SP SS ATP MLCK c- kit MTL SP SS P < 0.05P < 0.01 ATP MLCK c- kit P < 0.05P < 0.01 P < 0.05P < 0.01 R285.5 A

186 Traditional Chinese Drug Research & Clinical Pharmacology2015 MarchVol. 26 No. 2 MsPGN TGF- β 1/Smads/ILK TGF- β 1 Smad3 ILK Smad7 mrna MsPGN TGF- β 1/Smads/ ILK β β β β β 12 2 11. 4450002. 430065 SD 10.0 g kg - 1 10.0 g kg - 1 5.0 mg kg - 1 5 10 15 d 1 10 d 10 d MTL P 2014-11- 12

2015 3 26 2 187 SP SS ATP MLCK c- kit MTL SP SS P < 0.05P < 0.01 ATP MLCK c- kit P < 0.05P < 0.01 P < 0.05P < 0.01 R285.5 A 1003-2015 9783 02-0186- 06 d o i 10.3969/j.issn.1003-9783.2015.02.012 E ffect of Raw or Bran-fried Rhizoma Atractylodis on Gastrointestinal Motility of Experimental Spleen Deficiency Rats LIU Fen 1 2 LIU Yanju 2 TIAN Chunman1. 1 Hubei Institute for NationalitiesEnshi 445000 HubeiChina2. Hubei University of Chinese MedicineWuhan 430065 Hubei China AbstractObjective To observe the effect of raw or bran-fried Rhizoma Atractylodis RAon gastrointestinal motility of rats with experimental spleen deficiency syndromeand to explore the possible mechanism. Methods Fifty male SD rats were evenly randomized into normal groupmodel groupraw RA 10.0 g kg -1 group bran-fried RA 10.0 g kg -1 groupand domperidone5.0 mg kg -1 group. Except for the normal groupthe rats in other groups were fed with Xiaochengqi Decoction plus irregular diet to induce spleen deficiency syndromeand the modeling time covered 15 d. After the establishment of modelthe medication groups were given the corresponding medicineand the normal group and model group were given same volume of saline once a day for 10 days. After treatmentgastric emptying rate and intestinal propulsion proportion were evaluated after gastric gavage of Carbo Medicinalisblood was sampled from abdominal aortic to detect rat plasma motilin MTL substance PSP and somatostatinsswith radioimmunity. Colorimetric assay was used for the determination of ATP content of rat jejunum epithelial cellsand immunohistochemical method was used to detect the expression of myosin light chain kinasemlckin rat gastric antrum smooth muscle cells and c-kit in colonic tissues. Results Compared with the model groupthe gastric emptying rate was decreased and intestinal propulsion proportion was increased significantly in the normal groupraw RA groupbran-fried RA groupand domperidone groupwhile the plasma MTLSP and SS contents were increased to different degreesand ATP content of jejunum epithelial cellsmlck expression of rat gastric antrum smooth muscle cells and c-kit expression of colonic tissues were also increasedthe differences being statistically significantp < 0.05P < 0.01. Compared with raw RA group bran-fried RA group had more obvious effectand the differences between the two groups were statistically significantp < 0.05P < 0.01. Conclusion RA extract can exerts an overall improvement of rat gastrointestinal dysfunction induced by spleen deficiencyand the effect of bran-fried RA is superior to that of raw RA group. KeywordsRhizoma Atractylodis extract Fried with branspleen deficiency syndromegastrointestinal function Signal transduction

188 Traditional Chinese Drug Research & Clinical Pharmacology2015 MarchVol. 26 No. 2 1. 4 5 10.0 g kg - 1 10.0 g kg - 1 5.0 [1] mg kg - 1 10 [2] [3] 4 5 3 60 g kg - 1 1 1 15 d 1 1 10 d 1.1 SD 50 200± 1. 5 20 g 1. 5. 1 1 SKSF 20130105 1. 2 5 % 2 % CMC- Na 2.5 ml 500 g 30 min 1 % 1.2 80 % ml 6 ml 12 h 80 % 3 2 h 1 3 500 g L - 1 %= 20130104 100 % 20130508 CMC- Na [I 125 ] MTL PSP SS EDTA- 2Na 20130401201305062013030120130102 20130305 24 h - / 100 % ATP 1. 5. 2 MTL P SP smmlck Cajal SS MTL SP c- kit SP DAB SS 1. 5. 3 ATP 0.5 20130402201305012013060120130204 20130302 cm 0.5 cm 1. 3 GC- 1500 γ ATP UN- 500 1. 5. 4 DG5032 MLCK c- kit UT6062 5 mm 0.8~ 1 cm XSP- 1 4 % 300 - SABC Image- Pro Plus 5.1 Image- Pro Plus 5.1 Motic MLCK

2015 3 26 2 189 c- kit 5 ATP 5 P < 0.05 3 MLCK c- kit 2 M T L S P S S x ± s n = 1 0 1. 6 SPSS17.0 Table 2 Effect of Atractylodes rhizome extract before and after ± x ± s Fried with Bran on Plasma MTLSP and SS content of model rat 2 2. 1 P < ± ± 0.01 P < P < 0.05P < 0.01 0.05 P < 0.01 P > 0.05 P < 0.05 3 A T P x ± s n = 1 0 Table 3 Comparison of ATP content in the jejunum tissue of rats P < 0.05 1 in different groups 1 x ± s n = 10 Table 1 Comparison of gastric emptying and intestinal propulsion of rats in different groups ± ± ± ± ± ± ± ± ± ± ## P < 0.01 * P < 0.05 ** P < 0.01 P < 0.05 P < 2. 2 M T L S P S S MTL SP SS MLCK c- kit P < 0.01 P < 0.01 MLCK MTL SP SS c- kit P < 0.05P < 0.01 P < 0.05P < 0.01 MLCK c- kit SP P < 0.05 MTL SP P < 0.05P < 0.01 c- kit SS P < 0.05P < 0.01 P < 0.05 MTL SP SS MLCK c- kit P < 0.05 P < 0.05P < 0.01 2 1 2 4 2. 3 A T P ATP P < 3 0.01 ± ± ± ± ± ± ± ± ± ± ± ± ± ## P < 0.01 * P < 0.05 ** P < 0.01 P < 0.05 P < 0.01 μ ± ± ± ± ± ## P < 0.01 * P < 0.05 ** P < 0.01 P < 0.05 P < 0.01 P < 0.05 2. 4 M L C K c - k i t MLCK c- kit ATP P < 0.05P < 0.01 ATP P < 0.05P < 0.01

190 Traditional Chinese Drug Research Clinical Pharmacology 2015 March Vol 26 No 2 A. 正常组 图1 B. 模型组 C. 多潘立酮组 D. 生苍术组 E. 麸炒苍术组 大鼠胃窦平滑肌细胞内 M L C K 免疫组化染色图 200 Figure 1 The inimmunohistochemical staining results of MLCK in gastric antrum smooth muscle cells of rats 200 A. 正常组 图2 B. 模型组 C. 多潘立酮组 D. 生苍术组 E. 麸炒苍术组 大鼠结肠 c - k i t 免疫组化染色图 200 Figure 2 The inimmunohistochemical staining results of colonic c- kit of rats 200 表4 对大鼠胃窦平滑肌细胞内 M LC K 结肠组织中 c - k i t 表 达量的影响 x ± s n=10 Table 4 Comparison of the expression of MLCK in gastric antrum smooth muscle cells and c- kit in colon tissue of rats in different groups 剂量 ｇ ｋｇ １ ＭＬＣＫ ｃ ｋｉｔ 正常组 １１ ０２±３ ４９ １６ ２７±４ ０１ 模型组 组别 生苍术组 １０ ０ 麸炒苍术组 １０ ０ 多潘立酮组 ０ ００５ ５ ０７±１ ２９ ９ ０５±２ ６４ １０ ９９±３ １７ ９ １４±２ ７７ 功能低下性疾病都存在脾虚证的证候 运用补脾运脾 法多能奏效 因此 临床上对脾虚证的治疗其重要法 则就是调理脾胃 脾胃功能与胃肠运动关系密切 胃 肠运动对食物的消化和吸收意义重大 目前 文献记 载的制备脾虚证模型的方法很多 如情志失和法 苦 寒泻下法 过度疲倦法 饮食失节法 [6]等 根据李时 ５ ６９±１ ２１ 珍的论述 大黄因其性苦寒 能耗伤元气阴血 损伤 １２ ０７±２ ８７ 脾胃 因此 在大黄苦寒泻下基础上改进的破气苦降 １５ ９８±３ ８１ 加饥饱失常法是目前较普遍运用的一种建模方法 且 ９ ０１±２ ３２ 注 与正常组比较 ##P 0.01 与模型组比较 *P 0.05 **P 0.01 与多潘立酮组比较 P 0.05 P 0.01 与生苍术组比较 P 0.05 简单易行 有研究报道[7]该种造模方法所造成的脾虚 证模型症状与临床所见的脾虚症表现非常相似 本实 验亦利用此法 用含有大黄药味的小承气汤耗伤大鼠 脾胃元气以致脾虚 究方面开展的工作还很少 而临床研究却发现苍术具 研究[8]表明 脾虚证与胃肠炎症 胃肠道的神经 有很好的调理胃肠功能 促进胃肠动力 防治胃肠慢 肌肉调控系统 胃肠激素分泌障碍等多种病理生理改 性炎症及溃疡性胃肠疾病的作用 [4] 目前 临床应用 变都有关系 其主要临床表现为胃排空延迟和小肠传 得较多的是麸炒苍术 一般认为苍术麸炒后燥性会降 输功能障碍 本研究选取胃排空率及小肠推进比为指 低 而健脾作用则会增强 但尚缺乏对其确切机制的 标 观察苍术对脾虚大鼠胃肠功能的影响 实验中观 进一步研究 有文献[4]报道 麸炒法可改变包括挥发 察到脾虚模型大鼠胃排空及小肠推进率降低明显 油在内的苍术的多种化学成分 本研究从胃肠动力学 MTL 是一种很重要的胃肠激素 具有促进胃肠运动 方面对苍术麸炒前后防治脾虚证的作用进行比较 以 增强胃肠道收缩力和张力的作用 SP 是一种神经元 阐明麸炒法是否可增强苍术的健脾作用及其可能的作 递质 具有兴奋胃肠运动的功能 在整个胃肠道和肠 用机制 神经系统都有广泛的分布 对几乎所有的消化道平滑 脾虚证以运动 消化吸收功能障碍作为主要表 肌都具有刺激其收缩的作用 [9] SS 可调节胃肠蠕动 现 是一个多器官多系统功能障碍的综合性症候群 通过抑制胃肠道激素和乙酰胆碱而起到间接调整胃肠 其中 消化系统障碍是这一症候群的中心 有研究 [5] 道运动的作用[10] 三者均能很好的体现胃肠功能的变 显示 绝大部分的消化系统疾病以及能量代谢或免疫 化 实验表明 脾虚可使上述胃肠激素表达明显异

2015 3 26 2 191 ICC c- kit [16] ICC c- kit - c- kit [11] ATP ICC ATP [12] ATP ATP ATP ATP Ca 2+ CaM [13] Ca 2+ [14] Ca 2+ - CaM- MLCK MLCK MLCK MLCK Ca 2+ - CaM- MLCK Caja ICC ICC [15] ICC ICC C- kit ICC ICC