Ya-Ping Lin

Effects of electroacupuncture on gastric myoelectric activity and substance P in the dorsal vagal complex of rats

Acupuncture has been widely applied to treat gastrointestinal diseases in the clinic. However, its precise mechanism remains unknown. In the present study, effects of electroacupuncture (EA) at Zusanli acupoints (ST36) on gastric myoelectric activity and substance P (SP) in the dorsal vagal complex (DVC) were observed. EA at ST36 significantly increased the frequency of slow waves and the frequency of the spike potentials superimposed on the slow waves, which persisted for 90 min after cessation of EA and was completely abolished after bilateral vagotomy. Meanwhile, the mean optical density of SP immunoreactive fibers in the DVC increased following EA. The results suggested that EA at ST36 promoted the gastric myoelectric activity, which was regulated by the vagus, and SP in the DVC may be involved in the excitatory effects. Furthermore, EA had a relative long-term post-effect. We presumed that DVC may be a supraspinal center in the effect of EA on gastric function.

Effect of acupuncture plus mild hypothermia on MAPK/ERK pathway of brain tissues in rats with cerebral ischemia-reperfusion injury

ObjectiveTo observe the protective effect of acupuncture plus mild hypothermia on brain tissues in rats with cerebral ischemia-reperfusion injury (CIRI), and the influence on protein expression levels of phosphorylated Raf-1, MEK-2 and ERK1/2 in the mitogen-activated protein kinase (MAPK)/extracellular regulated protein kinases (ERK) pathway, and to explore the mechanism of acupuncture plus mild hypothermia therapy for the ischemic stroke.MethodsNinety Sprague-Dawley (SD) rats were randomly divided into a blank control group, a sham operation group, a model group, an acupuncture group, a mild hypothermia group and an acupuncture plus mild hypothermia group, 15 rats in each group. Except the rats in the blank control group, the remaining rats were used to prepare the middle cerebral artery occlusion (MCAO) models according to the modified occlusion method using lines, while only the occlusion lines were inserted without blocking the brain arteries of rats in the sham operation group. When the vital signs of rats were stable, rats in the blank control group did not receive any intervention; rats in the sham operation group and the model group received fastening without treatment; rats in the acupuncture group, the mild hypothermia group, and the acupuncture plus mild hypothermia group were treated with the corresponding therapeutic methods. 72 h later, observed neurologic injury score, evaluated infarction area ratio by 2,3,5-tripheyl tetrazolium chloride (TTC) staining, determined apoptosis by TUNEL assay, and measured the phosphorylated Raf-1, MEK-2 and ERK1/2 protein expression levels in rat ischemic hippocampal tissues by Western blot assay.ResultsCompared with the blank control group and the sham operation group, after modeling, the neurologic injury score, infarction area ratio and apoptotic cells were increased, and phosphorylated Raf-1, MEK-2 and ERK1/2 protein expression levels were significantly increased in the model group; the differences were statistically significant (P<0.05 or P<0.01). Compared with the model group, after acupuncture or mild hypothermia therapy, neurologic injury score and infarction area ratio were decreased; apoptotic cells and phosphorylated Raf-1, MEK-2 and ERK1/2 protein expression levels were significantly decreased; the differences were statistically significant (P<0.05 or P<0.01). Compared with the acupuncture group, neurologic injury score and phosphorylated Raf-1, MEK-2 and ERK1/2 protein expression levels were decreased in the acupuncture plus mild hypothermia group; differences between the groups were statistically significant (P<0.05 or P<0.01). Compared with the mild hypothermia group, phosphorylated Raf-1, MEK-2 and ERK1/2 protein expression levels decreased in the acupuncture plus mild hypothermia group, and differences were statistically significant (P<0.01).ConclusionAcupuncture or mild hypothermia therapy can improve neurologic injury, reduce infarction area and apoptosis, which brought about protective effect on the brain tissues, in the MCAO model. The protective effect of acupuncture plus mild hypothermia group is the strongest. The mechanism may involve the MAPK/ERK pathway, by reducing the phosphorylated Raf-1, MEK-2 and ERK1/2 protein expression levels.摘要目的观察针刺联合亚低温疗法对脑缺血再灌注大鼠脑组织的保护作用, 以及对丝裂原激活的蛋白激酶 (mitogen-activated protein kinase, MAPK)/细胞外调节蛋白激酶(extracellular regulated protein kinases, ERK)通路上磷酸化Raf-1、MEK-2、ERK1/2 蛋白表达水平的影响, 探讨针刺联合亚低温疗法干预缺血性中风的机制。方法将90 只Sprague-Dawley (SD)大鼠随机分为空白组、假手术组、模型组、针刺组、亚低温组和针刺联合亚低温组, 每 组15 只。随机挑选15 只为空白组, 其余大鼠根据线拴法并改良以复制大脑中动脉缺血(middle cerebral artery occlusion, MCAO)模型, 假手术组只插入线栓不阻断大脑中动脉。待大鼠生命体征稳定后, 空白组不做任何处理, 假手术组和模型组只捆绑不治疗, 针刺组、亚低温组、针刺联合亚低温组进行相应的处理。72 h 后, 观察神经功 能缺损评分, 氯化三苯基四氮唑(2,3,5-tripheyl tetrazolium chloride, TTC)染色检测梗死面积比, TUNEL法检测凋亡细 胞, Western blot 法检测大鼠缺血侧海马组织磷酸化Raf-1、MEK-2、ERK1/2 蛋白的表达水平。结果造模后, 模 型组大鼠神经功能缺损评分、梗死面积比值及凋亡细胞增多, 磷酸化Raf-1、MEK-2、ERK1/2 蛋白的表达水平明显 增高, 与空白组及假手术组差异有统计学意义(P<0.05 或P<0.01)。经针刺及亚低温治疗后, 各治疗组神经功能 缺损评分、梗死面积比值减少, 凋亡细胞及磷酸化Raf-1、MEK-2、ERK1/2 蛋白的表达水平均明显降低, 与模型组 差异均有统计学意义(P<0.05 或P<0.01)。与针刺组比较, 针刺联合亚低温组神经功能缺损评分, 磷酸化Raf-1 蛋 白激酶(Raf-1)、丝裂原活化蛋白激酶-2 (mitogen-activated protein kinase-2, MEK-2)、细胞外信号调节激酶1/2 (extracellular signal regulated kinase1/2, ERK1/2)蛋白的表达水平下降, 组间差异有统计学意义(P<0.05 或P<0.01)。 与亚低温组比较, 针刺联合亚低温组磷酸化Raf-1、MEK-2、ERK1/2 蛋白的表达水平下降, 组间差异有显著统计学 意义(P<0.01)。结论针刺及亚低温疗法均可改善MCAO 模型大鼠的神经功能缺损, 减少脑梗死面积及细胞凋亡, 对脑组织起保护作用, 并且针刺联合亚低温组的保护作用较强。其机制可能是通过MAPK/ERK 通路, 降低磷酸化 Raf-1、MEK-2、ERK1/2 蛋白的表达水平。

The Effects of Moxibustion on Serum Interleukins and T Cell Subset in H.polyri Infected Rats

Abstract Objective: to observe the effects of moxibustion on histomorphological changes of gastric mucosa, as well as on serum IL-6、IL-8、TNF-α，Hp IgG、CD3+、CD4+、CD8+ in helicobacter pylori (Hp) infected rats, so that to better understand how the moxibustion repairs the Hp- induced gastric mucosal injury. Methods: 40 SD rats were randomly assigned to four groups: group A (blank control), group B (Hp infection model), group C (moxibustion plus model), group D (electro-acupuncture plus model), 10 for each group. The “NaHCO3 plus Indometacin and Hp intragastric administration” method was employed to make gastritis model. Acupoints selected for “repair” purpose were Zu San Li (ST36), Zhong Wan (CV12), Guan Yuan (RN4), Pi Shu (BL20), Wei Shu(BL21). The histomorphological changes of gastric mucosa in rats were observed under light microscope after HE stain; IL-6, IL-8, TNF-α, Hp IgG values were evaluated by ELISA method; values of CD3+、CD4+、CD8+、CD4+/CD8+ were measured by flow cytometry method. Results: compared with group A, the values of IL-6, IL-8, TNF-α, Hp IgG and CD8+ in group B were increased（P＜0.01）, whereas the values of CD3+、CD4+、CD4+/CD8+ were decreased（P＜0.01）. Compared with group B, the values of IL-8（P＜0.05）,TNF-α（P＜0.05）, IL-6（P＜0.01）, Hp IgG（P＜0.01） and CD8+ （P＜0.05） in group C were decreased, whereas the values of CD3+（P＜0.05）,CD4+（P＜0.05）,CD4+/CD8+ （P＜0.05） were increased, meanwhile such values in group D had no significant changes. Compared with group D, the values of IL-6（P＜0.05）,IL-8 （P＜0.05）and Hp IgG （P＜0.01）in group C were decreased, whereas CD4+/CD8+（P＜0.05）were increased, all those changes had statistical significance. Conclusion: the preventive and therapeutic effects of moxibustion on Hp related gastritis might be realized by two ways- to inhibit the secretion of proinflammatory cytokines such as IL-6, IL-8 and TNF-α, or to regulate the production of immune factors (such as up-regulation of CD3+, CD4+ and down-regulation of CD8+).

Effect of moxibustion on the motility, absorption and activities of ATPase in small intestine of spleen-deficiency rats

ObjectiveTo observe the effect of moxibustion on intestinal propulsion rate, content of serum D-xylose, content of adenosine triphosphate (ATP) and activities of adenosine triphosphatases (ATPase) in jejunum tissues of spleen-deficiency rats, and to explore the effect of moxibustion on the production of ATP and activities of membrane proteins, for analyzing the mechanism of moxibustion in reinforcing the spleen-stomach.MethodsForty healthy Sprague Dawley rats were randomly divided into four groups, namely group A (blank control group), group B (model control group), group C (moxibustion group) and group D (herbs group). The animal model of spleen-deficiency was established by intragastric administration with Da Huang (Radix et Rhizoma Rhei) infusion. The rats in group C received moxibustion at Zusanli (ST 36), Zhongwan (CV 12), Guanyuan (CV 4), Pishu (BL 20) and Weishu (BL 21). The rats in group D received intragastric administration with Si Jun Zi decoction. The intestinal propulsion was measured by toner method, and the content of D-xylose in serum was detected by phloroglucinol method. Colorimetry method was used to detect the content of ATP and activities of Na+-K+-ATPase and Ca2+-Mg2+-ATPase in jejunum tissues.ResultsCompared with group A, the intestinal propulsion rate, content of D-xylose in serum, content of ATP, activities of Na+-K+-ATPase and Ca2+-Mg2+-ATP in jejunum tissues decreased significantly in group B (P<0.05 or P<0.01). Compared with group B, the intestinal propulsion rate, content of D-xylose in serum, content of ATP, activities of Na+-K+-ATPase and Ca2+-Mg2+-ATPase in jejunum tissues increased significantly in group C and D (P<0.05 or P<0.01), while there were no significant differences between group C and group D.ConclusionMoxibustion can increase the intestinal propulsion rate, content of D-xylose in serum and ATP in jejunum tissues, as well as the activities of Na+-K+-ATPase and Ca2+-Mg2+-ATPase of spleen-deficiency rats, which suggests that moxibustion can enhance the motility and absorption functions of small intestine by promoting the production of ATP in intestinal epithelial cells, increasing the content of membrane proteins, and improving transmembrane transport.