Study on Acupuncture and Moxibustion of the Notch Signaling Pathway in Bone Marrow Cells to Alleviate Cyclophosphamide-Induced Myelosuppression in Mice

Abstract

Objective: To evaluate the protein expression and distribution of Notch signaling-related, differentially expressed genes (DEGs) in the bone marrow cells of mice. Additionally, verification of acupuncture regulated protein expression of these DEGs to protect bone marrow stem cells and alleviate cyclophosphamide (CTX) chemotherapy-induced myelosuppression was examined as well.Methods: Thirty two Healthy male Kunming mice of clean grade were randomized into blank group (A), model group (B), acupuncture group (C) and moxibustion group (D). Mice in groups B, C, and D were intraperitoneally injected with 100mg/kg/d CTX for three successive days to generate the myelosuppressive model; while mice in group A were injected with an equivalent volume of sterile saline. After immobilizing the animals, mice in groups C and D received daily acupuncture and moxibustion in points "DU14", "BL17" " BL23" and " ST36"respectively,while those in group A and B were received no treatment. After five days of treatment,the mice were euthanized by cervical dislocation and their femurs collected for marrow harvest , Small-throughput cDNA microarray was used for initial DEG screening, followed by immunohistochemistry, realtime quantitative PCR (RT-qPCR), and Western Blot analysis to detect protein expression and quantity of the Notch signaling-related DEGs in the bone marrow cells.Results: Expression of numb proteins (i.e., numb1 and numb2) was found to be upregulated while notch2 and jag1 were down regulated in bone marrow cells of acupuncture-treated, tumor-bearing mice.Conclusion: Notch signaling pathway serves as an important signal transduction pathway for acupuncture and moxibustion to relieve CTX chemotherapy-induced myelosuppression. Regulation of associated DEGs in notch signaling pathway in marrow cells is the key mechanism by which acupuncture and moxibustion improve marrow hematopoiesis after chemotherapy.

Moxibustion for the Treatment of Cancer and its Complications: Efficacies and Mechanisms

“…It was found that both acupuncture and moxibustion increased the WBC numbers and serum CSF activity in mice with myelosuppression, with similar results observed in the sera of the corresponding clinical study subjects. 56,57 These pieces of evidence suggest that moxibustion has a good synergistic effect in cancer patients during chemotherapy, and may be safe and effective for chemotherapyinduced GI side effects and myelosuppression, but further clinical research is needed.…”

“…cm) from the skin, 5 times per week, 2 wk TM vs T Up: Th1 cells, CD4 CD25 T cells, Tregs Down: size, tumor area, tumor weight, VEGF score TM vs TC Up: CD4T cells, Th2 cells, Cd69, Cd86, Cd20 Down: net body weight, VEGF score Xue et al 28 Breast Cancer Housanli 6 mg, 3 pillars, once a day MPTG vs PTG Up: survival rate; Down: tumor volume MTG vs TG Up: thymus index, spleen index, WBC, IL-2, INF-γ Down: IL-10, TGF-β1, HIF-1α, VEGFA, PD-1, PD-L1 MPTG vs TG Up: IL-2, INF-γ Down: IL-10, TGF-β1, PD-1, PD-L1, CD34, HIF-1α, VEGFA Zhang et al 29 NSCLC ST36 2 mg, 5 pillars, once a day, 10 d TM vs T Up: the integral of survival state on day Down: tumor volume, IL-6 Wang et al 30 HCC BL18 5 mg, 0 cm from skin, 5 pillars, every other day, 20 d (prevention group 1); 40 d (prevention group 2) Prevention group 2 vs T Down: TSGF, TNF-α (serum) Lu et al 31 LLC DU14, BL17, BL23, ST36 Moxa stick: 250 × 4 (mm), 3 min, 2 cm from skin, half once a day, 3 d, the other half, once a day, 5 d CTX + M vs CTX Up: IL-12, TNF-α (spleen)Lu et al32 …”

Research progress on the mechanism of action of acupuncture in the treatment of gastric cancer

“…The cell cycle is the process that a cell undergoes through growth and division to form two cells, and the cell cycle includes interphase, which includes G1, S, and G2 phases and mitotic (M) phase, and cell proliferation in tumors is associated with cell regulation [36] , and Cyclin D1 binds to CDK4/6 and acts as a mitotic sensor to integrate extracellular mitotic signals and cell cycle progression. A study investigated the effect of acupuncture on the cell cycle of mouse bone marrow cells, the regulatory protein Cyclin D1 regulates the G1/S phase detection site, and the expression of the cell cycle regulatory protein Cyclin D1 was found to be down-regulated after acupuncture, Cyclin D1 inhibited cells from G1 phase into S phase, leading to increased cell genome stability and inhibiting the proliferation of gastric cancer cells [37][38] . If Cyclin D1 is highly expressed, it activates cyclin-dependent kinase (CDK) activity, and the recently identified established CDK/cyclin complex regulates the cell cycle and is involved in the regulation of gene expression through phosphorylation of key components of transcription and pre-mRNA processing mechanisms.…”