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ObjectiveTo investigate the protective effect of safranal against sepsis-related liver injury （SRLI） induced by lipopolysaccharide （LPS） in mice and its mechanism. MethodsA total of 32 experimental male C57BL/6 mice were divided into control group， single drug group， model group， and treatment group using the simple random method， with 8 mice in each group. The mice in the single drug group and the treatment group were intraperitoneally injected with safranal （60 mg/kg） for 7 days of pretreatment， and the mice in the model group and the treatment group were intraperitoneally injected with LPS （10 mg/kg） to induce acute liver injury. The activities of serum alanine aminotransferase （ALT） and aspartate aminotransferase （AST） were measured； HE staining was used to observe liver tissue sections； immunohistochemistry was used to analyze the expression of the downstream protein heme oxygenase-1 （HO-1） in the signal pathway； TUNEL was used to analyze the apoptosis of hepatocytes； Western blot was used to measure the expression of total proteins （nuclear factor erythroid 2-related factor 2 ［Nrf-2］ and HO-1） in liver tissue. The human liver cell line L02 was pretreated with safranal （100 μmol/L）， followed by induction of acute hepatocellular injury with LPS （100 ng/mL）， and DCFH-DA fluorescent labeling was used to detect reactive oxygen species （ROS）. ResultsAfter safranal pretreatment， the treatment group had significantly lower levels of ALT and AST than the model group （both P<0.001）， with a relatively intact pseudolobular structure and a smaller necrotic area in the liver. Compared with the model group， the treatment group had significant increases in the expression levels of Nrf2 and HO-1 in liver tissue after safranal+LPS treatment （both P<0.001）， and immunohistochemistry showed that safranal pretreatment increased the number of HO-1-positive cells. In the cell model of LPS-induced acute liver injury， the treatment group had a significant reduction in the production of ROS compared with the model group. ConclusionSafranal can exert a protective effect against SRLI induced by LPS in mice through the Nrf2/HO-1 pathway.
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ObjectiveTo investigate the protective effect of tanshinone I (T-I) on hepatic ischemia-reperfusion injury (HIRI) in mice. MethodsA total of 36 C57BL/6J mice were randomly divided into sham-operation group, ischemia-reperfusion (IR) group, IR+T-I (5 mg/kg) group, IR+T-I (10 mg/kg) group, IR+T-I (20 mg/kg) group, and IR+T-I (40 mg/kg) group, with 6 mice in each group. Each group was given intraperitoneal injection. The mice in the sham-operation group and the IR group were injected with an equal volume of the solvent olive oil; the mice in the IR+T-I groups were administered once a day for 7 consecutive days, a model of 70% HIRI was established at 2 hours after the last administration, and serum and liver samples were collected after 6 hours of reperfusion. Related kits were used to measure the serum level of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) and the content of superoxide dismutase (SOD), malondialdehyde (MDA), caspase-3, and reduced glutathione (GSH) in liver tissue; HE staining was used to observe liver histopathology; the TUNEL method was used to measure the level of hepatocyte apoptosis; immunohistochemistry was used to measure the protein expression of caspase-3 and heme oxygenase-1 (HO-1). A one-way analysis of variance was used for comparison of continuous data between multiple groups, and the least significant difference t-test was used for further comparison between two groups. ResultsCompared with the IR group, the IR+T-I (20mg/kg) group had significant reductions in the serum levels of ALT (192.48±23.67 U/L vs 336.90±41.52 U/L, P＜0.01) and AST (123.19±9.16 U/L vs 206.90±18.81 U/L, P＜0.01), and thus 20 mg/kg was determined as the optimal concentration. Compared with the IR group, the IR+T-I (20 mg/kg) group had significant reductions in MDA (1.34±0.21 μmol/mg vs 3.48±0.95 μmol/mg, P＜0.05) and caspase-3 (0.69±0.97 μmol/mg vs 1.04±0.35 μmol/mg, P＜0.05) and significant increases in SOD (274.47±30.53 U/mg vs 160.29±27.37 U/mg, P＜0.05) and GSH (2.12±0.27 μmol/mg vs 1.03±0.42 μmol/mg, P＜0.05). HE staining showed that the IR group had disordered structure of hepatic lobules and focal or extensive degeneration and necrosis of hepatocytes; compared with the IR group, the IR+T-I (20 mg/kg) group had a reduction in the area of hepatocyte necrosis and a basically complete structure of the liver. Immunohistochemistry showed that compared with the IR group, the IR+T-I (20 mg/kg) group had significant reductions in the number of apoptotic hepatocytes and the protein expression of caspase-3 and a significant increase in the protein expression of HO-1. ConclusionT-I exerts a protective effect against HIRI in mice by inhibiting liver oxidative stress response and hepatocyte apoptosis.
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ObjectiveTo investigate the effect of kaempferol on the proliferation, migration, invasion, and apoptosis of human hepatoma Bel-7402 cells and related molecular mechanism. MethodsHepatoma Bel-7402 cells cultured in vitro were randomly divided into control group and low-, middle-, and high-concentration experimental groups. The experimental groups were treated with low-, middle-, and high-concentration kaempferol (25, 50, and 100 μmol/L), and the control group was treated with an equal volume of dimethyl sulfoxide. CCK-8 assay was used to observe the effect of kaempferol on the viability of Bel-7402 cells; plate colony formation assay was used to evaluate the effect of kaempferol on cell colony formation ability; wound healing assay and Transwell chamber were used to observe the effect of kaempferol on cell migration and invasion; Western blot was used to measure the expression of apoptosis- and cycle-related proteins. A one-way analysis of variance was used for comparison between multiple groups, and the least significant difference t-test was used for further comparison between two groups. ResultsAfter 24 hours of treatment, the cell viability was 100.00%±2.72% in the control group and 75.70%±2.42%, 62.79%±2.45%, and 43.41%±2.11%, respectively, in the low-, middle-, and high-concentration experimental groups, and compared with the control group, the experimental groups had a significant reduction in cell viability (all P＜0.05). The number of cell colonies was 923.3±35.2 in the control group and 682.7±24.4, 464.0±22.0, and 327.3±14.0, respectively, in the low-, middle-, and high-concentration experimental groups, and compared with the control group, the experimental groups had a significant reduction in cell colony formation ability (all P＜0.05). After 24 hours of treatment, the relative migration rate was 100.00%±1.11% in the control group and 63.33%±1.16%, 51.72%±3.23%, and 37.18%±2.71%, respectively, in the low-, middle-, and high-concentration experimental groups, and the number of transmembrane cells was 212.0±3.0 in the control group and 134.0±2.0, 71.0±2.0, and 34.0±1.0, respectively, in the low-, middle-, and high-concentration experimental groups; compared with the control group, the experimental groups had significant reductions in relative migration rate and number of transmembrane cells (all P＜0.05). After 48 hours of treatment, compared with the control group, the low-, middle-, and high-concentration experimental groups had a significant reduction in the expression of the anti-apoptotic protein Bcl-2 (all P＜0.05), a significant increase in the expression of the pro-apoptotic protein Bax (all P＜0.05), and a significant reduction in the expression of C＜italic/＞yclinD1 (all P＜005). ConclusionKaempferol can inhibit the proliferation, migration, and invasion of human hepatoma Bel-7402 cells and promote the apoptosis of such cells, possibly by regulating the apoptosis proteins Bax and Bcl-2 and downregulating the expression of CyclinD1.
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ObjectiveTo investigate the protective effect of pinocembrin (PIN) in a mouse model of liver injury induced by acetaminophen (APAP). MethodsA total of 50 healthy male C57BL/6J mice were randomly divided into blank group, PIN (50 mg/kg) group, APAP (300 mg/kg) model group, PIN (30 mg/kg)+APAP (300 mg/kg) experimental group, and PIN (50 mg/kg)+APAP (300 mg/kg) experimental group, with 10 mice in each group. The mice in the blank group and the model group were given an equal volume of normal saline by gavage, and those in the PIN group and the PIN+APAP groups were given PIN by gavage once a day, for 7 consecutive days. At 2 hours after the last administration, the mice in the model group and the PIN+APAP groups were given intraperitoneal injection of APAP 300 mg/kg once, and those in the blank group and the PIN group were given intraperitoneal injection of an equal volume of normal saline. Serum samples were collected to measure the levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST); liver tissue homogenate was prepared to measure the biochemical parameters of malondialdehyde (MDA), superoxide dismutase (SOD), and glutathione (GSH); HE staining was used to observe liver histopathology. A one-way analysis of variance was used for comparison of continuous data between multiple groups, and the least significant difference t-test was used for further comparison between two groups. ResultsCompared with the blank group, the APAP (300 mg/kg) model group had significant increases in the activities of ALT and AST (P＜0.01), suggesting that a model was successfully established, while the PIN (30 mg/kg)+APAP (300 mg/kg) group and the PIN (50 mg/kg)+APAP (300 mg/kg) group had significant reductions in the levels of ALT and AST (P＜0.01). Compared with the blank group, the APAP (300 mg/kg) model group had a significant increase in the level of MDA and significant reductions in SOD activity and GSH level in the liver (P＜001); compared with the APAP (300 mg/kg) model group, the PIN (30 mg/kg)+APAP (300 mg/kg) group and the PIN (50 mg/kg)+APAP (300 mg/kg) group had a significant reduction in the level of MDA and significant increases in SOD activity and GSH level in the liver (P＜0.05). Histopathological observation showed that PIN significantly improved liver injury caused by APAP and helped to maintain normal liver histomorphology. ConclusionPIN exerts a marked protective effect on liver injury induced by APAP, possibly by inhibiting oxidative stress in the liver.
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ObjectiveTo investigate the protective effect of pinocembrin (PIN) in a mouse model of liver injury induced by acetaminophen (APAP). MethodsA total of 50 healthy male C57BL/6J mice were randomly divided into blank group, PIN (50 mg/kg) group, APAP (300 mg/kg) model group, PIN (30 mg/kg)+APAP (300 mg/kg) experimental group, and PIN (50 mg/kg)+APAP (300 mg/kg) experimental group, with 10 mice in each group. The mice in the blank group and the model group were given an equal volume of normal saline by gavage, and those in the PIN group and the PIN+APAP groups were given PIN by gavage once a day, for 7 consecutive days. At 2 hours after the last administration, the mice in the model group and the PIN+APAP groups were given intraperitoneal injection of APAP 300 mg/kg once, and those in the blank group and the PIN group were given intraperitoneal injection of an equal volume of normal saline. Serum samples were collected to measure the levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST); liver tissue homogenate was prepared to measure the biochemical parameters of malondialdehyde (MDA), superoxide dismutase (SOD), and glutathione (GSH); HE staining was used to observe liver histopathology. A one-way analysis of variance was used for comparison of continuous data between multiple groups, and the least significant difference t-test was used for further comparison between two groups. ResultsCompared with the blank group, the APAP (300 mg/kg) model group had significant increases in the activities of ALT and AST (P＜0.01), suggesting that a model was successfully established, while the PIN (30 mg/kg)+APAP (300 mg/kg) group and the PIN (50 mg/kg)+APAP (300 mg/kg) group had significant reductions in the levels of ALT and AST (P＜0.01). Compared with the blank group, the APAP (300 mg/kg) model group had a significant increase in the level of MDA and significant reductions in SOD activity and GSH level in the liver (P＜001); compared with the APAP (300 mg/kg) model group, the PIN (30 mg/kg)+APAP (300 mg/kg) group and the PIN (50 mg/kg)+APAP (300 mg/kg) group had a significant reduction in the level of MDA and significant increases in SOD activity and GSH level in the liver (P＜0.05). Histopathological observation showed that PIN significantly improved liver injury caused by APAP and helped to maintain normal liver histomorphology. ConclusionPIN exerts a marked protective effect on liver injury induced by APAP, possibly by inhibiting oxidative stress in the liver.