| Traditional Chinese Medicine seems the human body as a whole. Upper and lower,inside and outside, exterior and interior, zang-fu organs are connected by the channels in both physiology and pathology condition. The channel has the function to move qi and blood,reflect the disorder and treat the disease. The disorders of zang-fu organs could be detected by the changes of channels and acupoints through the pathway of organs-channel-acupoints’.Acupuncture could restore the balance of zang-fu organs by stimulating the acupoints.Certain acupuncture stimuli, in other word, the effective stimuli is needed in order to achieve the therapeutic effect. Acupuncture deqi is acupuncture induction of both doctor and patient that symbolize the achievement of good effect. Effective stimuli is a key to stimulate a good state of deqi and achieve therapeutic effect.Our previous animal and clinical studies have demonstrated that acupuncture has a good analgetic effect on primary dysmenorrhea. Sanyinjiao (SP6) has the best therapeutic effect which reflected the along meridian specificity of acupoints. Based on our previous research,this study observed the analgetic effect, change of uterus micro-circulation and temperatures of the points, line between the points, and the uterus cause by different acupuncture stimuli, to further study influence of different stimuli at SP6 on its therapeutic effect and related mechanism when treating dysmenorrheal.Objectives:Experiment A: To study the preparation of dysmenorrheal rat model of cold stagnation syndrome. Experiment B, C, and D are to observe the effects of acupuncture intervention with different stimuli on primary dysmenorrheal, and thereby discuss the mechanism.Methods:Experiment A: 60 three-month-old female SD rats during anestrus were divided randomly into the blank group, the saline group, dysmenorrheal model group, 5-day-frozen group,5-day-frozen with pharmaceutical intervention group, 10-day-frozen with pharmaceutical intervention group (10 in each group).Estradiol Benzoate injection was administrated each day to animals of dysmenorrheal model group, 5-day-frozen with pharmaceutical intervention group, 10-day-frozen with pharmaceutical intervention group by subcutaneous injection. The injection continued for 10 days with the dosages ranged from 0.5mg/each animal in the first day to 0.2 mg/each from the second to the ninth day, and to 0.5mg/each animal in the last day.1 hour later after the last time of Estradiol Benzoate injection, oxytocinum was given to animals in the dysmenorrheal model group, 5-day-frozen with pharmaceutical intervention group, and 10-day-frozen with pharmaceutical intervention group (2U/each animal) by intra-peritoneal injection. Animals in the saline group were injected with saline of the same quantum. After the injection,animals in the 5-day-frozen group, 5-day-frozen with pharmaceutical intervention group, and 10-day-frozen with pharmaceutical intervention group was placed into -25 ℃ freezer for 4 hours (with a 5 seconds interval between every 2 hours for ventilation). Animals in the blank group were fastened the same way with no intervention applied. At the 10th day after model preparation,rats’ scores on writhing response, cold syndrome scale, whole blood reduction viscosity, erythrocyte rigidity index, and erythrocyte aggregation index were observed. Experiments B, C, and D: 104 three-month-old female SD rats during anestrus was divided randomly into the saline group, cold stagnation dysmenorrheal group, A stimuli( ’Deqi’)group,and B stimuli(No ’Deqi’)group. Rats in A stimuli( ’Deqi’ ) group (deep insertion of thick needle with manipulation) were treated byΦ0.25×40mm needle insertion on Sanyinjiao (SP6) for 4-5mm with 30-second needle manipulation of even reinforcement and reduction at the beginning and 10 minutes after the insertion. Rats in B stimuli(No ’Deqi’ ) group (shallow insertion of thin needle with no manipulation) were treated by Φ0.18×13mm needle insertion on Sanyinjiao (SP6) for l-2mm with no needle manipulation. Needles were retained for 20 mins in both two groups. 40 rats were sacrificed after the observation of writhing response, for the measurements of uterus LTB4, plasma 6-keto-PGF1α and TXB2 contents. 32 rats were used for the observation of microcirculation change of the uterus. Another 32 rats were used for the observation of temperature changes of Sanyinjiao (SP6), Xuehai (SP10), the uterus, and the line between the above two points.Results:1. Model preparation of cold stagnation dysmenorrheal ratsCompared with that of the blank group, rats in the 5-day-frozen group, 5-day-frozen with pharmaceutical intervention group, and 10-day-frozen with pharmaceutical intervention group had significantly increased cold syndrome score (p<0.01); rats in the dysmenorrheal model group, 5-day-frozen group, and 5-day-frozen with- pharmaceutical intervention group had remarkably increased whole blood reduction viscosity (p<0.05); and rats of the 5-day-frozen with pharmaceutical intervention group had obviously increased erythrocyte rigidity index(p<0.05). Compared with that of the saline group, rats in the dysmenorrheal model group,5-day-frozen with pharmaceutical intervention group, and 10-day-frozen with pharmaceutical intervention group have remarkably increased writhing response score (p<0.05).2. Effects on pain and uterus LTB4 of cold stagnation dysmenorrheal rats by acupuncture intervention of different acupuncture stimuliCompared with that of the saline group, rats in cold stagnation dysmenorrheal group have drastically declined writhing latent period (p<0.01), sharply increased writhing times and score, and slightly increased uterus LTB4 content (p>0.05).In comparison to the cold stagnation dysmenorrheal group, rats in A stimuli( ’Deqi’ )group have remarkably increased writhing latent period (p<0.01), significantly decreased writhing times and score (p<0.01), and slightly decreased uterus LTB4content (p>0.05); rats in B stimuli(No ’Deqi’ ) group are of sharply decreased writhing score (p<0.05) and slightly increased uterus LTB4 content (p>0.05).Rats in A stimuli( ’Deqi’)group showed obviously prolonged writhing latent period(p<0.01), drastically decreased writhing times (p<0.01) and writhing score (p<0.05),compared with that of the B stimuli(No ’Deqi’ ) group.3. Effects of acupuncture with different acupuncture stimuli on temperatures of the points, lines between the points, and uterus of cold stagnation dysmenorrheal ratsCompared with that of the saline group, cold stagnation dysmenorrheal group, and the B stimuli(No ’Deqi’ ) group, rats in A stimuli( ’Deqi’ ) group showed drastically decreased point temperature at left Sanyinjiao (SP6) from the insertion of the needle to the 5th minute.Temperature of the same point on the right side increased remarkably from the 5th to the 10th minute (p<0.01).The animals also had significantly decreased temperature on right Xuehai (SP10) from the insertion to the 5th minute after (p<0.05, p<0.01). The temperature raised from the 5th to 10th minute (p<0.05, p<0.01), and went down again from the 10th to 20th minute (p<0.01). The same point on the left experienced no significant change in temperature at all time points(p>0.05).Compared with saline group, cold stagnation dysmenorrheal group, and B stimuli(No’Deqi’ ) group, A stimuli( ’Deqi’ ) group had a decreased point temperature of the right Sanyinjiao (SP6) from the insertion to the 5th minute (p<0.05, p<0.01). And the temperature increased from the 5th to the 10th minute (p<0.05, p<0.01). Temperature of the line between Sanyinjiao (SP6) and Xuehai (SP10) on the left had no drastic change at all time points(p>0.05).There was no significant difference among all groups on temperature of uterus area(p>0.05).4. Effects of different acupuncture stimuli intensities on uterus microcirculation and microcirculation substances of cold stagnation dysmenorrheal ratsCompared with that of the saline group, rats in the cold stagnation dysmenorrheal group had remarkably shrunken uterus capillary diameter at all time points from the 5th to the 30thminutes (p<0.01). They also had significantly reduced uterus micro-vessel diameter(p<0.01) and definition (p<0.01) from the 5th to the 40th minute, blood flow (p<0.01), and plasma 6-keto-PGF1α (p<0.05), and drastically increased TXB2/6-keto-PGF1α content(p<0.01).In contrast with the cold stagnation dysmenorrheal group, rats in the A stimuli( ’Deqi’)group had significantly higher uterus capillary diameter at all the time points from the 5th to the 30th minute (p<0.01 or p<0.05), significantly dilated uterus micro-vessel diameter at all the time points from the 5th to the 40th minute (p<0.01), and evidently improved blood flow(p<0.01). The definition of uterus micro-vessels of animals in the group were obviously improved at the 5th,10th, 30th, and 40th minute with remarkably increased plasma 6-keto-PGF1α(p<0.05) and significantly decreased TXB2/6-keto-PGF1α contents (p<0.05). Compared with the cold stagnation dysmenorrheal group, B stimuli(No ’Deqi’ ) group had significantly expanded micro-vessel diameter at the 20th and 30th minute, and drastically decreased plasma TXB2/6-keto-PGF1α.In comparison with the B stimuli(No ’Deqi’ ) group, A stimuli( ’Deqi’)group had obviously dilated capillary diameter at all the time points from the 5th to the 30th minute(p<0.05 or p<0.01), significantly dialated micro-vessel diameter at all time points from the 5th to the 40th minute (p<0.01), highly improved micro-vessel definition at the 30th and 40th minute (p<0.05), and remarkably improved micro-vessel blood flow at all the time points from the 5th to the 40th minute (p<0.01).Conclusions:1. The study, combining model preparation of cold stagnation syndrome with dysmenorrheal together, successfully set up dysmenorrheal rat models of cold stagnation syndrome. Disorder of uterus micro-circulation and circulation related substance was existed in model rats.2. Both the therapies of deep insertion of thick needle with manipulation and shallow insertion of thin needle with no manipulation have analgetic effects on dysmenorrheal rats of cold stagnation syndrome. Among the two, the former is far better. Different acupuncture stimuli perform different regulatory trends on rats’ uterus LTB4 concentration. The mechanism need to be further studied.3. Different acupuncture stimuli intensities have different effects on temperature of point area:shallow insertion of thin needle with no manipulation has no obverse effects on temperatures of Sanyinjiao (SP6), Xuehai (SP10), the line between the two points, and uterus region. While the therapy of deep insertion of thick needle with manipulation can cause drastic temperature changes of the two points and the line between them. The law of the temperature change is characterized by decreased in instant-5mins and 10-20mins but increased in 5-10mins and 20-40mins. This change may be the reflection of acupuncture ’deqi’ through the pathway of’zang-fu organs-channel-acuponts’.4. Acupuncture of both stimuli intensities performed benign regulatory effects on uterus microcirculation and microcirculation related substances. The therapy of deep insertion of thick needle with manipulation performed more obvious instant and after effects. On the contrary, the therapy of shallow insertion of thin needle with no manipulation is mainly marked by its cumulative after effects. The therapy of deep insertion of thick needle with manipulation has a more obvious after effects compared to that of shallow insertion of thin needle with no manipulation. |
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