针刺“相对穴”对MC形态变化及释放HIS、BK和SP影响的研究
发布时间：2019-01-24 12:59
【摘要】：目的“相对穴”是身体各部阴阳相对的两个腧穴,同时取用,发挥协同增效作用。本研究观察针刺大鼠“相对穴”阴陵泉与阳陵泉穴区局部肥大细胞(Mast cell,MC)趋经穴募集、脱颗粒情况、P物质(Substance P,SP)表达水平及MC释放到血清中SP、组织胺(Histamine,HIS)、缓激肽(Bradykinin,BK)的含量变化,初步探究“相对穴”配穴针刺效应产生的机制,为“相对穴”配穴理论的临床应用提供一定的实验基础。方法40只Wistar大鼠随机分成空白对照组(K组)、针刺阳陵泉穴组(A组)、针刺阴陵泉穴组(B组)、针刺“相对穴”阴陵泉与阳陵泉穴组(AB组)及针刺旁开阴陵泉与阳陵泉穴组(CD组)5组,每组各8只。空白对照组与其它各组同样的方法和时间固定,不给予其它任何刺激;针刺组给予电针刺激,隔日一次,第4次干预完成后2小时取材。采用酶联免疫检测技术(Enzyme-Linked Immunosorbent assay,ELISA)检测血清中的SP、HIS和BK的含量变化;穴区局部组织采用免疫组化技术(Immunohistochemistry,IHC),做冰冻切片观察穴区MC形态及结构的变化;做石蜡切片观察穴区局部组织SP的表达水平变化,并用Image-Pro Plus 6.0图像分析软件定量计算穴区局部SP的阳性表达光密度值。结果1各组大鼠穴区局部MC数量、脱颗粒率比较:各组大鼠的穴区局部MC数及脱颗粒率依次为:AB组最高,A组、B组次之,K组、CD组最低。与K组、CD组相比,其他各针刺组穴区局部MC数及脱颗粒率显著升高,其中AB组与各组比较差异最为显著(P0.01);A组与B组相比、K组与CD相比,差异没有统计学意义(P0.05)。2各组大鼠穴区局部和血清中SP含量比较:各组大鼠穴区局部和血清中SP含量依次为:AB组最多,A组、B组次之,K组、CD组最少。与K组相比,各针刺组穴区局部SP阳性表达水平和血清中含量均有不同程度的增多;各针刺组比较,CD组SP表达水平显著低于A、B、AB各针刺组(P0.01);A组与B组相比,差异没有统计学意义(P0.05)。3各组大鼠血清中HIS、BK的含量比较:HIS含量检测结果为,与K组相比,各针刺组含量均显著高于K组(P0.05);各针刺组相比较,AB组的HIS含量最高;A组的含量低于B组(P0.05);CD组的含量最低,各组之间相比,差异均有统计学意义(P0.05,P0.01);BK含量检测结果为:各针刺组BK含量均显著高于K组(P0.05),其中AB组的含量最高;A组与B组相比,无显著性差异(P0.05);CD组分别和A组、B组及AB组相比较,CD组的BK含量最低,差异有统计学意义(P0.05)。结论穴区局部MC趋经穴募集、脱颗粒率和SP表达水平升高及血清中SP、HIS、BK的含量升高是“相对穴”配穴协同增效的针刺效应机制的可能因素,同时提示我们可以将穴区MC募集、脱颗粒及释放HIS、BK含量的变化及SP的阳性表达水平作为量化针刺效应的参考指标之一,并为临床判断针效的指标选取提供一定的实验和理论基础。
[Abstract]:Objective "relative acupoints" are two acupoints of relative yin and yang in different parts of body, which can be used simultaneously and exert synergistic effect. The aim of this study was to observe the tendency of meridian recruitment and degranulation of mast cells (Mast cell,MC) in the "relative acupoints" of "relative points" and Yanglingquan acupoints in rats, the expression of substance P (Substance) SP and the release of MC into the serum of SP, histamine (Histamine,). The content of HIS), bradykinin (Bradykinin,BK) was changed and the mechanism of acupuncture effect of "relative acupoint" combined with acupoint was preliminarily explored, which provided a certain experimental basis for clinical application of "relative acupoint" point matching theory. Methods Forty Wistar rats were randomly divided into control group (group K), acupuncture group (group A) and acupuncture group (group B). There were 8 rats in each group in acupuncture group (group AB) and group B (CD group) with "relative acupoint" and "relative acupoint" (AB group) and 5 groups (group CD) with acupuncture at Kaiyingquan and Yanglingquan acupoints (group CD). The blank control group was fixed with the same method and time as the other groups, and the acupuncture group was given electroacupuncture stimulation once every other day, 2 hours after the 4th intervention. Enzyme linked immunosorbent assay (Enzyme-Linked Immunosorbent assay,ELISA) was used to detect the changes of SP,HIS and BK in serum, and the changes of MC morphology and structure were observed by using immunohistochemical technique (Immunohistochemistry,IHC). The expression level of SP in local tissue of acupoint was observed by paraffin section, and the optical density of positive expression of SP in local area of acupoint was calculated quantitatively by Image-Pro Plus 6.0 image analysis software. Results 1The number of local MC and the rate of degranulation: the number of local MC and the rate of degranulation were the highest in group AB, group A, group B, group K and group CD were the lowest. Compared with group K and group CD, the number of local MC and the rate of degranulation in other acupuncture groups were significantly higher than those in group K, and the difference between AB group and each group was most significant (P0.01). Compared with group A and group B, there was no significant difference between group K and CD (P0.05). 2 the content of SP in acupoint area and serum of rats in each group was higher than that in group A and group B (P0.05). The content of SP in group A was the highest in group A, and in group A, the content of SP in group A was the highest in group A. Group B followed, group K and group CD were the least. Compared with group K, the positive expression level of SP and the content of serum in each acupuncture group were increased in different degrees, and the SP expression level in CD group was significantly lower than that in each acupuncture group (P0.01). There was no significant difference between group A and group B (P0.05). 3 comparison of HIS,BK content in serum of rats in each group: the results showed that the content of HIS in group A was significantly higher than that in group K (P0.05). The content of HIS in group A was lower than that in group B (P0.05); CD), the difference was statistically significant (P0.05, P0.01). The BK content of each acupuncture group was significantly higher than that of K group (P0.05), the content of AB was the highest in group A and group B, there was no significant difference between group A and group B (P0.05). Compared with group A, group B and group AB, the content of BK in CD group was the lowest, and the difference was statistically significant (P0.05). Conclusion the increase of local MC recruitment, degranulation rate and SP expression level and the increase of SP,HIS,BK content in serum are the possible factors of acupuncture effect mechanism of "relative acupoint" combined with acupoints. At the same time, the changes of MC recruitment, degranulation and release of HIS,BK and the positive expression of SP in the acupoint area can be regarded as one of the reference indexes of the quantitative acupuncture effect. It also provides a certain experimental and theoretical basis for clinical evaluation of needle-effect index selection.
【学位授予单位】：华北理工大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R245
[Abstract]:Objective "relative acupoints" are two acupoints of relative yin and yang in different parts of body, which can be used simultaneously and exert synergistic effect. The aim of this study was to observe the tendency of meridian recruitment and degranulation of mast cells (Mast cell,MC) in the "relative acupoints" of "relative points" and Yanglingquan acupoints in rats, the expression of substance P (Substance) SP and the release of MC into the serum of SP, histamine (Histamine,). The content of HIS), bradykinin (Bradykinin,BK) was changed and the mechanism of acupuncture effect of "relative acupoint" combined with acupoint was preliminarily explored, which provided a certain experimental basis for clinical application of "relative acupoint" point matching theory. Methods Forty Wistar rats were randomly divided into control group (group K), acupuncture group (group A) and acupuncture group (group B). There were 8 rats in each group in acupuncture group (group AB) and group B (CD group) with "relative acupoint" and "relative acupoint" (AB group) and 5 groups (group CD) with acupuncture at Kaiyingquan and Yanglingquan acupoints (group CD). The blank control group was fixed with the same method and time as the other groups, and the acupuncture group was given electroacupuncture stimulation once every other day, 2 hours after the 4th intervention. Enzyme linked immunosorbent assay (Enzyme-Linked Immunosorbent assay,ELISA) was used to detect the changes of SP,HIS and BK in serum, and the changes of MC morphology and structure were observed by using immunohistochemical technique (Immunohistochemistry,IHC). The expression level of SP in local tissue of acupoint was observed by paraffin section, and the optical density of positive expression of SP in local area of acupoint was calculated quantitatively by Image-Pro Plus 6.0 image analysis software. Results 1The number of local MC and the rate of degranulation: the number of local MC and the rate of degranulation were the highest in group AB, group A, group B, group K and group CD were the lowest. Compared with group K and group CD, the number of local MC and the rate of degranulation in other acupuncture groups were significantly higher than those in group K, and the difference between AB group and each group was most significant (P0.01). Compared with group A and group B, there was no significant difference between group K and CD (P0.05). 2 the content of SP in acupoint area and serum of rats in each group was higher than that in group A and group B (P0.05). The content of SP in group A was the highest in group A, and in group A, the content of SP in group A was the highest in group A. Group B followed, group K and group CD were the least. Compared with group K, the positive expression level of SP and the content of serum in each acupuncture group were increased in different degrees, and the SP expression level in CD group was significantly lower than that in each acupuncture group (P0.01). There was no significant difference between group A and group B (P0.05). 3 comparison of HIS,BK content in serum of rats in each group: the results showed that the content of HIS in group A was significantly higher than that in group K (P0.05). The content of HIS in group A was lower than that in group B (P0.05); CD), the difference was statistically significant (P0.05, P0.01). The BK content of each acupuncture group was significantly higher than that of K group (P0.05), the content of AB was the highest in group A and group B, there was no significant difference between group A and group B (P0.05). Compared with group A, group B and group AB, the content of BK in CD group was the lowest, and the difference was statistically significant (P0.05). Conclusion the increase of local MC recruitment, degranulation rate and SP expression level and the increase of SP,HIS,BK content in serum are the possible factors of acupuncture effect mechanism of "relative acupoint" combined with acupoints. At the same time, the changes of MC recruitment, degranulation and release of HIS,BK and the positive expression of SP in the acupoint area can be regarded as one of the reference indexes of the quantitative acupuncture effect. It also provides a certain experimental and theoretical basis for clinical evaluation of needle-effect index selection.
【学位授予单位】：华北理工大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R245
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