化瘀止痛贴膏调节IL-6/STAT3信号通路对急性软组织损伤大鼠炎症反应的影响
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湖南省中医药管理局科研项目(B2023052)


Effect of Huayu Zhitong Plaster on inflammatory response in rats with acute soft tissue injury by regulating IL-6/STAT3 signal pathway
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    摘要:

    目的 探讨化瘀止痛贴膏调节白细胞介素-6(IL-6)/信号转导与转录激活子3(STAT3)信号通路对急性软组织损伤(ASTI)大鼠炎症反应的影响。方法 将SPF级SD大鼠随机分为对照组、模型组、化瘀止痛贴膏低、中、高剂量组、精制狗皮膏组。利用机械冲击挫伤法建立ASTI动物模型,建模成功后,各组贴敷相应药物,1次/d,每次给药8 h,连续7d。对大鼠进行ASTI损伤评分;采用血液流变检测仪检测血液流变学指标;苏木精和伊红(HE)染色法观察大鼠损伤部位肌肉组织病理变化;酶联免疫吸附法(ELISA)检测各组大鼠损伤部位肌肉组织中IL-6、IL-β、肿瘤坏死因子-α(TNF-α)水平;实时荧光定量PCR(qRT-PCR)法检测损伤部位肌肉组织中IL-6与STAT3 mRNA表达;蛋白免疫印迹(Western blot)法检测大鼠损伤部位肌肉组织中IL-6/STAT3信号通路蛋白表达。结果 与对照组比较,模型组大鼠软组织损伤评分、全血黏度、血浆黏度、红细胞压积、肌肉组织病理学评分、IL-6、IL-β、TNF-α水平、IL-6与STAT3 mRNA表达水平及IL-6、p-STAT3/STAT3蛋白表达水平升高(均P<0.05);与模型组比较,化瘀止痛贴膏低、中、高剂量组和狗皮膏药组大鼠软组织损伤评分、全血黏度、血浆黏度、红细胞压积、肌肉组织病理学评分、IL-6、IL-β、TNF-α水平、IL-6与STAT3 mRNA表达水平及IL-6、p-STAT3/STAT3蛋白表达水平降低(均P<0.05);化瘀止痛贴膏高剂量组与精制狗皮膏组大鼠以上指标差异均无统计学意义(P>0.05)。结论 化瘀止痛贴膏可能通过下调IL-6/STAT3信号通路缓解ASTI大鼠的炎症反应

    Abstract:

    Objective To investigate the effect of Huayu Zhitong Plaster on inflammatory response in rats with acute soft tissue injury (ASTI) by regulating the interleukin-6 (IL-6)/signal transducer and activator of transcription 3 (STAT3) signal pathway. Methods SPF grade SD rats were randomly grouped into control group, model group, low, medium and high dose Huayu Zhitong plaster groups, and Goupigao group. ASTI animal model was established by mechanical impact contusion method. After successful modeling, each group applied corresponding drugs once a day for 8 hours each time for 7 days. The rats were scored with ASTI damage; the hemorheology indexes were measured by hemorheology detector; the pathological changes of muscle tissue at the injured site were observed by hematoxylin and eosin (HE) staining; enzyme-linked immunosorbent assay (ELISA) was applied to detect the levels of interleukin (IL)-6, IL-β and tumor necrosis factor-α (TNF-α) in the muscle tissue of the injured rats in each group; the expression of IL-6 and STAT3 mRNA in muscle tissue of the injured site was detected by real-time fluorescence quantitative PCR (qRT-PCR); Western blot was applied to detect the expression of IL-6/STAT3 signal pathway protein in the injured muscle tissue of rats. Results Compared with the control group, the soft tissue injury score, whole blood viscosity, plasma viscosity, hematocrit, muscle histopathological score, the IL-6, IL-β, TNF-α levels, the IL-6 and STAT3 mRNA expression levels, and the IL-6 and p-STAT3/STAT3 protein expression levels in the model group were increased (P<0.05); compared with the model group, the soft tissue injury score, whole blood viscosity, plasma viscosity, hematocrit, muscle histopathological score, the IL-6, IL-β, TNF-α levels, the IL-6 and STAT3 mRNA expression levels, and the IL-6 and p-STAT3/STAT3 protein expression levels in the low, medium and high dose Huayu Zhitong Plaster groups and the Goupigao group were decreased (P<0.05); there was no obvious difference in the above indexes between the high dose Huayu Zhitong Plaster groups and the Goupigao group (P>0.05) Conclusion Huayu Zhitong Plaster may alleviate the inflammatory reaction of ASTI rats by down-regulating IL-6/STAT3 signal pathway

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  • 在线发布日期: 2024-06-18
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