独活寄生汤抑制白细胞介素-1β诱导的软骨细胞凋亡的作用机制
摘要
目的探讨独活寄生汤抑制白细胞介素(IL)-1β诱导的软骨细胞凋亡的作用机制。方法从8只SPF级SD大鼠膝关节分离获取软骨细胞并培养,采用Ⅱ型胶原免疫组化染色鉴定。噻唑蓝(MTT)检测不同浓度独活寄生汤对IL-1β诱导的软骨细胞凋亡的影响,从而优化独活寄生汤最佳干预浓度。将第2代软骨细胞随机分为空白组、模型组、独活寄生汤组,空白组以含10%胎牛血清(FBS)的DMEM培养液培养,模型组以10 ng/ml IL-1β和10%FBS的DMEM培养液培养,独活寄生汤组采用300μg/ml独活寄生汤、10 ng/ml IL-1β和10%FBS的DMEM培养液培养。干预48 h,逆转录PCR检测各组软骨细胞中miR-98的表达;Western印迹法检测各组软骨细胞中B细胞淋巴瘤(Bcl)-2、Bcl-2相关X蛋白(Bax)、Caspase-3、Caspase-9蛋白表达。结果Ⅱ型胶原免疫组化染色后阳性组软骨细胞胞质为棕黄色。100、200、300、400μg/ml独活寄生汤均能抑制IL-1β诱导的软骨细胞凋亡,以300μg/ml独活寄生汤最为明显(P<0.01)。与模型组比较,独活寄生汤组软骨细胞中miR-98表达明显下降(P<0.05)。与模型组比较,独活寄生汤组软骨细胞中的Bcl-2蛋白表达显著升高,而Bax、Caspase-3、Caspase-9蛋白表达明显下降(P<0.05)。结论独活寄生汤能够抑制IL-1β诱导的软骨细胞凋亡,可能是通过下调miR-98起作用的。
引文
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14 殷春明,潘晓华.膝骨关节炎患者关节液中miR-140-3p表达可反映骨关节炎进程[J].中国组织工程研究,2016;20(35):5277-83.
15 Wang J,Chen L,Jin S,et al.MiR-98 promotes chondrocyte apoptosis by decreasing Bcl-2 expression in a rat model of osteoarthritis[J].Acta Biochim Biophys Sin(Shanghai),2016;48(10):923-9.
2 Ge Q,Wang H,Xu X,et al.PDK1 promotes apoptosis of chondrocytes via modulating MAPK pathway in osteoarthritis[J].Tissue Cell,2017;49(6):719-25.
3 Wang J,Chen L,Jin S,et al.Altered expression of microRNA-98 in IL-1beta-induced cartilage degradation and its role in chondrocyte apoptosis[J].Mol Med Rep,2017;16(3):3208-16.
4 Huang Z,Zhou M,Wang Q,et al.Mechanical and hypoxia stress can cause chondrocytes apoptosis through over-activation of endoplasmic reticulum stress[J].Arch Oral Biol,2017;84:125-32.
5 荀培军,艾有利.独活寄生汤在骨科疾病治疗中的应用[J].中医正骨,2014;26(12):56-8.
6 李西海,张翼,叶蕻芝,等.独活寄生汤治疗风寒湿痹型膝骨关节炎的机制[J].中医正骨,2012;24(1):68-71.
7 周红梅,金亮,吴斌,等.独活寄生汤治疗骨关节炎疗效的Meta分析[J].中国中医急症,2015;24(12):2099-102.
8 Wang J,Chen L,Jin S,et al.Altered expression of microRNA-98 in IL-1beta-induced cartilage degradation and its role in chondrocyte apoptosis[J].Mol Med Rep,2017;16(3):3208-16.
9 刘广宇,张杰,张晓峰.内质网跨膜蛋白肌醇需求酶1α介导的凋亡通路在兔骨关节炎发病中的作用及机制[J].中国老年学杂志,2017;37(2):291-3.
10 杨建辉,吕建国,聂会勇,等.白藜芦醇对白细胞介素-1β诱导的软骨细胞凋亡及增殖能力的影响[J].中国老年学杂志,2014;34(21):6151-3.
11 Gu Y,Chen J,Meng Z,et al.Diazoxide prevents H2O2-induced chondrocyte apoptosis and cartilage degeneration in a rat model of osteoarthritis by reducing endoplasmic reticulum stress[J].Biomed Pharmacother,2017;95:1886-94.
12 Jiang L,Wang W,He Q,et al.Oleic acid induces apoptosis and autophagy in the treatment of Tongue Squamous cell carcinomas[J].Sci Rep,2017;7(1):11277.
13 Rao Z,Wang S,Wang J.Peroxiredoxin 4 inhibits IL-1beta-induced chondrocyte apoptosis via PI3K/AKT signaling[J].Biomed Pharmacother,2017;90:414-20.
14 殷春明,潘晓华.膝骨关节炎患者关节液中miR-140-3p表达可反映骨关节炎进程[J].中国组织工程研究,2016;20(35):5277-83.
15 Wang J,Chen L,Jin S,et al.MiR-98 promotes chondrocyte apoptosis by decreasing Bcl-2 expression in a rat model of osteoarthritis[J].Acta Biochim Biophys Sin(Shanghai),2016;48(10):923-9.