| 王厚东,张秀峰,裘建明,鲁振锋,沈忠.自体富血小板血浆用于肛瘘术后创面修复的实验研究[J].浙江中西医结合杂志,2023,33(6): |
| 自体富血小板血浆用于肛瘘术后创面修复的实验研究 |
| The Research on Autologous Platelet-rich Plasma Used to Repair Perianal Tissue Defect After Anal Fistulectomy |
| 投稿时间：2022-06-12 修订日期：2023-05-05 |
| DOI： |
| 中文关键词: 富血小板血浆 PRP 肛瘘 |
| 英文关键词:platelet-rich plasma PRP Anal Fistula |
| 基金项目:浙江省科技厅公益项目(LGF20H030002)；杭州市卫健委一般项目(2017A26)；杭州市科技局引导项目(20171226Y17);杭州市卫生科技计划一般项目(20210250)。 |
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| 中文摘要: |
| 目的 探索自体富血小板血浆(platelet-rich plasma, PRP)修复肛瘘术后创面的作用。方法 10只实验用猪采用挂线法进行肛瘘造模。38d后瘘管造影及病理检查评估瘘管形成。瘘管切除术后的左、右侧肛周创面随机分为PRP组与乏血小板血浆（platelet-poor plasma, PPP）组，PRP组创面予以富血小板血浆注射，PPP组创面予以乏血小板血浆注射。术后第12天取创面中央组织样本，酶联免疫吸附试验（Enzyme Linked Immunosorbent Assay，ELISA）检测组织样本中表皮细胞生长因子（Epidermal Growth Factor,EGF），促血管生成素Ⅱ (angiopoietin-2,Ang-2)、白介素-6（interleukin-6, IL-6）、白介素-12（interleukin-6, IL-12）含量。免疫组化检测CD34表达水平，免疫荧光检测波形蛋白（Vimentin）表达情况。观察创面愈合时间，超声评估创面愈合效果 结果 PRP中血小板浓度为(668.90±188.30)10^9/L 显著高于外周静脉血(194.10±44.44)10^9/L 及PPP (80.90±28.58)10^9/L (F=76.32，p＜0.01)。ELISA检测结果显示PRP注射组创面组织中EGF及Ang-Ⅱ含量均显著高于PPP注射组（293.72±21.21pg/mL vs 243.94±?38.14pg/mL，t=4.839，p＜0.01；87.29±9.46 pg/mL vs 71.10?±?8.06pg/mL，t=5.527，p＜0.01）。炎症因子IL-6及IL-12在PPP注射组的创面组织中含量均显著高于PRP注射组（639.53?±?70.34pg/mL vs 469.91±?51.16pg/mL，t=8.274，p＜0.01；194.84?±?21.81pg/mL vs 156.84±?18.52pg/mL，t=5.633，p＜0.01）。免疫组化显示PRP注射组的CD34水平显著高于PPP注射组（0.20±0.03 vs 0.15±0.03，t=3.181，p=0.01），免疫荧光检测结果显示Vimentin蛋白在PRP注射组的组织中的水平高于PPP注射组（61.30±6.57 vs 27.49±3.77，t=7.731，p＜0.01）。PRP组愈合时间为(16.9±2.0 d)明显低于PPP组 (21.2±2.1d)(t=-4.66, p＜0.01)。结论 PRP局部注射肛瘘术后创面可以提高创面组织中EGF及Ang-Ⅱ含量，从而促进上皮生长及新生血管形成，同时可以减轻组织炎症反应，促进肛瘘术后创面愈合。 |
| 英文摘要: |
| Objective To assess the effect of autologous platelet-rich plasma (PRP) for repair of perianal tissue defect after anal fistulectomy. Method Ten healthy pigs underwent rubber band ligation surgery. Fistulography and histopathological evaluation were performed on the 38th after surgery.After fistulectomy，left and right perianal defects were randomly divided into two groups.The experimental group underwent PRP injection, and the control group underwent platelet-poor plasma (PPP) injection. Tissue samples were taken from the central of wounds on day 12 after the surgery. The level of Epidermal Growth Factor (EGF), angiopoietin-2(Ang-2), interleukin-6, (IL-6) and interleukin-12, (IL-12) in wound tissues were detected by Enzyme Linked Immunosorbent Assay（ELISA）. Immunohistochemical analysis of CD34 expression and immunofluorescence assay of Vimentin expression in wound tissues were carried out at the same time. Wound healing time among the two groups were compared. Ultrasound was also used to evaluate the tissue healing. Results The platelet count in PRP (668.90±188.30) 10^9/L was higher than that in peripheral venous blood (194.10±44.44)10^9/L and PPP (80.90±28.58)10^9/L (F=76.32，p＜0.01). The results of ELISA showed that the level of EGF expression and Ang-Ⅱ expression in PRP group were significantly higher than the PPP group(293.72±21.21pg/mL vs 243.94±?38.14pg/mL, t=4.839, p＜0.01; 87.29±9.46 pg/mL vs 71.10?±?8.06pg/mL, t=5.527, p＜0.01). ELISA also revealed that the degree of IL-6 expression and IL-12 expression in PPP group were significantly higher than that in the PRP group(639.53?±?70.34pg/mL vs 469.91±?51.16pg/mL, t=8.274, p＜0.01; 194.84?±?21.81pg/mL vs 156.84±?18.52pg/mL, t=5.633, p＜0.01). Immunohistochemical analysis showed that the CD34 level in PRP group was significantly higher than that in PPP group (0.20±0.03 vs 0.15±0.03, t=3.181, p=0.01). Vimentin expression detected by immunofluorescence assay was higher in PRP group than PPP group (61.30±6.57 vs 27.49±3.77, t=7.731, p＜0.01). The wound healing time in the PRP group (16.9±2.0)d was significantly lower than that in the control group (21.2±2.1)d (t=-4.66, p＜0.01). The ultrasound scan showed that there was no pseudo healing between the two group.Conclusion Local injection of PRP could attenuate inflammatory response and increase the level of EGF and Ang-Ⅱ in wound tissue after anal fistulectomy, which could promote epithelial growth and neovascuarization, thus accelerating wound healing wound healing. |
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