[Protecting action of acupuncture serum on calcium over-loaded neurons of the hippocampus in rats of ischemia-reperfusion].

OBJECTIVE: To investigate the effect of acupuncture serum on Ca2+ content in the cultured nervous cells of hippocampus after ischemia-reperfusion, so as to probe into humoral factors in acupuncture treatment. METHODS: The neurons of the hippocampus from the new born rats were cultured for 9-11 days. Fluorescein-molecular probe Fluo-3 AM was used for staining of intracellular Ca2+. Fluorescent levels in the nervous cells cultured with the serum of the normal rats or the rats given electroacupuncture at "Baihui" (GV 20), "Zusanli" (ST 36), "Quchi" (LI 11) and "Sanyinjiao" (SP 6) for 2 weeks were determined by using a laser confocal microscope. RESULTS: After the normal serum was added, the intracellular Ca2+ fluorescent levels increased to 697 +/- 113 from 461 +/- 96, while after acupuncture serum was added, the Ca2+ fluorescent levels decreased to 584 +/- 103 from 673 +/- 108, indicating that after addition of acupuncture serum, the increased intracellular Ca2+ content could be decreased. CONCLUSION: There are some active substances in acupuncture serum which can obviously decrease intracellular Ca2+ content after ischemia-reperfusion, so as to provide a direct evidence for role of humoral factor in acupuncture treatment.

[Correlation between the therapeutic efficacy of electroacupuncture of ear-body acupoints and mast cells in experimental gastric ulcer rats].

OBJECTIVE: To study the interrelation between the therapeutic efficacy of electroacupuncture (EA) and mast cells (MC) in experimental gastric ulcer rats. METHODS: Thirty Wistar rats were randomly divided into normal control group, model group and EA group, with 10 cases in each. Experimental gastric ulcer model was established by injecting 20% glacial acetic acid (0.05 ml) into the tissue between the muscular layer and the placenta percreta of the gastric anterior wall in the gastric antrum under anesthesia. EA (2/15 Hz, 1 mA) was applied to bilateral "Wei" (Stomach, an otopoint) and "Zusanli" (ST 36) for 30 min, once daily for 7 days. The tissues of these two acupoints were sampled to be stained with acetylcholine esterase (AChE) and toluidine blue (TB) respectively (counterstaining) after section (15 microm). The gastric ulcer tissue was stained with H&E method RESULTS: Compared with control group, the number and degranulation ratios of MC in the dermis of "Wei"-point and ST36 region in model group and EA group increased significantly (P<0.01); while in comparison with model group, both the number and the degranulation ratio of MC in EA group decreased considerably (P<0.01). H&E stained tissues showed smooth gastric mucosal membrane in control group, ulcerative injury involving the muscular layer in model group, and mild injury in EA group. CONCLUSION: EA can suppress the degranulation of MC in "Wei"-point and ST36 regions and effectively relieve ulcerative injury in gastric ulcer rats. There exists a certain degree of interrelation among the degranulation ratio of MC, gastric ulcerative injury, and the therapeutic efficacy.

[Influence of Qingpeng plaster on blood vessels and mast cells in skin tissue of channel points of nude mice].

OBJECTIVE: To investigate the changes in the numbers of blood vessels and mast cells, which is relative to blood microcirculation in skin, during Qingpeng plaster (ZANG medicine) being applied to the skin surface of nude mice. METHOD: Fifteen healthy adult nude mice were divided into Qingpeng plaster group and the control group. The sites of skin tissues stuck with the plaster or with control plaster were selected after 7 days treated, and then all the slices stained and observed. RESULT: The numbers of the vessels and the mast cells increased in the dermal tissue of Qingpeng paster group. Although there were a few dilated blood vessels in the control group, but the numbers of the vessels and the mast cells were obviously less than those of the Qingpeng plaster group. Statistical analysis showed that there was a significant difference (P < 0.001) between tow groups. CONCLUSION: Qingpeng plaster can cause obvious vascular dilatation and promot mast cells aggregation in the Qingpeng plaster stuck parts of the skin tissue.

[Effect of electroacupuncture and moxibustion of "Dazhui" (GV14) on the number and distribution of degranulated mast cells in GV14 region].

OBJECTIVE: To compare the effects of electroacupuncture (EA) and moxibustion on the degranulated mast cell number and distribution in "Dazhui" (GV14) region of rats. METHODS: Fifteen Wistar rats were randomly divided into normal control, EA and moxibustion groups, with 5 cases in each group. "Dazhui" (GV14) was stimulated by EA (3/10 Hz, 2-4 V, 20 mm) or moxibustion (2 cones, 20 mm), thereafter, the tissues of GV14 region were sampled, quickly frozen, sectioned (20 microm) and stained with toluidine blue separately for observing the degranulated state of mast cells under microscope. RESULTS: The mast cells (MCs) distributed widely in GV14 region from the dermis, subcutaneous tissue to muscle layer. Compared with control group, MCs of moxibustion group were significantly fewer in the number (P<0.05), and those degranulated MCs in both EA and moxibustion groups were evidently more in the number (P<0.01), and the number and percentage of the degranulated MCs in moxibustion group were significantly more than those of EA group (P<0.05), suggesting that EA and moxibustion could promote the release of granules of mast cells in the stimulated acupoint region, and the effect of moxibustion is stronger. CONCLUSION: Both EA and moxibustion can promote the degranulation of MCs, and the effect of moxibustion is markedly stronger in potentiating the release of granules from MCs in GV14 in rats.

[Acupuncture-serum decreases Ca2+ content in cultured rat myocardial cells].

OBJECTIVE: To explore whether humoral factors play a role in the mechanisms of acupuncture. METHODS: Primary culture of myocardial cells of neonatal rats were carried out. Five days later, they were labeled by fluorescent molecular probe Fluo-3AM. Changes of Ca2+ contents in the cultured myocardial cells after addition of the normal rat serum or acupuncture-serum of the rat who received acupuncture at "Neiguan" (PC 6) and Jianshi (PC 5), were dynamically observed by a confocal laser scanning microscopy. RESULTS: After addition of normal serum of the rat, the intracellular Ca2+ level increased to a certain degree, and then gradually tended to stability, which was significantly decreased by addition of the acupuncture-serum (P < 0.01). CONCLUSION: The serum of the rat who received acupuncture at acupoints can decrease the Ca2+ level in cultured myocardial cells, which provides a direct evidence for serum factors involving in acupuncture mechanism.

[The chemical constituents of Breynia rostrata].

AIM: To study the chemical constituents of Breynia rostrata Merr. METHODS: Chromatography was used to isolate and purify the chemical constituents, their structures were identified by spectral analysis. RESULTS: Four glycosides were identified as 6-O-methylpropanoyl-alpha-D-glucopyranoside (1), 4"-phenolic-6-O-methylpropanoyl-beta-D-glucopyranoside (2), 1-O-galloyl-beta-D-glucopyranoside (3), arbutin (4). CONCLUSION: Compounds 1 and 2 are new compounds; 3 and 4 were isolated from Breynia rostrata Merr. for the first time.

[Preliminary study on effect of acupuncture serum on Ca2+ content in cultured neurons of cerebral cortex].

OBJECTIVE: To observe effect of serum of the rat after acupuncture on Ca2+ content of cultured nervous cells in vitro for exploring the role of humoral factors in acupuncture. METHODS: Neurons of the cerebral cortex from the new born rats were taken and were cultured in a medium, 7 - 10 days later, fluorescein-molecular probe Fluo-3AM was used for staining of intracellular Ca2+. The intracellular Ca2+ levels in the nervous cells cultured with the serum of the normal rats or the rats given acupuncture at "Baihui" (GV 20), "Zusanli" (ST 36), "Quchi" (LI 11) and "Sanyinjiao" (SP 6) for 2 weeks were determined by using a laser confocal microscope. RESULTS: After addition of the serum of.normal rats, the intracellular Ca2+ level increased and then gradually got stable, and when the acupuncture serum was added to the cultured medium, its level decreased to a certain extent. CONCLUSION: Acupuncture serum can significantly decrease Ca2+ level in cultured cells of the cerebral cortex, which provides a direct evidence for involvement of humoral factors in the role of acupuncture.

[Effect of acupuncture serum on intracellular Ca2+ concentration in cultured neurons].

OBJECTIVE: To study the humoral factor of acupuncture in improving functional activities of the organism. METHODS: Neurons of the cerebral cortex from the new born rats were taken to be cultured in a strictly controlled medium. Seven to ten days later, fluorescein-molecular probe Fluo-3AM was used for staining intracellular Ca(2+). Ca(2+) concentration and its changes after application of acupuncture serum were measured by using a laser confocal microscope. Normal serum and acupuncture serum were derived from the normal control rats and the rats undergoing electro-acupuncture of "Baihui (GV 20)", "Zusanli (ST 36)", "Quchi (LI 11)" and "Sanyinjiao (SP 6)" for 2 weeks. RESULTS: After application of normal serum, the intracellular Ca(2+) concentration increased strikingly immediately at the beginning, and gradually got stable afterwards. When acupuncture serum was added to the culture fluid, Ca(2+) concentration lowered significantly. CONCLUSION: These results suggest that bioactive substances in serum collected from rats undergoing acupuncture can affect the intracellular Ca(2+) concentration of the cerebral neurons, and the humoral factor may participate in the effect of acupuncture in regulating functional activities of the organism.