青娥方对去卵巢骨质疏松大鼠骨组织PI3K/Akt信号通路的影响  

Effect of Qing'e Formula on PI3K/Akt Signaling Pathway in Bone Tissueof Ovariectomized Osteoporosis Rats

作　　者：郭世明[1] 张嵩图 司在武 黄云梅 GUO Shiming;ZHANG Songtu;SI Zaiwu;HUANG Yunmei(Zhangzhou Traditional Chinese Medicine Hospital,Zhangzhou,Fujian 363000,China;Academy of Integrative Medicine,Fujian University of Traditional Chinese Medicine,Fuzhou,Fujian 350122,China;Fujian Key Laboratory of Integrative Medicine on Geriatrics,Fuzhou,Fujian 350122,China)

机构地区：[1]福建省漳州市中医院,福建漳州363000 [2]福建中医药大学中西医结合研究院,福建福州350122 [3]福建省中西医结合老年性疾病重点实验室,福建福州350122

出　　处：《福建中医药》2024年第5期16-20,共5页Fujian Journal of Traditional Chinese Medicine

基　　金：国家自然科学基金项目(81904228);福建省自然科学基金项目(2023J011853);福建省中医药科研项目(2021ZYJC24);漳州市自然科学基金项目(ZZ2023J14)。

摘　　要：目的 探讨青娥方对去卵巢骨质疏松大鼠骨组织磷脂酰肌醇3激酶/蛋白激酶B(PI3K/Akt)信号通路的影响。方法 采用随机数字表法将72只3月龄SPF级雌性SD大鼠分为假手术组、模型组和青娥方组,每组24只。大鼠均按40 mg/kg腹腔注射2%戊巴比妥钠溶液麻醉,于下腹部正中作切口,模型组和青娥方组均切除双侧卵巢,假手术组切除卵巢周围少量脂肪组织后立即关腹,缝合伤口。造模1周后,假手术组和模型组予2 mL0.9%氯化钠溶液灌胃,青娥方组按浸膏量0.42 g/(kg·d)予青娥方药液灌胃,每日1次,连续灌胃12周。3组分别于干预4、8、12周后各处死8只并取材。Masson染色法观察股骨颈形态;检测股骨骨密度和骨生物力学指标最大载荷;ELISA法检测血清PI3K、Akt含量;qPCR和Western blot检测腰椎PI3K、Akt和糖原合成酶激酶3β(GSK3β)mRNA相对表达水平和蛋白表达量。结果 与假手术组比较,模型组干预第4周腰椎Akt mRNA相对表达水平明显升高(P<0.05);第12周股骨最大载荷明显降低(P<0.05);第8、12周股骨骨密度明显降低(P<0.05),腰椎PI3K mRNA相对表达水平均明显升高(P<0.05),PI3K蛋白表达量明显升高(P<0.05);第4、8、12周血清PI3K、Akt含量均明显升高(P<0.05),腰椎Akt、GSK3β蛋白表达量均明显升高(P<0.05),GSK3β mRNA相对表达水平明显升高(P<0.05)。与模型组比较,青娥方组干预第4周腰椎Akt mRNA相对表达水平和蛋白表达量明显降低(P<0.05);第12周股骨最大载荷明显升高(P<0.05),血清Akt含量均明显降低(P<0.05),腰椎Akt蛋白表达量明显降低(P<0.05);第8、12周股骨骨密度明显升高(P<0.05),腰椎PI3K、GSK3β mRNA相对表达水平均明显降低(P<0.05),PI3K蛋白表达量明显降低(P<0.05);第4、8、12周血清PI3K含量明显降低(P<0.05)。结论青娥方可改善去卵巢骨质疏松模型大鼠骨密度及骨生物力学性能,其机制可能与抑制PI3K/Akt信号通路有关。Objective: To investigate the effect of Qing'e formula on the PI3K/Akt signaling pathway in bone tissue of ovariectomizedosteoporosis rats. Methods: 72 3-month-old SPF grade female SD rats were randomly divided into sham group, model groupand Qing'e formula group using a random number table method, with 24 rats in each group. Rats were anesthetized by intraperitonealinjection of 2% pentobarbital sodium solution at a dose of 40 mg/kg, and an incision was made in the middle of the lower abdomen.Both the model and Qing'e formula groups had bilateral ovaries removed, while the sham group had a small amount of adipose tissuearound the ovaries removed and immediately closed the abdomen and sutured the wound. After one week of modeling, the sham andmodel groups were given 2 mL of 0.9% sodium chloride solution by gavage, while the Qing'e formula group was given Qing'e formulasolution by gavage at a dose of 0.42 g/(kg·d), once a day for 12 consecutive weeks. After intervention for 4, 8, and 12 weeks, 8mice were euthanized and samples were taken from each of the 3 groups. The Masson staining method was used to observe the morphologyof the femoral neck;femoral bone mineral density and maximum load of the femur were detected;ELISA method was usedto detect serum PI3K and Akt level;qPCR and western blot were used to detect the mRNA relative expression level and proteinexpression of PI3K, Akt, and glycogen synthase kinase 3β (GSK3β) in the lumbar spine. Results: Compared with the sham group,the mRNA relative expression level of Akt in the model group significantly increased in the lumbar spine at week 4 of intervention(P<0.05), while the maximum load of the femur significantly decreased in the 12th week (P<0.05);at weeks 8 and 12, the femoralbone mineral density significantly decreased (P<0.05), and the mRNA relative expression level of PI3K in the lumbar spine significantlyincreased (P<0.05), while the protein expression of PI3K significantly increased (P<0.05);at weeks 4, 8, and 12, the level ofserum PI3K and Akt significa

关 键 词：骨质疏松症 去卵巢大鼠 青娥方 骨密度 生物力学 PI3K AKT 

分 类 号：R285.5[医药卫生—中药学]