[Effect of Acupuncture and Moxibustion Intervention on Contents of Serum Interleukin-7 and In- terleukin-18 in Cyclophosphamide-induced Immune Suppression in Tumor-bearing Mice].

OBJECTIVE: To observe the effect of acupuncture and moxibustion intervention on the contents of serum IL-7 and IL-18 and white blood cell counts in cyclophosphamide (CTX)-induced hypofunction of immunity in tumor-bearing mice, so as to explore its mechanism underlying improvement of immune suppression caused by chemotherapy. METHODS: Sixty-four male Kunming mice were implanted with S 180 sarcoma at the left armpit (for making tumor-bearing model) , and then, randomized into control group, model group, acupuncture group and moxibustion group( 16 mice in each group). The mice of the latter 3 groups were injected with CTX (i. p. , 150 mg · kg(-1) · d(-1)), and those of the control group were injected (i. p.) with normal saline. Filiform acupuncture needle or moxibustion was applied to "Dazhui" (GV 14) , unilateral "Geshu" (BL 17) , "Shenshu"(BL 23) and "Zusanli"(ST 36) once daily for 3 or 5 days. Contents of serum IL-7 and IL-18 were detected by ELISA. RESULTS: In comparison with the control group, the plasma leukocyte counts from day 2 to day 5, the spleen index and serum IL-18 contents on day 3 and 5 after administration of CTX were significantly decreased, and serum IL-7 contents on day 3 and 5 after CTX remarkably increased in the model group (P < 0.05). Following the treatment, the spleen index on day 3 and serum IL-18 content on day 5 in the moxibustion group, serum IL-7 and IL-18 contents on day 3 and 5 in the acupuncture group were significantly up-regulated (P < 0.05). The effect of acupuncture was markedly superior to that of moxibustion in up-regulating serum IL-18 contents on day 3 and 5 after the treatment (P < 0.05). No significant differences were found between the acupuncture and moxibustion groups in the leukocyte counts and spleen index after the treatment, and in serum IL-7 contents on day 3 (P > 0.05). CONCLUSION: Acupuncture and moxibustion can up-regulate the serum IL-7 and IL-18 and white blood cell levels in immuno-suppression mice, wnicn may contribute to their effects in relieving immune hypofunction caused by alkylating agent CTX.

[Effects of acupuncture and moxibustion on serum contents of hematopoietic growth factor in mice with marrow inhibition].

OBJECTIVE: To observe serum contents of granulocyte-macrophage colony-stimulating factor (GM-CSF) and granulocyte colony-stimulating factor (G-CSF) in mice with marrow inhibition before and after acupuncture and moxibustion treatment, so as to discuss the molecular biological mechanisms of acupuncture and moxibustion on improving marrow inhibition and increasing white cells after chemotherapy. METHODS: Eighty clean-grade male Kunming mice were selected and randomly divided into a normal group, a model group, an acupuncture group and a moxibustion group according to the weight, 20 cases in each one. Mice in the model group, acupuncture group and moxibustion group were injected with cyclophosphamide (CTX) to establish mice models of marrow inhibition, while mice in the normal group received intraperitoneal injection of 0.9% NaCl. Four hours after model establishment, mice in the acupuncture group and moxibustion group were treated with acupuncture or moxibustion at "Dazhui" (GV 14), "Geshu" (BL 17), "Shenshu" (BL 23) and "Zusanli" (ST 36), respectively. Mice in the normal group and model group were immobilized without any treatment. All the treatment was given once a day for consecutive 5 days. Mice blood samples were collected from caudal vein. With manual examination, the white blood cells in peripheral blood were measured on each day from model establishment to end of treatment. Enzyme linked immunosorbent assay (ELISA) method was used to measure the serum contents of GM-CSF and G-CSF 3 days and 5 days after treatment. RESULTS: Compared with the normal group, the white cells in the model group were all reduced at each time point (all P<0.05), and the serum contents of GM-CSF and G-CSF were significantly reduced (all P<0.05). Three days after treatment, compared with the model group, the white cells in the acupuncture group and moxibustion group were increased, and the difference in acupuncture group was significant (P<0.05); the serum contents of GM-CSF and G-CSF were significantly lifted (P<0.05). Four days after treatment, compared with the model group, the white cells in the acupuncture group and moxibustion group were increased (both P<0.05). Five days after treatment, compared with the model group, the white cells in the acupuncture group and moxibustion group were increased and close to the normal level; the serum contents of GM-CSF and G-CSF were significantly lifted (all P<0.05). CONCLUSION: Through increasing serum contents of GM-CSF and G-CSF in CTX mice, acupuncture and moxibustion could prompt maturation and proliferation of myeloid hematopoietic cells, which is benefit to the reconstruction of hematopoietic function and relieve the marrow inhibition caused by CTX, and thus lift peripheral white blood cells.

[Effects of acupuncture-moxibustion on contents of IL-12 and TNF-α in spleen of cyclophosphamide- induced cancer-bearing mice].

OBJECTIVE: To explore the molecular biology mechanism of acupuncture on improving immune function damage induced by chemotherapy in cancer-bearing mice. METHODS: Seventy-two mice (36 mice in 3-day treatment and 5-day treatment, respectively) which were successfully made into cancer-bearing model were divided into a blank group, a model group, an acupuncture group and a moxibustion group by stratified randomization method, 9 mice in each one. Except for the mice in the blank group, the remaining mice were treated with intra-peritoneal injection of cyclophosphamide (CTX, 150 mg/kg), to establish the cancer-bearing mice of CTX. The mice in the blank group were treated with intraperitoneal injection of 0.9% NaCl (identical dose as other groups). After 4 h, the mice in the acupuncture group and moxibustion group were treated with acupuncture and moxibustion at "Dazhui" (GV 14), "Geshu" (BL 17), "Shenshu" (BL 23), "Zusanli" (ST 36), once a day. The mice in the blank group and model group were treated with immobilization and fixation during the same time. On the next day of the end of 3-day and 5-day treatment, the sample was collected. The ELISA method was applied to measure the contents of IL-12 and TNF-α in spleen of all the mice. RESULTS: After 3-day and 5-day treatment, compared with the blank group, the contents of IL-12 and TNF-α in spleen in the model group were reduced (all P < 0.05); compared with the model group, the contents of IL-12 and TNF-α in spleen in the acupuncture group and moxibustion group were increased (all P < 0.05), but the content of IL-12 and TNF-α in the acupuncture group was not different from that in the moxibustion group (both P > 0.05). CONCLUSION: Acupuncture and moxibustion could effectively increase the contents of IL-12 and TNF-α in spleen of CTX cancer-bearing mice, which could relieve chemotherapy-induced immune function damage to improve immune function.

[Study on dynamic effect of acupuncture on marrow cell cycle regulatory protein cyclin D1 expression and cell cycle in mice with cyclophosphamide induced myelosuppression].

OBJECTIVE: To explore the molecular biological mechanism of acupuncture and moxibustion (A&M) in reducing chemotherapy-related toxicity, relieving myelosuppression and increasing peripheral white blood cells (WBC). METHODS: Two hundred and twenty-four male Kunming mice of clean grade were randomized equally into 4 groups, the blank control group (A), the model group (B), the acupuncture group (C), and the moxibustion group (D). Except those in Group A, mice were duplicated into myelosuppression model with cyclophosphamide (CTX) using the accepted method. After being modeled, mice in Group C and D were treated with acupuncture and moxibustion respectively, once a day for 7 successive days, while those in Group A and B were dealt with the same actions (seizing and fixing) every day but no therapy was given. From day 2 to day 7 of the treatment, 8 mice were taken from every group per day and killed in batches. Their peripheral WBC was counted and bone marrow for detecting Cyclin D1 expression and percentages of bone marrow cells in different cycle stages using immunohistochemistry and flow cytometer respectively. RESULTS: WBC count restored to exceed the baseline in group C and D at day 5 of the treatment, being one day earlier than that in group B. Cyclin D1 expression in the bone marrow raised in Group C and D, and reached the peak at day 4, showing significant difference as compared with that in Group B (P < 0.01). The phase G1 marrow cell percentages in Group C and D was lower than that in Group B at all days of detection, showing statistical significance at day 2-4 (P < 0.05 or P < 0.01); while the percentages of phase S and G2-M cells in the two treated groups was higher than that in group B all the times. CONCLUSIONS: While CTX damaged marrow cells, it intervened the cell cycle regularity and reduced the DNA content to cause myelosuppression and leucocytopenia. A&M therapy could improve the Cyclin D1 expression, speed up the cell transition from phase G1 to phase S and increase the synthesis of DNA. At the mean time, taking advantage of the block at phase G2, it can repair the injured DNA, speed up cell mitosis for turning into multiplication, improve the anti-injury and repairing ability of cells to protect bone marrow cells, and relieve the myelosuppression.

[Effects of acupuncture and moxibustion on DNA excision repair-related proteins of bone marrow cell in cyclophosphamide-induced mice].

OBJECTIVE: To explore the molecular biological mechanism of acupuncture and moxibustion for relieving myelosuppression and increasing white blood cells. METHODS: Two hundred and twenty-four clean male Kunming mice were randomly divided into a control group, a model group, an acupuncture group and a moxibustion group, 56 mice in each group. The model of myelosuppression was made with Cyclophosphamide. In the acupuncture group and the moxibustion group, acupoints "Dazhui" (GV 14), "Geshu" (BL 17), "Shenshu" (BL 23) and "Zusanli" (ST 36) were used for treatment with acupuncture and moxibustion, respectively, while, in the control group and the model group, there were no treatment carried out except catching and fixing. The changes of bone marrow cell DNA pol beta and XPD between the 2nd and 7th day were examined with immunohistochemical method. RESULTS: Acupuncture and moxibustion markedly up-regulated the expression of bone marrow cell DNA pol beta and XPD, and promoted the base excision repair and nucleotide excision repair, which leads to the relieving Cyclophosphamide-induced myelosuppression and increasing the number of white blood cells. CONCLUSION: For acupuncture and moxibustion, one of the bone major mechanisms in relieving post-chemotherapy myelosuppression, protecting hemopoietic function and increasing the white blood cells is that it can promote the repair of the bone marrow cell DNA excision and protect hemopoietic cells from injury by chemical drugs.

[The original meaning of concept of Shichen in acupoint selection according to time].

OBJECTIVE: To explore the original meaning of concept of shichen in acupoint selection according to time of "needling methods of midnight-noon ebb-flow (Ziwu Liuzhu)" and "eight methods of the intelligent turtle (Linggui Bafa)", so as to provide more scientific and accurate calculating method for acupoint selection according to time in clinical acupuncture and moxibustion. METHODS: Based on the theory of "Heaven-people corresponding", use modern calculating time method to return to the ancient traditional concept of shichen. Compared ancient and modern calculating time methods of shichen and referred to sundial time-calculating method, in combination with changes of day and night time in 24 solar terms, use modern time-calculating method to calculate, so as to deny the mechanical time-calculating method of heavenly stems and earthly branches, in which each shichen is equal to 2 hours and it has been used from the last stage of the Ming dynasty and the initial stage of the Qing dynasty up to now. RESULTS: Return to the original meaning of shichen in acupoint selection according to time, and time of each shichen should change along with changes of day and night time in the four seasons of one year. CONCLUSION: The concrete calculating method of shichen for different stages of four seasons in one year should be different.