Objective: To compare the effects of acupuncture and moxibustion alone or in combination on the number of mast cells and expression levels of cytoketatin 18 (CK18) and CK19 (marker of Meckel cells), and calcitonin gene-related peptide (CGRP), neuropeptide-Y (NPY) and bradykinin (BK) in the local acupoint area of rats with chronic atrophic gastritis (CAG).
Methods: Fifty male SD rats were randomly divided into normal, CAG model, moxibustion, acupuncture and acupuncture+moxi-bustion groups (10 rats in each group). The CAG model was established by gavage of 1-methyl-3-nitro-1-nitrosoguanidine (170 μg/mL,1 mL/100 g, once a week) and 40% ethanol solution (twice a week) for 12 consecutive weeks. After successful establishment of CAG model, moxibustion, manual acupuncture or acupuncture+moxibustion was applied to bilateral "Zusanli" (ST36) and "Zhongwan"(CV12) for 15 min, once daily for 14 consecutive days. At the end of the experiment, the gastric mucosal tissues were collected for observing histopathological changes of gastric mucosa after H.E. staining, and the tissues of the stimulated ST36 region collected for detecting the expression levels of CK18, CK19, CGRP, NPY and BK and the number of mast cells in the local ST36 region by immunohistochemistry.
Results: Compared with the normal group, the number of mast cells, the expression levels of CK19, NPY and BK in the ST36 area were significantly increased (P<0.05), and the expression level of CGRP was apparently decreased (P<0.05) in the model group. In comparison with the model group, the number of mast cells and the expression levels of CGRP and NPY in the moxibustion group, the expression of CGRP in the acupuncture group, and the number of mast cells, as well as the expression levels of CK18, CK19 and CGRP in the acupuncture+moxibustion group were significantly up-re-gulated (P<0.05). The effect of acupuncture combined with moxibustion was obviously superior to that of moxibustion or acupuncture in up-regulating the expression of CK18 and CK19 (P<0.05) and superior to that of moxibustion in down-regulating BK expressio level (P<0.05). No significant changes were found in the expression of CK18 after modeling (vs the normal group), in the expression levels of CK18, CK19 and BK after moxibustion and acupuncture (vs the model group), in the number of mast cells and expression of NPY after acupuncture (vs the model group), and in the expression levels of NPY and BK after acupuncture+moxibustion (vs the model group) (P>0.05). H.E. staining showed infiltration of many lymphocytes in the gastric mucosa and submucosal layers, atrophy and necrosis of lots of main cells with vacuole-like changes, and disordered arrangement of the atrophic glands in the model group, which was milder particularly in the acupuncture + moxibustion group.
Conclusion: Acupuncture combined with moxibustion of ST36 can up-regulate the levels of local CK18, CK19 and CGRP proteins and number of mast cells, moxibustion may up-regulate the levels of CGRP and NPY and number of mast cells, while acupuncture may up-regulate the expression of CGRP in the local stimulated area in CAG rats.
目的：观察针刺与艾灸单独或联合使用对慢性萎缩性胃炎（CAG）大鼠穴区局部组织的梅克尔细胞标记物(CK18、CK19)、肥大细胞及相关神经肽表达的影响。方法：SD大鼠随机分为正常组、模型组、艾灸组、针刺组、针刺+艾灸组, 每组10只。采用1-甲基-3-硝基-1-亚硝基胍联合多因素复合法连续12周复制CAG模型。造模成功后分别对艾灸组、针刺组、针刺+艾灸组大鼠“足三里”“中脘”进行艾灸、针刺、针刺+艾灸干预, 15 min/次, 1次/d, 连续干预14 d。用HE染色法观察各组大鼠胃黏膜组织病理情况, 免疫组织化学法观察“足三里”穴区组织CK18、CK19、肥大细胞、降钙素基因相关肽(CGRP)、神经肽-Y(NPY)、缓激肽的表达。结果：正常组大鼠胃黏膜偶见散在炎性细胞, 腺体结构清晰, 排列整齐；模型组胃黏膜及黏膜下层有大量淋巴细胞浸润, 腺体萎缩且结构排列紊乱；艾灸组有大量淋巴细胞浸润, 黏膜出血点较多, 腺体结构排列紊乱；针刺组有炎性细胞浸润, 散在出血点, 腺体萎缩明显；针刺+艾灸组黏膜结构相对完整, 有少量炎性细胞, 偶见萎缩。与正常组比较, 模型组局部穴区CK19、NPY、缓激肽表达及肥大细胞计数均上升(P<0.05), CGRP表达下降(P<0.05)；与模型组比较, 艾灸组肥大细胞计数及CGRP、NPY表达均上升(P<0.05), 针刺组CGRP表达升高(P<0.05), 针刺+艾灸组CK18、CK19、CGRP表达及肥大细胞计数均上升(P<0.05)；与针刺+艾灸组比较, 艾灸组、针刺组CK18、CK19的表达降低(P<0.05), 艾灸组缓激肽的表达升高(P<0.05)。结论：针刺与艾灸联合使用可增强CAG大鼠“足三里”穴区CK18、CK19、CGRP的表达及肥大细胞的数量, 且联合使用与针刺、艾灸单独使用对于梅克尔细胞、肥大细胞及神经肽表达的影响有差异。.
Keywords: Chronic atrophic gastritis; Manual acupuncture; Moxibustion; Zusanli(ST36).