Drug-induced liver injury (DILI) is considered to be a significant cause of drug wastage. To mitigate clinical DILI risks, assessing drugs using human liver models is crucial since animal studies may fall short due to species-specific liver pathway variations. Cell-based preclinical hepatotoxicity testing is often pertinent. In the present study, cells from a human liver cancer line (HepG2 and HepaRG) were cultured in both formats of 2D and 3D spheroids to explore their responses to drugs. Liver-specific marker expressions across cell lines and culture formats were also examined to assess disparities in DILI marker expressions. After treating each cell with the drugs, cytotoxicity and liver injury markers aspartate aminotransferase and alanine aminotransferase were increased. In addition, liver specific markers albumin and urea decreased in a drug concentration-dependent manner. These findings were consistent with drug sensitivity. Additionally, mRNA expression levels of cytochrome P450 enzymes (CYPs) involved in hepatocellular drug metabolism were compared following treatment with enzyme inducers. CYP1A2 and CYP2C9 were not epxressed in HepG2 cells. HepaRG cells exhibited significantly increased expression of CYP1A2, 2C9, and 3A4 post-treatment. Notably, enzyme expression was notably higher in 3D cultures than in 2D cultures. Collectively, these findings suggest that HepaRG cells and 3D cultures hold promise for evaluating DILI during early-stage drug development.

본 연구는 스트렙토조토신에 의해 유도한 당뇨병, 알코올성 간 손상 실험용 랫드에서 혈액 지질 감소, 당뇨병 및 알코올성 간 손상 예방에 미치는 지장김치 추출물의 경구투여 효과를 조사하였다. 당뇨병 모델동물에서, 혈액 지질 프로파일, ALT, AST 수준은 DC (당뇨병 대조군)와 비교할 때 김치추출물 투여군에서 낮아졌으며 혈당은 DCJK (DC+지장김치 추출물 경구투여군)가 DCCK (DC+일반김치 추출물 경구투여군)에 비해 낮았다. 인슐린 수준은 NC (정상대조군), DCJK > DCCK > DC 순서로 높게 나타났다. 알코올성 간 손상 모델동물에서, 혈액 ALT, AST, 빌리루빈 농도는 AC (알코올 대조군, 일일 소주 1병 섭취군) > ACCK (AC+일반김치 추출물 경구투여군) > ACJK (AC+지장김치 추출물 경 구투여군) > NC 순서로 낮았다. 간 조직의 임상병리학적 소견은 AC에서 가장 심각한 손상을 보였으나 소주1병과 김치 추출물 특히, 지장김치 추출물 경구투여군은 일반김치 추출물 경구투여군에 비해서 회복되는 속도가 개선되었다. 결과는 당뇨병 및 알코올성 간 손상 모델동물에서 지장김치의 섭취가 혈액 지질 프로파일, 혈당, ALT, AST 수준을 낮추고 인슐린 분비능력을 개선하여 줌으로써 항당뇨 및 알코올성 간 손상 보호효과를 갖는다는 사실을 보여준다.

Oxidative stress is one of common cause of fatty changes in the liver. Antioxidant capacity was confirmed in various vegetables including black radish (Raphanus sativus L. var niger). Fermentation of vegetables using Lactobacillus plantarum has been known to generate bioactive components. This study was conducted to determine if fermented black radish (FBR) ameliorates oxidative liver injury induced by CCl4 in rats. To accomplish this, FBR (250 and 500 mg/ kg) was orally administered to rats for 7 consecutive days, single CCl4 (1.5 mL/kg) treatment or no treatment orally. Serum chemistry at 24 hours after CCl4 injury showed that FBR (500 mg/kg) significantly reduced the level of both alanine aminotransferase and aspartate aminotransferase in CCl4 exposed rats. Moreover, FBR treatment significantly increased radical-scavenging effects in livers with the reduction of lipid peroxidation in CCl4 exposed rats. Histopathologic findings including Kupffer cell activation in the liver of each group matched those of serum chemistry. Collectively, black radish, through fermentation, exerts hepatoprotective capacity in CCl4 induced liver injury in rats through anti-oxidation.

Lentinus tigrinus (L. tigrinus), a white-rot fungus that grows naturally on rotten hardwood during spring and summer in China, is an edible and medicinal mushroom containing a valuable combination of nutrients including high amino acid concentrations and low sugar levels. However, no reports have isolated and characterized FIP genes from L. tigrinus to date. In our study, two novel fungal immunomodulatory proteins (FIPs) from Lentinus tigrinus were identified and named Fip-lti1 and Fip-lti2. The bioactive characteristics of Fip-lti1 and Fip-lti2 were compared to a well-known FIP (LZ-8 from Ganoderma lucidum) to investigate the effect of Fip-lti1 and Fip-lti2 expression on Concanavalin A (ConA)-induced liver injury. Both Fip-lti1 and Fip-lti2 protected livers from ConA-induced necrosis, as evidenced by decreased serum aminotransferase levels (AST, ALT) and relieved liver histology. Levels of proinflammatory cytokines (TNF-α, IL-1β, and IL-6) and oxidative stress (SOD, MDA) were shown to be reduced by expressing Fip-lti1 and Fip-lti2. In addition, the hepatoprotective effect of Fip-lti1, Fip-lti2, and LZ-8 correlated with ameliorating the imbalance of Th1/Th2 (IFN-γ/IL-4). The observed liver protection of Fip-lti1 and Fip-lti2 was mechanistically explored. Treatments with Fip-lti1 and Fip-lti2 regulated GATA3/T-bet expression, activated the decreased Nrf-2/HO-1 pathway, and countered the upregulated NLRP3/ASC/NF-κBp65 signaling in ConA-stimulated liver injury. In conclusion, we identified two fungal proteins (Fip-lti1 and Fip-lti2) that can protect liver from ConA-induced liver injury.

Lipopolysaccharide (LPS) is the major of outer membrane of gram-negative bacteria and one of the most potent microbial initiators of inflammation. From the previous study showed that exposure to a low dose of LPS renders animals tolerant to a lethal dose of LPS, and protects against the toxicity of various chemicals. However, the effects of LPS treatment in thioacetamide (TA) - induced liver injury remain largely unknown. Liver injury caused by various toxic chemicals such as carbon tetrachloride, alcohol, dimethylnitrosamine. Here, we induced rat liver injury by intraperitoneal injection of TA, a representative hepatic fibrosis inducer. In this study, we investigated the effects of LPS in TA group, LPS group, LPS/TA group and vehicle control group on Sprague-Dawely rats (five rats for each group). All rats at the end of the experiment were sacrificed, and liver and serum were obtained. Serological analysis and hematoxylin and eosin staining showed that LPS/TA co-treatment was associated with decrease of aspartate aminotransferase (AST), alanine transaminase (ALT) and totalbilirubin and fibrosis than in TA-treated rats. RT-PCR showed that the levels of IL-6 and Cox2 mRNA were lower in the liver of LPS/TA-cotreated rats than in TA-treated rats. There were no significant differences ALT, ALP, AST, total-bilirubin, IL-6 and Cox2 between vehicle control and LPS-treated rats. These results imply that LPS/TA cotreatment partially alleviates the TA-induced liver injury of rats by reducing inflammatory response.

This study was performed to evaluate the biological activity and protective effect of Cassia tora ethanol extracts against D-galactosamine induced hepatotoxicity in rats. Male Sprague-Dawley rats were grouped into normal group, D-galactosamine treated group(control), D-galactosamine plus 0.25% Cassia tora extracts treated group and D-galactosamine plus 0.5% Cassia tora extracts treated group. Normal and control group were fed control diet and Cassia tora extracts treated groups were fed experimental diets containing 0.25% or 0.5% Cassia tora ethanol extracts for 5 weeks. Body weight gain and liver weight of rats were not significantly different between groups. Cholesterol and triglyceride concentrations in serum and liver were significantly lower in rats treated only with D-galactosamine compared to normal group, and improved in Cassia tora extracts supplemented rats. D-galactosamine treatment significantly increased serum aspartate transaminase, alanine transaminase, gamma glutamyl transferase and alkaline phosphatase, however, the activities of aspartate transaminase and alanine transaminase were significantly decreased in Cassia tora extracts supplemented rats when compared with D-galactosamine treated control group. Cassia tora extracts significantly suppressed the D-galactosamine-induced elevation of liver thiobarbituric acid reactive substances(TBARS) contents. Superoxide dismutase activity was decreased by D-galactosamine treatment, however by the supplementation of Cassia tora ethanol extracts, significantly increased in dose-dependent manner. Glutathione peroxidase activity in rats fed diets containing Cassia tora extracts was decreased compared to control. Based on these results, we concluded that Cassia tora ethanol extracts may prevents the D-galactosamine-induced hepatotoxicity probably via an antioxidant mechanism.

발효에 의한 헛개열매의 기능성 상승 정도를 검토하고자, 헛개열매열수추출물을 발효시킨 후 급성 및 만성 알코올 투여 간손상 동물모델을 통하여 체중감량 억제, 알코올 분해 및 간기능 개선 효능을 검증하였다. 급성 알코올 투여 동물모델에서 헛개열매발효군(ET-FHWE)은 알코올대조군(ET)에 비하여 혈청 알코올 농도가 유의적으로 감소되었고, 특히 알코올 투여 3시간 후의 알코올 농도는 헛개열매추출액발효물에 의해 46.1%, 헛개열매열수추출물에 의해 19.1% 감소된 것으로 나타났다. 또한 알코올 투여에 의해 증가된 혈청 아세트알데하이드 농도는 헛개열매추출액발효물에 의해 알코올 투여 3시간 후에는 48.7%, 5시간 후에는 39%로 알코올대조군(ET)보다 유의적으로 감소하였고, 이는 헛개열매열수추출물은 발효에 의해 알코올 및 아세트알데하이드 분해능이 증가하는 것으로 사료되었다. 만성 알코올 투여 간손상 동물모델 실험에서 알코올 투여에 의해 상승된 혈청 알코올 농도는 헛개열매열수추출물과 헛개열매추출액발효물 투여에 의해 각각 31%, 41% 유의적으로 감소하였다. 혈청 아세트알데하이드 농도와 γ-GTP 활성도는 헛개열매열수추출물과 헛개열매추출액발효물 투여에 의해 알코올대조군(ET)보다 유의적으로 감소되었으며, 장기간 알코올 투여에 의한 체중 감소 억제 및 간조직 지질수준의 유의적 감소를 나타내었다. 또한 헛개열매추출액발효물은 장기간의 알코올 투여로 인해 감소된 혈당을 유의적으로 증가시키는 것으로 나타났다. 본 연구 결과, 급성 알코올 투여 동물모델에서 헛개열매열수추출물은 발효에 의해 알코올 및 아세트알데하이드 분해능이 증진되었고, 만성알코올 투여 모델을 통한 실험에서는 발효에 의해 헛개열매의 간기능 개선효능이 유지됨과 동시에 일부 효능(혈청 지질 및 혈당 수준 개선능)은 증가하는 것으로 나타났다. 따라서 헛개열매추출액발효물은 급성 및 만성 알코올성 간손상 억제에 있어서 헛개열매열수추출물보다 더욱 강력한 기능성 물질로 활용될 수 있을 것으로 기대된다.

시로미 열매 추출물의 라디칼 소거 활성과 간 손상 동물모델에서의 항산화 효소 활성 측정을 통해 추출물의 항산화 효과를 평가하였다. In vitro 실험에서, 시로미 열매 추출물은 12.0 ug/ml의 농도에서 DPPH 라디칼을 50% 소거하는 활성을 보였고, 1 mg/ml의 농도에서 56.1%의 지질과산화 억제 활성을 나타내었다. 그리고 CCl4로 유도한 간 손상 동물모델에서 serum ALT 및 AST의 수치를 감소시켜 간보호 효능을 나타내었고, 간 조직의 지질 과산화를 최대 73% 억제하였으며, 항산화 효소인 SOD 및 catalase의 활성을 유의적으로 증가시켰다.

We previously reported that purified hepatocyte-like cells derived from human embryonic stem cell (hESC) promoted the liver tissue recovery not only by cell replacement, but also by delivering proteins (secretome) that enhance endogenous host liver regeneration. In this study, we investigated possible therapeutic effects of secretomes obtained from undifferentiated hESC and mesenchymal stem cell (hMSC), and explored the underlying mechanism in a mouse model of chronic liver injury. Mice pre-intoxicated with dimethylnitrosamine (DMN) were treated with single intraperitoneal injection of secretome or medium used to support the growth of hESCs or hMSCs. Both hESC- and MSC-secretomes induced robust host liver regeneration, as determined by biochemical and histological analyses. The expression of MMP2 was significantly increased in the liver that received hESC- or hMSC-secretome, compared to control groups. In contrast, expression of α-SMA, a hallmark of activated hepatic stellate cells, was profoundly decreased after administration of both secretomes. These results suggest that hESCs and MSCs may release soluble factors that support the host tissue regeneration of chronically injured liver.