Abstract：
OBJECTIVE: To explore the mechanism of moxibustion in the treatment of asthmatic inflammation from the point of short-chain fatty acids (SCFAs) in rats with asthma.
METHODS: A total of 48 SD rats (half male and half female) were randomly divided into 4 groups： normal, model, lung treatment and joint-treatment of lung and intestine (joint-treatment), with 12 rats in each group. The asthma model was made by subcutaneous (bilateral back and inguinal regions) and intraperitoneal injection of mixture solution of ovalbumin and aluminium hydroxide gel (on day 1 and 8) and followed by inhalation of atomized 1% ovalbumin (20 min from day 15, once daily for one week). Moxibustion was applied to bilateral \"Feishu\" (BL13) for rats of the lung treatment group or bilateral \"Feishu\" (BL13) and \"Tianshu\" (ST25) for rats of the joint treatment group. One hour after the intervention, the rats in the later three groups were separately given atomized 1% ovalbumin solution inhalation for 20 min. The treatment was conducted for 30 min, once daily for 14 consecutive days. At the end of the intervention, the percentage of inflammatory cells in blood was detected by biochemical method and histopathological changes of the lung were observed after H.E. staining. The inflammatory cells in the bronchoalveolar lavage fluid (BALF) were counted after Wright-Giemsa staining. The mRNA expressions of interleukin (IL)-4, IL-5, IL-13, IL-17, IL-33, leukotriene (LT), thymic stromal lymphopoietin (TSLP) and prostaglandin D2 (PGD2) were detected by real-time PCR, and the contents of SCFAs in rats\' feces were detected by gas chromatography-mass spectrometry.
RESULTS: Relevant to the normal group, the model group had an obvious increase in the percentages of neutrophils, lymphocytes and eosinophils in the blood, the percentages of neutrophils and eosinophils in the BALF, and in the expression levels of PGD2, TSLP, LT, IL-4, IL-5, IL-13, IL-17 and IL-33 mRNAs in the lung tissues (P<0.01, P<0.05), and a marked decrease in the contents of acetic acid, propionic acid, isobutyric acid, butyric acid and valeric acid in feces (P<0.05, P<0.01). After the treatment, the percentages of neutrophils and lymphocytes in the peripheral blood, eosinophils in the BALF, and the expression levels of PGD2, TSLP, LT, IL-4, IL-17, IL-33 mRNAs in the lung tissues in both the lung treatment and joint treatment groups, as well as neutrophils of BALF, and expression of IL-5 and IL-13 mRNAs in the joint treatment group were significantly down-regulated (P<0.01, P<0.05), while the contents of acetic acid, propionic acid and valerate in the lung treatment group, and acetic acid, propionic acid, isobutyric acid, butyric acid and valeric acid in the joint treatment group were all strikingly increased (P<0.05, P<0.01). The effect of the joint treatment was superior to that of lung treatment in down-regulating the expressions of LT and IL-5 mRNAs (P<0.05, P<0.01) and up-requlating the content of propionic acid (P<0.05). Results of H.E. staining showed thickened alveolar wall, infiltration of a large number of inflammatory cells and interstitial fibrous tissue hyperplasia around the bronchus and scattered arrangement of cells of the lung tissue in the model group, which was relatively milder in both lung treatment and joint treatment groups, particularly the later.
CONCLUSIONS: Joint treatment of asthma from the lung and intestine can better regulate the contents of intestinal SCFAs and alleviate the inflammatory response of asthmatic model rats, thus, intestinal SCFAs may be involved in the process of moxibustion in improving inflammatory response.
目的：从对肠道短链脂肪酸（SCFAs）的调控角度探讨艾灸改善哮喘炎性反应的可能作用机制。方法：48只SD大鼠随机分为正常组、模型组、从肺论治组和肺肠同治组，每组12只。采用卵清白蛋白（OVA）致敏和雾化激发法制备哮喘大鼠模型。从肺论治组艾灸大鼠双侧“肺俞”30 min；肺肠同治组艾灸大鼠双侧“肺俞”“天枢”共30 min。模型组、从肺论治组、肺肠同治组大鼠上述干预结束1 h后，分别予以1% OVA溶液雾化20 min，均每天1次，连续14 d。同血细胞分析仪检测外周血中炎性细胞百分比，HE染色法观察肺组织病理形态学改变，瑞氏-吉姆萨染色法计数肺泡灌洗液（BALF）中炎性细胞，实时荧光定量PCR法检测肺组织中白细胞介素（IL）-4、IL-5、IL-13、IL-17、IL-33、前列腺素D2（PGD2）、胸腺基质淋巴细胞生成素（TSLP）和白三烯（LT） mRNA的表达，气相色谱-质谱联用法检测粪便中SCFAs的含量。结果：与正常组相比，模型组大鼠肺组织结构重度异常，肺泡壁增厚，支气管周围可见大量炎性细胞浸润；外周血中性粒细胞、淋巴细胞、嗜酸性粒细胞百分比，BALF中嗜酸性粒细胞、中性粒细胞百分比，肺组织中IL-4、IL-5、IL-13、IL-17、IL-33、PGD2、TSLP、LT的mRNA表达均显著升高（P<0.01，P<0.05）；粪便中乙酸、丙酸、异丁酸、丁酸、戊酸的含量均显著降低（P<0.05，P<0.01）。与模型组相比，从肺论治组和肺肠同治组大鼠肺组织病理形态学损伤均显著改善；外周血中性粒细胞和淋巴细胞百分比，BALF中嗜酸性粒细胞百分比，肺组织中IL-4、IL-17、IL-33、PGD2、TSLP、LT的mRNA表达均显著降低（P<0.01，P<0.05）；粪便中乙酸、丙酸、戊酸的含量均显著升高（P<0.05，P<0.01）。除此之外，与模型组相比，肺肠同治组大鼠BALF中中性粒细胞百分比，肺组织中IL-5、IL-13的mRNA表达也均显著降低（P<0.05，P<0.01），粪便中异丁酸、丁酸含量也均显著升高（P<0.05，P<0.01）。与从肺论治组相比，肺肠同治组大鼠肺组织中IL-5、LT的mRNA表达均显著降低（P<0.01，P<0.05），粪便中丙酸含量显著升高（P<0.05）。结论：艾灸“肺肠同治”可以更好地调节肠道SCFAs的含量，缓解哮喘模型大鼠炎性反应；肠道SCFAs可能参与了艾灸改善哮喘大鼠炎性反应的作用过程。.
METHODS: A total of 48 SD rats (half male and half female) were randomly divided into 4 groups： normal, model, lung treatment and joint-treatment of lung and intestine (joint-treatment), with 12 rats in each group. The asthma model was made by subcutaneous (bilateral back and inguinal regions) and intraperitoneal injection of mixture solution of ovalbumin and aluminium hydroxide gel (on day 1 and 8) and followed by inhalation of atomized 1% ovalbumin (20 min from day 15, once daily for one week). Moxibustion was applied to bilateral \"Feishu\" (BL13) for rats of the lung treatment group or bilateral \"Feishu\" (BL13) and \"Tianshu\" (ST25) for rats of the joint treatment group. One hour after the intervention, the rats in the later three groups were separately given atomized 1% ovalbumin solution inhalation for 20 min. The treatment was conducted for 30 min, once daily for 14 consecutive days. At the end of the intervention, the percentage of inflammatory cells in blood was detected by biochemical method and histopathological changes of the lung were observed after H.E. staining. The inflammatory cells in the bronchoalveolar lavage fluid (BALF) were counted after Wright-Giemsa staining. The mRNA expressions of interleukin (IL)-4, IL-5, IL-13, IL-17, IL-33, leukotriene (LT), thymic stromal lymphopoietin (TSLP) and prostaglandin D2 (PGD2) were detected by real-time PCR, and the contents of SCFAs in rats\' feces were detected by gas chromatography-mass spectrometry.
RESULTS: Relevant to the normal group, the model group had an obvious increase in the percentages of neutrophils, lymphocytes and eosinophils in the blood, the percentages of neutrophils and eosinophils in the BALF, and in the expression levels of PGD2, TSLP, LT, IL-4, IL-5, IL-13, IL-17 and IL-33 mRNAs in the lung tissues (P<0.01, P<0.05), and a marked decrease in the contents of acetic acid, propionic acid, isobutyric acid, butyric acid and valeric acid in feces (P<0.05, P<0.01). After the treatment, the percentages of neutrophils and lymphocytes in the peripheral blood, eosinophils in the BALF, and the expression levels of PGD2, TSLP, LT, IL-4, IL-17, IL-33 mRNAs in the lung tissues in both the lung treatment and joint treatment groups, as well as neutrophils of BALF, and expression of IL-5 and IL-13 mRNAs in the joint treatment group were significantly down-regulated (P<0.01, P<0.05), while the contents of acetic acid, propionic acid and valerate in the lung treatment group, and acetic acid, propionic acid, isobutyric acid, butyric acid and valeric acid in the joint treatment group were all strikingly increased (P<0.05, P<0.01). The effect of the joint treatment was superior to that of lung treatment in down-regulating the expressions of LT and IL-5 mRNAs (P<0.05, P<0.01) and up-requlating the content of propionic acid (P<0.05). Results of H.E. staining showed thickened alveolar wall, infiltration of a large number of inflammatory cells and interstitial fibrous tissue hyperplasia around the bronchus and scattered arrangement of cells of the lung tissue in the model group, which was relatively milder in both lung treatment and joint treatment groups, particularly the later.
CONCLUSIONS: Joint treatment of asthma from the lung and intestine can better regulate the contents of intestinal SCFAs and alleviate the inflammatory response of asthmatic model rats, thus, intestinal SCFAs may be involved in the process of moxibustion in improving inflammatory response.
目的：从对肠道短链脂肪酸（SCFAs）的调控角度探讨艾灸改善哮喘炎性反应的可能作用机制。方法：48只SD大鼠随机分为正常组、模型组、从肺论治组和肺肠同治组，每组12只。采用卵清白蛋白（OVA）致敏和雾化激发法制备哮喘大鼠模型。从肺论治组艾灸大鼠双侧“肺俞”30 min；肺肠同治组艾灸大鼠双侧“肺俞”“天枢”共30 min。模型组、从肺论治组、肺肠同治组大鼠上述干预结束1 h后，分别予以1% OVA溶液雾化20 min，均每天1次，连续14 d。同血细胞分析仪检测外周血中炎性细胞百分比，HE染色法观察肺组织病理形态学改变，瑞氏-吉姆萨染色法计数肺泡灌洗液（BALF）中炎性细胞，实时荧光定量PCR法检测肺组织中白细胞介素（IL）-4、IL-5、IL-13、IL-17、IL-33、前列腺素D2（PGD2）、胸腺基质淋巴细胞生成素（TSLP）和白三烯（LT） mRNA的表达，气相色谱-质谱联用法检测粪便中SCFAs的含量。结果：与正常组相比，模型组大鼠肺组织结构重度异常，肺泡壁增厚，支气管周围可见大量炎性细胞浸润；外周血中性粒细胞、淋巴细胞、嗜酸性粒细胞百分比，BALF中嗜酸性粒细胞、中性粒细胞百分比，肺组织中IL-4、IL-5、IL-13、IL-17、IL-33、PGD2、TSLP、LT的mRNA表达均显著升高（P<0.01，P<0.05）；粪便中乙酸、丙酸、异丁酸、丁酸、戊酸的含量均显著降低（P<0.05，P<0.01）。与模型组相比，从肺论治组和肺肠同治组大鼠肺组织病理形态学损伤均显著改善；外周血中性粒细胞和淋巴细胞百分比，BALF中嗜酸性粒细胞百分比，肺组织中IL-4、IL-17、IL-33、PGD2、TSLP、LT的mRNA表达均显著降低（P<0.01，P<0.05）；粪便中乙酸、丙酸、戊酸的含量均显著升高（P<0.05，P<0.01）。除此之外，与模型组相比，肺肠同治组大鼠BALF中中性粒细胞百分比，肺组织中IL-5、IL-13的mRNA表达也均显著降低（P<0.05，P<0.01），粪便中异丁酸、丁酸含量也均显著升高（P<0.05，P<0.01）。与从肺论治组相比，肺肠同治组大鼠肺组织中IL-5、LT的mRNA表达均显著降低（P<0.01，P<0.05），粪便中丙酸含量显著升高（P<0.05）。结论：艾灸“肺肠同治”可以更好地调节肠道SCFAs的含量，缓解哮喘模型大鼠炎性反应；肠道SCFAs可能参与了艾灸改善哮喘大鼠炎性反应的作用过程。.
摘要：
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