Inhibitory effect of Patrinia on BRL-3A cell apoptosis through the TLR4/PI3K/AKT/GSK3β and TLR4/P38/JNK signaling pathways

Chen,Xiaoli; Yan,Xiuping; Guo,Lu

doi:10.3892/mmr.2018.8466

Inhibitory effect of Patrinia on BRL-3A cell apoptosis through the TLR4/PI3K/AKT/GSK3β and TLR4/P38/JNK signaling pathways

Authors:
- Xiaoli Chen
- Xiuping Yan
- Lu Guo
View Affiliations

Affiliations: Hepatopathy Department, Sixth People's Hospital of Qingdao, Qingdao, Shandong 266033, P.R. China
Published online on: January 22, 2018 https://doi.org/10.3892/mmr.2018.8466
Pages: 5344-5349

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

The present study investigated the inhibitory effect of Patrinia on lipopolysaccharide (LPS)-induced apoptosis of rat liver BRL‑3A cells. A Cell Counting Kit‑8 assay was performed to measure the effect of Patrinia on BLR‑3A cell activities. A biochemical assay was employed to detect the release of lactate dehydrogenase (LDH) in BRL‑3A cells induced by different doses of LPS. Based on the release rate of LDH, drug concentrations were set at 0.5, 1 and 2 g/l. Apoptotic morphology of cells was observed via Hoechst 33342 staining and flow cytometry was performed to detect apoptosis rates. Western blotting was performed to detect the expression of toll‑like receptor 4 (TLR4), protein kinase B (AKT), phosphorylated (P)‑AKTSer473, glycogen synthase kinase 3β (GSK3β), P‑GSK3βSer9, P38, P‑P38, c‑Jun N‑terminal kinase (JNK), P‑JNK, B‑cell lymphoma‑2 (Bcl‑2), Bcl‑2 associated X protein (Bax) and active‑caspase‑3 proteins. The translocation of GSK3β was observed by immunofluorescence staining. Results revealed that Patrinia increases cell activities and inhibits apoptosis. The expression levels of TLR4, P‑P38 and P‑JNK were reduced, whereas the expression of P‑AKTSer473 and P‑GSK3βSer9 were increased. Patrinia significantly reduced GSK3β nuclear translocation induced by LPS, and significantly decreased the mRNA expression levels of Bax/Bcl‑2 and caspase‑3 in BRL‑3A cells induced by LPS. In conclusion, Patrinia may significantly reduce apoptosis of BRL‑3A induced by LPS via the TLR4/PI3K/AKT/GSK3β and TLR4/P38/JNK signaling pathways, providing evidence for its potential use in liver disease therapy.

View Figures

View References

1	Lopez M, Kopec AK, Joshi N, Geddings JE, Cline H, Towery KL, Rockwell CE and Mackman N: Fas-induced apoptosis increases hepatocyte tissue factor procoagulant activity in vitro and in vivo. Toxicol Sci. 141:453–464. 2014. View Article : Google Scholar : PubMed/NCBI
2	Jiang X and Wang X: Cytochrome c promotes caspase-9 activation by inducing nucleotide binding to Apaf-1. J Biol Chem. 275:31199–31203. 2000. View Article : Google Scholar : PubMed/NCBI
3	Tait SW and Green DR: Mitochondria and cell death: Outer membrane permeabilization and beyond. Nat Rev Mol Cell Biol. 11:621–632. 2010. View Article : Google Scholar : PubMed/NCBI
4	Fulda S: Modulation of mitochondrial apoptosis by PI3K inhibitors. Mitochondrion. 13:195–198. 2013. View Article : Google Scholar : PubMed/NCBI
5	Johnson-Farley NN, Patel K, Kim D and Cowen DS: Interaction of FGF-2 with IGF-1 and BDNF in stimulating Akt, ERK, and neuronal survival in hippocampal cultures. Brain Res. 1154:40–49. 2007. View Article : Google Scholar : PubMed/NCBI
6	Tournier C, Hess P, Yang DD, Xu J, Turner TK, Nimnual A, Bar-Sagi D, Jones SN, Flavell RA and Davis RJ: Requirement of JNK for stress-induced activation of the cytochrome c-mediated death pathway. Science. 288:870–874. 2000. View Article : Google Scholar : PubMed/NCBI
7	Zhang YF: The research progress of Patrinia. J Chin Med Mat. 1:148–152. 2009.(In Chinese).
8	Du C, Fang M, Li Y, Li L and Wang X: Smac, a mitochondrial protein that promotes cytochrome c-dependent caspase activation by eliminating IAP inhibition. Cell. 102:33–42. 2000. View Article : Google Scholar : PubMed/NCBI
9	Mohammadi A, Tajik N, Shah-Hosseini A, Alavian SM, Sharifi Z and Jarahi L: FAS and FAS-ligand promoter polymorphisms in hepatitis B virus infection. Hepat Mon. 15:e264902015. View Article : Google Scholar : PubMed/NCBI
10	Muzio M, Chinnaiyan AM, Kischkel FC, O'Rourke K, Shevchenko A, Ni J, Scaffidi C, Bretz JD, Zhang M, Gentz R, et al: FLICE, a novel FADD-homologous ICE/CED-3-like protease, is recruited to the CD95 (Fas/APO-1) death-inducing signaling complex. Cell. 85:817–827. 1996. View Article : Google Scholar : PubMed/NCBI
11	Flanagan L, Sebastià J, Tuffy LP, Spring A, Lichawska A, Devocelle M, Prehn JH and Rehm M: XIAP impairs Smac release from the mitochondria during apoptosis. Cell Death Dis. 1:e492010. View Article : Google Scholar : PubMed/NCBI
12	Chen HC, Kanai M, Inoue-Yamauchi A, Tu HC, Huang Y, Ren D, Kim H, Takeda S, Reyna DE, Chan PM, et al: An interconnected hierarchical model of cell death regulation by the BCL-2 family. Nat Cell Biol. 17:1270–1281. 2015. View Article : Google Scholar : PubMed/NCBI
13	Gross A: BCL-2 family proteins as regulators of mitochondria metabolism. Biochim Biophys Acta. 1857:1243–1246. 2016. View Article : Google Scholar : PubMed/NCBI
14	Martinou JC and Youle RJ: Mitochondria in apoptosis: Bcl-2 family members and mitochondrial dynamics. Dev Cell. 21:92–101. 2011. View Article : Google Scholar : PubMed/NCBI
15	Ha H, Han D and Choi Y: TRAF-mediated TNFR-family signaling. Curr Protoc Immunol Chapter. 11:Unit11.9D. 2009. View Article : Google Scholar
16	Weichhaus M, Chung ST and Connelly L: Osteoprotegerin in breast cancer: Beyond bone remodeling. Mol Cancer. 14:1172015. View Article : Google Scholar : PubMed/NCBI
17	Guicciardi ME, Malhi H, Mott JL and Gores GJ: Apoptosis and necrosis in the liver. Compr Physiol. 3:977–1010. 2013.PubMed/NCBI
18	Yarza R, Vela S, Solas M and Ramirez MJ: c-Jun N-terminal kinase (JNK) signaling as a therapeutic target for alzheimer's disease. Front Pharmacol. 6:3212016. View Article : Google Scholar : PubMed/NCBI
19	Kim YR, Kang TW, To PK, Xuan Nguyen NT, Cho YS, Jung C and Kim MS: HOXB13-mediated suppression of p21WAF1/CIP1 regulates JNK/c-Jun signaling in prostate cancer cells. Oncol Rep. 35:2011–2016. 2016. View Article : Google Scholar : PubMed/NCBI
20	Hengartner MO: The biochemistry of apoptosis. Nature. 407:770–776. 2000. View Article : Google Scholar : PubMed/NCBI
21	Ma M, Wang L, Ma Y, Yang Y, Chen B and Zhu X: Effects of norepinephrine on proliferation and apoptosis of neonatal cardiac fibroblasts in rats. Zhonghua xin xue guan bing za zhi. 43:542–547. 2015.(In Chinese). PubMed/NCBI