Genista cephalantha Spach. protects against acetaminophen-induced liver failure via preserving the glutathione redox system, reducing inflammatory response, and inhibiting hepatocyte death in rats

Abstract

Objectives: The current study was conducted to assess the protective mechanisms of n-BuOH fraction from the aerial part of Genista cephontala (BEGC) on APAP-induced liver injury compared to necrostatine-1 (Nec-1).

Materials and methods: A model of APAP-induced hepatotoxicity was created in male rats by injecting a single dose; 1000 mg/kg APAP, the protective effect was performed with (200 mg/kg; 10 days) BEGC compared to Nec-1, (1.8 mg/kg).

Results: BEGC or NeC-1 pretreatment significantly abolished impaired effects in APAP-rats, by decreasing the generation of TBARS and ROS in mitochondrial and cytosolic fractions and maintaining liver function activities. A marked response was observed in the levels of both GSH and GSH-system enzymes in liver homogenates and mitochondrial fractions to BEGC. BEGC/ Nec-1 successfully regulated the inflammatory mediators (IL-β, TNF-α, HMGB1, and acHMGB1) and MPO levels. During APAP treatment, no caspase-3 or -8 activity was detected, and the level of fk18; M30 was higher than the levels of cck18; M65. Moreover, RIPK3 and MLKL levels were increased in the APAP group. These results suggested that necroptosis predominates during the APAP liver injury model. Interestingly, these necroptotic factors were significantly down-regulated by BEGC treatment. Both biochemical and histopathological findings were consistent with each other.

Conclusion: From all these findings, the hepatoprotective effect of BEGC could be due to the abundance of polyphenols identified by LC-MS/MS analysis, as well as the synergistic interactions of all contents.

Keywords: Acetaminophen; Cell death; Hepatoprotection; Inflammation; LC-MS/MS; Necroptosis; Necrostatin-1; Oxidative stress.

Figure 1

LC-MS/MS profile of n-BuOH extract of Genista cephalantha (BEGC)

Figure 2

Effect of Genista cephalantha (BEGC)(200 mg/Kg) on hepatic markers (AST, ALT, LDH, and γ-GT activities)

Figure 3

A: Effect of Genista cephalantha (BEGC)(200 mg/Kg) on cytosolic and mitochondrial ROS levels (nmol/mg protein)

Figure 4

A, B, C, and D: Effect of Genista cephalantha (BEGC)(200 mg/Kg) on cytosolic glutathione and glutathione-metabolizing enzymes in liver tissues and on mitochondrial glutathione and glutathione-metabolizing enzymes against APAP-induced hepatotoxicity

Figure 5

A: Effect of Genista cephalantha (BEGC)(200 mg/Kg) on plasma caspase-3 and caspase-8 levels (RFU/hr /µl) from rats exposed to APAP, B: Effect of BEGC (200 mg/Kg) on plasma CK 18 levels (M30, M65)(U/L) from rats exposed to APAP

Figure 6

Effect of Genista cephalantha (BEGC)(200 mg/Kg) on necroptosis markers; RIPK3 (µg/mg protein) and MIKL (pg/mg protein) in liver tissues against APAP-induced hepatotoxicity

Figure 7

Effect of Genista cephalantha (BEGC)(200 mg/Kg) on proinflammatory markers TNF-α (pg/mg protein) and IL-β (pg/mg protein) in liver tissues against APAP-induced hepatotoxicity

Figure 8

Effect of Genista cephalantha (BEGC)(200 mg/Kg) on sera levels of HMGB1 (ng/ml) and ac-HMGB1 (ng/ml) from rats exposed to APAP

Figure 9

Effect of Genista cephalantha (BEGC)(200 mg/Kg) on MPO activity in liver tissues against APAP-induced hepatotoxicity

Figure 10

Micrograph of histopathological examination of liver tissues against APAP-induced hepatotoxicity (H&E, 400 xs)