昆明山海棠对CIA大鼠足爪组织MMP-13蛋白表达及血清和足爪组织中IL-12、IL-23和IL-37水平的影响

目的:探讨昆明山海棠(Tripterygium hypoglaucum Hutch,THH)对胶原性关节炎(collagen-induced arthritis,CIA)大鼠的干预作用及可能的机制。方法:SD大鼠50只,随机分为正常组和胶原性关节炎模型制作组,用鸡Ⅱ型胶原诱导制作CIA大鼠模型。将造模成功的36只大鼠随机分为模型组、地塞米松治疗组与THH 200 mg/kg、400 mg/kg干预组。ELISA法检测CIA大鼠血清和足爪组织中促炎因子IL-12、IL-23和抑炎因子IL-37的含量;HE染色观察各组大鼠足爪组织病理学变化;Western blot法检测足爪组织MMP-13蛋白的表达;荧光标记法检测大鼠足爪组织中MMP-13的活性。结果:较模型组比较,THH 400 mg/kg治疗组大鼠体重下降明显减轻(P0.01),血清中IL-12和IL-23的水平分别降低了28.31%和41.57%(P0.01),足爪组织中IL-12和IL-23的含量分别下降了30.78%和39.46%,而血清和足爪组织中IL-37的水平显著升高了79.43%和75.78%(P0.01),足爪皮下组织病理变化明显减轻,足爪组织中MMP-13蛋白表达降低了49.22%(P0.01),且MMP-13活性明显下降。而THH 200 mg/kg治疗组大鼠上述指标无明显改善。结论:THH对CIA大鼠炎症的明显抑制作用可能与其降低促炎因子IL-12和IL-23、增高抑炎因子IL-37含量,抑制炎性细胞浸润及血管增生、下调MMP-13蛋白表达、降低MMP-13活性有关。

Effect of Tripterygium hypoglaucum Hutch on MMP-13 protein expression and IL-12, IL-23 and IL-37 levels in serum and foot paws of rats with collagen-induced arthritis

AIM: To investigate the effect of tripterygium hypoglaucum Hutch (THH) on collagen-induced arthritis (CIA) in rats and its possible mechanism. METHODS: SD rats were randomly divided into normal group and CIA group. The rat model of type Ⅱ CIA was successfully established and the model rats were randomly divided into 4 different groups: model group, dexamethasone group, THH (200 mg/kg) group, and THH (400 mg/kg) group. The contents of IL-12, IL-23 and IL-37 in the serum and foot paws of the CIA rats were detected by ELISA. The histopathological changes of the skin of the food paws were observed by HE staining. The protein expression of MMP-13 was determined by Western blot. The MMP-13 activity in the foot paws was detected by fluorescence labeling method. RESULTS: Compared with CIA group, THH at dose of 400 mg/kg significantly reduced the weight loss in type Ⅱ CIA rats (P<0.01). THH at dose of 400 mg/kg obviously decreased the contents of IL-12 by 28.31%, IL-23 by 41.57% in the serum and IL-12 by 30.78%, IL-23 by 39.46% in the foot paws, while IL-37 was significantly increased by 79.43% in the serum and 75.78% in the foot paws (P<0.01). The pathological changes of the subcutaneous tissues were improved by treating with THH (400 mg/kg). The protein expression of MMP-13 was significantly decreased by 31.82% (P<0.01), and the MMP-13 activity was also reduced. THH at dose of 200 mg/kg had no obvious improvement on the above indexes. CONCLUSION: THH has significant inhibitory effect on rat CIA by reducing the content of proinflammatory cytokines IL-12 and IL-23, increasing the content of anti-inflammatory factor IL-37, inhibiting inflammatory cell infiltration and vascular proliferation, and attenuating the protein expression of MMP-13 and MMP-13 activity in rats.