酒花提取物调控OPG/RANKL信号通路对大鼠成骨细胞增殖作用的影响  

Effect of Humulus lupulus Linn.Extract Regulating OPG/RANKL Signaling Pathway on the Proliferation of Rat Osteoblasts

作　　者：李敏[1] 扎克艳·地力夏提 胡君萍[1] 杨建华[1,2] LI Min;ZHAKEYAN Dilixiati;HU Junping;YANG Jianhua(College of Pharmacy,Xinjiang Medical University,Urumqi 830011 Xinjiang,China;Department of Pharmacy,The First Affiliated Hospital of Xinjiang Medical University,Urumqi 830054 Xinjiang,,China)

机构地区：[1]新疆医科大学药学院,新疆乌鲁木齐830011 [2]新疆医科大学第一附属医院药学部,新疆乌鲁木齐830054

出　　处：《中药新药与临床药理》2023年第8期1047-1052,共6页Traditional Chinese Drug Research and Clinical Pharmacology

基　　金：新疆天然药物活性组分与释药技术重点实验室项目(XJDX1713)。

摘　　要：目的探讨酒花提取物调控OPG/RANKL信号通路对大鼠成骨细胞增殖作用的影响。方法采用Ⅱ型胶原酶消化法提取新生SD大鼠颅骨成骨细胞,并采用ALP染色、HE染色、茜素红染色分别对大鼠成骨细胞进行鉴定。将大鼠成骨细胞分为空白对照组、阳性对照组(1×10^(-6)mol·L^(-1)雌激素)及酒花提取物低、中、高剂量组(10、20、40μg·mL^(-1)),干预24 h。采用CCK-8法检测大鼠成骨细胞的增殖情况;采用RT-PCR及Western Blot法检测成骨细胞NF-κB受体活化因子配体(RANKL)、骨保护素(OPG)的mRNA及蛋白表达水平。结果培养的细胞符合成骨细胞的典型特征,细胞纯度较高,可用于后续实验。与空白对照组比较,阳性对照组和酒花提取物各剂量组的大鼠成骨细胞增殖率显著升高(P<0.01);阳性对照组和酒花提取物中剂量组大鼠成骨细胞OPG mRNA表达显著上调(P<0.01);阳性对照组和酒花提取物各剂量组大鼠成骨细胞RANKL mRNA表达明显下调(P<0.05,P<0.01),OPG蛋白表达显著上调(P<0.01),RANKL蛋白表达显著下调(P<0.01),mRNA及蛋白表达的OPG/RANKL比值均显著升高(P<0.05,P<0.01)。结论酒花提取物可以促进大鼠成骨细胞的增殖活性,可能与调控OPG/RANKL信号通路有关。Objective To investigate the effect of Humulus lupulus Linn.extract in regulating the OPG/RANKL signaling pathway on the proliferation of rat osteoblasts.Methods Cranial osteoblasts(COB)of newborn SD rats were extracted by type Ⅱ collagenase digestion,and identified by ALP staining,HE staining and alizarin red staining,respectively.The rat osteoblasts were divided into blank control group,positive control group(1×10^(-6)mol·L^(-1)estrogen)and low-,medium-and high-dose groups(10,20 and 40μg·mL^(-1))of Humulus lupulus Linn.extract,intervened for 24 hours.The proliferation of rat osteoblasts was detected by CCK-8 method,and the mRNA and protein expression levels of receptor activator of NF-κB ligand(RANKL)and osteoprotegerin(OPG)were detected by RTPCR and Western Blot methods.Results The cultured cells conformed to the typical characteristics of osteoblasts,and the cells were of high purity and could be used for subsequent experiments.Compared with the blank control group,the proliferation rate of rat osteoblasts was significantly increased in the positive control group and each dose group of Humulus lupulus Linn.extract(P<0.01);the mRNA expression of OPG in rat osteoblasts was significantly upregulated in the positive control group and the medium-dose group of Humulus lupulus Linn.extract(P<0.01);the mRNA expression of RANKL in rat osteoblasts was significantly down-regulated in the positive control group and each dose group of Humulus lupulus Linn.extract(P<0.05,P<0.01),OPG protein expression was significantly upregulated(P<0.01)and RANKL protein expression was significantly down-regulated(P<0.01)in the positive control and Humulus lupulus Linn.extract groups,and the mRNA and protein expressions of OPG/RANKL ratio were significantly increased(P<0.05,P<0.01).Conclusion Humulus lupulus Linn.extract can promote the proliferative activity of rat osteoblasts,which may be related to the regulation of the OPG/RANKL signaling pathway.

关 键 词：酒花提取物 大鼠成骨细胞 骨质疏松 OPG/RANKL信号通路 

分 类 号：R285.5[医药卫生—中药学]