目的 探討藏藥經(jīng)典方肺熱普清散對(duì)小鼠肝炎病毒A59（MHV-A59）感染小鼠模型的影響及其對(duì)炎癥因子的作用。方法 以雄性昆明種小鼠為實(shí)驗(yàn)對(duì)象，利用 MHV-A59 滴鼻操作建立小鼠冠狀病毒感染模型，設(shè)置對(duì)照組、模型組、陽性（利巴韋林）組和肺熱普清散低、中、高劑量（25、50、100 mg·kg^-1）3組，每天ig給藥1次，連續(xù)給藥10 d，考察記錄造模期內(nèi)小鼠體質(zhì)量變化，處死后稱體質(zhì)量并計(jì)算小鼠肺臟指數(shù)和肝臟指數(shù)；使用Elisa試劑盒測定小鼠血清中炎癥因子白細(xì)胞介素-1β（IL-1β）和白細(xì)胞介素-18（IL-18）含量；利用實(shí)時(shí)熒光定量PCR（qRT-PCR）方法檢測小鼠肝臟中IL-1β、IL-18 的mRNA表達(dá)水平和血清中MHV-A59病毒載量；通過蘇木素-伊紅（HE）染色，檢測肺熱普清散對(duì)肝臟和肺臟細(xì)胞壞死及炎癥細(xì)胞浸潤的影響。結(jié)果 與對(duì)照組比較，模型組的體質(zhì)量增長緩慢（P＜0.01）；肺熱普清散 3 個(gè)劑量組的體質(zhì)量每日遞增，中、高劑量組在第5～8天均與模型組有顯著差異（P＜0.01）；各組小鼠肺臟指數(shù)組間無顯著性差異，肺熱普清散中劑量組肝臟指數(shù)與模型組有顯著性差異（P＜0.05）；模型組病毒感染后血清中炎癥因子 IL-1β 和 IL-18 顯著性升高（P＜0.001），與模型組比較，肺熱普清散 3個(gè)劑量組均可降低血清中 IL-18水平（P＜0.001、0.01）；模型組肝臟組織中 IL-1β和IL-18 的 mRNA 表達(dá)水平均顯著升高 （P＜0.001）；與模型組比較，肺熱普清散高劑量組肝臟中 IL-1β轉(zhuǎn)錄水平顯著降低（P＜0.01）；與模型組比較，肺熱普清散 3組實(shí)驗(yàn)動(dòng)物肝臟組織中 IL-18水平未出現(xiàn)顯著性變化；MHV載量測定結(jié)果顯示，與模型組比較，利巴韋林組和肺熱普清散高劑量組血清中MHV載量顯著性降低（P＜0.01）；病理切片結(jié)果顯示，模型組肝臟細(xì)胞出現(xiàn)廣泛壞死和變性，并出現(xiàn)炎癥細(xì)胞浸潤，氣球樣變等現(xiàn)象，肺熱普清散高劑量和利巴韋林組出現(xiàn)點(diǎn)狀病灶、炎癥細(xì)胞浸潤等明顯減少現(xiàn)象；模型組肺組織肺泡明顯間隔增厚，間質(zhì)水腫，肺泡被壓縮，有炎癥細(xì)胞浸潤，毛細(xì)血管擴(kuò)張充血等現(xiàn)象，而肺熱普清散中、高劑量組和陽性組肺組織肺泡間隔較窄，肺泡舒展，局部有炎性浸潤和出血，肺泡腔內(nèi)的分泌物明顯減少。結(jié)論 肺熱普清散對(duì)MHV-A59感染小鼠模型具有保護(hù)作用，其作用機(jī)制與抑制病毒復(fù)制和抑制感染后炎癥因子有關(guān)。

Objective To explore the effects and inflammatory factors of the Tibetan medicine Feire Puqing Powder (FPP) on the mouse model of hepatitis coronavirus A59 (MHV-A59) infection.Methods Male Kunming mice were used as experimental subjects treated with MHV-A59 by nose-dropping, and six groups were set up: the control, the model, the positive (ribavirin), and the FPP's low-, medium-, and high- dosage (25, 50, and 100 mg·kg^-1) groups, and the FPP was ig administered once a day for 10 consecutive days, and the body weight of the mice during the modeling period was examined and recorded, and the lung and liver indices of the mice were examined after execution; The serum levels of IL-1β and IL-18 were determined using the Elisa kit; The mRNA of IL-1β and IL-18 in the liver, and the viral load in the serum were detected using the qRT-PCR; The pathological sections of liver and lung were stained with hematoxylin-eosin (HE), and the necrotic and inflammatory cell infiltration of the liver and lung in each experimental group were observed and recorded under the microscope.Results Compared with the control group, the body weight of the model group increased slowly (P < 0.01); The body weight of the three dose groups of FPP increased daily, and there was a significant difference between middle and high dose groups with the model group on days 5-8 (P < 0.01); There was a significant difference at the liver index of the middle dose group of FPP vs the model group (P < 0.05), while others were no significant difference on the lung index of the mice in each group; The serum inflammatory factors IL-1β and IL-18 were significantly elevated (P < 0.001), and compared with the model group, all three dose groups of FPP reduced the serum level of IL-18, (P < 0.001, 0.01); The IL-1β and IL-18 were significantly elevated in liver tissues of the model group (P < 0.001); Compared with the model group, the transcript level of IL-1β in the liver treated by FPP high-dose group was significantly reduced (P < 0.01); The level of IL-18 in the liver tissues of experimental animals in FPP triple-dose group did not show significant changes; The serum MHV load in each group of mice showed that, compared with the model group, there was a significant reduction of MHV load in the serum in the ribavirin group and the FPP high-dose group (P < 0.01); Liver histopathological sections showed that: Model group showed extensive necrosis and degeneration, with inflammatory cell infiltration and ballooning, while punctate foci and inflammatory cell infiltration were reduced in the FPP high-dose and ribavirin groups; Histopathological sections of lung tissue showed that: lung tissue of the model group showed obvious thickening of alveolar septa, mesenchymal edema, compressed alveoli, with inflammatory cell infiltration, and capillary dilation and congestion, capillary dilatation and congestion, while the alveolar intervals of lung tissues in the middle-dose group, high-dose group and ribavirin group of FPP were narrower, the alveoli were stretched out, and there were still inflammatory infiltration and hemorrhage locally, and secretion in the alveolar lumen was significantly reduced.Conclusion FPP has a protective effect on the MHV-A59 mouse infection, and the mechanism is related to the inhibition of viral replication and the inhibition of inflammatory factor increase.

R29；R51；R37

西藏自治區(qū)科技廳科技發(fā)展項(xiàng)目（XZ202202YD0020C）；日喀則市區(qū)域科技協(xié)同創(chuàng)新專項(xiàng)（QYXTZX-RKZ2023-01）