红姜提取物保护早期膝骨关节炎模型大鼠的关节软骨

背景:研究报道,联合使用姜提取物降低血清促炎细胞因子白细胞介素1β、白细胞介素6、肿瘤坏死因子α等水平与膝骨关节炎中软骨损伤的减轻有关。目的:观察红姜提取物灌胃对早期膝骨关节炎大鼠关节软骨保护情况及血清白细胞介素1β、白细胞介素6、肿瘤坏死因子α和软骨Col2α1 mRNA水平表达的影响,探讨红姜提取物对早期膝骨关节炎大鼠关节软骨保护作用及可能机制。方法:将50只SPF级SD大鼠随机分为空白组、模型组、红姜低剂量组、红姜高剂量组、阳性对照组,每组各10只。除空白组外,其余40只大鼠膝关节腔注射4%木瓜蛋白酶0.2 mL+0.03 mol/L的L-半胱氨酸混合溶液,建造膝骨关节炎模型。空白组与模型组常规饲养;红姜低剂量组、红姜高剂量组、阳性对照组分别予50 mg/kg的红姜提取物水溶液、100 mg/kg红姜提取物水溶液、18 mg/kg的塞来昔布胶囊水溶液灌胃,所有干预每日1次,共持续4周。治疗4周后取大鼠膝关节软骨进行番红O-固绿染色,并对关节软骨行Mankin评分,检测血清中白细胞介素1β、白细胞介素6、肿瘤坏死因子α及软骨中Col2α1 mRNA表达水平。实验方案经广州中医药大学动物实验伦理委员会批准,批准号:20190917002。结果与结论:①膝关节软骨的病理切片显示,模型组及各治疗组均有软骨基质流失,各治疗组Mankin评分均比空白组评分高(P<0.05),比模型组评分低(P<0.05),其中红姜高剂量组与阳性对照组评分差异无显著性意义(P>0.05),均显著低于红姜低剂量组(P<0.05);②血清白细胞介素1β、白细胞介素6、肿瘤坏死因子α结果显示,阳性对照组、红姜高剂量组、红姜低剂量组均比空白组表达上调(P<0.05),均比模型组表达下调(P<0.05),且各治疗组间水平阳性对照组<红姜高剂量组<红姜低剂量组(P<0.05);③软骨中Col2α1 mRNA结果显示,空白组与红姜高剂量组、阳性对照组的Col2α1 mRNA表达差异无显著性意义(P>0.05),模型组和红姜低剂量组Col2α1 mRNA表达相较其他3组均显著上调(P<0.05);④结果说明,红姜提取物可能主要通过抑制白细胞介素1β、白细胞介素6、肿瘤坏死因子α等炎症因子的表达,达到了保护膝骨关节炎关节软骨作用,从而延缓膝骨关节炎的发展进程;且相对于低剂量组,红姜提取物高剂量组的抗炎效果更好。

Protective effect of red ginger extract on articular cartilage of rats with early knee osteoarthritis

BACKGROUND: Studies have reported that the combined use of ginger extract to reduce the levels of serum pro-inflammatory cytokines, including interleukin-1β, interleukin-6, tumor necrosis factor-α, is related to the reduction of cartilage injury in knee osteoarthritis.OBJECTIVE: To observe the protective effect of red ginger extract on articular cartilage and the expression of serum interleukin-1β, interleukin-6, tumornecrosis factor-α and cartilage tissue type II collagen α1 mRNA in rats with early knee osteoarthritis, and to explore the protective effect of red ginger extracton articular cartilage of rats with early knee osteoarthritis and its possible mechanism.METHODS: Fifty SPF Sprague-Dawley rats were randomly divided into blank group, model group, low-dose red ginger, high-dose red ginger and positivecontrol group (n=10 per group). Except for the blank group, the rats in the other four groups were used to prepare knee osteoarthritis models byintraarticular injection of 4% papain 0.2 mL+0.03 mol/L L-cysteine mixed solution. The rats in the blank and model groups were fed routinely, and the lowdosered ginger, high-dose red ginger and positive control groups were given 50 mg/kg red ginger extract aqueous solution, 100 mg/kg red ginger extractaqueous solution and 18 mg/kg celecoxib capsule aqueous solution respectively. All the interventions were conducted once a day, for 4 continuous weeks.Four weeks after treatment, the rats in each group were killed and the knee joints were stained with safranin O-fast green. The articular cartilage wasscored by Mankin scoring. The expression levels of interleukin-1β, interleukin-6, tumor necrosis factor-α in serum and type II collagen α1 mRNA in cartilagewere detected. The study protocol was approved by the Animal Ethics Committee of Guangzhou University of Chinese Medicine, with an approval No.20190917002.RESULTS AND CONCLUSION: The pathological section of knee cartilage showed that there was cartilage matrix loss in the model and each treatment group,and the Mankin score of each treatment group was significantly higher than that of the blank group (P < 0.05) and lower than that of the model group (P < 0.05).There was no significant difference between the high-dose group and the positive control group (P > 0.05), but the scores of the two groups were lower thanthat of the low-dose group. The levels of interleukin-1β, interleukin-6, and tumor necrosis factor-α were upregulated in the positive control group, high-dosered ginger group and low-dose red ginger group compared with the blank group and down-regulated compared with the model group (P < 0.05). Moreover,and the levels of these cytokines were ranked as follows: positive control group < high-dose red ginger group < low-dose red ginger group (P < 0.05). The levelof type II collagen α1 mRNA in cartilage showed no significant difference between the blank group and the high-dose red ginger group and the positive controlgroup (P > 0.05), whereas the expression of type II collagen α1 mRNA was significantly increased in the model group and low-dose red ginger group comparedwith the other three groups (P < 0.05). To conclude, red ginger extract may protect the articular cartilage of knee osteoarthritis by inhibiting interleukin-1β,interleukin-6, and tumor necrosis factor-α, thereby delaying the development of knee osteoarthritis. Compared with the low-dose group, high-dose red gingerextract has better anti-inflammatory effect.