石榴籽油对骨性关节炎大鼠软骨和软骨下骨元素及血清氧化应激指标的影响及意义-现代医学

石榴籽油对骨性关节炎大鼠软骨和软骨下骨元素及血清氧化应激指标的影响及意义

作者：张涛¹ 许文胜² 刘雄伟² 王志³ 郑丽娜³ 张伟松³ 刘吉明³ 赵东³ 赵欣磊³

单位：1. 内蒙古科技大学包头医学院医学免疫学教研室, 内蒙古包头 014010;
2. 内蒙古科技大学包头医学院第一附属医院骨三科, 内蒙古包头 014010;
3. 内蒙古科技大学包头医学院, 内蒙古包头 014060

分类号：R684.3

出版年·卷·期（页码）：2019·47·第十二期（1476-1481）

摘要：

目的： 研究不同剂量的石榴籽油对骨性关节炎（OA）大鼠股骨髁软骨和软骨下骨钙（Ca）、磷（P）等元素含量，血清碱性磷酸酶（ALP）、超氧化物歧化酶（SOD）活性及丙二醛（MDA）含量的影响，找到促进软骨生成和有效抑制关节退变的实验依据。方法： 8周SD大鼠108只，分成空白组、模型对照组、实验1组、实验2组和实验3组，其中空白组20只，模型对照组和实验1组、实验2组、实验3组各22只。除空白组实施假手术外，其余4组均通过改良Hulth法建立OA动物模型，驱赶4周后每组随机挑选2只大鼠断颈处死，通过MicroCT参数分析确认建模成功。除喂食普通饲料外，每天给予模型对照组大鼠1.59 g·kg^-1纯玉米油灌胃，实验1、2、3组大鼠分别给予0.95、1.27和1.59 g·kg^-1纯石榴籽油（PSO）灌胃，各组大鼠均在相同条件下喂养，自由饮水和进食，每7 d称重1次，以调整PSO剂量。继续喂养12周后用1%的戊巴比妥钠按90 mg·kg^-1剂量给大鼠腹腔注射，使大鼠麻醉致死，取腹腔静脉血6 ml，3 000 r·min^-1离心10 min后取上清液用原子吸收分光光度计分别检测大鼠血清MDA水平及ALP、SOD活性。取出股骨髁标本纵切，干燥后用EDX 4500H能量色散X荧光光谱仪和JEM-4000EX能量选择电子显微镜观察切面，根据元素对电子束的吸收光谱计算Ca、P等元素的含量。结果： 经玉米油干预后的模型对照组大鼠软骨和软骨下骨Ca、P含量以及血清MDA、ALP、SOD活性与空白组相比差异无统计学意义（P>0.05）；经PSO干预后的3个实验组与模型对照组相比除SOD活性升高外其余指标均降低，差异有统计学意义（P<0.05）。与实验1组相比，实验2、3组Ca含量以及血清MDA活性降低，SOD活性升高，差异有统计学意义（P<0.05）。结论： 玉米油对OA软骨退变及软骨下骨硬化增生无抑制作用。PSO能有效降低OA大鼠软骨及软骨下骨Ca、P含量，减弱软骨转化活性，对软骨退变及软骨下骨硬化、增生具有抑制作用；同时PSO能降低ALP活性，提示其具有雌激素活性，可阻碍ALP活性升高而改善软骨高转化状态；另外PSO能有效减弱氧化应激反应代谢产物MDA活性，对氧自由基产生淬灭作用。

Objective: To studythe effect of different doses of pomegranate seed oil(PSO) on chondral and subchondral bone elements calcium(Ca),phosphorus(P) content in the condyle cartilage layer of femur and serum alkaline phosphatase(ALP),peroxidase dismutase(SOD) activity and malondialdehyde(MDA) content in osteoarthritis(OA)rats, and reveal the experimental basis for promoting cartilage repairation and inhibiting joint degeneration. Methods: Experimental animals were divided into 5 groups:blank group, model control group, low dose group(experimental group 1), middle dose group(experimental group 2) and high dose group(experimental group 3). Rats in the blank group were performed sham operations, rats in the other four groups wereestablished OA models by the classic Hulth method.After driven 4 weeks,two rats were randomly selected from each group (except the blank group) and killed with broken necks, and MicroCT parameter analysis were used to confirm the models were successful. Rats in 5 groups were fed withnormal diet, besides,rats in the model control group were fed with 1.59 g·kg^-1 pure corn oil by gavage every day,rats in the experimental group 1,2,3 were fed with 0.95, 1.27 and 1.59 g·kg^-1 pure PSO by gavage. Ratsin each group were fed under the same conditions, and were allowed to drink and eat freely. All the rats were weighed every 7 days to adjust the dose of PSO. Continue feeding for 12 weeks, the rats were injected intraperitoneally with 1% pentobarbital sodium at 90 mg·kg^-1 to anesthetize them to death, and 6 ml abdominal venous blood was taken and centrifugated with 3 000 r·min^-1 for 10 min. The levels of serum MDA and the activity of ALP, SOD were measured with an atomic absorption spectrophotometer. The femoral condyle specimens were cut longitudinally, and after drying, the sections were observed with EDX 4500H energy dispersive X-ray fluorescence spectrometer and JEM-400 0EX energy selective electron microscope. Ca and P contents were calculated according to the absorption spectra of the elements on the electron beam.Results: There was no significant difference in the content of Ca and P in cartilage and subchondral bone and the activity of MDA, ALP and SOD in serum between the model control group and the blank group after corn oil intervention (P>0.05).After PSO intervention, the other indexes of the three experimental groups were lower than those of the model control group except for the increase of SOD activity,the difference was statistically significant(P<0.05). Compared with experimental group1, the content of Ca and the activity of MDA in serum decreased while the activity of SOD increased in experimental group 2,3,the difference was statistically significant(P<0.05). Conlusion: Corn oil has no inhibitory effect on OA cartilage degeneration and subchondral osteosclerosis. PSO can effectively reduce the content of Ca and P in cartilage and subchondral bone of OA rats, weaken the activity of cartilage transformation, inhibit the cartilage degeneration, subchondral bone sclerosis and proliferation. At the same time, PSO can reduce the activity of ALP, suggesting that it has estrogen activity, can block the increase of ALP activity and improve the high transformation state of cartilage. In addition, PSO can effectively reduce the activity of MDA,has quenching effect onoxygen free radicals.

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