Shi Hong, Cheng Bin, Li Jiang-Hui, Chen Shu-Li, Tan Qi-Wen, Jin Zhi-Gao, Jing Xiang-Hong
Institute of Acu-moxibustion, China Academy of Chinese Medical Sciences, Beijing 100700, China.
Zhen Ci Yan Jiu. 2010 Oct;35(5):323-9.
To observe changes of mast cells (MCs) number and morphology, and substance P (SP) expression in Evans blue (EB) extravasated region around acupoint "Pishu" (BL 20) and "Weishu" (BL 21) after acute gastric mucosal injury (AGMI) so as to investigate the mechanism underlying visceral problems-induced acupoint activation.
Thirty adult Wistar rats were randomly divided into normal control (n = 15) and AGMI groups (n = 15). AGMI model was duplicated by perfusing the rats with 0.5 mol/L HCl (1 mL/100 g) after fasting for 20 h. Five hours after AGMI, the rats were treated by tail-intravenous injection of EB dye (5 mg/100 g, 50 mg/mL in normal saline) for inducing dye-plasma extravasation in the skin around BL 20, BL 21 regions, etc. at the back. The rats of the normal control group were treated with tail-intravenous injection of 0.9% NaCl. The skin and subcutaneous tissues (2 mmx 2 mm) of extravasated EB dye points (BL 20 or BL 21 region) and those 2 mm lateral to the extravasated EB dye points in the model group and the corresponding points in the normal control group were sampled (followed by fixing them in 4% paraformaldehyde), sectioned and stained by toluidine blue (for labeling MCs). The expression of SP in the extravasated EB dye skin and subcutaneous tissues was detected by immunohistochemistry (n = 5) and western blot (n = 5) respectively. The number of MCs in these samples was counted and the degranulation rate of MCs calculated.
The total number of MCs and the number of degranulated MCs were significantly more in the EB extravasation points (corresponding to BL 20/BL 21 area) of AGMI group than those in the control spots of AGMI group and than those in the normal control group (P < 0.05, P < 0.001). The degranulation rate of MCs was significantly higher in the EB extravasation points of AGMI group than those in the control spots of AGMI group and in the normal control group (P < 0.01). In comparison with normal control group, the SP expression level was increased consideraly in the control spots of AGMI group and AGMI group (P < 0.01).
After AGMI, the numbers of MCs and the degranulated MCs, and the SP expression level in BL 20/BL 21 area were increased significantly, suggesting an involvement of MCs and SP in the process of AGMI-induced activation of acupoints.
观察急性胃黏膜损伤（AGMI）后，大鼠“脾俞”（BL 20）和“胃俞”（BL 21）穴位周围伊文思蓝（EB）外渗区域肥大细胞（MCs）数量及形态变化，以及P物质（SP）表达情况，以探讨内脏病变诱发穴位激活的机制。
将30只成年Wistar大鼠随机分为正常对照组（n = 15）和AGMI组（n = 15）。禁食20 h后，给大鼠灌胃0.5 mol/L盐酸（1 mL/100 g）复制AGMI模型。AGMI后5 h，经尾静脉注射EB染料（5 mg/100 g，用生理盐水配制成50 mg/mL），诱导大鼠背部BL 20、BL 21等区域皮肤染料-血浆外渗。正常对照组大鼠经尾静脉注射0.9%氯化钠溶液。取模型组EB染料外渗点（BL 20或BL 21区域）及其外侧2 mm处的皮肤和皮下组织（2 mm×2 mm），以及正常对照组相应部位组织（用4%多聚甲醛固定），切片，甲苯胺蓝染色（标记MCs）。分别采用免疫组化（n = 5）和蛋白质免疫印迹法（n = 5）检测EB染料外渗皮肤和皮下组织中SP的表达。计数这些样本中MCs的数量，并计算MCs的脱颗粒率。
AGMI组EB外渗点（对应BL 20/BL 21区域）的MCs总数和脱颗粒MCs数量显著多于AGMI组的对照点及正常对照组（P < 0.05，P < 0.001）。AGMI组EB外渗点的MCs脱颗粒率显著高于AGMI组的对照点及正常对照组（P < 0.01）。与正常对照组相比，AGMI组的对照点及AGMI组的SP表达水平均显著升高（P < 0.01）。
AGMI后，BL 20/BL 21区域的MCs数量、脱颗粒MCs数量及SP表达水平均显著增加，提示MCs和SP参与了AGMI诱发的穴位激活过程。
