Hepatopoietin Cn (HPPCn) Generates Protective Effects on Acute Liver Injury
Hepatopoietin Cn (HPPCn) Generates Protective Effects on Acute Liver Injury
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肝生成素 Cn (HPPCn) 对急性肝损伤产生保护作用
DOI：
10.3389/fphar.2019.00646
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发表时间：
2019-07-04
影响因子：
5.6
通讯作者：
Zhang, Da-Jin
中科院分区：
文献类型：
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作者：
Li, Na;Liu, Feng-Jiao;Zhang, Da-Jin
目的：观察肝细胞生成素Cn（HPPcn）对急性肝损伤的保护作用。研究方法：10 mmol/L CCl 4、150 mmol/L乙醇或0.6 mmol/L H2 O2处理SMMC 7721人肝癌细胞6 h后，分别与10、100或200 ng/ml重组人HPPCn蛋白（rhHPPCn）孵育24 h。使用CCK-8测定分析细胞存活率。流式细胞仪检测CCl 4诱导SMMC 7721细胞凋亡的作用。检测SMMC 7721细胞裂解液和细胞培养上清中谷草转氨酶（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）的含量。用不同浓度的rhHPPCn（0、10和100 ng/ml）处理SMMC 7721细胞。免疫组化法检测细胞增殖指数（BrdU掺入和PCNA表达）。以20%CCl4按5 ml/kg体重一次性腹腔注射建立小鼠急性肝损伤模型。注射CCl 4后1小时，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GOT和GPT水平。评价肝脏的病理学变化。免疫组化法检测PCNA表达水平。结果：rhHPPCn能提高SMMC 7721细胞的存活率，抑制化学毒性诱导的细胞凋亡。rhHPPCn处理后，细胞上清液中GOT、GPT、MDA和LDH水平显著降低，GSH-PX和SOD水平显著升高。BrdU掺入和PCNA表达以浓度依赖性方式增加，表明rhHPPCn促进细胞增殖。结果表明，rhHPPCn能显著降低CCl 4诱导的急性肝损伤小鼠血清GOT和GPT水平。rhHPPCn可减轻急性损伤后的组织损伤，增加PCNA表达，促进细胞增殖。结论：rhHPPCn在体内外均能通过促进肝细胞增殖和抑制肝细胞凋亡来保护肝细胞免受化学毒素的损伤。本研究为急性肝损伤的临床治疗提供了新的思路。
Objective: To observe the protective role of hapatopoietin Cn (HPPcn) on acute liver injury. Methods: Six hours after 10 mmol/L CCl4, 150 mmol/L ethanol, or 0.6 mmol/L H2O2 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 CCl4-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% CCl4 at a volume of 5 ml/kg body weight. One hour after CCl4 injection, 1.25 or 2.5 mg rhHPPCn/12 h/kg body weight was injected via 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. Results: 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 CCl4-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 CCl4-induced acute liver injury mice. rhHPPCn alleviated the tissue damage and increased PCNA expression, indicating the promotion of proliferation after acute injury. Conclusion: rhHPPCn protects hepatocytes from chemical toxins by promoting proliferation and inhibiting apoptosis in vivo and in vitro. Our study provides new insights for the clinical treatment of acute liver injury.