摘要
目的探讨羽扇豆醇(LUP)通过调控磷脂酰肌醇3-激酶(PI3K)/丝氨酸-苏氨酸激酶(AKT)/核因子κB(NF-κB)信号通路对骨质疏松(OP)大鼠炎性反应的影响。方法60只SD大鼠随机选择12只作为对照(NC)组,其余大鼠构建OP模型后,随机分为模型组(OP组)、羽扇豆醇组(LUP组,50 mg/kg)、PI3K激活剂组(740 Y-P组,10 mg/kg)、LUP+740 Y-P组。Micro-CT检测大鼠骨小梁微结构变化[包括骨密度(BMD)、骨体积分数(BV/TV)、骨小梁厚度(Tb.Th)、骨小梁数量(Tb.N)];测量骨干质量、灰质量、磷和钙水平;HE染色观察股骨病理形态;ELISA法以及实时荧光定量PCR(qPCR)检测血清肿瘤坏死因子-α(TNF-α)、白细胞介素-4(IL-4)、IL-17、IL-10、碱性磷酸酶(ALP)、Ⅰ型前胶原氨基端原肽(PINP)、抗酒石酸酸性磷酸酶5b(TRACP-5b)、Ⅰ型胶原交联羧基末端肽(CTX-Ⅰ)水平;Western blot检测股骨组织PI3K/AKT/NF-κB通路蛋白表达。结果与NC组相比,OP组大鼠BMD、BV/TV、Tb.N、Tb.Th、干质量、灰质量、钙和磷含量、IL-10、IL-4、ALP、PINP水平显著降低(P<0.05),TNF-α、IL-17、TRACP-5b、CTX-Ⅰ水平和p-PI3K/PI3K、p-AKT/AKT、p-NF-κB/NF-κB比值显著上升(P<0.05);与OP组相比,LUP组BMD、BV/TV、Tb.N、Tb.Th、干质量、灰质量、钙和磷含量、IL-10、IL-4、ALP、PINP水平显著升高(P<0.05),TNF-α、IL-17、TRACP-5b、CTX-Ⅰ水平和p-PI3K/PI3K、p-AKT/AKT、p-NF-κB/NF-κB比值显著降低(P<0.05);740 Y-P组呈现出与LUP组相反的趋势,且PI3K激活剂740 Y-P可明显减弱LUP大鼠的上述保护作用。结论LUP可减轻OP大鼠的炎症反应和骨质流失,其作用可能与抑制PI3K/AKT/NF-κB通路活化有关。
This study was performed to investigate the impacts of lupeol(LUP)on inflammatory responses in osteoporosis(OP)rats by regulating phosphatidylinositol 3-kinase(PI3K)/serine-threonine kinase(AKT)/nuclear factorκB(NF-κB)signaling pathway.Twelve of 60 SD rats were randomly selected as the control(NC)group.The other rats were randomly divided into model group(OP group),lupeol group(LUP group,50 mg/kg),PI3K activator group(740 Y-P group,10 mg/kg),and LUP+740 Y-P group after OP model was constructed.Micro CT was used to detect the changes of bone trabecular microstructure in rats(bone mineral density(BMD),bone volume fraction(BV/TV),bone trabecular thickness(Tb.Th),bone trabecular number(Tb.N));the bone dry weight,ash weight,phosphorus,calcium levels were measured;HE staining was applied to observe the pathological morphology of the femur;ELISA and real-time fluorescent quantitative PCR(qPCR)were applied to detect serum levels of tumor necrosis factor-α(TNF-α),interleukin-4(IL-4),IL-17,IL-10,alkaline phosphatase(ALP),procollagen I Nterminal propeptide(PINP),tartrate-resistant acid phosphatase 5b(TRACP-5b)and cross-linked carboxyterminal telopeptide of typeⅠcollagen(CTX-Ⅰ)level;Western blot was applied to detect the expression of PI3K/AKT/NF-κB pathway proteins in femoral tissue.Compared with NC group,the levels of BMD,BV/TV,Tb.N,Tb.Th,dry weight,ash weight,contents of calcium and phosphorus,IL-10,IL-4,ALP and PINP in OP group were significantly decreased(P<0.05),while TNF-α,IL-17,TRACP-5b,CTX-Ⅰlevels and P-PI3K/PI3K,P-AKT/AKT,p-NF-κB/NF-κB ratios were significantly increased(P<0.05).Lupeol could reverse these changes in OP rats(P<0.05),while PI3K activator group 740 Y-P could promote these changes.In LUP+740 Y-P group,PI3K activator 740 Y-P significantly weakened the above protective effect of lupeol.In conclusion,lupeol can alleviate the inflammation and bone loss in OP rats,and its effect may be related to the inhibition of PI3K/AKT/NF-κB pathway activation.
作者
方红育
周少怀
黄涛
李嘉琼
张莹
胡长青
FANG Hongyu;ZHOU Shaohuai;HUANG Tao;LI Jiaqiong;ZHANG Ying;HU Changqing(Department of Orthopedics,Wuhan Third Hospital,Wuhan 430060,China)
出处
《免疫学杂志》
CAS
CSCD
北大核心
2023年第7期608-615,共8页
Immunological Journal
基金
武汉市医学科研项目(WX19D14)。
