贝母素乙通过ADAMTS-5调控PPARγ/c-SKI信号通路影响大鼠膝骨关节炎的研究OA
Research on knee osteoarthritis rats by regulating the PPARγ/c-SKI signaling pathway by ADAMTS-5
目的 探讨贝母素乙(PEI)通过含血小板蛋白结合基序的解聚蛋白样金属蛋白酶-5(ADAMTS-5)调控过氧化物酶体增殖物激活受体γ(PPARγ)/细胞型禽肉瘤病毒癌基因同源物(c-SKI)信号通路影响膝骨关节炎(KOA)大鼠的作用机制.方法 通过关节腔内注射碘乙酸钠构建大鼠KOA模型.将大鼠分为对照组、模型组、实验组(腹腔注射3.5 mg·kg-1 PEI)和ADAMTS-5过表达组(尾静脉注射oe-ADAMTS-5质粒).用机械刺激法检测大鼠疼痛阈值;用苏木精-伊红染色法检测大鼠膝关节病理损伤情况;用蛋白质印迹法检测大鼠膝关节组织中的ADAMTS-5、基质金属蛋白酶(MMP)-3、MMP-13和PPARγ/c-SKI信号通路蛋白水平;用免疫组化法检测大鼠膝关节组织中的Ⅱ型胶原蛋白α1链(Col2α1)蛋白水平;用相应试剂盒检测大鼠血清中的炎症因子水平.结果 对照组、模型组和实验组的疼痛阈值分别为(19.15±2.87)、(7.10±1.03)和(13.69±1.27)g,ADAMTS-5蛋白相对表达水平分别为1.00±0.16、1.77±0.21 和 1.35±0.18;对照组、模型组、实验组和 ADAMTS-5过表达组的Col2α1相对表达水平分别为1.00±0.10、0.28±0.04、0.76±0.13和 0.58±0.06,MMP-3 相对表达水平分别为 1.00±0.15、2.11±0.36、1.62±0.22和 1.83±0.25,MMP-13 相对表达水平分别为 1.00±0.18、1.98±0.28、1.27±0.16 和 1.45±0.19,肿瘤坏死因子-α(TNF-α)水平分别为(62.74±6.93)、(159.37±26.40)、(98.08±14.09)和(124.43±17.79)pg·mL-1,白细胞介素(IL)-4 水平分别为(75.62±11.54)、(19.56±2.59)、(53.28±6.34)和(41.67±5.31)pg·mL-1,IL-6 水平分别为(42.65±9.77)、(133.84±13.22)、(84.19±11.71)和(105.24±18.19)pg·mL-1,IL-10 水平分别为(98.06±24.50)、(23.42±10.61)、(64.72±20.96)和(38.09±12.86)pg·mL-1,PPARγ蛋白相对表达水平分别为1.00±0.13、0.39±0.07、0.73±0.09和0.51±0.08,c-SKI蛋白相对表达水平分别为1.00±0.17、0.32±0.05、0.86±0.15和0.44±0.09,模型组的上述指标与对照组比较,实验组的上述指标与模型组比较,ADAMTS-5过表达组的上述指标与实验组比较,在统计学上差异均有统计学意义(均P＜0.05).结论 PEI抑制KOA大鼠炎症因子水平,减轻骨纤维化,改善骨损伤,从而缓解关节疼痛,可能是通过下调ADAMTS-5表达激活PPARγ/c-SKI信号通路来实现的.
Objective To investigate the mechanism of peiminine(PEI)affects pain and cartilage injury in rats with knee osteoarthritis(KOA)by regulating the peroxisome proliferat or-activated receptor gamma(PPARγ)/cellular sloan kettering institute(c-SKI)signaling pathway through a disintegrin and metalloproteinase with thrombospondin motifs 5(AD AMTS-5).Methods The KOA model was established in rats by intra-articular injection of sodium iodoacetate.The rats were divided into control group,model group,experimental group(intraperitoneal injection of 3.5 mg·kg-1 PEI)and ADAMTS-5 overexpression group(tail vein injection of oe-ADAMTS-5 plasmid).The pain threshold of rats was measured using the mechanical stimulation method;the pathological injury of the rat knee joint was examined by hematoxylin-eosin(HE)staining;the protein levels of ADAMTS-5,matrix metalloproteinase(MMP)-3,MMP-13 and proteins in the PPARγ/c-SKI signaling pathway in rat knee joint tissues were detected by Western blot;the protein level of collagen type Ⅱ alpha 1 chain(Col2α1)in rat knee joint tissues was determined by immunohistochemistry;the levels of inflammatory factors in rat serum were measured using corresponding commercial kits.Results The pain thresholds in control group,model group and experimental group were(19.15±2.87),(7.10±1.03)and(13.69±1.27)g,respectively;the relative expression levels of ADAMTS-5 were 1.00±0.16,1.77±0.21 and 1.35±0.18,respectively;Col2α1 levels in control,model,experimental,and ADAMTS-5 overexpression groups were 1.00±0.10,0.28±0.04,0.76±0.13 and 0.58±0.06,respectively;the relative expression levels of MMP-3 were 1.00±0.15,2.11±0.36,1.62±0.22 and 1.83±0.25,respectively;the relative expression levels of MMP-13 were 1.00±0.18,1.98±0.28,1.27±0.16 and 1.45±0.19,respectively;the tumor necrosis factor-alpha(TNF-α)levels were(62.74±6.93),(159.37±26.40),(98.08±14.09)and(124.43±17.79)pg·mL-1,respectively;the interleukin(IL)-4 levels were(75.62±11.54),(19.56±2.59),(53.28±6.34)and(41.67±5.31)pg·mL-1,respectively;IL-6 levels were(42.65±9.77),(133.84±13.22),(84.19±11.71)and(105.24±18.19)pg·mL-1,respectively;IL-10 levels were(98.06±24.50),(23.42±10.61),(64.72±20.96)and(38.09±12.86)pg·mL-1,respectively;the relative expression levels of PPARγ were 1.00±0.13,0.39±0.07,0.73±0.09 and 0.51±0.08,respectively;the relative expression levels of c-SKI were 1.00±0.17,0.32±0.05,0.86±0.15 and 0.44±0.09,respectively.The above indicators in model group were statistically significantly different from those in control group,those in experimental group were statistically significantly different from those in model group,and those in ADAMTS5 overexpression group were statistically significantly different from those in experimental group(all P＜0.05).Conclusion PEI inhibits inflammatory cytokine levels,alleviates bone fibrosis,and improves bone injury in KOA rats,thereby relieving joint pain,possibly by downregulating ADAMTS-5 expression to activate the PPARγ/c-SKI signaling pathway.
陈红涛;吴金海;曲军;胡润武
南阳市第一人民医院 急诊创伤外科,河南南阳 473000南阳市第一人民医院 急诊医学科,河南南阳 473000南阳市第一人民医院 急诊创伤外科,河南南阳 473000南阳市第一人民医院 急诊创伤外科,河南南阳 473000
医药卫生
贝母素乙膝骨关节炎含血小板蛋白结合基序的解聚蛋白样金属蛋白酶5过氧化物酶体增殖物激活受体γ/细胞型禽肉瘤病毒癌基因同源物c-SKI信号通路软骨损伤
peiminineknee osteoarthritisdisintegrin and metalloproteinase with thrombospondin motifs 5peroxisome proliferator-activated receptor gamma/cellular sloan kettering institute signaling pathwaycartilage injury
《中国临床药理学杂志》 2026 (5)
693-699,7
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