針刺“相對(duì)穴”對(duì)MC形態(tài)變化及釋放HIS、BK和SP影響的研究
發(fā)布時(shí)間：2019-01-24 12:59
【摘要】：目的“相對(duì)穴”是身體各部陰陽(yáng)相對(duì)的兩個(gè)腧穴,同時(shí)取用,發(fā)揮協(xié)同增效作用。本研究觀察針刺大鼠“相對(duì)穴”陰陵泉與陽(yáng)陵泉穴區(qū)局部肥大細(xì)胞(Mast cell,MC)趨經(jīng)穴募集、脫顆粒情況、P物質(zhì)(Substance P,SP)表達(dá)水平及MC釋放到血清中SP、組織胺(Histamine,HIS)、緩激肽(Bradykinin,BK)的含量變化,初步探究“相對(duì)穴”配穴針刺效應(yīng)產(chǎn)生的機(jī)制,為“相對(duì)穴”配穴理論的臨床應(yīng)用提供一定的實(shí)驗(yàn)基礎(chǔ)。方法40只Wistar大鼠隨機(jī)分成空白對(duì)照組(K組)、針刺陽(yáng)陵泉穴組(A組)、針刺陰陵泉穴組(B組)、針刺“相對(duì)穴”陰陵泉與陽(yáng)陵泉穴組(AB組)及針刺旁開陰陵泉與陽(yáng)陵泉穴組(CD組)5組,每組各8只�？瞻讓�(duì)照組與其它各組同樣的方法和時(shí)間固定,不給予其它任何刺激;針刺組給予電針刺激,隔日一次,第4次干預(yù)完成后2小時(shí)取材。采用酶聯(lián)免疫檢測(cè)技術(shù)(Enzyme-Linked Immunosorbent assay,ELISA)檢測(cè)血清中的SP、HIS和BK的含量變化;穴區(qū)局部組織采用免疫組化技術(shù)(Immunohistochemistry,IHC),做冰凍切片觀察穴區(qū)MC形態(tài)及結(jié)構(gòu)的變化;做石蠟切片觀察穴區(qū)局部組織SP的表達(dá)水平變化,并用Image-Pro Plus 6.0圖像分析軟件定量計(jì)算穴區(qū)局部SP的陽(yáng)性表達(dá)光密度值。結(jié)果1各組大鼠穴區(qū)局部MC數(shù)量、脫顆粒率比較:各組大鼠的穴區(qū)局部MC數(shù)及脫顆粒率依次為:AB組最高,A組、B組次之,K組、CD組最低。與K組、CD組相比,其他各針刺組穴區(qū)局部MC數(shù)及脫顆粒率顯著升高,其中AB組與各組比較差異最為顯著(P0.01);A組與B組相比、K組與CD相比,差異沒有統(tǒng)計(jì)學(xué)意義(P0.05)。2各組大鼠穴區(qū)局部和血清中SP含量比較:各組大鼠穴區(qū)局部和血清中SP含量依次為:AB組最多,A組、B組次之,K組、CD組最少。與K組相比,各針刺組穴區(qū)局部SP陽(yáng)性表達(dá)水平和血清中含量均有不同程度的增多;各針刺組比較,CD組SP表達(dá)水平顯著低于A、B、AB各針刺組(P0.01);A組與B組相比,差異沒有統(tǒng)計(jì)學(xué)意義(P0.05)。3各組大鼠血清中HIS、BK的含量比較:HIS含量檢測(cè)結(jié)果為,與K組相比,各針刺組含量均顯著高于K組(P0.05);各針刺組相比較,AB組的HIS含量最高;A組的含量低于B組(P0.05);CD組的含量最低,各組之間相比,差異均有統(tǒng)計(jì)學(xué)意義(P0.05,P0.01);BK含量檢測(cè)結(jié)果為:各針刺組BK含量均顯著高于K組(P0.05),其中AB組的含量最高;A組與B組相比,無(wú)顯著性差異(P0.05);CD組分別和A組、B組及AB組相比較,CD組的BK含量最低,差異有統(tǒng)計(jì)學(xué)意義(P0.05)。結(jié)論穴區(qū)局部MC趨經(jīng)穴募集、脫顆粒率和SP表達(dá)水平升高及血清中SP、HIS、BK的含量升高是“相對(duì)穴”配穴協(xié)同增效的針刺效應(yīng)機(jī)制的可能因素,同時(shí)提示我們可以將穴區(qū)MC募集、脫顆粒及釋放HIS、BK含量的變化及SP的陽(yáng)性表達(dá)水平作為量化針刺效應(yīng)的參考指標(biāo)之一,并為臨床判斷針效的指標(biāo)選取提供一定的實(shí)驗(yàn)和理論基礎(chǔ)。
[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.
【學(xué)位授予單位】：華北理工大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：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.
【學(xué)位授予單位】：華北理工大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R245
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