电针不同穴位对实验性类痛经大鼠镇痛效应及其机理的研究
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摘要
电针不同穴位对实验性类痛经大鼠镇痛效应及其机理的研究
针灸学认为刺激穴位可特异性地对相应脏腑器官产生调治作用,这种作用与非穴和其它经穴比较存在差异。这种差异就是经穴效应的特异性。经穴效应特异性是经络学说的核心内容,是针灸临床循经取穴的重要依据。而且大量临床实践及实验研究都证实穴位效应确实存在特异性。
现代神经科学对针灸机理的研究结果已显示经穴效应存在特异性。但这种特异性的产生主要是由于支配穴位所在部位的神经节段的不同而引起的。它主要与穴位所在的部位有关,而与经典针灸理论所强调的穴位所处的“经”关系不大。即处于同神经节段的不同经脉上的穴位、相同经脉上的不同穴位以及与非穴位间的效应不存在差异。那么,经穴效应特异性到底为何?同神经节段内非穴位以及不同经脉上的穴位间是否存在效应特异性?相同经脉上不同穴位间的效应又如何呢?
本研究以实验性类痛经大鼠模型为研究对象,从电针镇痛作用及机制入手,观察电针同神经节段支配的胞宫相关经穴、非相关经穴以及非经非穴对实验性类痛经大鼠的行为学反应、子宫平滑肌收缩力、子宫组织局部致痛物质以及中枢痛觉调制系统的影响,探讨同神经节段支配的经穴与非经穴、相关经穴与非相关经穴,以及同一经脉不同穴位间是否存在经穴效应特异性以及经穴效应特异性是否具有一定的规律。为经穴效应特异性的研究提供系统有力的实验依据。
实验中所选经穴为：相关经穴为足太阴脾经三阴交、血海穴；非相关经穴为足少阳胆经悬钟穴；非经非穴为胆经与胃经之间平悬钟穴处的非经非穴点。
实验选用动情间期SD雌性大鼠156只,按照随机数字法分为盐水组、模型组、电针三阴交组、电针血海组、电针悬钟组、电针非穴组6组,每组26只。除盐水组外,其余各组大鼠均皮下注射苯甲酸雌二醇连续10天,第1、10天每只大鼠皮下注射0.5 mg,第2-9天每只均皮下注射0.2mg。末次给药1h后,腹腔注射缩宫素2u／只。盐水组每日给予同等剂量的生理盐水。根据Schmauss行为学评分标准,记录20分钟内扭体反应的潜伏期、扭体评分及扭体次数；应用BL-420E+生物机能实验系统记录仪记录20分钟内大鼠子宫收缩次数和强度；采用放射免疫法检测子宫前列腺素E2(PGE2)、前列腺素F2α(PGF2α)含量；采用免疫组化法检测脊髓背角内K-阿片受体表达；采用ELISA法定量检测中脑中央导水管周围灰质(PAG)中脑啡肽(ENK)、β内啡肽(β-EP)、强啡肽(Dyn)、内吗啡肽(EM)的含量。实验结果如下：
1电针不同经穴对大鼠行为学反应的影响
模型组与盐水组相比：扭体潜伏期明显缩短,扭体评分、扭体次数明显增高,均有统计学意义(P＜0.01)；说明模型制备成功。
三阴交组扭体潜伏期与模型组相比明显延长,差异有统计学意义(P＜0.05),其余各组扭体潜伏期与模型组相比亦延长,但差异无统计学意义(P＞0.05)；扭体评分和次数各组与模型组相比均明显减少,差异有统计学意义(P＜0.01)。但三阴交组扭体评分与盐水组相比差异亦无统计学意义(P＞0.05)。说明电针不同穴位后,可减轻实验性类痛经大鼠的扭体反应,其中电针三阴交穴的效应最佳,而血海穴、悬钟穴与非穴间的效应无明显差异。
2电针不同经穴对大鼠子宫收缩程度的影响
模型组与盐水组相比：子宫收缩次数明显增加,子宫收缩强度明显增强,差异均有统计学意义(P＜0.01)；说明造模后子宫平滑肌出现痉挛性收缩。
三阴交组和血海组的子宫收缩次数与模型组相比均减少,差异有统计学意义(P＜0.05),悬钟组和非穴组的子宫收缩次数与模型组相比亦减少,但差异无统计学意义(乃0.05);三阴交组的子宫收缩强度与模型组相比明显降低,差异有统计学意义(P＜0.05),其余各组的子宫收缩强度与模型组相比亦降低,但差异无统计学意义(P＞0.05)。除非穴组外,其余各组的子宫收缩强度与盐水组相比,差异均无统计学意义(P＞0.05)。结果说明：电针穴位后,可调节实验性类痛经大鼠子宫痉挛性收缩程度,从而缓解痛反应。其中,电针三阴交穴的效应最佳,血海穴和悬钟穴亦有一定的缓解效应,血海穴较悬钟穴效应稍佳。非穴的效应无明显改变。
3电针不同经穴对大鼠子宫PGE2、PGF2α含量及PGE2／PGF2α比值的影响
模型组与盐水组相比：PGF2α含量明显增高,PGE2含量明显降低,比值明显升高,差异均有统计学意义(P<0.01)；说明造模后,子宫组织中致痛物质含量升高。
各电针组PGF2α含量与模型组相比均明显降低,差异有统计学意义(三阴交：P<0.05；其余各组：P<0.01)。三阴交组PGE2含量与模型组相比明显升高,差异有统计学意义(P<0.01),且与盐水组相比差异无统计学意义(P>0.05),三阴交组PGE2含量比其它各组均明显升高,差异有统计学意义(P＜0.01)；其余各组PGE2含量与模型组相比均无明显升高,差异无统计学意义(P＞0.05)。各电针组比值与模型组相比均明显降低,差异有统计学意义(P＜0.01),且三阴交和悬钟组比值与盐水组相比差异亦无统计学意义(P＞0.05)。结果说明：电针穴位后,可通过调节实验性类痛经大鼠子宫组织致痛物质的含量而达到镇痛作用。综合效应来看,三阴交组的调节作用较强,其余各组间效应无明显差异。
4电针不同经穴对大鼠各节段脊髓背角K-阿片受体表达的影响
针刺不同穴位及非穴对各脊髓节段K-阿片受体表达的影响不同。T13节段：
模型组与盐水组相比：IOD值升高,但差异无统计学意义(P＞0.05)
三阴交和血海组的IOD值与模型组相比明显升高,差异有统计学意义(P<0.01)；悬钟和非穴组的IOD值与模型组相比亦有升高,但差异无统计学意义(P＞0.05)。三阴交和血海组的IOD值与非穴相比明显升高,差异有统计学意义(P＜0.01)。
L1节段：
模型组与盐水组相比：IOD值升高,但差异无统计学意义(P＞0.05)
三阴交和血海组的IOD值与模型组相比明显升高,差异有统计学意义(P<0.01)；悬钟和非穴组的IOD值与模型组相比亦有升高,但差异无统计学意义(P＞0.05)。三阴交和血海组的IOD值与悬钟组和非穴组相比明显升高,差异有统计学意义(P＜0.01)
L2节段：
模型组与盐水组相比：IOD值升高明显,差异有统计学意义(P＜0.01)
三阴交和血海组的IOD值与模型组相比明显升高,差异有统计学意义(P<0.01)；悬钟和非穴组的IOD值与模型组相比亦有升高,但差异无统计学意义(P＞0.05)。三阴交组的IOD值与非穴组相比明显升高,差异有统计学意义(P＜0.01)。三阴交组的IOD值较血海和悬钟组也明显升高,差异有统计学意义(P＜0.01)。
L6节段：
模型组与盐水组相比：IOD值升高,但差异无统计学意义(P＞0.05)
三阴交、血海和非穴组的IOD值与模型组相比明显升高,差异有统计学意义(P＜0.01);悬钟组的IOD值与模型组相比亦有升高,但差异无统计学意义(P＞0.05)。三阴交组的IOD值与非穴组相比升高,差异有统计学意义(P＜0.05)。三阴交和血海组的IOD值均较悬钟组明显升高,差异有统计学意义(P＜0.01)。
S1节段：
模型组与盐水组相比：IOD值升高,但差异无统计学意义(P＞0.05)
三阴交、血海和悬钟组的IOD值与模型组相比明显升高,差异有统计学意义(三阴交：P＜0.01,血海、悬钟：P＜0.05)；非穴组的IOD值与模型组和盐水组相比差异均无统计学意义(P＞0.05)。三阴交组的IOD值与非穴相比明显升高,差异有统计学意义(P＜0.01)。三阴交组的IOD值较血海组亦明显升高,差异有统计学意义(P＜0.05)。
以上结果说明：电针可调节各脊髓节段背角内κ-受体的表达,但不同穴位对不同节段调节的强度不同。三阴交和血海穴在各节段都有调节作用,三阴交穴的调节作用较血海穴明显；悬钟穴和非穴也有一定的调节作用,但作用均较三阴交和血海穴组弱。悬钟和非穴间无明显差异。
5电针不同经穴对PAG内阿片肽物质的影响
5.1电针不同经穴对大鼠PAG内ENK含量的影响：
模型组与盐水组相比：PAG内ENK含量有所升高,但差异无统计学意义(P＞0.05)。说明实验性类痛经模型使大鼠PAG内ENK含量升高不明显。
三阴交和悬钟组的ENK含量与模型组相比明显升高,差异有统计学意义(三阴交：P＜0.01；悬钟组：P＜0.05)；血海和非穴组的ENK含量与模型组相比亦有升高,但差异无统计学意义(P＞0.05)。三阴交组的ENK含量与血海、悬钟和非穴组相比明显升高,差异有统计学意义(血海、非穴：P＜0.01,悬钟：P＜0.05)。悬钟与血海的组间差异无统计学意义(P＞0.05)。说明电针穴位可调节大鼠PAG内ENK含量的水平,但调节程度不同。三阴交和悬钟穴可明显调节PAG内ENK含量,其中以三阴交穴较好。而电针血海穴和非穴无明显变化。
5.2电针不同经穴对大鼠PAG内β-EP含量的影响：
模型组与盐水组相比：PAG内β-EP含量有所升高,但差异无统计学意义(P＞0.05)。说明实验性类痛经模型使大鼠PAG内β-EP含量升高不明显。
三阴交和悬钟组的β-EP含量与模型组相比明显升高,差异有统计学意义(P＜0.01)；血海和非穴组的β-EP含量与模型组相比亦有升高,但差异无统计学意义(P＞0.05)。三阴交和悬钟组β-EP含量与非穴相比均明显升高,差异有统计学意义(三阴交：P＜0.01；悬钟组：P＜0.05)。三阴交与血海和悬钟组相比差异均有统计学意义(P＜0.01)。悬钟与血海组间差异无统计学意义(乃0.05)。说明电针穴位可调节大鼠PAG内β-EP含量的水平,但调节程度不同。三阴交和悬钟穴可明显调节PAG内β-EP含量,其中以三阴交穴较好。而电针血海穴和非穴无明显变化。
5.3电针不同经穴对大鼠PAG内Dyn含量的影响：
模型组与盐水组相比：PAG内Dyn含量有所升高,但差异无统计学意义(P＞0.05)。说明实验性类痛经模型使大鼠PAG内Dyn含量升高不明显。
三阴交和悬钟组的Dyn含量与模型组相比明显升高,差异有统计学意义(P＜0.01)；而血海和非穴组的Dyn含量与模型组相比亦有升高,但差异无统计学意义(P＞0.05)。三阴交组Dyn含量与非穴组相比明显升高,差异有统计学意义(P＜0.01)。三阴交与血海和悬钟组相比差异均有统计学意义(P＜0.01)。悬钟与血海组间差异无统计学意义(P＞0.05)。说明电针穴位可调节大鼠PAG内Dyn含量的水平,但调节程度不同。三阴交和悬钟穴可明显调节PAG内Dyn含量,其中以三阴交穴较好。而电针血海穴和非穴无明显变化。
5.4电针不同经穴对大鼠PAG内EM含量的影响：
模型组与盐水组相比：PAG内EM含量有所升高,但差异无统计学意义(P＞0.05)。说明实验性类痛经模型使大鼠PAG内EM含量升高不明显。
各组EM含量与模型组相比均有升高,但差异无统计学意义(P＞0.05)。说明：针刺各穴位及非穴均无明显调节大鼠PAG内EM含量的作用。
综上所述：电针不同经穴可引起实验性类痛经大鼠PAG内阿片肽含量发生变化,说明针刺可通过调节脑内阿片肽的水平而起到镇痛作用。但电针不同经穴及非穴对PAG内阿片肽含量的影响不同。在电针同神经节段的三个穴位中,三阴交和悬钟穴均可使ENK、β-EP和Dyn的含量升高,三阴交穴的升高效应较明显,悬钟次之,而血海和非穴无明显调节作用；四个穴位对EM的含量均无明显调节作用,说明EM可能不参与电针对内脏痛的调节。
结论
电针同神经节段支配的穴位与非穴位对内脏痛均有镇痛作用,但它们的镇痛效应不同,相关经脉上的特定穴镇痛效应最佳,相关经脉上的非特定穴与非相关经穴效应相近,并且与非穴的效应也相近。针刺穴位可通过调节子宫功能状态和中枢内痛觉调制系统来达到对内脏痛的镇痛作用。并且不同穴位对各部位的调节作用不同。相关经脉上的穴位较之非相关经脉上的穴位以及非穴有更明显的脊髓节段性调节作用,其中以相关经脉中的特定穴作用最强；相关经脉和非相关经脉上的特定穴对脊髓上中枢均有调节作用,但相关经脉上特定穴的调节作用更明显；相关经脉上的非特定穴和非穴无明显调节作用。因此,穴位不同,其调节机制不同,从而产生的效应就不同。由此认为：经穴效应具有特异性,这种特异性是相对的,而不是绝对的。经穴效应特异性虽与其所处的神经节段密切相关,但与其所在的“经”可能有更加密切的联系。
Electroacupuncture Analgesia on Experimental Dysmenorrhea Model Rat and Its Mechanism
The theory of acupuncture considers that stimulating acupoints can regulate the functions of the corresponding viscera specifically, of which is different comparing to the functions regulated by stimulating non-acupoint or acupoints on other meridians. This difference is specificity of the acupoint effect, which is the core content of the meridian theory and an important basis of locating acupoints along meridian in clinical. A large amount of clinical research and experimental study have confirmed the existence of specificity.
The research results of modern neuroscience on acupuncture mechanism also showed the existence of specificity of acupoint effect, which was mainly caused by different nerve segments innervating acupoint area. And it was mostly related to the locations of acupoints but had nothing to do with the meridians. That meant there was no difference found among acupoint effect of different meridians innervated by the same nerve segment or difference between acupoints on the same meridian and non acupoints. And what is specificity of acupoint effect in the end? Is there effect specificity among the different meridian acupoints and non-acupoint innervated by the same nerve segment? How about the effect of different acupoints on the same meridian?
The aim of the present study was to explore whether the specificity of acupoint effects exists and has certain principles or not by comparing the effect and possible mechanism of electroacupuncture analgesia on experimental dysmenorrhea model rat at different acupoints innervated by the same nerve segment.
Selected acupoints for the experiment were as follows:Sanyinjiao(SP 6), Xuehai(SP 10) on the Spleen meridian of Foot Taiyin as relevant acupoints; Xuanzhong(GB 39) on Gallbladder Meridian of Foot Shaoyang as non-relevant acupoints; and non-acupoints located in the middle acupoint between the gallbladder meridian and the stomach meridian at the horizontal line of Xuanzhong(GB 39).
The experiments used 156 female Sprague-Dawley (SD) rats which were during their diestrus. SD rats were randomly divided into 6 groups, which included saline control group, model control group, EA Sanyinjiao group, EA Xuehai group, EA Xuanzhong group, EA non-acupoint group, and 26 rats in each group. Except the saline control group, the rest of groups were injected estradiol benzoate for 10 days, the first and 10th day 0.5 mg for each, and from 2th to 9th days 0.2mg for each.1 hour after the last injection, Oxytocin was given to each by intraperitoneal injection with 2u. The saline control group was given for the same dose of saline everyday. According to the standard of Schmauss behavior score, we recorded latency of writhing, writhing times and writhing score in 20 minutes; applied BL-420E+logger of experimental system to record times and intensity of uterine contraction in 20 minutes; used radio immunoassay to detect the content of PGE2, PGF2a in uterine; observed expression ofκ-opioid receptor in the dorsal horn of the spinal cord by immunohistochemical staining; and detected ENK, B-EP, Dyn, EM content in the periaqueductal gray (PAG) of midbrain by ELISA. The results were as follows:
1 Effect of Electroacupuncture at different acupoints on writhing response in rats
Compared to the saline control group, latency of writhing, writhing times and writhing score of model control group significantly changed (P<0.01), that meant the model was successfully prepared.
All the EA groups comparing to the model control group:writhing latency of EA Sanyinjiao group was significantly prolonged (p<0.05), writhing latency of the other groups was also prolonged, but there was no statistical difference (p>0.05); writhing score and writhing times decreased significantly in each group.(p<0.01).but the writhing score of Sanyinjiao group had no statistical difference comparing to the saline control group (p>0.05).
The results above showed that:After electro-acupuncture treatment on different acupoints, the writhing response of experimental dysmenorrheal model in rats could be relieved at different degrees. There was the best effect in EA Sanyinjiao group, and the differences were not significant among EA Xuehai group, EA Xuanzhong group and EAnon acupoint group.
2 Effect of electroacupuncture on different acupoints about the uterine contraction of rats
Model control group comparing to the saline control group:the time and intensity of uterine contraction significantly increased (p<0.01) which meant the smooth muscle of uterus was induced spasmodic contraction.
the model was successfully prepared.
All the EA groups comparing to model control group:time of uterine contractions were reduced significantly in Sanyinjiao group and Xuehai group (p <0.05); and the intensity of uterine contraction in Sanyinjiao group had a better effect than other groups (p<0.05).
All the EA groups comparing to the saline control group, intensity of uterine contraction hadn't a significant difference in other groups except non-acupoint group (p>0.05).
The results above showed that:acupuncture could improve spasmodic contraction of uterus of experimental dysmenorrhea rat to alleviate pain in the uterus. As a conclusion, the EA Sanyinjiao group had the best effect than others, Xuehai group and Xuanzhong group also had a certain effect, with a better effect in Xuehai group. The effect of non-acupoint hadn't significantly changed.
3 Effect of electroacupuncture on different acupoints on the content of PGE2, PGF2a and the content ratio of PGE2/PGF2a in the rat uterus
Model control group comparing to the saline control group:the content of PGF2a significantly increased, and the content of PGE2 decreased significantly. The both ratio increased significantly (p<0.01); which implied that the content of pain-producing substance in uterine tissue increased after modeling.
All the EA groups comparing to model control group:PGF2a content decreased significantly (Sanyinjiao group:p<0.05; the other groups:p<0.01); PGE2 content in Sanyinjiao group increased more significantly than other groups (p<0.01); and the ratio of PGE2/PGF2a in EA groups significantly decreased (p< 0.01).
All the EA groups comparing to the saline control group:PGE2 content of all groups except Sanyinjiao group significantly decreased (Xuehai group:p<0.05; the other groups:p<0.01); the ratio of PGE2/PGF2a between Sanyinjiao group and Xuanzhong group showed no statistical differences (p>0.05)
PGE2 content in Sanyinjiao group were significantly increased than the other groups (p<0.01).
The results above indicated that:Acupuncture could regulate the content of pain-producing substance in uterus tissue of experimental dysmenorrhea rat to relieve pain. According to the comprehensive effect, Sanyinjiao group achieved best effect than the other groups.
4 Effect of electroacupuncture on different acupoints on expression of theκ-opioid receptor in dorsal horn of spinal cord of different segments of rat spinal cord
Different acupoints and non-acupoint have different effects on the expression ofκ-receptor in the spinal cord segments. T13 segment:
Model control group comparing to the saline control group:The integral optical density (IOD) of the K-receptors positive expression increased, but had no statistical differences (P>0.05).
All the EA groups comparing to model control group:the IOD in Sanyinjiao group and Xuehai group both increased significantly (P<0.01); the IOD in Xuanzhong group and non-acupoint group increased, but had no statistical differences (P>0.05).
The acupoint groups comparing to non-acupoint group:the IOD in Sanyinjiao group and Xuehai group both increased significantly (P<0.01); the IOD in Xuanzhong group hadn't showed any statistical differences (P>0.05). L1 segment:
Model control group comparing to the saline control group:the IOD increased, and the difference was not significant (P> 0.05).
All the EA groups comparing to model control group:the IOD in Sanyinjiao group andXuehai group both increased significantly (P<0.01); the IOD in Xuanzhong group and non-acupoint group also increased, but had no statistical differences (P>0.05).
Comparing to non-acupoint group and Xuanzhong group, the IOD in Sanyinjiao group andXuehai group significantly increased (P<0.01). L2 segment:
Model control group comparing to the saline control group:the IOD significantly increased, and had statistical difference(P<0.01).
All the EA groups comparing to model control group:the IOD in Sanyinjiao group and Xuehai group both increased significantly (P<0.01); the IOD in Xuanzhong group and non-acupoint group also increased, but had no statistical differences (P>0.05).
Comparing to Xuehai group, Xuanzhong group non-acupoint group, the IOD in Sanyinjiao group significantly increased (P<0.01). L6 segment:
Model control group comparing to the saline control group:the IOD increased, and the difference was not significant (P>0.05).
All the EA groups comparing to model control group:the IOD in Sanyinjiao group, Xuehai group and non-acupoint group increased obviously (P<0.01); the IOD in Xuanzhong group also increased, but had no statistical differences (P>0.05).
Comparing to non-acupoint group:the IOD in Sanyinjiao group significantly increased (P<0.01).there were more significant statistical differences in increasing of the IOD in Sanyinjiao group and Xuehai group than that in Xuanzhong group (P <0 .01). S1 segment:
Model control group comparing to the saline control group:the IOD increased, and the difference was not significant (P>0.05).
All the EA groups comparing to model control group:the IOD in Sanyinjiao group, Xuehai group and Xuanzhong group increased obviously (Sanyinjiao:P<0.01, Xuehai group, Xuanzhong group:P<0.05); the IOD in non-acupoint group had no statistical differences (P>0.05).
Comparing to Xuehai group and non-acupoint group, the IOD in Sanyinjiao group significantly increased (P<0.01).
Comparison among the acupoint groups, the IOD in Sanyinjiao group increased more significantly than that in Xuehai group (P<0.05).
These results show that:EA can adjust K-receptor expression of dorsal horn of spinal cord in all segments, but different acupoints have different effect on the different segments. Sanyinjiao group and Xuehai group can regulate all the segments, and Sanyinjiao group has a better effect; Xuanzhong group and non-acupoint group just have a certain obvious effect in individual segment, and there are no significant differences among Xuanzhong group and non-acupoint group.
5 Effect of electroacupuncture on different acupoints on endopioid peptide in PAG
5.1 Effect of electroacupuncture on different acupoint on ENK content in PAG:
Model control group comparing to the saline control group:ENK content in PAG increased, but had no statistical differences (P>0.05), which meant that in the rat model of experimental dysmenorrhea, the ENK content in PAG didn't increase significantly.
All the EA groups comparing to model control group:the ENK content in Sanyinjian group and Xuanzhong group significantly increased (Sanyinjiao group:P <0.01; Xuanzhong group:P<0.05); but there were no significant differences in increasing of ENK content between Xuehai group and non-acupoint group (P>0.Q5).
Comparing to the Xuanzhong group, Xuehai group and non-acupoint group, the ENK content in Sanyinjian group significantly increased, (Xuehai and non-acupoint group:P<0.01, Xuanzhong group:P<0.05); there were no significant differences on increasing of ENK content between Xuehai group and Xuanzhong group (P>0.05).
These results suggest that:Acupuncture could regulate the levels of ENK in PAG of rats with different effects. Sanyinjiao group and Xuanzhong group could significantly adjust ENK content in PAG, and Sanyinjiao group had a better effect. And the Xuehai group and non-acupoint group had no significant changes.
5.2 Effect of electro-acupuncture on different acupoints on B-EP content in the PAG:
Model control group comparing to the saline control group:the B-EP content in PAG increased, but had no statistical differences (P>0.05), which meant that the B-EP content in PAG didn't increase significantly in the rat model of experimental dysmenorrhea.
All the EA groups comparing to model control group:the B-EP content in Sanyinjian group and Xuanzhong group had increased significantly (P<0.01); and there were no statistical differences in increasing of B-EP content in Xuehai group and non-acupoint group (P>0.05).
Comparing to the non-acupoint group, the B-EP content in Sanyinjian group and Xuanzhong group significantly increased(Sanyinjian group:P<0.01, Xuanzhong group:P<0.05).
There were statistical differences between Sanyinjiao group and Xuehai group, and also between Sanyinjiao group and Xuanzhong group (P<0.01); and there were no significant differences between Xuanzhong group and Xuehai group (P>0.05).
The results above suggest that:Acupuncture could adjust the B-EP content in PAG of rat with different effects. Sanyinjiao group and Xuanzhong group could significantly regulate the B-EP content in PAG, and Sanyinjiao group showed a better effect. But the Xuehai group and non-acupoint group hadn't changed significantly.
5.3 Effect of electro-acupuncture on different acupoints on the Dyn content in PAG:
Model control group comparing to the saline control group:the Dyn content in PAG increased, but had no statistical differences (P> 0.05), which meant that the Dyn content in PAG didn't increase significantly in the rat model of experimental dysmenorrhea.
All the EA groups comparing to model control group:the Dyn content in Sanyinjian group and Xuanzhong group significantly increased (P<0.01); and there were no statistical differences in increasing of Dyn content in Xuehai group and non-acupoint group (P>0.05).
Comparing to the non-acupoint group, the Dyn content in Sanyinjian group significantly increased (P<0.01).
There were statistical differences between Sanyinjiao group and Xuehai group, Sanyinjiao group and Xuanzhong group (P<0.01); and there were no significant differences between Xuanzhong group and Xuehai group (P>0.05).
The results above showed that:Acupuncture could adjust the Dyn content in PAG of the rat with different effects. Sanyinjiao group and Xuanzhong group could both regulate the Dyn content in PAG significantly, and Sanyinjiao group had a better effect. The Xuehai group and non-acupoint group showed no significant changes.
5.4 Effect of electro-acupuncture on different acupoints on the EM content in PAG:
Model control group comparing to the saline control group:the EM content in PAG increased, but had no statistical differences (P> 0.05), which meant that increasing of the EM content in PAG wasn't significant in the rat model of experimental dysmenorrhea.
All the EA groups comparing to the model control group:EM content of each group all increased, but differences were not significant (P> 0.05).
The experiment results above suggest that:Acupuncture on acupoints and non-acupoints could not improve the EM content in PAG of rats significantly.
Conclusion
Behavior response in rats showed that electroacupuncture on acupoints and non-acupoints innervated by the same nerve segment, all of which could alleviate dysmenorrhea reaction in rats with different effects. Sanyinjiao showed the best effect than Xuehai, Xuanzhong and non-acupoint, and the three groups had no significant differences. EA could adjust contraction of uterine smooth muscle and the content of pain-producing substances in uterus and the central pain modulation system to play an analgesic effect. However, acupuncture on different acupoints played different roles. Comparing to EA on Xuanzhong as non-relevant acupoint and non-acuppoint, EA on Sanyinjiao and Xuehai as relevant acupoints had more significant effect onκ-opioid receptor expression of dorsal horn of spinal cord in all segments, and Sanyinjiao has a best effect. Sanyinjiao and Xuanzhong as the special acupoints of relevant and non-relevant meridian could regulate the contents of endopioid peptide in PAG, and Sanyinjiao has a better effect. All of this implied that the existence of specificity of acupoint effect, the effect of acupoints was not only closely related to innervation of the nerve segment, but also "meridian" still played a key role, and the specificity of acupoint effect is relative, not absolute.
针灸学认为刺激穴位可特异性地对相应脏腑器官产生调治作用,这种作用与非穴和其它经穴比较存在差异。这种差异就是经穴效应的特异性。经穴效应特异性是经络学说的核心内容,是针灸临床循经取穴的重要依据。而且大量临床实践及实验研究都证实穴位效应确实存在特异性。
现代神经科学对针灸机理的研究结果已显示经穴效应存在特异性。但这种特异性的产生主要是由于支配穴位所在部位的神经节段的不同而引起的。它主要与穴位所在的部位有关,而与经典针灸理论所强调的穴位所处的“经”关系不大。即处于同神经节段的不同经脉上的穴位、相同经脉上的不同穴位以及与非穴位间的效应不存在差异。那么,经穴效应特异性到底为何?同神经节段内非穴位以及不同经脉上的穴位间是否存在效应特异性?相同经脉上不同穴位间的效应又如何呢?
本研究以实验性类痛经大鼠模型为研究对象,从电针镇痛作用及机制入手,观察电针同神经节段支配的胞宫相关经穴、非相关经穴以及非经非穴对实验性类痛经大鼠的行为学反应、子宫平滑肌收缩力、子宫组织局部致痛物质以及中枢痛觉调制系统的影响,探讨同神经节段支配的经穴与非经穴、相关经穴与非相关经穴,以及同一经脉不同穴位间是否存在经穴效应特异性以及经穴效应特异性是否具有一定的规律。为经穴效应特异性的研究提供系统有力的实验依据。
实验中所选经穴为：相关经穴为足太阴脾经三阴交、血海穴；非相关经穴为足少阳胆经悬钟穴；非经非穴为胆经与胃经之间平悬钟穴处的非经非穴点。
实验选用动情间期SD雌性大鼠156只,按照随机数字法分为盐水组、模型组、电针三阴交组、电针血海组、电针悬钟组、电针非穴组6组,每组26只。除盐水组外,其余各组大鼠均皮下注射苯甲酸雌二醇连续10天,第1、10天每只大鼠皮下注射0.5 mg,第2-9天每只均皮下注射0.2mg。末次给药1h后,腹腔注射缩宫素2u／只。盐水组每日给予同等剂量的生理盐水。根据Schmauss行为学评分标准,记录20分钟内扭体反应的潜伏期、扭体评分及扭体次数；应用BL-420E+生物机能实验系统记录仪记录20分钟内大鼠子宫收缩次数和强度；采用放射免疫法检测子宫前列腺素E2(PGE2)、前列腺素F2α(PGF2α)含量；采用免疫组化法检测脊髓背角内K-阿片受体表达；采用ELISA法定量检测中脑中央导水管周围灰质(PAG)中脑啡肽(ENK)、β内啡肽(β-EP)、强啡肽(Dyn)、内吗啡肽(EM)的含量。实验结果如下：
1电针不同经穴对大鼠行为学反应的影响
模型组与盐水组相比：扭体潜伏期明显缩短,扭体评分、扭体次数明显增高,均有统计学意义(P＜0.01)；说明模型制备成功。
三阴交组扭体潜伏期与模型组相比明显延长,差异有统计学意义(P＜0.05),其余各组扭体潜伏期与模型组相比亦延长,但差异无统计学意义(P＞0.05)；扭体评分和次数各组与模型组相比均明显减少,差异有统计学意义(P＜0.01)。但三阴交组扭体评分与盐水组相比差异亦无统计学意义(P＞0.05)。说明电针不同穴位后,可减轻实验性类痛经大鼠的扭体反应,其中电针三阴交穴的效应最佳,而血海穴、悬钟穴与非穴间的效应无明显差异。
2电针不同经穴对大鼠子宫收缩程度的影响
模型组与盐水组相比：子宫收缩次数明显增加,子宫收缩强度明显增强,差异均有统计学意义(P＜0.01)；说明造模后子宫平滑肌出现痉挛性收缩。
三阴交组和血海组的子宫收缩次数与模型组相比均减少,差异有统计学意义(P＜0.05),悬钟组和非穴组的子宫收缩次数与模型组相比亦减少,但差异无统计学意义(乃0.05);三阴交组的子宫收缩强度与模型组相比明显降低,差异有统计学意义(P＜0.05),其余各组的子宫收缩强度与模型组相比亦降低,但差异无统计学意义(P＞0.05)。除非穴组外,其余各组的子宫收缩强度与盐水组相比,差异均无统计学意义(P＞0.05)。结果说明：电针穴位后,可调节实验性类痛经大鼠子宫痉挛性收缩程度,从而缓解痛反应。其中,电针三阴交穴的效应最佳,血海穴和悬钟穴亦有一定的缓解效应,血海穴较悬钟穴效应稍佳。非穴的效应无明显改变。
3电针不同经穴对大鼠子宫PGE2、PGF2α含量及PGE2／PGF2α比值的影响
模型组与盐水组相比：PGF2α含量明显增高,PGE2含量明显降低,比值明显升高,差异均有统计学意义(P<0.01)；说明造模后,子宫组织中致痛物质含量升高。
各电针组PGF2α含量与模型组相比均明显降低,差异有统计学意义(三阴交：P<0.05；其余各组：P<0.01)。三阴交组PGE2含量与模型组相比明显升高,差异有统计学意义(P<0.01),且与盐水组相比差异无统计学意义(P>0.05),三阴交组PGE2含量比其它各组均明显升高,差异有统计学意义(P＜0.01)；其余各组PGE2含量与模型组相比均无明显升高,差异无统计学意义(P＞0.05)。各电针组比值与模型组相比均明显降低,差异有统计学意义(P＜0.01),且三阴交和悬钟组比值与盐水组相比差异亦无统计学意义(P＞0.05)。结果说明：电针穴位后,可通过调节实验性类痛经大鼠子宫组织致痛物质的含量而达到镇痛作用。综合效应来看,三阴交组的调节作用较强,其余各组间效应无明显差异。
4电针不同经穴对大鼠各节段脊髓背角K-阿片受体表达的影响
针刺不同穴位及非穴对各脊髓节段K-阿片受体表达的影响不同。T13节段：
模型组与盐水组相比：IOD值升高,但差异无统计学意义(P＞0.05)
三阴交和血海组的IOD值与模型组相比明显升高,差异有统计学意义(P<0.01)；悬钟和非穴组的IOD值与模型组相比亦有升高,但差异无统计学意义(P＞0.05)。三阴交和血海组的IOD值与非穴相比明显升高,差异有统计学意义(P＜0.01)。
L1节段：
模型组与盐水组相比：IOD值升高,但差异无统计学意义(P＞0.05)
三阴交和血海组的IOD值与模型组相比明显升高,差异有统计学意义(P<0.01)；悬钟和非穴组的IOD值与模型组相比亦有升高,但差异无统计学意义(P＞0.05)。三阴交和血海组的IOD值与悬钟组和非穴组相比明显升高,差异有统计学意义(P＜0.01)
L2节段：
模型组与盐水组相比：IOD值升高明显,差异有统计学意义(P＜0.01)
三阴交和血海组的IOD值与模型组相比明显升高,差异有统计学意义(P<0.01)；悬钟和非穴组的IOD值与模型组相比亦有升高,但差异无统计学意义(P＞0.05)。三阴交组的IOD值与非穴组相比明显升高,差异有统计学意义(P＜0.01)。三阴交组的IOD值较血海和悬钟组也明显升高,差异有统计学意义(P＜0.01)。
L6节段：
模型组与盐水组相比：IOD值升高,但差异无统计学意义(P＞0.05)
三阴交、血海和非穴组的IOD值与模型组相比明显升高,差异有统计学意义(P＜0.01);悬钟组的IOD值与模型组相比亦有升高,但差异无统计学意义(P＞0.05)。三阴交组的IOD值与非穴组相比升高,差异有统计学意义(P＜0.05)。三阴交和血海组的IOD值均较悬钟组明显升高,差异有统计学意义(P＜0.01)。
S1节段：
模型组与盐水组相比：IOD值升高,但差异无统计学意义(P＞0.05)
三阴交、血海和悬钟组的IOD值与模型组相比明显升高,差异有统计学意义(三阴交：P＜0.01,血海、悬钟：P＜0.05)；非穴组的IOD值与模型组和盐水组相比差异均无统计学意义(P＞0.05)。三阴交组的IOD值与非穴相比明显升高,差异有统计学意义(P＜0.01)。三阴交组的IOD值较血海组亦明显升高,差异有统计学意义(P＜0.05)。
以上结果说明：电针可调节各脊髓节段背角内κ-受体的表达,但不同穴位对不同节段调节的强度不同。三阴交和血海穴在各节段都有调节作用,三阴交穴的调节作用较血海穴明显；悬钟穴和非穴也有一定的调节作用,但作用均较三阴交和血海穴组弱。悬钟和非穴间无明显差异。
5电针不同经穴对PAG内阿片肽物质的影响
5.1电针不同经穴对大鼠PAG内ENK含量的影响：
模型组与盐水组相比：PAG内ENK含量有所升高,但差异无统计学意义(P＞0.05)。说明实验性类痛经模型使大鼠PAG内ENK含量升高不明显。
三阴交和悬钟组的ENK含量与模型组相比明显升高,差异有统计学意义(三阴交：P＜0.01；悬钟组：P＜0.05)；血海和非穴组的ENK含量与模型组相比亦有升高,但差异无统计学意义(P＞0.05)。三阴交组的ENK含量与血海、悬钟和非穴组相比明显升高,差异有统计学意义(血海、非穴：P＜0.01,悬钟：P＜0.05)。悬钟与血海的组间差异无统计学意义(P＞0.05)。说明电针穴位可调节大鼠PAG内ENK含量的水平,但调节程度不同。三阴交和悬钟穴可明显调节PAG内ENK含量,其中以三阴交穴较好。而电针血海穴和非穴无明显变化。
5.2电针不同经穴对大鼠PAG内β-EP含量的影响：
模型组与盐水组相比：PAG内β-EP含量有所升高,但差异无统计学意义(P＞0.05)。说明实验性类痛经模型使大鼠PAG内β-EP含量升高不明显。
三阴交和悬钟组的β-EP含量与模型组相比明显升高,差异有统计学意义(P＜0.01)；血海和非穴组的β-EP含量与模型组相比亦有升高,但差异无统计学意义(P＞0.05)。三阴交和悬钟组β-EP含量与非穴相比均明显升高,差异有统计学意义(三阴交：P＜0.01；悬钟组：P＜0.05)。三阴交与血海和悬钟组相比差异均有统计学意义(P＜0.01)。悬钟与血海组间差异无统计学意义(乃0.05)。说明电针穴位可调节大鼠PAG内β-EP含量的水平,但调节程度不同。三阴交和悬钟穴可明显调节PAG内β-EP含量,其中以三阴交穴较好。而电针血海穴和非穴无明显变化。
5.3电针不同经穴对大鼠PAG内Dyn含量的影响：
模型组与盐水组相比：PAG内Dyn含量有所升高,但差异无统计学意义(P＞0.05)。说明实验性类痛经模型使大鼠PAG内Dyn含量升高不明显。
三阴交和悬钟组的Dyn含量与模型组相比明显升高,差异有统计学意义(P＜0.01)；而血海和非穴组的Dyn含量与模型组相比亦有升高,但差异无统计学意义(P＞0.05)。三阴交组Dyn含量与非穴组相比明显升高,差异有统计学意义(P＜0.01)。三阴交与血海和悬钟组相比差异均有统计学意义(P＜0.01)。悬钟与血海组间差异无统计学意义(P＞0.05)。说明电针穴位可调节大鼠PAG内Dyn含量的水平,但调节程度不同。三阴交和悬钟穴可明显调节PAG内Dyn含量,其中以三阴交穴较好。而电针血海穴和非穴无明显变化。
5.4电针不同经穴对大鼠PAG内EM含量的影响：
模型组与盐水组相比：PAG内EM含量有所升高,但差异无统计学意义(P＞0.05)。说明实验性类痛经模型使大鼠PAG内EM含量升高不明显。
各组EM含量与模型组相比均有升高,但差异无统计学意义(P＞0.05)。说明：针刺各穴位及非穴均无明显调节大鼠PAG内EM含量的作用。
综上所述：电针不同经穴可引起实验性类痛经大鼠PAG内阿片肽含量发生变化,说明针刺可通过调节脑内阿片肽的水平而起到镇痛作用。但电针不同经穴及非穴对PAG内阿片肽含量的影响不同。在电针同神经节段的三个穴位中,三阴交和悬钟穴均可使ENK、β-EP和Dyn的含量升高,三阴交穴的升高效应较明显,悬钟次之,而血海和非穴无明显调节作用；四个穴位对EM的含量均无明显调节作用,说明EM可能不参与电针对内脏痛的调节。
结论
电针同神经节段支配的穴位与非穴位对内脏痛均有镇痛作用,但它们的镇痛效应不同,相关经脉上的特定穴镇痛效应最佳,相关经脉上的非特定穴与非相关经穴效应相近,并且与非穴的效应也相近。针刺穴位可通过调节子宫功能状态和中枢内痛觉调制系统来达到对内脏痛的镇痛作用。并且不同穴位对各部位的调节作用不同。相关经脉上的穴位较之非相关经脉上的穴位以及非穴有更明显的脊髓节段性调节作用,其中以相关经脉中的特定穴作用最强；相关经脉和非相关经脉上的特定穴对脊髓上中枢均有调节作用,但相关经脉上特定穴的调节作用更明显；相关经脉上的非特定穴和非穴无明显调节作用。因此,穴位不同,其调节机制不同,从而产生的效应就不同。由此认为：经穴效应具有特异性,这种特异性是相对的,而不是绝对的。经穴效应特异性虽与其所处的神经节段密切相关,但与其所在的“经”可能有更加密切的联系。
Electroacupuncture Analgesia on Experimental Dysmenorrhea Model Rat and Its Mechanism
The theory of acupuncture considers that stimulating acupoints can regulate the functions of the corresponding viscera specifically, of which is different comparing to the functions regulated by stimulating non-acupoint or acupoints on other meridians. This difference is specificity of the acupoint effect, which is the core content of the meridian theory and an important basis of locating acupoints along meridian in clinical. A large amount of clinical research and experimental study have confirmed the existence of specificity.
The research results of modern neuroscience on acupuncture mechanism also showed the existence of specificity of acupoint effect, which was mainly caused by different nerve segments innervating acupoint area. And it was mostly related to the locations of acupoints but had nothing to do with the meridians. That meant there was no difference found among acupoint effect of different meridians innervated by the same nerve segment or difference between acupoints on the same meridian and non acupoints. And what is specificity of acupoint effect in the end? Is there effect specificity among the different meridian acupoints and non-acupoint innervated by the same nerve segment? How about the effect of different acupoints on the same meridian?
The aim of the present study was to explore whether the specificity of acupoint effects exists and has certain principles or not by comparing the effect and possible mechanism of electroacupuncture analgesia on experimental dysmenorrhea model rat at different acupoints innervated by the same nerve segment.
Selected acupoints for the experiment were as follows:Sanyinjiao(SP 6), Xuehai(SP 10) on the Spleen meridian of Foot Taiyin as relevant acupoints; Xuanzhong(GB 39) on Gallbladder Meridian of Foot Shaoyang as non-relevant acupoints; and non-acupoints located in the middle acupoint between the gallbladder meridian and the stomach meridian at the horizontal line of Xuanzhong(GB 39).
The experiments used 156 female Sprague-Dawley (SD) rats which were during their diestrus. SD rats were randomly divided into 6 groups, which included saline control group, model control group, EA Sanyinjiao group, EA Xuehai group, EA Xuanzhong group, EA non-acupoint group, and 26 rats in each group. Except the saline control group, the rest of groups were injected estradiol benzoate for 10 days, the first and 10th day 0.5 mg for each, and from 2th to 9th days 0.2mg for each.1 hour after the last injection, Oxytocin was given to each by intraperitoneal injection with 2u. The saline control group was given for the same dose of saline everyday. According to the standard of Schmauss behavior score, we recorded latency of writhing, writhing times and writhing score in 20 minutes; applied BL-420E+logger of experimental system to record times and intensity of uterine contraction in 20 minutes; used radio immunoassay to detect the content of PGE2, PGF2a in uterine; observed expression ofκ-opioid receptor in the dorsal horn of the spinal cord by immunohistochemical staining; and detected ENK, B-EP, Dyn, EM content in the periaqueductal gray (PAG) of midbrain by ELISA. The results were as follows:
1 Effect of Electroacupuncture at different acupoints on writhing response in rats
Compared to the saline control group, latency of writhing, writhing times and writhing score of model control group significantly changed (P<0.01), that meant the model was successfully prepared.
All the EA groups comparing to the model control group:writhing latency of EA Sanyinjiao group was significantly prolonged (p<0.05), writhing latency of the other groups was also prolonged, but there was no statistical difference (p>0.05); writhing score and writhing times decreased significantly in each group.(p<0.01).but the writhing score of Sanyinjiao group had no statistical difference comparing to the saline control group (p>0.05).
The results above showed that:After electro-acupuncture treatment on different acupoints, the writhing response of experimental dysmenorrheal model in rats could be relieved at different degrees. There was the best effect in EA Sanyinjiao group, and the differences were not significant among EA Xuehai group, EA Xuanzhong group and EAnon acupoint group.
2 Effect of electroacupuncture on different acupoints about the uterine contraction of rats
Model control group comparing to the saline control group:the time and intensity of uterine contraction significantly increased (p<0.01) which meant the smooth muscle of uterus was induced spasmodic contraction.
the model was successfully prepared.
All the EA groups comparing to model control group:time of uterine contractions were reduced significantly in Sanyinjiao group and Xuehai group (p <0.05); and the intensity of uterine contraction in Sanyinjiao group had a better effect than other groups (p<0.05).
All the EA groups comparing to the saline control group, intensity of uterine contraction hadn't a significant difference in other groups except non-acupoint group (p>0.05).
The results above showed that:acupuncture could improve spasmodic contraction of uterus of experimental dysmenorrhea rat to alleviate pain in the uterus. As a conclusion, the EA Sanyinjiao group had the best effect than others, Xuehai group and Xuanzhong group also had a certain effect, with a better effect in Xuehai group. The effect of non-acupoint hadn't significantly changed.
3 Effect of electroacupuncture on different acupoints on the content of PGE2, PGF2a and the content ratio of PGE2/PGF2a in the rat uterus
Model control group comparing to the saline control group:the content of PGF2a significantly increased, and the content of PGE2 decreased significantly. The both ratio increased significantly (p<0.01); which implied that the content of pain-producing substance in uterine tissue increased after modeling.
All the EA groups comparing to model control group:PGF2a content decreased significantly (Sanyinjiao group:p<0.05; the other groups:p<0.01); PGE2 content in Sanyinjiao group increased more significantly than other groups (p<0.01); and the ratio of PGE2/PGF2a in EA groups significantly decreased (p< 0.01).
All the EA groups comparing to the saline control group:PGE2 content of all groups except Sanyinjiao group significantly decreased (Xuehai group:p<0.05; the other groups:p<0.01); the ratio of PGE2/PGF2a between Sanyinjiao group and Xuanzhong group showed no statistical differences (p>0.05)
PGE2 content in Sanyinjiao group were significantly increased than the other groups (p<0.01).
The results above indicated that:Acupuncture could regulate the content of pain-producing substance in uterus tissue of experimental dysmenorrhea rat to relieve pain. According to the comprehensive effect, Sanyinjiao group achieved best effect than the other groups.
4 Effect of electroacupuncture on different acupoints on expression of theκ-opioid receptor in dorsal horn of spinal cord of different segments of rat spinal cord
Different acupoints and non-acupoint have different effects on the expression ofκ-receptor in the spinal cord segments. T13 segment:
Model control group comparing to the saline control group:The integral optical density (IOD) of the K-receptors positive expression increased, but had no statistical differences (P>0.05).
All the EA groups comparing to model control group:the IOD in Sanyinjiao group and Xuehai group both increased significantly (P<0.01); the IOD in Xuanzhong group and non-acupoint group increased, but had no statistical differences (P>0.05).
The acupoint groups comparing to non-acupoint group:the IOD in Sanyinjiao group and Xuehai group both increased significantly (P<0.01); the IOD in Xuanzhong group hadn't showed any statistical differences (P>0.05). L1 segment:
Model control group comparing to the saline control group:the IOD increased, and the difference was not significant (P> 0.05).
All the EA groups comparing to model control group:the IOD in Sanyinjiao group andXuehai group both increased significantly (P<0.01); the IOD in Xuanzhong group and non-acupoint group also increased, but had no statistical differences (P>0.05).
Comparing to non-acupoint group and Xuanzhong group, the IOD in Sanyinjiao group andXuehai group significantly increased (P<0.01). L2 segment:
Model control group comparing to the saline control group:the IOD significantly increased, and had statistical difference(P<0.01).
All the EA groups comparing to model control group:the IOD in Sanyinjiao group and Xuehai group both increased significantly (P<0.01); the IOD in Xuanzhong group and non-acupoint group also increased, but had no statistical differences (P>0.05).
Comparing to Xuehai group, Xuanzhong group non-acupoint group, the IOD in Sanyinjiao group significantly increased (P<0.01). L6 segment:
Model control group comparing to the saline control group:the IOD increased, and the difference was not significant (P>0.05).
All the EA groups comparing to model control group:the IOD in Sanyinjiao group, Xuehai group and non-acupoint group increased obviously (P<0.01); the IOD in Xuanzhong group also increased, but had no statistical differences (P>0.05).
Comparing to non-acupoint group:the IOD in Sanyinjiao group significantly increased (P<0.01).there were more significant statistical differences in increasing of the IOD in Sanyinjiao group and Xuehai group than that in Xuanzhong group (P <0 .01). S1 segment:
Model control group comparing to the saline control group:the IOD increased, and the difference was not significant (P>0.05).
All the EA groups comparing to model control group:the IOD in Sanyinjiao group, Xuehai group and Xuanzhong group increased obviously (Sanyinjiao:P<0.01, Xuehai group, Xuanzhong group:P<0.05); the IOD in non-acupoint group had no statistical differences (P>0.05).
Comparing to Xuehai group and non-acupoint group, the IOD in Sanyinjiao group significantly increased (P<0.01).
Comparison among the acupoint groups, the IOD in Sanyinjiao group increased more significantly than that in Xuehai group (P<0.05).
These results show that:EA can adjust K-receptor expression of dorsal horn of spinal cord in all segments, but different acupoints have different effect on the different segments. Sanyinjiao group and Xuehai group can regulate all the segments, and Sanyinjiao group has a better effect; Xuanzhong group and non-acupoint group just have a certain obvious effect in individual segment, and there are no significant differences among Xuanzhong group and non-acupoint group.
5 Effect of electroacupuncture on different acupoints on endopioid peptide in PAG
5.1 Effect of electroacupuncture on different acupoint on ENK content in PAG:
Model control group comparing to the saline control group:ENK content in PAG increased, but had no statistical differences (P>0.05), which meant that in the rat model of experimental dysmenorrhea, the ENK content in PAG didn't increase significantly.
All the EA groups comparing to model control group:the ENK content in Sanyinjian group and Xuanzhong group significantly increased (Sanyinjiao group:P <0.01; Xuanzhong group:P<0.05); but there were no significant differences in increasing of ENK content between Xuehai group and non-acupoint group (P>0.Q5).
Comparing to the Xuanzhong group, Xuehai group and non-acupoint group, the ENK content in Sanyinjian group significantly increased, (Xuehai and non-acupoint group:P<0.01, Xuanzhong group:P<0.05); there were no significant differences on increasing of ENK content between Xuehai group and Xuanzhong group (P>0.05).
These results suggest that:Acupuncture could regulate the levels of ENK in PAG of rats with different effects. Sanyinjiao group and Xuanzhong group could significantly adjust ENK content in PAG, and Sanyinjiao group had a better effect. And the Xuehai group and non-acupoint group had no significant changes.
5.2 Effect of electro-acupuncture on different acupoints on B-EP content in the PAG:
Model control group comparing to the saline control group:the B-EP content in PAG increased, but had no statistical differences (P>0.05), which meant that the B-EP content in PAG didn't increase significantly in the rat model of experimental dysmenorrhea.
All the EA groups comparing to model control group:the B-EP content in Sanyinjian group and Xuanzhong group had increased significantly (P<0.01); and there were no statistical differences in increasing of B-EP content in Xuehai group and non-acupoint group (P>0.05).
Comparing to the non-acupoint group, the B-EP content in Sanyinjian group and Xuanzhong group significantly increased(Sanyinjian group:P<0.01, Xuanzhong group:P<0.05).
There were statistical differences between Sanyinjiao group and Xuehai group, and also between Sanyinjiao group and Xuanzhong group (P<0.01); and there were no significant differences between Xuanzhong group and Xuehai group (P>0.05).
The results above suggest that:Acupuncture could adjust the B-EP content in PAG of rat with different effects. Sanyinjiao group and Xuanzhong group could significantly regulate the B-EP content in PAG, and Sanyinjiao group showed a better effect. But the Xuehai group and non-acupoint group hadn't changed significantly.
5.3 Effect of electro-acupuncture on different acupoints on the Dyn content in PAG:
Model control group comparing to the saline control group:the Dyn content in PAG increased, but had no statistical differences (P> 0.05), which meant that the Dyn content in PAG didn't increase significantly in the rat model of experimental dysmenorrhea.
All the EA groups comparing to model control group:the Dyn content in Sanyinjian group and Xuanzhong group significantly increased (P<0.01); and there were no statistical differences in increasing of Dyn content in Xuehai group and non-acupoint group (P>0.05).
Comparing to the non-acupoint group, the Dyn content in Sanyinjian group significantly increased (P<0.01).
There were statistical differences between Sanyinjiao group and Xuehai group, Sanyinjiao group and Xuanzhong group (P<0.01); and there were no significant differences between Xuanzhong group and Xuehai group (P>0.05).
The results above showed that:Acupuncture could adjust the Dyn content in PAG of the rat with different effects. Sanyinjiao group and Xuanzhong group could both regulate the Dyn content in PAG significantly, and Sanyinjiao group had a better effect. The Xuehai group and non-acupoint group showed no significant changes.
5.4 Effect of electro-acupuncture on different acupoints on the EM content in PAG:
Model control group comparing to the saline control group:the EM content in PAG increased, but had no statistical differences (P> 0.05), which meant that increasing of the EM content in PAG wasn't significant in the rat model of experimental dysmenorrhea.
All the EA groups comparing to the model control group:EM content of each group all increased, but differences were not significant (P> 0.05).
The experiment results above suggest that:Acupuncture on acupoints and non-acupoints could not improve the EM content in PAG of rats significantly.
Conclusion
Behavior response in rats showed that electroacupuncture on acupoints and non-acupoints innervated by the same nerve segment, all of which could alleviate dysmenorrhea reaction in rats with different effects. Sanyinjiao showed the best effect than Xuehai, Xuanzhong and non-acupoint, and the three groups had no significant differences. EA could adjust contraction of uterine smooth muscle and the content of pain-producing substances in uterus and the central pain modulation system to play an analgesic effect. However, acupuncture on different acupoints played different roles. Comparing to EA on Xuanzhong as non-relevant acupoint and non-acuppoint, EA on Sanyinjiao and Xuehai as relevant acupoints had more significant effect onκ-opioid receptor expression of dorsal horn of spinal cord in all segments, and Sanyinjiao has a best effect. Sanyinjiao and Xuanzhong as the special acupoints of relevant and non-relevant meridian could regulate the contents of endopioid peptide in PAG, and Sanyinjiao has a better effect. All of this implied that the existence of specificity of acupoint effect, the effect of acupoints was not only closely related to innervation of the nerve segment, but also "meridian" still played a key role, and the specificity of acupoint effect is relative, not absolute.
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