目的 探討亞精胺促進(jìn)線粒體自噬緩解急性胰腺炎的療效及作用機(jī)制。方法 將C57BL/6J小鼠隨機(jī)分為對(duì)照組、模型組、亞精胺組、亞精胺＋環(huán)孢菌素A組，每組5只。通過每小時(shí)向小鼠腹膜內(nèi)注射1次50 μg/kg的蛙皮素構(gòu)建急性胰腺炎模型。對(duì)照組按照同模型組的處理方式注射等體積無菌生理鹽水；亞精胺組腹膜內(nèi)注射100 mmol的亞精胺；亞精胺＋環(huán)孢菌素A組在給予亞精胺治療的同時(shí)，腹膜內(nèi)注射10 mg/kg的環(huán)孢菌素A。蘇木精–伊紅（HE）染色評(píng)估小鼠胰腺組織損傷程度。TUNEL染色評(píng)估胰腺腺泡細(xì)胞凋亡情況。ELISA法測(cè)定小鼠血清炎癥因子白細(xì)胞介素-6（IL-6）、腫瘤細(xì)胞壞死因子-α（TNF-α）、白細(xì)胞介素-1β（IL-1β）、C反應(yīng)蛋白（CRP）的水平。免疫組織化學(xué)染色（IHC）評(píng)估髓過氧化物酶（MPO）水平。JC-1染料測(cè)定胰腺組織提取的總線粒體的線粒體膜電勢(shì)水平。Western blotting法測(cè)定胰腺組織細(xì)胞質(zhì)基質(zhì)型LC3（LC-3Ⅰ）、微管相關(guān)蛋白1輕鏈3Ⅱ（LC-3Ⅱ）、泛素結(jié)合蛋白（p62）、NOD樣受體熱蛋白結(jié)構(gòu)域相關(guān)蛋白3（NLRP3）、半胱氨酸天冬氨酸蛋白酶（Caspase-1）、cleaved-Caspase-1、IL-1β、PTEN誘導(dǎo)假定激酶1（PINK1）、磷酸化Parkin（p-Parkin）和Parkin的表達(dá)水平。結(jié)果 與模型組相比，亞精胺組胰腺組織幾乎不存在中性粒細(xì)胞的大量浸潤，腺泡細(xì)胞腫脹程度明顯減弱，基本無腺泡細(xì)胞壞死；TUNEL染色陽性細(xì)胞數(shù)和MPO IHC陽性染色細(xì)胞數(shù)明顯降低；小鼠血清IL-6、TNF-α、IL-1β、CRP的水平明顯降低；胰腺組織LC-3Ⅰ/Ⅱ、NLRP3、IL-1β、cleaved-Caspase-1、PINK1和p-Parkin的表達(dá)水平明顯降低，p62的表達(dá)水平明顯增強(qiáng)；胰腺組織總線粒體膜電勢(shì)水平明顯增強(qiáng)（P＜0.05）。與亞精胺組相比，亞精胺＋環(huán)孢菌素A組完全逆轉(zhuǎn)了亞精胺的治療效果。結(jié)論 亞精胺可以促進(jìn)線粒體自噬緩解急性胰腺炎。

Objective To explore the effect and mechanism of spermidine promoting mitochondrial autophagy in alleviating acute pancreatitis. Methods C57BL/6J mice were randomly divided into control group, model group, spermidine group, and spermidine + cyclosporin A group, with 5 mice in each group. Acute pancreatitis model was established by intraperitoneal injection of 50 μg/kg of frogskin into mice every hour. Control group was injected with equal volume sterile saline according to the treatment method of the same model group. The spermidine group was given intraperitoneal injection dose of 100 mmol spermidine, Spermidine + cyclosporine A group was given 10 mg/kg cyclosporine A by intraperitoneal injection at the same time of spermidine treatment. Hematoxylin-eosin (HE) staining was used to evaluate the degree of pancreatic tissue injury in mice. The apoptosis of pancreatic acinar cells was evaluated by TUNEL staining. Serum levels of inflammatory factor IL-6, TNF-α, IL-1β, and CRP were determined by ELISA. Myeloperoxidase (MPO) levels were evaluated by immunohistochemical staining (IHC). The level of mitochondrial membrane potential of total mitochondria extracted from pancreatic tissue was determined by JC-1 dye. LC-3Ⅰ, LC-3Ⅱ, p62, NLRP3, Caspase-1, cleaved- Caspase-1, IL-1β, PINK1, p-Parkin, Parkin were determined by Western blotting method. Results Compared with the model group, there was almost no large infiltration of neutrophils in the pancreatic tissue of spermidine group, the swelling degree of acinous cells was significantly reduced, and there was basically no acinous cell necrosis. The number of TUNEL positive cells and MPO IHC positive cells decreased significantly. Serum levels of IL-6, TNF-α, IL-1β, and CRP were significantly decreased. The expression levels of LC-3-Ⅰ/Ⅱ, NLRP3, IL-1β, cleaved Caspase-1, PINK1, and p-Parkin in pancreatic tissue were significantly decreased, and the expression level of p62 was significantly increased. The total mitochondrial membrane potential of pancreatic tissue was significantly increased (P < 0.05). Compared with the spermidine group, spermidine + cyclosporine A group completely reversed the therapeutic effect of spermidine. Conclusion Spermidine can promote mitophagy and relieve acute pancreatitis.

新疆維吾爾自治區(qū)自然科學(xué)基金資助項(xiàng)目（2022D01C107）