福氏完全佐剂注入兔双后膝关节腔复制RA模型。艾灸组动物采用艾粒施灸两侧"肾俞"穴各5壮,每日1次,6d为一疗程,共治疗3个疗程。治疗后观测各动物左、右后膝关节周长,运用免疫组化法检测细胞因子信号抑制因子1(SOCS 1)、SOCS 3的表达。结果:造模后各个时间点模型组动物的左、右后膝关节周长均较对照组明显增加(P<0.01),治疗后艾灸组动物左、右后膝关节周长均较模型组降低(P<0.05)。模型组与对照组比较,滑膜细胞SOCS 1、SOCS 3蛋白含量显著升高(P<0.01);艾灸组与模型组比较,滑膜细胞SOCS 1、SOCS 3蛋白含量降低(P<0.01)。结论:艾灸对实验性RA具有抗炎效应,艾灸对滑膜细胞JAK-STAT信号通路中相关负反馈调节因子SOCS 1、SOCS 3表达的调节可能是其实现抗炎效应的机制之一。     

Effects of different courses of moxibustion treatment on intestinal flora and inflammation of a rat model of knee osteoarthritis

ObjectiveThis study was done to determine the effects of different courses of moxibustion on a rat knee osteoarthritis (KOA) model, and explore the dose–effect relationship of moxibustion on KOA from the perspectives of intestinal flora and inflammatory factors.MethodsWistar rats were randomly divided into five groups: normal, model, moxibustion for 2 weeks, moxibustion for 4 weeks and moxibustion for 6 weeks groups (n = 5 each group). A KOA rat model was induced by monosodium iodoacetate, and moxibustion intervention was performed at the acupoints “Dubi” (ST35) and “Zusanli” (ST36), once every other day. Pathologic changes in the cartilage of rat knee joints were assessed after intervention, and fecal samples were subjected to 16S rRNA high-throughput sequencing for microbial diversity analysis.ResultsDamage to the knee articular cartilage was obvious in the model group, which also had increased levels of pro-inflammatory factors, decreased levels of anti-inflammatory factors, and intestinal flora disorders with decreased diversity. The degree of cartilage damage in the 4 and 6 weeks of moxibustion groups was significantly improved compared with the model group. The 4 and 6 weeks of moxibustion groups also demonstrated reduced levels of interleukin-1β and tumor necrosis factor-α and increased levels of interleukin-10 (P < 0.05). Both the abundance and diversity of the intestinal flora were increased, approaching those of the normal group. Abundances of probiotics Eubacterium coprostanoligenes group and Ruminococcaceae UCG-014 increased, while that of the pathogenic bacteria Lachnospiraceae NK4A136 group decreased (P < 0.05). Although the abundance of Lachnospiraceae NK4A136 group decreased in the 2 weeks of moxibustion group compared with the model group (P < 0.05), there was no statistically significant difference in serum inflammatory factors, flora species diversity or degree of pathological damage compared with the model group.ConclusionMoxibustion treatment led to significant improvements in the intestinal flora and inflammatory factors of rats with KOA. Moxibustion treatment of 4 and 6 weeks led to better outcomes than the 2-week course. Moxibustion for 4 and 6 weeks can regulate intestinal flora dysfunction with increased probiotics and reduced pathogenic bacteria, reduce pro-inflammatory factors and increase anti-inflammatory factors. No significant differences were seen between the effects of moxibustion for 4 weeks and 6 weeks.     

喉疾灵口含片抗炎抑菌作用实验研究

目的：对喉疾灵口含片进行抗炎、抑菌作用实验研究。方法：采用二甲苯致小鼠耳壳肿胀、角叉菜致大鼠足趾肿胀、大鼠棉球肉芽肿等炎症模型,以及含药血清体外抑菌实验模型,评价喉疾灵口含片的抗炎、抑菌作用。结果：与空白对照比较,喉疾灵口含片明显抑制了小鼠耳壳肿胀（P〈0．05或P〈0．01）、大鼠足趾肿胀以及大鼠棉球肉芽肿的形成（P〈0．05）;其含药血清对金黄色葡萄球菌、铜绿假单胞菌及白色念珠菌具有明显的抑制作用（P〈0．05或P〈0．01）。结论：上述结果表明,喉疾灵口含片对化学刺激所致炎症有明显的抑制作用,其含药血清具有显著的体外抑菌作用。     

艾灸预处理对大鼠应激性胃黏膜损伤的保护作用

目的:探讨艾灸预处理对大鼠应激性胃黏膜损伤保护作用及抗炎症损伤机制。方法:将48只健康SD大鼠随机分为4组,即空白组、模型组、艾灸穴位组、艾灸非穴组。束缚冷应激法制作应激性胃黏膜损伤大鼠模型,按Guth法计算胃黏膜损伤指数(UI),光镜下观察大鼠胃黏膜组织学改变,放射免疫法测定血清IL-1β,TNF-α和IL-10的含量。结果:与艾灸非穴组比较,艾灸足三里、中脘等穴位可使应激性胃黏膜损伤大鼠UI明显下降(P0.01)、血清IL-1β含量降低(P0.05)、TNF-α含量降低(P0.01)、IL-10含量升高(P0.05)。结论:艾灸足三里、中脘等穴位预处理可促进束缚水浸应激所造成大鼠胃黏膜损伤的修复、减轻急性炎症反应,该保护作用可能是通过抑制细胞炎症反应的免疫促炎因子IL-1β、TNF-α和促进抗炎因子IL-10而达到其抗胃黏膜损伤作用。     

Purified active lotus plumule (Nelumbo nucifera Gaertn) polysaccharides exert anti-inflammatory activity through decreasing Toll-like receptor-2 and -4 expressions using mouse primary splenocytes

Ethnopharmacological significance

Lotus plumule is widely used as traditional Chinese medicine. Among the active components in lotus plumule, polysaccharides exhibit promising potential for its potent anti-inflammatory effects. However, the anti-inflammatory mechanism of purified polysaccharides from lotus plumule remains unknown.To evaluate their anti-inflammatory potential and possible mechanisms of purified polysaccharides in lotus plumule, two active lotus plumule polysaccharides, fractions F1 and F2, were subjected to assay their anti-inflammatory potential and possible mechanisms using murine primary splenocytes in the absence or presence of lipopolysaccharide (LPS).

Materials and methods

Two purified active lotus plumule polysaccharides, F1 and F2, were cultured independently with murine primary splenocytes in the absence or presence of LPS under four different experiment models in vitro. Changes in pro-inflammatory IL-1β, IL-6 and TNF-α, as well as anti-inflammatory IL-10 cytokines secreted by the treated splenocytes were determined using an enzyme-linked immunosorbent assay (ELISA). The amount of Toll-like receptor (TLR)-2 and TLR-4 mRNA expression levels in the cells were quantitated using a two-step real-time polymerase chain reaction (PCR) assay.

Results

The results showed that F1 and F2 treatments alone, particularly F2, significantly (P<0.05) decreased pro-/anti-inflammatory (IL-1β/IL-10 and TNF-α/IL-10) cytokine secretion ratios dose-dependently. F1 and F2 treatments in the presence of LPS significantly decreased TLR-2 and/or TLR-4 mRNA expression levels in the splenocytes under inflammatory and repair experiment models.

Conclusions

The present study proved that F1 and F2 had strong anti-inflammatory effects through inhibiting TLR-2 and/or TLR-4 expressions in the splenocytes in normal, inflammatory and repair situations. Our results further suggest that F2, which is a glycoprotein with low molecular weight of 25.7 kDa, may serve as a promising lead for the development of selective TLR antagonistic agents for inflammatory diseases.     

艾灸预处理对应激性胃黏膜损伤大鼠血清IL-1β、TNF-α和IL-10的影响

目的观察艾灸预处理对应激性胃黏膜损伤大鼠白细胞介素-1β（IL-1β）、肿瘤坏死因子-α（TNF-α）和白细胞介素-10（IL-10）的影响,探讨艾灸预处理抗胃黏膜炎症损伤的作用机制。方法将SD大鼠随机分为空白组、模型组、艾灸穴位组、艾灸非穴点对照组。束缚冷应激法制作应激性胃黏膜损伤模型,按Guth法计算胃黏膜损伤指数（UI）,放射免疫法测定血清IL-1β、TNF-α和IL-10的含量。结果与模型组和艾灸非穴点对照组比较,艾灸足三里、中脘等穴可使应激性胃黏膜损伤UI明显下降、血清IL-1β含量降低、TNF-α含量降低、IL-10含量升高。结论艾灸足三里、中脘等穴预处理可促进大鼠胃黏膜损伤的修复、减轻急性炎症反应,可能是通过抑制细胞炎症反应的免疫促炎因子IL-1β、TNF-α和促进抗炎因子IL-10而达到其抗胃黏膜损伤作用。