补肾生血药调控JAK2-STATs通路纠正造血细胞增殖-凋亡失衡改善化疗后骨髓抑制的机制研究

Chemotherapy is a common treatment of malignant tumors, bone marrow suppression is its common toxic side effects. Bone marrow suppression patients with low blood index, are often due to anemia, infection and bleeding to affect the anti-tumor treatment, so it should to be actively controlled. Bone marrow suppression after chemotherapy is mainly due to the influence of chemotherapy drugs on bone marrow cells DNA, which can inhibit bone marrow cells to proliferate, cause them to apoptosis, and lead to an imbalance between the two. The JAK2-STATs pathway can regulate the expression of different types of STATs to coordinate the proliferation-apoptosis balance of the hematopoietic system. Previous studies have shown that Bushen Shengxue Medicine (BSSX) not only could improve the bone marrow suppression after chemotherapy, and promote hematopoietic cell proliferation, but also could regulate the expression of JAK2, STAT5. This indicate that the mechanism of BSSX improving bone marrow suppression after chemotherapy may be related to regulating JAK2-STATs pathway and correcting the hematopoietic cell proliferation-apoptotic imbalance. To verify this hypothesis, the study intends to use cyclophosphamide induced bone marrow suppression mouse model, and use BSSX to intervene. The apoptosis and proliferation of bone marrow cells at different time points will be examed, and the expression of JAK2-STATs pathway be detected to carry out the initial verification. Then primary culture the mice bone marrow cells of chemotherapeutic drugs in vivo and in vitro injuryed. Using BSSX drug serum and pathway inhibitors to intervene, and to further verify the role of BSSX on the regulation of JAK2-STATs pathway. The findings of this study can provide some scientific proof for using supply kidney method to treat bone marrow suppression after chemotherapy.

化疗是恶性肿瘤常用治疗手段，骨髓抑制为其常见毒副反应，患者常因贫血、感染、出血而影响抗癌治疗，故应积极防治。骨髓抑制主要是化疗药损伤骨髓细胞DNA，使其凋亡增加、增殖受抑二者失衡。JAK2-STATs通路能调控STATs表达来协调造血系统增殖-凋亡平衡。前期研究表明，补肾生血药能改善化疗后骨髓抑制，促造血细胞增殖，并能调控JAK2、STAT5表达，提示补肾生血药改善化疗后骨髓抑制的机制可能与调控JAK2-STATs通路，纠正造血系统增殖-凋亡失衡有关。为验证该假说，本研究拟采用环磷酰胺致骨髓抑制小鼠模型，以补肾生血药进行干预，通过检测不同时间点骨髓细胞凋亡、增殖情况及JAK2-STATs通路关键分子表达进行初步验证。对化疗药体内、外损伤后的小鼠骨髓细胞进行培养，以补肾生血药血清、通路抑制剂进行干预，进一步验证补肾生血药对该通路的调控作用。研究结果可为“从肾论治”化疗后骨髓抑制提供科学依据。

化疗药常常引发血液毒性，造成骨髓抑制，表现为骨髓细胞凋亡增加、增殖受抑，导致造血系统增殖-凋亡失衡。JAK2-STATs通路是调控骨髓造血的主要通路，可影响造血细胞增殖和凋亡。本研究分别采用体内、体外骨髓抑制模型，通过对器官、细胞、分子等不同层次指标的检测，观察补肾生血药对化疗后骨髓细胞凋亡与增殖的影响，以及对JAK2-STATs通路的调控作用。结果显示，补肾生血药可保护骨髓细胞形态结构，减轻骨髓有核细胞减少，降低骨髓细胞凋亡率，增加造血干/祖细胞比例，降低造血负调控因子TNF-α、IFN-γ、TGF-β含量，升高造血生长因子IL-3含量，增加造血祖细胞集落形成单位数，促进骨髓细胞进入增殖周期，升高外周血白细胞数，从而改善化疗药引发的骨髓细胞凋亡-增殖失衡。其机制与抑制P53/Bax线粒体凋亡途径，上调细胞周期调控分子cyclinD1表达，调控JAK2-STATs通路相关。综合上述研究结果，本研究为临床“从肾论治”化疗后骨髓抑制提供了科学依据。项目资助发表核心期刊论文5篇，录用1篇，培养硕士研究生8人，其中5名已经取得硕士学位，3名在读。项目投入经费53万元，支出47.2649万元，各项支出基本与预算相符。剩余经费5.7351万元，将继续用于本项目研究后续支出。