[Effects of electroacupuncture at points of a traditional acupuncture formula that induces labor with different parameters on uterine contractility in late-stage pregnant rats].

OBJECTIVE: To select the most effective parameters of electroacupuncture (EA) at Hegu (LI4) and Sanyinjiao (SP6), a traditional acupuncture formula that induces labor, by comparing its effects on uterine contraction in late-stage pregnant rats, so as to improve the effects of acupuncture on induction of labor and its clinical maneuverability and provide the basis for further mechanism research. METHODS: One hundred and eight female healthy adult Wistar rats were randomly divided into eight groups including one non-pregnancy control group (normal control group) and seven pregnant groups. Pregnant rats were prepared by free mating and vaginal plugs observation method and were randomly divided into six EA groups and one no EA group (model group) according to the requirements of a 2×3 factorial design. EA parameters included 2 types of waves (continuous wave and sparse-dense wave) and 3 types of frequency (15, 30 and 50 Hz). Rats in treatment groups received EA on bilateral LI4 for 20 min and then bilateral SP6 for 5 min with certain parameters accordingly. Each acupoint and a nearby non-acupoint at the proximal end (2 to 3 mm from the acupoint) were connected with two leads of one EA line. The current intensity of EA was 0.2 to 0.3 mA. A muscle tension transducer and the BL-420 Biological Experimental System were placed on the lower uterine segment of rats for detecting contractility, frequency and lasting-time of contraction wave. RESULTS: Compared with non-pregnancy control group, the amplitude, frequency and lasting time of contraction waves of uterine contraction in all EA groups and model group were significantly increased. Compared with control group, amplitudes of contraction waves of uterine contraction of both EA1 group (15 Hz continuous wave) and EA6 group (2/50 Hz sparse-dense wave) were significantly increased (P<0.01 or P<0.05); frequency of contraction waves of uterine contraction of EA6 group was significantly increased (P<0.01 or P<0.05); lasting time of contraction waves of uterine contraction of EA1 group, EA5 group (2/30 Hz sparse-dense wave) and EA6 group were significantly increased (P<0.01 or P<0.05). Comparisons between EA6 group and the other EA groups showed that, EA6 group had significantly higher levels of amplitude and frequency of contraction waves of uterine contraction than the other EA groups (P<0.01 or P<0.05) except EA1 group, there was no significant difference between EA1 group and EA6 group (P>0.05), but the mean value of EA6 group was higher than that of EA1 group; EA6 group had significantly higher levels of lasting time of contraction waves of uterine contraction than the other EA groups (P<0.01 or P<0.05) except EA1 group and EA5 group, there was no significant difference between EA6 group and EA1 group or EA5 group (P>0.05), but the mean value of EA6 group was higher than EA1 group and EA5 group. CONCLUSION: EA with sparse-dense wave (2 Hz sparse wave and 50 Hz dense wave, alternately) at LI4 and SP6 was more effective than other EA parameters for increasing uterine contraction amplitude, frequency, and lasting-time of contraction waves in late-stage pregnant rats. This combination of electric wave and frequency for EA at LI4 and SP6, a traditional acupuncture formula for inducing labor, is recommended to be applied in clinical practice for parturition as well as for treatment of uterine atony.

Experimental study on acupuncture activating the gonadotropin-releasing hormone neurons in hypothalamus.

OBJECTIVE: To probe into the most effective site, extra-ordinary point, acupoint and channel for regulating reproductive endocrine function by means of the study on acupuncture activating the gonadotropin-releasing hormone neurons (GnRH) in hypothalamus. METHODS: Female SD rats of reproductive age were used, and the in vivo study on GnRH neurons in hypothalamus was made with mimic sexual stimulation and feedback regulation. The neuron-activating effects of the acupoints on the channels pertaining to the zang- and fu-organs related with reproductive endocrine and the extra-ordinary points in different regions were studied using the discharge of the neuron as index, and then the neurons were labeled with horseradish peroxidase (HRP). RESULTS: Acupuncture was given at two acupoints each on the three yin channels of foot, the three yang channels of foot, the Conception Vessel and the Governor Vessel. The order of the mean increasing percentage in the hypothalamic GnRH neuron electric activity was: the Gallbladder Channel > the Spleen Channel > the Stomach Channel/the Bladder Channel/the Conception Vessel > the Liver Channel > the Kidney Channel > the Governor Vessel; for different acupoints, it was: Guanyuan (CV 4) > Sanyinjiao (SP 6) > Zusanli (ST 36) > Daimai (GB 26)/Yanglingquan (GB 34) > Shenshu (BL 23) > Weizhong (BL 40) > Yaoyangguan (GV 3)/Liangmen (ST 21)/Fujie (SP 14) > Qimen (LR 14)/Yingu (KI 10) > Tangzhong (CV 17)/Zhiyang (GV 9); for different positions, it was: the lower abdominal part/the lower limb part > the thoracodorsal part; for the extra-ordinary points, it was: Zigong (EX-CA1) > Dannang (EX-LE6)/Yaoyan (EX-B7) > Baichongwo (EX-LE3)/Qianzheng > Jingbi/Bizhong/Taiyang (EX-HN5) > Erbai (EX-UE2)/ Dingchun; and for the distribution sites of the extra-ordinary points: lower abdominal region > the lower limb region > the craniofacial region > the upper limb region/the thoracodorsal region. CONCLUSION: For regulating the reproductive endocrine function, the acupoints located at the same neural segment with the reproductive organ should be selected as the main points, and it is necessary to combine with syndrome differentiation of the viscera and channels.

[Effect of electroacupuncture of "Hegu" (LI4) and "Zusanli" (ST36) on intestinal sensitivity and motility in irritable bowel syndrome rats].

OBJECTIVE: To observe the effect of electroacupuncture (EA) of "Hegu"(LI4) and "Zusanli"(ST36)on changes of intestinal sensitivity and colonic motility and expression of colonic 5-hydroxytryptamine 3A receptor (5-HT3AR) in irritable bowel syndrome (IBS) rats, so as to reveal its mechanism underlying improvement of IBS. METHODS: A total of 40 neonatal Wistar rats were randomly and equally divided into normal control, model, LI4 and ST36 groups (n＝10). The IBS model was induced by mother-infant separation, acetic acid enema and colorectal distension (CRD). EA (2 Hz/100 Hz, a tolerable strength) was applied to bilateral LI4 and ST36 for 20 min, once every other day for 5 times. The Bristol stool form scale was used to assess the gastrointestinal function, and the latency and number of abdominal muscular contraction waves of abdominal withdrawal reflex (AWR) were used to evaluate the intestinal sensitivity and motility respectively. The immunoactivity of 5-HT3AR of the colon tissue was detected by immunohistochemistry. RESULTS: After modeling, the score of Bristol fecal form scale, number of muscular contraction waves and expression levels of colonic 5-HT3AR in the myometrium and mucosal layers were significantly increased (P<0.01), and the latency of muscular initial contraction wave was obviously shortened in the model group relevant to the normal control group (P<0.01). After the intervention, the increased Bristol fecal form score, number of muscular contraction waves and expression levels of 5-HT3AR in the myometrium and mucosal layers as well as the decreased latency of muscular contraction were reversed in both LI4 and ST36 groups (P<0.01, P<0.05). The effect of EA of ST36 was significantly superior to those of EA-LI4 in lowering Bristol fecal scale score and 5-HT3AR expression in the muscular layer (P<0.01), but obviously inferior to those of EA-LI4 in increasing the latency of of muscular initial contraction wave and down-regulating muscular contraction waves and 5-HT3AR expression in the mucosal layer (P<0.05, P<0.01). CONCLUSION: Both EA-LI4 and EA-ST36 can significantly improve the symptoms of abdominal pain and diarrhea, but EA-LI4 is better in suppressing intestinal high sensitivity, and EA-ST36 is better in promoting intestinal motility, suggesting a specificity of effect of acupoints of different meridians.

[Perpendicular and transverse needling of "Sanyinjiao" (SP6) relieves abdominal pain by regulating arginine vasopressin and its receptor expression levels in uterus and hypothalamus in cold-stasis type dysmenorrhea rats].

OBJECTIVE: To observe the effect of perpendicular and subcutaneous transverse needling at "Sanyinjiao" (SP6) on visceral pain behavior, arginine vasopressin (AVP) content in the serum, uterine tissues, spinal cord and hypothalamus and expression of AVP receptors AVPR1A and AVPR1B in the uterine tissues, spinal cord and hypothalamus in cold-stasis (stasis due to pathogenic cold) type dysmenorrhea rats, so as to explore their mechanisms underlying pain relief. METHODS: Forty female SD rats were randomly divided into blank control, model, perpendicular needling and transverse needling groups, with 10 rats in each group. The cold-stasis dysmenorrhea rat model was established by exposure in a freezer (-25 ℃) for 4 h, once daily for 5 days, and subcutaneous injection of estradiol benzoate (once daily for 10 days) and intra-abdominal injection of oxytocin injection (once). For rats of the two acupuncture groups, acupuncture needles were inserted into the bilateral SP6 perpendicularly or transversely to a depth of about 4-5 mm, and retained for 20 min. The abdominal pain behavior was assessed by recording the writhing latency and scaling the rats' writhing reactions after modeling. The contents of AVP in the serum, uterus, spinal cord and hypothalamus tissues were assayed using ELISA and the expression of AVPR1A and AVPR1B in the uterus, spinal cord and hypothalamus was measured by using Western blot and quantitative real time-PCR, respectively. RESULTS: After mode-ling and compared with the blank control group, the writhing latency was significantly shortened (P<0.05), and the writhing score in the first 20 min was significantly increased (P<0.01) in the model group. After the intervention, the writhing latency was significantly prolonged (P<0.01), and the writhing scores in 20 min were significantly decreased (P<0.01) in the two needling groups. The AVP contents were obviously increased in the serum and uterine tissue (P<0.05, P<0.01) but decreased appa-rently in the spinal cord and hypothalamus tissues (P<0.01, P<0.05), and the expression levels of AVPR1A or AVPR1B protein and mRNA were markedly increased in the uterine tissues (P<0.01, P<0.05), and significantly decreased in the spinal cord and hypotha-lamus in the model group relevant to the control group (P<0.05, P<0.01). Following the intervention, The AVP content in the serum of the perpendicular needling group (P<0.05) and that in the uterus of the two needling groups were significantly decreased (P<0.01), as well as that in the hypothalamus was obviously increased in the two needling groups (P<0.05). The expression levels of AVPR1A protein and mRNA in the uterus were significantly down-regulated in the two needling groups (P<0.01, P<0.05) and AVPR1B protein in the hypothalamus of the perpendicular needling group was up-regulated (P<0.05). Moreover, no significant differences were found between the two needling groups in regulating the related indexes mentioned above (P>0.05). CONCLUSION: Both perpendicular and subcutaneous transverse needling at SP6 have an immediate analgesic effect in cold-stasis type dysmenorrhea rats, which may be related to their effects in regulating AVP levels and its receptor expression in the uterine and hypothalamus.

Waggle needling wields preferable neuroprotective and anti-spastic effects on post-stroke spasticity rats by attenuating γ-aminobutyric acid transaminase and enhancing γ-aminobutyric acid.

Waggle needling, a classical anti-spastic needling technique characterized by combination of acupuncture with joint movement, has gained increasing popularity of spasticity treatment in China. This study was designed to compare the anti-spastic effect of waggle needling to the routine needling and to explore its underlying mechanism. We established post-stroke spasticity model based on ischemia stroke operation (middle cerebral artery occlusion). Rats were divided into six groups: normal control group, sham-operated control group, ischemia stroke model group, waggle needling group, routine needling group and baclofen group. Neurological function and muscle tone were assessed by the Zea Longa score and modified Ashworth scale, respectively. Indirect muscle tone was testified with electrophysiological recording. Cerebral infarction was measured by 2,3,5-triphenyltetrazolium chloride staining. The concentrations and expressions of γ-aminobutyric acid transaminase (GABAT) and γ-aminobutyric acid (GABA) were detected by enzyme-linked immunosorbent assay and western blot assay. Waggle needling markedly alleviated neurological deficits, decreased cerebral infarction and eased muscle tone; simultaneously, attenuated GABAT and enhanced GABA expression in the cortical infarct regions in comparison with the routine needling (P < 0.01), yet showed similar therapeutic effect to the baclofen group (P > 0.05). These results preliminary supported that waggle needling as a potential promising non-pharmacological intervention for the treatment of cerebral ischemia and spasticity.

Effect of Electroacupuncture at Different Acupoints on the Expression of NMDA Receptors in ACC and Colon in IBS Rats.

OBJECTIVE: To observe the effects of electroacupuncture (EA) at different acupoints on the expression of N-methyl-D-aspartate receptor (NMDA receptor ) and behaviors in irritable bowel syndrome (IBS) rats. METHODS: Wistar rats were randomly divided into blank control group (blank group, n=10) and model preparation group (n=50); experimental rat model of IBS was established by the "neonatal maternal separation and acetic acid enema" combined with "colorectal distension stimulation" method. A total of 50 IBS rats were randomly assigned to five groups of 10 each: model group, Yintang (GV29) group, Neiguan (PC6) group, Tianshu (ST25) group, and Zusanli (ST36) group. Rats in four treatment groups, aged 9 weeks old, were treated with EA by HANS with a sparse-dense wave with a frequency of 2/100 Hz, current of 0.1-0.3mA, and 20 min/stimulation, every other day for a total of 5 sessions. After treatment, the abdominal visceral sensitivity was evaluated by abdominal withdrawal reflex (AWR), and the psychological and emotional behavior of rats were evaluated by the open-field test (OFT). The expression of NMDA receptors in anterior cingulate cortex (ACC) was detected by Quantitative Real-time PCR, and the positive expression of NMDA receptors in colon was detected by immunohistochemistry. RESULTS: The IBS rat's abdomen is more sensitive and irritable; NR1, NR2A, and NR2B in ACC and NR1 and NR2B in colon of rats significantly increased in the model group versus the normal group (P<0.01) and were inhibited in all treatment groups (P<0.01, P<0.05). Additionally, NR2A and NR2B in ACC reduced more in GV29 group (P<0.01) than in other treatment groups (P all<0.05) compared with the model group. The expression of NR2B in colon was significantly inhibited in ST36 group (P<0.01) and inhibited in GV29 group and ST25 group (P all <0.05) compared with the model group. And the expression of NR2B in colon was more inhibited in ST36 group than in PC6 group (P<0.01). CONCLUSIONS: EA at different acupoints could obviously relieve abdominal pain and abnormal behaviors in IBS rats in different degrees of effects. The effect of abdominal pain-relief, from greatest to least, is ST25, ST36, GV29, and PC6, while the effect of relieving abnormal behaviors caused by IBS, from greatest to least, is GV29, PC6, ST36, and ST25. There are significant differences in the expressions of NMDA receptors in ACC and colon among different acupoints. This difference should be related to the location distribution and indications of acupoints.

[Comparison of effects of electroacupuncture at "Tianshu" (ST25) and "Dachangshu" (BL25) on intestinal sensitivity and expression of muscarinic M3R and 5-HT3AR in irritable bowel syndrome rats].

OBJECTIVE: To observe and compare the effect of electroacupuncture (EA) of "Tianshu" (ST25) and "Dachangshu" (BL25) on intestinal sensitivity and the expression of M3 muscarinic acetylcholine receptor (M3R) and 5-hydroxytryptamine 3A receptor (5-HT3AR) in the colon tissue in irritable bowel syndrome (IBS) rats, so as to explore its mechanism underlying improvement of IBS. METHODS: Forty newborn Wistar rats were randomly divided into blank control, model, ST25 and BL25 groups (n＝10 rats per group). The IBS model was established by joint application of maternal separation, acetic acid enema plus colorectal distension from day 8 to 21 after birth. At the age of 9 weeks, EA (2 Hz/100 Hz, 0.1－0.3 mA) was applied to ST25 and BL25 for 20 min, once every other day for 5 times. The modified method was used to record the abdominal withdrawal reflex (AWR), so as to evaluate the visceral sensitivity by referring to the modified Al-Chaer's and colleagues' methods. The threshold (initial wave), the number and peak-peak value of contraction waves (AWR) within 90 s were recorded. The immunoactivity of M3R and 5-HT3AR in the colon tissue was detected by immunohistochemistry. RESULTS: After modeling, the latency was obviously shortened, the number and peak-peak values of contraction waves of intestine were significantly increased, and the immunoactivity of M3R and 5-HT3AR was notably up-regulated in the model group compared with the blank control group (P<0.01). After the EA intervention, the latency was obviously prolonged, the peak-peak value in both ST25 and BL25 groups, and the number of contraction waves in the ST25 group were significantly decreased (P<0.01), and the expression levels of M3 and 5-HT3A R were considerably down-regulated in both ST25 and BL25 groups relevant to the model group (P<0.01). The therapeutic effects were significantly better in the ST25 group than in the BL25 group in prolonging the latency and reducing the contraction wave number, and in down-regulating the expression of colonic M3R and 5-HT3AR (P<0.05, P<0.01). CONCLUSION: EA at ST25 and BL25 can reduce the intestinal sensitivity in IBS rats, which is probably associated with its effects in down-regulating the expression of M3R and 5-HT3AR in the colon. The therapeutic effects of ST25 were obviously better than those of BL25 in suppressing intestinal sensitivity and expression of colonic M3R and 5-HT3AR.

[Electroacupuncture at "Zusanli" (ST36) and "Chize" (LU5) of mother rats exposed to nicotine during pregnancy and lactation has a protective effect on development of lung function and morphology in neonatal rats].

OBJECTIVE: To compare the different effect of electroacupuncture (EA) at "Zusanli" (ST36) and "Chize" (LU5) of mother rats exposed to Nicotine during pregnancy and lactation on lung function and morphological changes in offspring rats, so as to explore the most effective acupoint for improving the development of lung in neonatal rats. METHODS: A total of 24 female pregnancy SD rats were randomly divided into 4 groups: normal control, model, EA-ST36 and EA-LU5 (n＝6 rats in each group). Rats of the normal group were treated by subcutaneous injection of normal saline, and those of the other 3 groups treated by subcutaneous injection of nicotine (1 mg•kg－1•d－1) beginning from the 6th day to about the 21st day of pregnancy (childbirth day) for nicotine exposure during pregnancy and lactation. The daily EA treatment (2 Hz /15 Hz，1 mA) was applied to bilateral ST36 and LU5 for 20 min, beginning from the 6th day of pregnancy to the 21st day (childbirth day). The lung function of the offspring rats including the peak inspiratory flow (PIF), peak expiratory flow (PEF), lung resistance (RL), exhalation resistance (RE)and lung dynamic compliance (Cdyn) was detected by using a lung function analysis system. Histopathological changes (severity of alveolarization) of the offspring rats' lung tissue were observed under microscope after H．E. stain. RESULTS: Compared with the normal group, the PIF, RL and RE values were significantly increased (P<0.01), and PEF and Cdyn values significantly decreased in the model group (P<0.01). The alveolar diameter in the model group was evidently increased relevant to the normal group (P<0.01). Following the intervention, modeling induced increase of PIF, RL, RE and alveolar diameter and decrease of PEF and Cdyn values in the EA-ST36 group, and the increased PIF, RL and RE levels in the EA-LU5 group were obviously suppressed relevant to the model group (P<0.01, P<0.05). Additionally, modeling induced obvious congestion and edema of the alveolar wall, alveolar deformation, rupture and fusion, and reduction of the number of the pulmonary alveoli were evidently milder in both EA-ST36 and EA-LU5 groups. No significant differences were found between the EA-ST36 and EA-LU5 groups in the abovementioned 5 indexes of pulmonary function and alveolar diameter (P>0.05)．. CONCLUSION: EA of ST36 and LU5 of mother rats experiencing nicotine exposure during pregnancy and lactation can improve the lung function and morphological changes in neonatal rats, and the effect of ST36 is relatively better.

[Effect of electroacupuncture at "Zusanli" (ST 36) and "Yanglingquan" (GB 34) on perinatal nicotine-exposure-induced lung function and morphology of neonatal rats].

OBJECTIVE: To compare the effects of electroacupuncture (EA) at "Zusanli" (ST 36) versus "Yanglingquan" (GB 34) in the pregnant rats on perinatal nicotine-exposure-induced lung function and morphology of newborn rats and explore the rule of acupoint effect in EA for the prevention from lung dysplasia in newborn rats. METHODS: A total of 24 female SD rats were randomized into a normal saline group (S group), a nicotine group (N group), a nicotine-ST 36 group (N + ST 36 group) and a nicotine-GB 34 group (N+GB 34 group), 6 rats in each one. Starting at the 6th day of pregnancy, 0.9% sodium chloride solution was injected subcutaneously in the S group, 1 mg/kg; and in the rest 3 groups, nicotine of the same dose was injected through to the 21st postnatal day to establish the perinatal nicotine-exposure model. Simultaneously, during model preparation, EA was applied at "Zusanli" (ST 36) and "Yanglingquan" (GB 34) in the N+ST 36 group and the N+GB 34 group respectively, once a day, through to the 21st postnatal day. The lung function analytic system for small animal was adopted to observe the changes in lung function indicators in newborn rats, such as peak inspiratory flow (PIF), peak expiratory flow (PEF), expiratory resistance (RE), inspiratory resistance (RI) and dynamic compliance (Cdyn). HE staining was used to observe the morphological changes of lung, such as alveolar fusion and rupture. RESULTS: Compared with the S group, PEF and Cdyn were lower and PIF, RI and RE higher in the N group (all P<0.01), additionally, alveoli were fused and ruptured, alveolar wall thickened, the numbers of alveoli reduced, the interspace of alveoli enlarged and the diameter increased (P<0.01). Compared with the N group, in the N+ST 36 group, PEF and Cdyn were increased, PIF, RI and RE reduced (P<0.05, P<0.01), the alveolar fusion and rupture relieved, the numbers of alveoli increased, alveolar wall thinner, the interpsace of alveoli became normal and the diameter was reduced significantly (P<0.01). In the N+GB 34 group, the changes of lung function and morphological indicators were not significant (P>0.05). CONCLUSION: Electroacupuncture at "Zusanli" (ST 36) in the pregnant rats significantly improves the perinatal nicotine-exposure-induced lung function and morphology of newborn rats than electroacupuncture at "Yanglingquan" (GB 34).

[Effect of Electroacupuncture at "Neiguan" (PC 6) and "Tianshu" (ST 25) for Colonic Motility and D 2 Receptor in Irritable Bowel Syndrome Rats].

OBJECTIVE: To observe and compare the effects of electroacupuncture (EA) at "Tianshu" (ST 25) and "Neiguan" (PC 6) for colonic motility and the expression of colon dopamine D 2 in irritable bowel syndrome (IBS) rats, and to explore the specificity of different meridians and different acupoints. METHODS: Forty Wistar newborn rats were randomly divided into blank, model, Tianshu and Neiguan groups. Separation of mother and child and acetic acid coloclyster combined with colorectal distension were used to establish IBS model in the model, Tianshu and Neiguan groups. At the age of 9 weeks, EA at bilateral ST 25 and PC 6 were applied in the corresponding groups 5 times, once every other day. After the intervention, the Bristol fecal score, the latent period of abdominal retraction reflex and the number of contraction waves were recorded. The expression of dopamine D 2 receptor was detected by immunohistochemistry. RESULTS: Compared with the blank group, the Bristol fecal score of the model group was higher (P<0.01), the 1st contraction wave latent period was shorter (P<0.01), the number of contraction waves in 90 s increased (P<0.01), the immunoreactive expression of D 2 receptor in colon decreased (P<0.01). Compared with the model group, the Bristol fecal scores of the Tianshu and Neiguan groups decreased (P<0.01), the 1st contraction wave latent periods were longer (P<0.01), the numbers of contraction waves in 90 s decreased (P<0.01), the positive expressions of D 2 receptor in colon increased (P<0.01, P<0.05). Compared with the Tianshu group, the immunoreactive expression of D 2 receptor in the Neiguan group decreased (P<0.01). CONCLUSION: EA at ST 25 and PC 6 can improve the symptoms of colonic motility in IBS rats. The effect of EA at ST 25 is better, which indicates that different meridians and different acupoints play specific effects.

[Effect of Electroacupuncture at "Yintang" (GV 29) and "Tianshu" (ST 25) on the Ethology and the Expression of TRPV 1 Receptor in Colon of Irritable Bowel Syndrome Rats].

OBJECTIVE: To observe the ethology and expression of transient receptor potential vanilloid-1(TRPV 1) in colon of irritable bowel syndrome (IBS) rats treated by electroacupuncture (EA) at "Yintang" (GV 29) and "Tianshu" (ST 25), so as to explore the different effectiveness of different acupoints and its related mechanism underlying improvement of the symptoms of somatopsychic illness. METHODS: Thirty-two neonatal Wistar rats were randomly divided into blank control group, model group, Yintang (GV 29) group, and Tianshu (ST 25) group (n=8 in each group). IBS model was established by neonatal maternal separation and acetic acid enema combined with colon and rectum expansion stimulation. Rats of the GV 29 and ST 25 groups were given EA treatment at age of 9 weeks old, 20 minutes, once every other day for 5 times. Latency of the 1st time contraction wave and numbers of contractive wave in 90 s were recorded to evaluate abdominal visceral sensitivity by abdominal withdrawal reflex. Horizontal and vertical movements were observed to assess the emotional and psychological behavior of rats by open field test. TRPV 1 expression in colon was detected by immunohistochemistry method. RESULTS: Compared with the black control group, latency of the 1st time contractive wave was significantly shortened and the wave numbers in 90 s were increased significantly in the model group (P<0.01), while the numbers of vertical and horizontal movements were also significantly reduced (P<0.01), accompanied with increased TRPV 1 expression in the colon (P<0.05). Compared with the model group, After EA intervention, the levels of the above-mentioned 5 indexes were significantly reversed in the GV 29 and ST 25 groups (P<0.01,P<0.05), except the number of horizontal movement in the ST 25 group (P>0.05). The expression level of TRPV 1 was significantly lower in the ST 25 group than in the GV 29 group (P<0.01). CONCLUSIONS: EA at GV 29 and ST 25 can alleviate the abdominal pain and improve the mental and emotional disorders in IBS rats. GV 29 has a better effect on relieving the depressive-like psychoemotional behavior of IBS rats, while ST 25 is more effective in treating abdominal pain. There is significantly different influence on TRPV 1 expression in colon between these two treatment groups, which may contribute to their different effect in pain relieving.

The Effect of Acupuncture to SP6 on Skin Temperature Changes of SP6 and SP10: An Observation of "Deqi".

Background. Deqi sensation is a complex but an important component for acupuncture effect. In this study, we tried to observe the relationship between Deqi and skin temperature changes and whether there was some relativity between Deqi and needle stimulations on cold congealing and dysmenorrhea rat model. Thirty-two female Sprague Dawley (SD) rats were randomly divided into four groups (Saline Control Group, Model Group, Group A with strong stimulation, and Group B with small stimulation). Group A and Group B were performed with different stimulations. We found that, compared with saline control group, model group, and Group B, Group A showed that the skin temperature changes on right acupoint SP6 and SP10 increased significantly at 5 min-10 min interval. The skin temperature changes on left SP6 decreased at instant-5 min interval. The skin temperature changes on right SP10 decreased significantly at instant-5 min interval and 10 min-20 min interval. Thermogenic action along Spleen Meridian of Foot Greater Yin was manifested as simultaneous skin temperature increase on right SP6 and SP10 at 5 min-10 min interval after needling SP6, which was helpful to illustrate the relationship between the characteristic of Deqi and needle stimulations.

[Effect of acupuncture at different time points on kidney function and oxygen free radical metabolism in rats with simulated weightlessness].

OBJECTIVE: To explore the effects of pre-acupuncture and immediate acupuncture on kidney function and oxygen free radical metabolism in rats with simulated weightlessness. METHODS: Twenty male clean-grade Wister rats were randomly divided into a normal control group, a model group, a pre-acupuncture group and an immediate acupuncture group, 5 rats in each one. The rats in the normal control group did not receive any treatment but free activities for 4 weeks. The rats in the rest groups received 4-week tail suspension to establish the model of simulated weightlessness. One week before the tail suspension, the rats in the pre-acupuncture group were treated with electroacupuncture at "Shenshu" (BL 23), "Pishu" (BL 20) and "Sanyinjiao" (SP 6) for 30 min per treatment, once a day for 7 days. The rats in the immediate acupuncture group received tail suspension and acupuncture at the same time; during the tail suspension, the electroacupuncture was applied at "Shenshu" (BL 23), "Pishu" (BL 20) and "Sanyinjiao" (SP 6) for 30 min per treatment, once every other day for 14 days. The colorimetric method was used to measure the content of blood urea nitrogen (BUN) in serum as well as activity of superoxide dismutase (SOD) and glutathione peroxidase (GSH-PX) and content of malonaldehyde (MDA) in renal tissue in each group. RESULTS: Compared with the normal control group, the content of BUN in the model group was increased significantly (P<0.01), the activity of SOD and GSH-PX in nephridial tissue was significantly reduced (both P<0.01), and the content of MDA was increased significantly (P<0.05). Compared with the model group, the content of BUN in the pre-acupuncture group and immediate acupuncture group was significantly reduced (P<0.01, P<0.05), the activity of GSH-PX in the pre-acupuncture group was obviously increased (P<0.05) and the content of MDA in the immediate acupuncture group was increased significantly (P<0.01). Compared with the immediate acupuncture group, the content of MDA in the pre-acupuncture group was lower (P<0.01). CONCLUSION: The pre-acupuncture and immediate acupuncture both have the capacity to improve the kidney function and anti-oxygen free radical injury in rats with simulated weightlessness, however, the capacity to increase the protection ability of the kidney and eliminate free radical in the pre-acupuncture group is superior to that in the immediate acupuncture group, which is likely to be related with improving antioxidant ability of kidney.

Effects of acupuncture applied to sanyinjiao with different stimuli on uterine contraction and microcirculation in rats with dysmenorrhea of cold coagulation syndrome.

In this study, we try to evaluate the effects of acupuncture stimulation with different amounts at Sanyinjiao (SP6) on uterine contraction and uterus microcirculation in rats with dysmenorrhea of cold coagulation syndrome and to explore whether there is direct relativity between "De qi" and needle stimulus intensity. Diestrus female rats were randomly divided into 4 groups, including saline control group, model control group, "A" stimulus group (with strong stimulus), and "B" stimulus group (with weak stimulus). We found that dysmenorrhea rats of the cold coagulation syndrome present a high intensity in uterine tension and high contraction of microvascular diameter. Acupuncture applied with two different stimuli could relieve the symptoms, but, compared with "B" stimulus, "A" stimulus leads to better outcomes on reducing uterine contraction and increasing diameter of uterine microvascular; moreover, hand manipulation during needling mediates the curative effect on the microvascular diameter. Our finding indicates that using thick needles and deep insertion with hand manipulation are more effective and achieve desired level of "De qi" in dysmenorrhea rats.

Effects of preventive acupuncture and moxibustion on fat accumulation, blood lipid, and uterus e 2 of menopause rats.

Objective. To observe the effect of preventive acupuncture and moxibustion on blood lipid of menopause rats. Methods. Seventy 10-month-old SD rats with estrous cycle disorders were divided into three control groups and four treatment groups (n = 10/group) and another ten 3.5-month-old female SD rats were chosen as young control group. Preventive acupuncture and moxibustion were applied at Guanyuan (CV 4). Body weight growth rate has been recorded. Plasma total cholesterol (TC), triglyceride (TG), low density lipoprotein (LDL), and high density lipoprotein (HDL) levels and uterus E 2 level were measured. Results. Compared to young control group, plasma TC and LDL increased and uterus E 2 reduced significantly in 12-month-old control group. Compared to 12-month-old control group, plasma TC and LDL level and body weight growth rate decreased while HDL level increased remarkably in preventive acupuncture 12-month-old group. Compared to 14-month-old control group, plasma TC level and body weight growth rate decreased remarkably in preventive moxibustion 14-month-old group. Conclusions. Preventive acupuncture and moxibustion can significantly decrease the plasma TG and LDL, increase the plasma HDL, and prevent fat accumulation. Our finding suggests that preventive acupuncture and moxibustion have beneficial effects on blood lipid. Different treatment effects were found between preventive acupuncture and preventive moxibustion.

[Briefing on the 61st academic conference of Japanese Acupuncture Association].

The main content of the 61st Academic Conference of Japanese Acupuncture Association is introduced and summarized in this article. Different from former conferences, it focuses on the academic activity which take patient into the major consideration, and on the feasibility study of aerospace acupuncture treatment. Meanwhile, study on Japanese technique of shallow insertion with fine needle and its results are also introduced. The developing tendency of Japanese Acupuncture Association is discussed at the meeting as well.

Acupuncture De-qi: From Characterization to Underlying Mechanism.

De-qi refers to the participant's subjective sensations and objective body responses as well as the acupuncturist's perceptions while the acupuncturist needles certain acupoints in the participant's body. In recent years, De-qi is getting increasing attention of the researchers and many efforts have been made to understand its mechanism. By the broad literature survey, this paper explores the subjective De-qi sensation of the patients, its influencing factors, and the resulting physiological responses. The purpose of this paper is expected to find out a possible mechanism of De-qi and to provide certain scientific evidence for acupuncture fundamental research and clinical practice.

[Influence of electroacupuncture stimulation of delivery-facilitating acupoints with different stimulating parameters on serum endocrine hormones of late-stage pregnant rats].

OBJECTIVE: To observe the influence of electroacupuncture (EA) stimulation of "Hegu" (LI 4) and "Sanyinjiao"(SP 6; a classical delivery-facilitating acupoint recipe) at different parameters on endocrine hormone levels of late-stage pregnant rats, so as to find the better stimulation parameters for delivery facilitation. METHODS: Ninety-six female Wistar rats were randomly divided into eight groups: normal control (normal), pregnancy (model), EA-15 Hz, EA-30 Hz, EA-50 Hz, EA-2 Hz/ 15 Hz, EA-2 Hz/30 Hz and EA-2 Hz/50 Hz, with 12 rats in each group except EA-30 Hz group (n = 10). EA stimulation (0.2 to 0. 3 mA) was delivered to bilateral LI 4 for 20 min and then bilateral SP 6 for 5 min (25 min). The contents of serum estradiol (E2), progesterone (P) and prostaglandin E2 (PGE2) were detected by ELISA. RESULTS: Compared with the normal group, serum E2, P and PGE2 contents were significantly increased in the pregnancy model group (P<0.01), and E2/ P level was downregulated mildly (P>0.05). Compared with the model group, the contents of serum E2 in the EA-15 Hz, EA-30 Hz, EA-50 Hz, EA-2 Hz/15 Hz and EA-2 Hz/50 Hz groups, serum PGE2 contents in the EA-15 Hz, EA-30 Hz and EA-2 Hz/50 Hz groups, and E2/P levels in the EA-15 Hz, EA-30 Hz, EA-50 Hz and EA-2 Hz/50 Hz groups were significantly increased (P<0.01, P<0.05), whereas serum P contents in the EA-15 Hz, EA-30 Hz, EA-50 Hz and EA-2 Hz/50 Hz groups were significantly down-regulated (P<0. 01, P<0. 05). The effects of EA-2 Hz/50 Hz group were significantly superior to those of EA-2 Hz/15 Hz and EA-2 Hz/ 30 Hz groups in raising serum E2 and E2/P levels (P<0.05, P<0.01). No significant differences were found among the EA-15 Hz, EA-30 Hz, EA-50 Hz and EA-2 Hz/50 Hz groups in raising serum E2, PGE2 and E2/P levels, and among the EA-30 Hz, EA-50 Hz and EA-2 Hz/50 Hz groups in lowering serum P levels (P>0. 05). It displayed that the effects of EA at 15 Hz, 30 Hz and 2 Hz/50 Hz on the abovementioned 4 indexes were relatively better. CONCLUSION: EA of LI 4 and SP 6 at 15 Hz, 30 Hz and 2 Hz/ 50 Hz can effectively up-regulate serum E2, PGE2 and E2/P levels and down-regulate serum P content in pregnancy rats, which may contribute to its effect in promoting labor.

Effect of electro-acupuncture at different acupoints on neuropeptide and somatostatin in rat brain with irritable bowel syndrome.

OBJECTIVE: To compare the regulatory effects of electro-acupuncture (EA) at acupoints Zusanli (ST36) and Hegu (LI4) on the visceral hyper-sensitivity in the rat model of irritable bowel syndrome (IBS), and to explore the acting targets and specialty of acupoints. METHODS: Except 8 rats of the normal control group, the rest 32 rats were prepared to set up the IBS models. IBS animal model was prepared by enema with acetic acid. Model rats were divided into three groups. Except for rats in the model group for control, those in the other two groups were treated 20 min by EA on ST36 (EA-ST36) and LI4 (EA-LI4) respectively for 2 weeks to observe the effect on behavior response of viscera sensitivity. The changes of neuropeptide (NPY), the somatostatin (SS) levels in blood and tissues of brain and intestine were monitored as well. RESULTS: The volume thresholds for abdomen uplifting and back hunching were obviously increased after EA-ST36 (P<0.05), but showed insignificant change after EA-LI4. NPY contents lowered and SS contents increased in model rats; both EA-ST36 and EA-LI4 could raise the level of thalamic NPY (P<0.01 and P<0.05, respectively), but showed insignificant effects on NPY in colonic tissue. As for SS content, its colonic level could be reduced by EA-S36 and EA-LI4 (P<0.01 and P<0.05, respectively), however, its blood level was affected only by EA-ST36 (P<0.05). CONCLUSIONS: EA-ST36 or EA-LI4 could regulate the NPY in thalamus and SS in colonic tissue, the former could affect blood level of SS as well. It is deemed that NPY and SS may be the key substances for regulating the action of acupuncture in the brain-intestinal axis; their different levels could be regarded as an indicator for the functional difference between the acupoints.

[Effects of electroacupuncture on pain reactions, expression of spinal kappa-opioid receptor and contents of enkephalin and beta-endorphin in periaqueductal gray of midbrain in dysmenorrhea model rats].

OBJECTIVE: To observe effects of electroacupuncture (EA) on the expression of kappa-opioid receptor in the dorsal horn of the spinal cord and contents of enkephalin(ENK) and beta-endorphin (beta-EP) in the periaqueductal gray (PAG) of midbrain in dysmenorrheal rats, so as to reveal its underlying mechanism in relieving dysmenorrhea. METHODS: A total of 80 female SD rats were randomized into saline control (control), model, Sanyinjiao (SP 6), Xuanzhong (GB 39), non-acupoint groups (16 rats/group). Dysmenorrhea model was established by subcutaneous injection of estradiol benzoate (0.5 mg/rat on the 1st day and 10th day, 0.2 mg/rat from the 2nd day to the 9th day). One hour after the last injection, oxytocin (2 U/rat) was given intraperitoneally, for rats of the control group, the same dose of saline was given (i. p.). On the 10th day, EA (2 Hz/100 Hz, 0.1-0.3 mA) was applied to "Sanyinjiao" (SP 6), "Xuanzhong" (GB 39) and non-acupoint (the mid-point between the Stomach Meridian and Gallbladder Meridian, and in parallel with GB 39) for 20 min, respectively. Latency and number of writhing response, and writhing score (according to Schmauss's and Yaksh's method) were recorded. The expression of kappa-opioid receptor (kappa-OR) in T13, L1 , L2, L6 and S1 segments of spinal cord was determined by immunohistochemistry, and the contents of ENK and beta-EP in the midbrain PAG were assayed by ELISA. RESULTS: (1) Compared with the saline control group, the writhing latency of the model group was significantly shortened (P < 0.01), while the writhing times and writhing score of the model group were increased significantly (P < 0.01). Compared with the model group, the writhing latency of SP 6 group was significantly prolonged (P < 0.05), while the writhing scores and writhing times of the SP 6, GB 39 and the non-acupoint groups decreased significantly (P < 0.01). (2) In comparison with the control group, kappa-OR expression in the dorsal horn of L2 segment of spinal cord was upregulated significantly in the model group (P < 0.05). Compared to the model group, kappa-OR expression levels in the dorsal horns (DHs) of spinal T13, L1, L2, L6 and S1 segments in the SP 6 group were upregulated significantly (P < 0.01). ENK and beta-EP contents of PAG in the SP 6 and GB 39 groups were increased considerably (P < 0.01, P < 0.05). The effects of SP 6 were significantly superior to those of GB 39 in upregulating kappa-OR expression of spinal L1, L2 and L6 DHs and in upregulating beta-EP content of PAG; and superior to non-acupoint in upregulating kappa-OR expression of spinal T13, L1, L2, L6 and S1 DHs and in increasing both ENK and beta-EP contents of PAG (P < 0.01, PF < 0.05). No significant differences were found between the non-acupoint group and the model group in writhing latency, kappa-OR expression levels of spinal T13, L1, L2 and S1 DHs, and in ENK and beta-EP contents of PAG (P > 0.05). CONCLUSION: EA of SP 6 can significantly alleviate pain reactions in dysmenorrhea rats, which is closely associated with its functions in upregulating spinal kappa-OR expression and ENK and beta-EP contents in PAG. EA of SP 6, GB 39 and non-acupoint has some different degrees of efficacies in relieving dysmenorrhea and in upregulating spinal K-OR expression.