[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.

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.

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.

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.

[Correlation between acupoints and zang-fu organs based on clinical trials].

A systematic analysis was made on the correlation between acupoints and pathological changes of the zang-fu organs through reviewing of literatures in the database of CNKI from 1959--2011 and the database of Pubmed of the past 10 years. The result showed that specificity was found on the pathological changes of zang-fu organs when acupoints were stimulated. And the pathological changes of the internal organs can be perceived from acupoints on the correspondent meridians, mainly regarding the heart, the stomach, the intestines, the lung and the liver, etc. And most of the researches focused on the correlation between acupoints of the heat meridian and the heart. It was also discovered that a lot of acupoints on various meridians could manifest the pathological changes of the same organ. Different effects of the same acupoints on different times could be found on the same internal organ. Therefore, it is concluded that there is a relative specificity between acupoints and pathological changes of the zang-fu organs. However, it is worth to study the regularity of specificity further.

[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.

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.

[Effect of electroacupuncture at different acupoints on plasma TXB2 and 6-keto-PGF(1alpha), contents in dysmenorrhea rats].

OBJECTIVE: To observe the effect of electroacupuncture (EA) on plasma thromboxane B2(TXB2) and 6-keto-prostaglandin F1alpha (6-keto-PGF1alpha) levels in dysmenorrhea rats in order to investigate its mechanism underlying relief of primary dysmenorrhea and specificity of acupoint efficacy. METHODS: Female SD rats with diestrus were randomly divided into saline control (control), model, EA Sanyinjiao (SP 6), EA Xuehai (SP 10), EA Xuanzhong (GB 39) and EA non-acupoint (NAP) groups, with 10 rats in each. Dysmenorrhea model was established by subcutaneous injection of Estradiol Benzoate (0.5 mg/rat on the 1st and 10th day, and 0.2 mg/rat from the 2nd to the 9th day) and intraperitoneal injection of Oxytocin (0.2 mL/rat, 1 h after last injection of Estradiol Benzoate on the 10th day). EA was applied to bilateral SP 6, SP 10, GB 39, and non-acupoint (the mid-point between the Gallbladder and Stomach meridian at the GB 39 level) for 20 min. The latency and score of writhing were recorded for 20 min. Plasma TXB2 and 6-keto-PGF1alpha contents were detected by radioimmunoassay. RESULTS: Compared with the control group, the latency of writhing in the model group was shortened considerably (P < 0.01), and the writhing score was increased significantly (P < 0.01). In comparison with the model group, the writhing latency was increased significantly only in the EA-SP 6 group (P < 0.05), and the writhing scores in the EA-SP 6, EA-SP 10, EA-GB 39 and EA-NAP groups were reduced remarkably (P < 0.01). Plasma TXB2 content and the ratio of TXB2/6-keto-PGF1alpha. were significantly higher in the model group than in the control group (P < 0.01). Compared to the model group, plasma TXB2 levels and the ratios of TXB2/6-keto-PGF1alpha. in the EA-SP 6, EA-SP 10, EA-GB 39 and EA-NAP groups were downregulated markedly (P < 0.05, P < 0.01), while plasma 6-keto-PGF1alpha was upregulated strikingly only in the EA-SP 6 group (P < 0.05). No significant differences were found among the EA-SP 6, EA-SP 10, EA-GB 39 and EA-NAP groups in the writhing latency and writhing score, plasma TXB2 and 6-keto-PGF1alpha, levels (P > 0.05). CONCLUSION: EA can relieve pain reaction in dysmenorrhea rats, which may be closely associated with its effects in downregulating plasma TXB2, upregulating plasma 6-keto-PGF1alpha, content, and balancing plasma TXB2/6-keto-PGF1alpha. The effect of EA of SP 6 is relatively better.

[Effect of electroacupuncture on the uterine microcirculation in dysmenorrhea rats].

UNLABELLED: OBJECTIVE To observe the effect of electroacupuncture (EA) on uterine microcirculation in dysmenorrhea rats so as to investigate its mechanism underlying relieving primary dysmenorrheal (PD). METHODS: Female SD rats undergoing diestrus were randomly divided into saline control, model, Sanyinjiao(SP 6), Xuanzhong(GB 39) and non-acupoint groups, with 6 rats in each. PD model was established by subcutaneous injection of estradiol benzoate for 10 days and intra-peritoneal injection of oxytocin (2 U) 1 h after the last estradiol injection. EA (2 Hz/100 Hz, 1- 1.5 mA) was applied to the above-mentioned acupoints and non-acupoint area for 20 minutes. Numbers and diameters of the uterine microvessels (11-100 microm) and capillaries (< or =10 microm), and the state of the uterine microcirculation were observed by using a Cold Light Microcirculation Detector. RESULTS: In comparison with the saline control group, the numbers and diameters of uterine microvessels and capillaries at the corresponding time-points 5 min, 10 min and 20 min were decreased significantly in the model group (P<0. 05,P<0. 01). While compared with the model group, the numbers and diameters of uterine microvessels and capillaries at 20 min after EA in the SP 6 group were increased significantly (P<0. 05). The diameter of uterus capillaries of SP 6 group was significantly bigger than that of the non-acupoint group at the time-point 20 min (P<0. 05). No significant differences were found among the SP 6, GB 39 and non-acupoint groups in the diameter of uterine microvessels (P>0. 05). CONCLUSION: EA of SP 6 can effectively increase the number of uterine microvessels and capillaries and the diameter of the uterine microvessels and capillaries in PD rats, which may contribute to its effect in relieving dysmenorrhea by improving uterine microcirculation.

[Effect of moxibustion on hepatic glycogen and ultrastructure of exercise-induced fatigue rats].

OBJECTIVE: To observe the effect of moxibustion of "Zusanli" (ST 36) on hepatic glycogen-level and ultrastructure changes in exercise-induced fatigue rats. METHODS: Thirty-three SD rats were randomly and equally divided into control, model and moxibustion groups. The fatigue model was established by forcing the rat to have a loaded exhaustion swim, once daily for 21 days. Moxibustion was applied to bilateral "Zusanli" for about 30 min, once every other day for 11 times. The hepatic glycogen content was detected by chromatometry and the hepatic ultrastructure was observed by using transmission electron microscope. RESULTS: The hepatic glycogen content in the fatigue model group decreased significantly compared with the control group(P<0. 01), and that in the moxibustion group was increased significantly compared with the model group(P<0. 05). Under transmission electron microscope , it was found that the glycogen in the hypatocytes of the model was decreased and the ultrastructure of mitochondria and rough endoplasmic reticulum was unclear in outline. In moxibustion group, more glycogen granules were found in hepatocytes, and the structure of mitochondria and rough endoplasmic reticulum was basically clear. CONCLUSION: Moxibustion may increase hepatic glycogen content and improve ultrastructure of hypatocytes in fatigue rats, which may be part of its mechanism underlying relieving exercise-induced fatigue.

[Effects of instant electroacupuncture at the different acupoints on IP3 in the uterus tissue of dysmenorrhea model rats].

OBJECTIVE: To compare the effects of instant electroacupuncture (EA) at the different acupoints on IP3 in the uterus tissue of dysmenorrhea model rats so as to investigate the specificity of acupoints. METHODS: Fifty female SD rats were randomly divided into a saline group, a model group, a Sanyinjiao (SF 6) group, a Xuehai (SP 10) group and a Hegu (LI 4) group, 10 rats in each group. The rats were given subcutaneous injection of Estradiol Beozoate injection for 10 consecutive days except those in the saline group, and intraperitoneal injection of 2U Oxytocin at 1 h after the last administration to create the dysmenorrhea rats model, and the saline group was given the same dose of saline every day. On the 10th day the rats in each EA group were given EA 20 min, and the rats in the saline group and model group were bound 20 min, and the writhing response was observed at the same time. The uterine IP3 contents were detected with enzyme-linked immunosorbent assay method. RESULTS: (1) Compared with (0.311+/- 0.253) in the saline group, the writhing scores per minute of (5.867 +/- 3.442) in the model group and (2.311 +/- 0.957) in the Xuehai (SP 10) group were both increased significantly (P < 0.01, P < 0.05), and (1.833 +/- 1.355) in the Sanyinjiao (SP 6) group and (0.743 +/- 0.306) in the Hegu (LI 4) group showed no significant differences (P > 0.05). Compared with that in the model group, the writhing scores per minute decreased significantly (all P < 0.01) in all the EA groups, with no significant differences among all the EA groups (all P > 0.05). (2) Compared with (2.698 +/- 1.491) ng/mg in the saline group, IP3 contents of the uterus of (0.813 +/- 0.899) ng/mg in the model group, (1.740 +/- 0.375) ng/mg in the Sanyinjiao (SP 6) group and (0.692 +/- 0.212) ng/mg in the Hegu (LI 4) group were all lower significantly (P < 0.05, P < 0.01), and (0.743+/- 0.306) ng/mg in the Xuehai (SP 10) group showed no significant differences (P > 0.05). Compared with that in the model group, IP3 content of the uterus in the Hegu (LI 4) group showed no significant difference (P > 0.05), and those in the Sanyinjiao (SP 6) group and in the Xuehai (SP 10) group increased significantly (both P < 0 05), which were significantly higher than that in the Hegu (II 4) group (P < 0.05, P < 0.01). CONCLUSION: There are no significant differences among the instant EA groups in improving the dysmenorrhea symptoms, but there is obvious specificity of acupoint effects in the regulation of IP3. Electroacupuncture at "Sanyinjiao (SP 6) " Xuehai (SP 10)" has more marked effect in dysmenorrhea model rats.

[Comparative study on the analgesic effect of electroacupuncture at "Sanyinjiao" (SP 6), "Hegu" (LI 4) and "Xuehai" (SP 10) in dysmenorrhea rats].

OBJECTIVE: To observe and compare the analgesic effect of electroacupuncture (EA) at "Sanyinjiso" (SP 6), "Hegu" (LI 4) and "Xuehai" (SP 10) in dysmenorrhea rats so as to investigate the functional specificity of acupoint. METHODS: One hundred SD female rats aged three-month-old in the diestrus examined by the vaginal smear screening were randomly divided into a saline group, a model group, a Sanyinjiao (SF 6) group, a Hegu (LI 4) group and a Xuehai (SF 10) group, 20 rats in each group. The rats in the last four groups were given Estradiol Benzoate combined with Oxytocin to prepare the models of dysmenorrhea and in the saline group were given the same dose of saline. Last three groups were treated with electroacupuncture (EA) at Sanyinjiao" (SF 6), "Hegu" (LI 4) and 'Xuehai" (SF 10) respectively. The writhing behaviors of the rats in each group were observed within 20 min performing electroacupuncture from the start of the pain and the waveforms of uterine contraction were recorded. RESULTS: In terms of the writhing behaviors, the writhing latency in the Hegu (LI 4) group was extended significantly (P < 0.05, P < 0.01) and the writhing numbers in the Hegu (LI 4) group were decreased significantly (both P < 0.05) as compared with those in the Sanyinjiao (SF 6) group and in the Xuehai (SF 10) group. In terms of the waveforms of uterine contraction, compared with that in the model group, the amplitude and the activity of uterine contraction in both Sanyinjiao (SF 6) group and Hegu (LI 4) group were decreased significantly (all P < 0.05), and with no significant difference between the Xuehai (SF 10) and the model group (P > 0.05). CONCLUSION: For comprehensive assessment of the analgesic effect of electroacupuncture at different acupoints by using the writhing behaviors and the waveforms of uterine contraction, "Hegu" (LI 4) is the best, "Sanyinjiao" (SP 6) is the next and "Xuehai" (SP 10) is the third. showing a relatively functional specificity of the acupoint.

[Effect of preventive moxibustion on heat shock proteins and stress hormones in natural menopausal rats].

OBJECTIVE: To investigate the effect of preventive moxibustion at the acupoint of 'Guanyuan" (CV 4) on the stress-related hormones, proteins and genes in natural menopausal rats, and explore its protective mechanism. METHODS: One hundred and twenty eight healthy female SD rats were used in this study, in which 16 rats aged 4 months were used as normal control group, the rest 10-month-old rats with disorder of estrus cycle were randomly divided into 7 groups, including 4 control groups at the age of 10, 12, 14 and 16 months, and 3 preventive moxibustion groups at the age of 12, 14 and 16 months, 16 cases in each group. The 10-month-old rats were treated with preventive moxibustion at the acupoint of "Guanyuan" (CV 4), twice a week for 8 weeks consistently, then observed on the following 12, 14 and 16 months. Its effects on the level of heat shock protein 70 (HSP 70), heat shock protein 70 mRNA (HSP 70 mRNA), corticotropin-releasing hormone (CRH), beta-endorphin (beta-EP), adrenocorticotrophic hormone (ACTH) in the different nuclei of hypothalamus were examined with radio-immunity, immunohistochemistry or in situ hybridization methods. RESULTS: In the control groups, the expression of HSP 70 and HSP 70 mRNA was increased in paraventricular, arcuate and supraoptic nucleus, the level of CRH, beta-EP was showed with declining trend in volatility, while the content of ACTH was increased. In comparison to the same-aged control groups, the preventive moxibustion groups were showed that the expression of HSP 70 in the 14-month-old group was lower and the expression of HSP 70 mRNA in the 12 and 16-month-old group was significantly higher in the paraventricular nucleus (all P < 0.05), the expression of HSP 70 in the 12 and 14-month-old groups and HSP 70 mRNA in the 12-month-old group was significantly increased in the arcuate nucleus (P < 0.05, P < 0.01), while the expression of HSP 70 in the 14-month-old group and HSP 70 mRNA in the 12 and 16-month-old group was significantly decreased in the supraoptic nucleus (all P < 0.05), the content of CRH and ACTH decreased significantly in 14-month group (P < 0.05, P < 0.01), but the content of beta-EP obviously went up in the 12-month-old group (P < 0.05). CONCLUSION: Preventive moxibustion at the acupoint of "Guanyuan" (CV 4) can regulate the expression of stress proteins, genes and hormones in the hypothalamus for the natural menopausal rats, which might be playing a protective role.

[Effects of electroacupuncture of "Sanyinjiao" (SP 6) on contents of MDA and beta-EP, and expression of HSP 70 in the uterus in rats with dysmenorrhea].

OBJECTIVE: To investigate the effect of electroacupuncture (EA) of "Sanyinjiao" (SP 6) on the uterus in dysmenorrhea rats so as to study its underlying analgesic mechanism. METHODS: A total of 48 SD rats during diestrus were randomized into normal saline (control) group, model group and acupuncture group according to a random number table, with 16 rats in each group. Dysmenorrhea model was established by subcutaneous injection of Estradiol benzoate (0.5 mg/d on the 1st and 10th day, and 0.2 mg/d from day 2 to day 9, once daily for 10 days) and oxytocin (2 U/rat, once on day 10). Malondialdehyde (MDA) and beta-endorphin (beta-EP) contents in the uterus were detected by radioimmunoassay, and the heat shock protein 70 (HSP 70) immunoactivity of the uterus was detected by immunohistochemistry. RESULTS: In comparison with the control group, MDA content in the uterus was increased significantly in the model group (P < 0.01), while the beta-EP level and the immunoactivity of HSP 70 immune-reaction (IR) positive products in the uterus decrease significantly (P < 0.01) and moderately, respectively in the model group. In comparison with the model group, uterine MDA content in the EA group was decreased significantly (P < 0.01), while uterine beta-EP level increased considerably (P < 0.01) and HSP 70 expression was upregulated to a certain degree. CONCLUSION: EA of "Sanyinjiao" (SP 6) can reduce MDA content and upregulate beta-EP level of the uterus in rats with dysmenorrhea, which may contribute to its analgesic effect in relieving dysmenorrhea by clearing away oxygen free radicals and raising analgesic substance in the uterus.

[Influence of electroacupuncture of "Dazhui" (GV 14) "Mingmen" (GV 4) and non-acupoint on the inflammation and immune reactions in adjuvant arthritis rats].

OBJECTIVE: To observe the effect of electroacupuncture (EA) on the levels of lymphocyte proliferation rate, spleen and serum interleukin-2 (IL-2), serum tumor necrosis factor-alpha (TNF-alpha), so as to analyze its immuno-regulative function in adjuvant arthritis (AA) rats. METHODS: A total of 40 Wistar rats were randomized into normal control, model, Dazhui (GV 14), Mingmen (GV 4) and non-acupoint groups. Adjuvant arthritis model was established by subcutaneous injection of Freund's complete adjuvant into the paw. EA was applied to "Dazhui" (GV 14), "Mingmen" (GV 4) and non-acupoint (mid-tail) for 20 min, once every other day, 8 times altogether. Paw volume (swelling degree of the voix pedis) was detected; T and B lymphocyte transformation rates (LTR), and IL-2 activity of the spleen tissue were assayed with MTT chromatometry, and serum IL-2 and TNF-alpha contents were determined by radioimmunoassay. RESULTS: Compared to normal group, the paw volume and serum TNF-alpha content of model group increased significantly (P < 0:01, P < 0.05), and T- and B-lymphocyte proliferation rates and spleen IL-2 activity in model group decreased considerably (P < 0.01). In comparison with model group, paw volumes of GV 14, GV 4, and non-acupoint groups and serum TNF-alpha content of GV 14 group lowered significantly (P < 0.01, P < 0.05); while T- and B-lymphocyte proliferation rates, spleen IL-2 activity in GV 14, GV 4, and non-acupoint groups, as well as serum IL-2 content in GV 14 and GV 4 groups increased remarkably (P < 0.01). Paw volumes of both GV 14 and GV 4 groups were significantly lower than that of non-acupoint group (P < 0.05); T- and B-lymphocyte proliferation rates and IL-2 activity of GV 4 group were significantly higher than those of non-acupoint group (P < 0.05). No significant differences were found between GV 14 and GV 4 groups in paw volume, T- and B-lymphocyte proliferation rates, spleen IL-2 activity, serum IL-2 and TNF-alpha contents, and between GV 14 and non-acupoint groups in T- and B-lymphocyte proliferation rates, as well as among GV 14, GV 4 and non-acupoint groups in serum IL-2 and TNF-alpha contents (P > 0.05). CONCLUSION: Electroacupuncture of GV 14 and GV 4 induced immunoregulation on lymphocyte, IL-2 and TNF-alpha may contribute to its anti-inflammation effect in AA rats.

[Effect of preventive acupuncture and moxibustion at "Guanyuan" (CV 4) on the expression of HSP 70 and HSP 70 mRNA in spleen and the contents of serum IL-2, TNF-alpha in menopausal rats].

OBJECTIVE: To observe the influence of preventive acupuncture (PA) and preventive moxibustion (PM) at "Guanyuan" (CV 4) on the immune function in natural climacteric rats. METHODS: A total of 160 female SD rats were randomized into control, PA and PM groups, the former one group was further divided into 10 month (mon), 12 mon, 14 mon and 16 mon subgroups, and the later two groups were further divided into 12 mon, 14 mon and 16 mon subgroups, with 16 rats in each. In addition, other 16 female SD rats aged 3.5 mon were used as the young control (YC) group. "Guanyuan" (CV 4) was punctured with an acupuncture needle and the needle was retained for 20 min, or given with one ignited moxa-cone from the age of 10 mon on. The treatment was conducted twice every week, 8 weeks altogether. The expression of HSP 70 and HSP 70 mRNA of the spleen tissue was detected by using immunohistochemistry and in situ hybridization respectively, and serum IL-2 and TNF-alpha contents were assayed by using radio-immunoassay. RESULTS: In comparison with YC group, 1) the expression of spleen HSP 70 and HSP 70 mRNA increased significantly in 10 mon control (mon-C), 12 mon-PM and 12 mon-PA groups, and 14 mon-PA group (only HSP 70 mRNA) (P < 0.05, P < 0.01); 2) HSP 70 expression decreased remarkably in 14 mon-C, 16 mon-C and 16 mon-PA groups (P < 0.05, P < 0.01); 3) IL-2 contents decreased evidently in 12 mon-C and 14 mon-C groups, and TNF-alpha contents increased obviously in 12 mon-PM, 12 mon-PA and 16 mon-C groups (P < 0.05). In comparison with the corresponding same age control groups, HSP 70 and HSP 70 mRNA expression increased significantly in 12 mon-PM and 12 mon-PA groups, 14 mon-PM and 16 mon-PM (HSP 70 only), 14 mon-PA (HSP 70 mRNA only) groups (P < 0.05, P < 0.01); IL-2 level of 12 mon-PM group, and TNF-alpha contents of 12 mon-PM and 12 mon-PA groups increased markedly (P < 0.05, P < 0.01). No significant differences were found between PM and PA groups in most different age groups (P > 0.05). CONCLUSION: Both preventive acupuncture and preventive moxibustion can upregulate the expression of spleen HSP 70 and HSP 70 mRNA and serum IL-2 and TNF-alpha levels, which may contribute to their effects in enhancing the immune function in menopausal rats.

[Effect of electroacupuncture of different acupoints on blood pressure, plasma angiotensin II, aldosterone and atrial natriuretic peptide in spontaneous hypertension rats].

OBJECTIVE: To observe the effect of electroacupuncture (EA) of different acupoints on blood pressure (BP), plasma angiotensin (Ang) II, aldosterone (ALD) and atrial natriuretic peptide (ANP) of SHR, so as to observe the relative specificity of effects of acupoints. METHODS: A total of 60 SHR were randomly divided into model, Quchi (LI 11), Zusanli (ST 36), Shenmen (HT 7), Jianyu (LI 15) and non-acupoint groups, with 10 cases in each. Other 10 Wistar rats were composed of normal group. EA (2 Hz, 2 mA) was applied to LI 11, ST 36, HT 7, LI 15 and non-acupoint for 30 min, followed by measuring BP with BP-6 system. Plasma Ang II, ALD and ANP contents were assayed by radioimmunoassay. RESULTS: Compared with control group and pre-treatment, both systolic pressure (SP) and diastolic pressure (DP) of Quchi (LI 11), Zusanli (ST 36) and Shenmen (HT 7) groups lowered significantly (P<0.05). No significant differences were found in the SP and DP of Jianyu (LI 15) and non-acupoint groups (P>0.05). After EA, the SP of Quchi (LI 11) and Shenmen (HT 7) groups were significantly lower than that of Jianyu (LI 15) and non-acupoint groups (P<0.05); and the DP of Quchi (LI 11), Zusanli (ST 36) and Shenmen (HT 7) groups were markedly lower than that of Jianyu (LI 15) and non-acupoint groups (P<0.05). Compared with normal group, the contents of plasma Ang II and ANP of model group decreased significantly, and plasma ALD level of model group increased remarkably (P<0.05). Compared with model group, plasma AngII levels in Quchi (LI 11), Zusanli (ST 36) and Shenmen (HT 7) groups and ALD in Zusanli (ST 36) and Shenmen (HT 7) groups lowered significantly (P<0.05), and plasma ANP content in Shenmen (HT 7) group increased obviously (P<0.05). CONCLUSION: EA of LI 11, HT 7 and ST 36 can effectively lowered both DP and SP in SHR, which may be closely related to its effects in regulating the contents of Ang II , ALD and ANP.

[Observation on the protective effect of preventive acupuncture and moxibustion on uterus in ovariectomized rats].

OBJECTIVE: To observe the effect of preventive acupuncture and moxibustion of "Guanyuan" (CV 4) on uterus in ovariectomized (OVX) rats. METHODS: A total of 80 female SD rats were randomly divided into 5 groups: normal control, sham-operation (sham), model, preventive acupuncture (PA) and preventive moxibustion (PM) groups, with 16 cases in each. PA (with the needle retained for 20 min after insertion) and PM (one moxa-cone/rat) were applied to "Guanyuan" (CV 4) separately before ovariectomy, two times a week, 4 weeks altogether. Then, ovariectomy was performed on rats of model, PA and PM groups. Uterus tissue was taken under anesthesia for homogenate (10 rats/group) or sectioning (5-6 microm, 6 rats/group). Uterus estradiol (E2), progestone (P) were detected with radioimmunoassay; superoxide dismutase (SOD), nitric oxide synthase (NOS), and malondialdehyde (MDA) were assayed with immunoturdidimetry. Partial uterus slices were stained with H & E method for observing morphological changes, or stained with immunohistochemical method or with in situ hybridization method for displaying estrogen receptor (ER)-alpha and ER-alpha mRNA expression separately. RESULTS: After ovariectomy, the rat's uterus presented obvious squamous metaplasia, hyperplasy, and thickening of the endomembrane, decrease in glands and blood vessels and increase in fibrous connective tissue, etc; while the situation was evidently better in PA and PM groups. In comparison with normal control group, E2, P contents and SOD, NOS activities of uterus tissue in model group decreased significantly (P < 0.01), while MDA in model group increased evidently (P < 0.01). Compared with model group, uterus P content in PA and PM groups increased obviously (P < 0.05), while uterus MDA decreased apparently (P < 0.01). Compared with normal control group, the expression of ER-alpha and ER-alpha mRNA in model group was downregulated markedly (P < 0.01); after PA and PM, both ER-alpha and ER-alpha mRNA expression increased obviously (P < 0.01). No significant differences were found between PA and PM groups in the above mentioned 7 indexes (P > 0.05). CONCLUSION: Preventive acupuncture and moxibustion of CV4 can postpone the structural degeneration of uterus in OVX rats, which may be related to their effects in modulating the secretion of uterus E2 and P, upregulating the expression of ER-a and ER-alpha mRNA, and improving the anti-oxidative ability.

[Experimental study on changes of expressions of ER-alpha and ER-alpha mRNA in different hypothalamic nuclei and the effect of "preventive acupuncture and moxibustion" in ovariectomy rats].

OBJECTIVE: To explore the mechanism of "preventive acupuncture and moxibustion" for regulating hypothalamic function in ovariectomy rats. METHODS: Thirty female SD rats, aged 3.5 months, were randomly divided into normal group, sham operation group, ovariectomy model group, preventive acupuncture group and preventive moxibustion group. After "preventive acupuncture and moxibustion" were previously given to the preventive acupuncture group and the preventive moxibustion group for one month, respectively. At the same time, ovarietomy was made in the preventive acupuncture group, the preventive moxibustion group and the ovariectomy model group. For the sham operation group, only a little fat around the ovary was removed without ovariectomy, and nothing was not applied to the normal group. Immunohistochemical and in situs hybridization methods were used to investigate changes of expressions of ER-alpha and ER-alpha mRNA in paraventricular nucleus, supraoptic nucleus and arcuate nucleus. RESULTS: In the model group the expressions of ER-alpha and ER-alpha mRNA in paraventricular nucleus and the expression of ER-alpha mRNA in supraoptic nucleus increased significantly (P<0.01), but the expression of ER-alpha dropped significantly in both supraoptic nucleus and arcuate nucleus (P<0.01). After preconditioning with acupuncture or moxibustion, the expression of ER-alpha mRNA in paraventricular nucleus significantly reduced (P<0.05), and the expression of ER-alpha had no significant change (P>0.05); the expression of ER-alpha in supraoptic nucleus and arcuate nucleus significantly increased (P<0.01 or P<0.05), and the expression of ER-alpha mRNA in paraventricular nucleus had no significant change (P>0.05). CONCLUSION: "Preventive acupuncture and moxibustion" at "Guanyuan" (CV 4) have an effect on the expressions of ER-alpha and ER-alpha mRNA in different hypothalamic nuclei in ovariectomy rats, which possibly is one of the ways for modulating the hypothalamic activity.