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肝生成素Cn(HPPCn)对急性肝损伤具有保护作用。

Hepatopoietin Cn (HPPCn) Generates Protective Effects on Acute Liver Injury.

作者信息

Li Na, Liu Feng-Jiao, Li Dan-Dan, Sun Chun-Xia, Li Jian, Qu Mei-Hua, Cui Chun-Ping, Zhang Da-Jin

机构信息

School of Pharmacy, Key Laboratory of Applied Pharmacology, Weifang Medical University, Wei Fang, China.

Center for Basic Medical Sciences, Sixth Medical Center of PLA General Hospital, Beijing, China.

出版信息

Front Pharmacol. 2019 Jul 4;10:646. doi: 10.3389/fphar.2019.00646. eCollection 2019.

Abstract

To observe the protective role of hapatopoietin Cn (HPPcn) on acute liver injury. Six hours after 10 mmol/L CCl, 150 mmol/L ethanol, or 0.6 mmol/L HO treatment, SMMC7721 human hepatoma cells were incubated with 10, 100, or 200 ng/ml recombinant human HPPCn protein (rhHPPCn) for an additional 24 h. The cell survival rate was analyzed using the CCK-8 assay. The CCl-induced apoptosis of SMMC7721 cells was detected by flow cytometry. Then, the levels of glutamic oxaloacetic transaminase (GOT), glutamic-pyruvic transaminase (GPT), malondialdehyde (MDA), lactate dehydrogenase (LDH), glutathione peroxidase (GSH-PX), and superoxide dismutase (SOD) in SMMC7721 cell lysates and cell culture supernatant were detected. SMMC7721 cells were treated with different concentrations of rhHPPCn (0, 10, and 100 ng/ml). The cell proliferation indexes (BrdU incorporation and PCNA expression) were detected by immunohistochemistry (IHC). An acute liver injury mouse model was established by a one-time intraperitoneal injection of 20% CCl at a volume of 5 ml/kg body weight. One hour after CCl injection, 1.25 or 2.5 mg rhHPPCn/12 h/kg body weight was injected the tail vein. The serum levels of GOT and GPT were detected at different time points. Pathological changes in the liver were evaluated. PCNA expression levels were observed by IHC. rhHPPCn increased the survival rate of SMMC7721 cells and inhibited chemical toxicity-induced cell apoptosis. The levels of GOT, GPT, MDA, and LDH in the cell supernatant were significantly reduced, while GSH-PX and SOD were significantly increased after rhHPPCn treatment in the CCl-treated SMMC7721 cells. BrdU incorporation and PCNA expression increased in a concentration-dependent manner, indicating that rhHPPCn promotes cell proliferation. The results showed that rhHPPCn significantly reduced the serum levels of GOT and GPT in CCl-induced acute liver injury mice. rhHPPCn alleviated the tissue damage and increased PCNA expression, indicating the promotion of proliferation after acute injury. rhHPPCn protects hepatocytes from chemical toxins by promoting proliferation and inhibiting apoptosis and . Our study provides new insights for the clinical treatment of acute liver injury.

摘要

观察肝生成素Cn(HPPcn)对急性肝损伤的保护作用。在用10 mmol/L四氯化碳、150 mmol/L乙醇或0.6 mmol/L过氧化氢处理6小时后,将SMMC7721人肝癌细胞与10、100或200 ng/ml重组人HPPCn蛋白(rhHPPCn)再孵育24小时。使用CCK - 8法分析细胞存活率。通过流式细胞术检测四氯化碳诱导的SMMC7721细胞凋亡。然后,检测SMMC7721细胞裂解物和细胞培养上清液中谷草转氨酶(GOT)、谷丙转氨酶(GPT)、丙二醛(MDA)、乳酸脱氢酶(LDH)、谷胱甘肽过氧化物酶(GSH - PX)和超氧化物歧化酶(SOD)的水平。用不同浓度的rhHPPCn(0、10和100 ng/ml)处理SMMC7721细胞。通过免疫组织化学(IHC)检测细胞增殖指数(BrdU掺入和PCNA表达)。通过一次性腹腔注射5 ml/kg体重的20%四氯化碳建立急性肝损伤小鼠模型。在注射四氯化碳1小时后,通过尾静脉注射1.25或2.5 mg rhHPPCn/12 h/kg体重。在不同时间点检测血清中GOT和GPT的水平。评估肝脏的病理变化。通过IHC观察PCNA表达水平。rhHPPCn提高了SMMC7721细胞的存活率并抑制了化学毒性诱导的细胞凋亡。在经四氯化碳处理的SMMC7721细胞中,rhHPPCn处理后细胞上清液中GOT、GPT、MDA和LDH的水平显著降低,而GSH - PX和SOD显著升高。BrdU掺入和PCNA表达呈浓度依赖性增加,表明rhHPPCn促进细胞增殖。结果表明,rhHPPCn显著降低了四氯化碳诱导的急性肝损伤小鼠血清中GOT和GPT的水平。rhHPPCn减轻了组织损伤并增加了PCNA表达,表明在急性损伤后促进了增殖。rhHPPCn通过促进增殖和抑制凋亡保护肝细胞免受化学毒素的损伤。我们的研究为急性肝损伤的临床治疗提供了新的见解。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/86af/6620608/f1e259497f9a/fphar-10-00646-g001.jpg

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