Yinchenhao decoction attenuates obstructive jaundice-induced liver injury and hepatocyte apoptosis by suppressing protein kinase RNA-like endoplasmic reticulum kinase-induced pathway

Figure 1 Yinchenhao decoction improves the obstructive jaundice induced histopathological changes in liver tissue. The liver tissue in group B showed an unclear texture, and the hepatic lobule structures were disordered. Moreover, cholestasis and fatty degeneration were observed. Group A: Control group; Group B: Obstructive jaundice model group; Group C: Yinchenhao decoction-treated group.

Figure 2 Yinchenhao decoction relieves the hepatic injury induced by obstructive jaundice. A: Total bilirubin levels in the serum; B: Direct bilirubin levels in the serum; C: Alanine aminotransferase levels in the serum; D: Aspartate transaminase levels in the serum; E: Gamma-glutamyl transpeptidase levels in the serum; F: Alkaline phosphatase levels in the serum. Group A: Control group; Group B: Obstructive jaundice model group; Group C: Yinchenhao decoction;-treated group. ^aP < 0.05, ^bP < 0.01, ^cP < 0.001. TBIL: Total bilirubin; DBIL: Direct bilirubin; ALT: Alanine aminotransferase; AST: Aspartate transaminase; GGT: Gamma-glutamyl transpeptidase; ALP: Alkaline phosphatase.

Figure 3 Yinchanhao decoction regulates the protein kinase RNA-like endoplasmic reticulum kinase-induced apoptotic signaling pathway in rat liver tissue. A: Representative Western blot images of protein kinase RNA-like endoplasmic reticulum kinase (PERK), CCAAT/enhancer-binding protein homologous protein (CHOP), growth arrest and DNA damage-inducible protein 34 (GADD34), B cell lymphoma/leukemia-2 gene related X protein (Bax), and B cell lymphoma/leukemia-2 gene (Bcl-2) proteins; B: The protein expression levels of PERK, CHOP, GADD34, Bax, and Bcl-2; C: The gene expression levels of PERK, CHOP, GADD34, Bax, and Bcl-2. The mRNA levels were normalized to β-actin expression. Group A: Control group; Group B: Obstructive jaundice model group; Group C: Yinchenhao decoction-treated group. ^aP < 0.05, ^bP < 0.01, ^cP < 0.001. PERK: Protein kinase RNA (PKR)-like endoplasmic reticulum kinase; CHOP: CCAAT/enhancer-binding protein homologous protein; GADD34: Growth arrest and DNA damage-inducible protein 34; Bcl-2: B cell lymphoma/leukemia-2 gene; Bax: Bcl-2 related X protein.

Figure 4 Yinchenhao decoction alleviates BRL-3A rat hepatocyte injury. A: Alanine aminotransferase levels in cell culture medium; B: Aspartate transaminase levels in cell culture medium. Group S: BCL-3A rat hepatocytes incubated in rat serum of the G1 group; group O: BCL-3A rat hepatocytes incubated in rat serum of the G2 group; group M: BCL-3A rat hepatocytes incubated in rat serum of the G2 + G3 group. ^aP < 0.05, ^bP < 0.01, ^cP < 0.001. ALT: Alanine aminotransferase; AST: Aspartate transaminase.

Figure 5 Yinchenhao decoction inhibits BRL-3A rat hepatocyte apoptosis. A: TUNEL results of each group; B: Apoptosis index of each group at 6 h, 24 h, and 48 h. Group S: BCL-3A rat hepatocytes incubated in rat serum of the G1 group; group O: BCL-3A rat hepatocytes incubated in rat serum of the G2 group; group M: BCL-3A rat hepatocytes incubated in rat serum of the G2 + G3 group. ^aP < 0.05, ^bP < 0.01, ^cP < 0.001.

Figure 6 Yinchanhao decoction regulates the protein kinase RNA-like endoplasmic reticulum kinase-induced apoptotic signaling pathway in BRL-3A rat hepatocytes. A: Representative Western blots of protein kinase RNA-like endoplasmic reticulum kinase, CCAAT/enhancer-binding protein homologous protein, growth arrest and DNA damage-inducible protein 34, B cell lymphoma/leukemia-2 gene related X protein, and B cell lymphoma/leukemia-2 gene proteins at 6 h, 24 h, and 48 h; B: Expression levels of the above proteins at 6 h; C: Expression levels of the corresponding genes at 6 h; D: Expression levels of the above proteins at 24 h; E: Expression levels of the corresponding genes at 24 h; F: Expression levels of the above proteins at 48 h; G: Expression levels of the corresponding genes at 48 h. The mRNA levels were normalized to GAPDH expression. Group S: BCL-3A rat hepatocytes incubated in rat serum of the G1 group; group O: BCL-3A rat hepatocytes incubated in rat serum of the G2 group; group M: BCL-3A rat hepatocytes incubated in rat serum of the G2 + G3 group. ^aP < 0.05, ^bP < 0.01, ^cP < 0.001. GADPH: Glyceraldehyde 3-phosphate dehydrogenase; PERK: Protein kinase RNA (PKR)-like endoplasmic reticulum kinase; CHOP: CCAAT/enhancer-binding protein homologous protein; GADD34: Growth arrest and DNA damage-inducible protein 34; Bcl-2: B cell lymphoma/leukemia-2 gene; Bax: B cell lymphoma/leukemia-2 gene related X protein.