补肾化痰方含药血清对卵巢切除大鼠骨髓间充质干细胞保护作用机制研究  

Protective mechanism of Bushen Huatan Decoction containing serum on bone marrow mesenchymal stem cells in ovariectomized rats

作　　者：周广文[1] 朱伟[2] 张妍 车希隆 李佳[1] 熊勇[1] ZHOU Guang-wen;ZHU Wei;ZHANG Yan;CHE Xi-long;LI Jia;XIONG Yong(Hubei University of Chinese Medicine,Wuhan Hubei 430065,China;Wuhan University,Wuhan Hubei430072,China)

机构地区：[1]湖北中医药大学,湖北武汉430065 [2]武汉大学,湖北武汉430072

出　　处：《时珍国医国药》2021年第12期2839-2842,共4页Lishizhen Medicine and Materia Medica Research

基　　金：国家自然科学基金(81904267);湖北省武汉市卫生健康委员会武汉市医学科研项目(WZ20C28)。

摘　　要：目的观察补肾化痰方对卵巢切除大鼠骨髓间充质干细胞保护作用。方法 3月龄SD雌性大鼠36只,随机分为6组,除假手术组外,其余各组均以卵巢切除术(OVX)造模,术后分组分笼常规饲养10周。各组大鼠分别给予生理盐水、中药药液、骨化三醇药液灌胃给药7天后制备血清。将假手术组及卵巢切除大鼠F3代骨髓间充质干细胞(bone marrow derived mesenchymal stem cells, BMSC)复苏培养后制备成单细胞悬液,依次分为对照组(Control),卵巢切除组(OVX),卵巢切除+补肾化痰含药血清高剂量组(OVX+H-BS),卵巢切除+补肾化痰含药血清中剂量组(OVX+M-BS),卵巢切除+补肾化痰含药血清低剂量组(OVX+L-BS),卵巢切除+骨化三醇含药血清组(OVX+A-S)。分别采用β-半乳糖苷酶活性染色、细胞计数试剂盒(Cell Counting Kit-8,CCK8)评估细胞增殖能力,采用ELISA试剂盒检测白细胞介素-6(interleukin-6,IL-6)、白细胞介素-10(interleukin-10,IL-10)、白细胞介素-1α(interleukin-1α,IL-1α)、白细胞介素-1β(interleukin-1β,IL-1β)、肿瘤坏死因子-α(Tumor Necrosis Factor, TNF-α)等炎症因子的表达。结果与对照组(Control)相比,OVX大鼠β-半乳糖苷酶活性染色阳性细胞数量明显增多,细胞增殖能力明显下降,IL-6、IL-10、IL-1α、IL-1β、TNF-α等炎症因子明显增加;与卵巢切除组(OVX)组相比,中药及西药组大鼠β-半乳糖苷酶活性染色阳性细胞数量显著性下降,细胞增殖能力升高,IL-6、IL-10、IL-1α、IL-1β、TNF-α等炎症因子减少,其中高剂量组(OVX+H-BS)骨化三醇含药血清组(OVX+AS)差异有统计学意义(P<0.05)。结论补肾化痰方能够减缓BMSC细胞的衰老,增强细胞增殖能力,减轻细胞炎症反应,从而减少BMSC的凋亡,起到防治骨质疏松的作用。Objective To observe the protective effect of Bushen Huatan Recipe on bone marrow mesenchymal stem cells in ovariectomized rats.Methods Thirty-six 3-month-old female SPF SD rats were randomly divided into 6 groups. In addition to the sham operation group, other groups underwent oophorectomy to make models. After 10 weeks,the rats in each group were given normal saline, traditional Chinese medicine solution and calcitriol solution by gavage for 7 days to prepare serum.The bone marrow mesenchymal stem cells(BMSC) of F3 passage of ovariectomized rats and control group were resuscitated and cultured to prepare single cell suspension, then were divided into Control group, OVX group, OVX+H-BS group, OVX+M-BS group, OVX+L-BS group and OVX+A-S group.The cell proliferation was evaluated by β-galactosidase activity staining and cell counting kit-8(CCK8). The levels of interleukin-6(IL-6), interleukin-10(IL-10),interleukin-1α(IL-1α)、 interleukin-1β(IL-1β) and tumor necrosis factor-α(TNF-α)were detected by ELISA kit.Results Compared with Control group, the number of β-galactosidase positive cells increased while the proliferation of cells decreased significantly, and inflammatory factors such as IL-6, IL-10, IL-1α, IL-1β and TNF-α increased significantly in rats with OVX;Compared with OVX group, the number of β-galactosidase positive cells decreased while the proliferation of cells increased significantly, and inflammatory factors such as IL-6, IL-10, IL-1α, IL-1β and TNF-α increased in traditional Chinese medicine and Western medicine group. There was a significant difference in OVX + H-BS and calcitriol containing serum group(P<0.05).Conclusion Bushen Huatan Decoction can slow down the aging of BMSC cells, enhance the proliferation of cells, reduce the inflammatory reaction of cells, thus reducing the apoptosis of BMSC, and play a role in the prevention and treatment of osteoporosis.

关 键 词：骨髓间充质干细胞 炎性衰老 补肾化痰方 绝经后骨质疏松症 

分 类 号：R285.5[医药卫生—中药学]