和厚朴酚对D-半乳糖致腰椎间盘退变大鼠血清炎症因子及细胞线粒体凋亡通路的影响  

Effects of honokiol on serum inflammatory factors and the mitochondrial apoptosis pathway in rats with D-galactose-induced intervertebral disc degeneration

作　　者：张超[1] 吴俊学[1] 王冶 谢海洋[1] 彭玲 ZHANG Chao;WU Junxue;WANG Ye;XIE Haiyang;PENG Ling(Department of Orthopedics,Affiliated Hospital of North Sichuan Medical College,Nanchong 637000,China;Department of Clinical Nutrition,Affiliated Hospital of North Sichuan Medical College,Nanchong 637000)

机构地区：[1]川北医学院附属医院骨外科,四川南充637000 [2]川北医学院附属医院临床营养科,四川南充637000

出　　处：《中国比较医学杂志》2021年第11期62-68,75,共8页Chinese Journal of Comparative Medicine

基　　金：南充市科技项目(18SXHZ0341)。

摘　　要：目的探索和厚朴酚对D-半乳糖致腰间盘退变大鼠血清炎症因子含量变化,细胞外基质标记物表达水平及凋亡通路相关蛋白表达水平的影响。方法 45只SD大鼠随机分为对照组(Control)、D-半乳糖(D-galactose, D-gal)模型组(Model)、和厚朴酚低剂量组(2 mg/kg)、和厚朴酚中剂量组(4 mg/kg)、和厚朴酚高剂量组(8 mg/kg),每组9只。模型组和和厚朴酚给药组采用皮下注射D-gal方法建立大鼠椎间盘退变模型,对照组皮下注射等体积生理盐水。造模成功后,给药组大鼠分别灌胃和厚朴酚2、4、8 mg/kg,其余各组大鼠灌胃生理盐水,连续给药4周后处死大鼠。HE和番红O染色观察比较各组腰椎间盘形态组织学;qRT-PCR检测大鼠椎间盘组织细胞外基质标记物的mRNA水平;ELISA检测大鼠血清中白细胞介素IL-1β、IL-6、IL-10和肿瘤坏死因子α(TNF-α)含量的变化;TUNEL染色观察大鼠椎间盘的组织凋亡情况;Western blot检测B细胞淋巴瘤-2(Bcl-2)/Bcl-2相关X蛋白(Bax)、胱天蛋白酶-9(Caspase-9)、Caspase-3、c-Myc的蛋白表达。结果模型组和和厚朴酚低、中剂量组椎间盘组织形态学评分较对照组显著升高(P<0.05)。和厚朴酚中、高剂量组椎间盘组织形态学评分较模型组显著降低(P<0.05)。模型组和和厚朴酚低剂量组软骨受损严重,和厚朴酚中、高剂量组软骨受损较轻。和厚朴酚能显著升高SRY相关HMG盒9(SOX-9)、II型胶原(Collagen II)和蛋白聚糖(Aggrecan)的表达水平(P<0.05)。模型组和和厚朴酚低、中、高剂量组IL-1β、IL-6、IL-10和TNF-α的含量较对照组均显著升高(P<0.05)。与模型组比较,和厚朴酚中、高剂量组IL-1β、IL-6和TNF-α含量均显著降低(P<0.05);IL-10的含量显著升高(P<0.05)。与对照组相比,模型组和和厚朴酚低、中剂量组凋亡细胞数显著升高(P<0.05),Bcl-2/Bax比值显著降低(P<0.05),Caspase-9、Caspase-3、c-Myc的蛋白水平均显著升高(P<0.05)。与模型组相比,�Objective To explore the effect of honokiol on serum inflammatory factors, extracellular matrix component expression, and apoptotic pathway-related protein expression in rats with D-galactose-induced intervertebral disc degeneration. Methods Forty-five SD rats were randomly divided into control(Control), D-galactose(D-gal) model(Model), honokiol low dose(2 mg/kg), honokiol medium dose(4 mg/kg) and honokiol high dose(8 mg/kg) groups, with 9 rats in each group. The rat model of intervertebral disc degeneration was established by subcutaneous injection of D-galactose in model and honokiol groups, and the Control group was subjected to subcutaneous injection of normal saline. After successful modeling, administration group rats were gavaged with magnolol at 2, 4 and 8 mg/kg, and the remaining rats were gavaged with normal saline in each group. Rats were sacrificed after 4 weeks of continuous administration. HE and safranin O staining was used to observe and compare the morphology and histology of intervertebral discs in each group. The mRNA levels of extracellular matrix components were measured by qRT-PCR in rat intervertebral discs. Changes of interleukin IL-1β, IL-6, tumor necrosis factor-α(TNF-α) and IL-10 in rat serum were detected by ELISAs. TUNEL staining was performed to observe apoptosis of rat intervertebral disc. Protein expression of B-cell lymphoma protein 2(Bcl-2)/Bcl-2 associated X protein(Bax), Caspase-9, Caspase-3 and c-Myc was detected by Western blot. Results The scores of intervertebral disc histomorphology in the D-galactose group and honokiol low and medium dose groups were significantly higher than that in the control group(P<0.05). Compared with that in the D-galactose group, the scores of intervertebral disc histomorphology in medium and high dose honokiol groups were significantly lower(P<0.05). Cartilage damage in the model group and the honokiol low dose group was severe, and that in honokiol medium and high dose groups was slightly damaged. Honokiol significantly increased the expressi

关 键 词：和厚朴酚 腰椎间盘退变 炎症因子 凋亡 

分 类 号：R-33[医药卫生]