YQFM Alleviates Side Effects Caused by Dasatinib through the ROCK/MLC Pathway in Mice

Abstract

Dasatinib, as a second-generation broad-spectrum tyrosine kinase inhibitor, presents an antitumor effect by inhibiting tyrosine kinases. However, dasatinib causes serious side effects, such as gastrointestinal bleeding and liver toxicity, possibly through the activation of ROCK kinase and MLC phosphorylation. At present, there is no effective prevention and treatment method. Previous research studies have shown that YQFM (YiQiFuMai powder injection) protects the blood-brain barrier by inhibiting the ROCK/MLC signaling pathway; whether YQFM can alleviate the side effects of dasatinib is unknown. In this study, dasatinib was injected (i.p. 70 mg/kg) and YQFM (i.p. 0.336 g/kg, 0.672 g/kg, 1.342 g/kg) was given in advance for 3 days to mice, to explore the effect of YQFM on side effects induced by Dasatinib. The results confirmed that YQFM significantly decreased Evans blue leakage in the small intestine and increased intestinal blood flow, increased the expression of ZO-1, Occludin, and VE-cadherin, and reduced the contents of D-lactic acid, s-VE-cadherin, Alanine aminotransferase (ALT), and Aspartate aminotransferase (AST) in serum. Finally, YQFM inhibited the expression of ROCK-1 and phosphorylation of MLC induced by Dasatinib. These findings suggested that YQFM could improve the side effects caused by Dasatinib linked with the ROCK/MLC signaling pathway, as shown in the graphical abstract.

Figure 1

Effect of YQFM on intestinal blood flow and histopathological changes of intestinal sections in mice with Dasatinib. YQFM (0.336, 0.672, 1.342 g/kg, i.p.) was injected intraperitoneally 1 h before being administered with dasatinib (70 mg/kg for 3 d, i.p.). (a) The representative images of intestinal blood flow in different groups. The magnitude of IBF is represented by different colors, with blue to red denoting low to high. (b) Quantitative analysis of IBF in different groups. Data are expressed as the mean ± SD, n = 6. ^##P < 0.01 vs. control group; ^∗P < 0.05 vs. model group. (c) Hematoxylin-eosin-stained slides of mouse intestinal sections in different groups were examined under a light microscope, and representative stained sections showed that YQFM improves Dasatinib-induced bleeding. Scale bar = 50 μm. n = 3.

Figure 2

Effect of YQFM on Dasatinib-induced intestinal vascular leakage and contents of S-VE-cadherin and D-lactic acid. YQFM (0.336, 0.672, 1.342 g/kg, i.p.) was injected intraperitoneally 1 h before being administered with dasatinib (70 mg/kg for 3 d, i.p.). (a) Representative gross appearance of the EB-stained intestine of a mouse. Data are expressed as the mean ± SD, n = 6. ^##P < 0.01 vs. control group; ^∗P < 0.05 vs. model group. (c) Content of S-VE-cadherin was analysed by ELISA. (d) Content of D-lactic acid was analysed by ELISA. Data are expressed as the mean ± SD. ^##P < 0.01 vs. control group; ^∗P < 0.05 vs. model mice, ^∗∗P < 0.01 vs. model group.

Figure 3

Effect of YQFM on the expression of connexin in mice with Dasatinib. YQFM (0.336, 0.672, 1.342 g/kg, i.p.) was injected intraperitoneally 1 h before being administered with dasatinib (70 mg/kg for 3 d, i.p.). (a–c) Representative western blots and the quantitative analysis of the ratio of ZO-1, occludin, and VE-cadherin. Data are expressed as the mean ± SD, n = 3. ^###P < 0.001 vs. control mice; ^##P < 0.01 vs. control mice; ^#P < 0.05 vs. control mice; ^∗∗∗P < 0.001 vs. model group; ^∗∗P < 0.01 vs. model mice; ^∗P < 0.05 vs. model group.

Figure 4

Effect of YQFM on the contents of ALT and AST in mice with Dasatinib. YQFM (0.336, 0.672, 1.342 g/kg, i.p.) was injected intraperitoneally 1 h before being administered with dasatinib (70 mg/kg for 3 d, i.p.). (a, b) Content of ALT and AST were analysed by using kits. Data are expressed as the mean ± SD, n = 6. ^##P < 0.01 vs. control mice; ^#P < 0.05 vs. control group; ^∗∗P < 0.01 vs. model mice; ^∗P < 0.05 vs. model group.

Figure 5

Effect of YQFM on the ROCK-1/MLC signaling pathway in mice with Dasatinib. YQFM (0.336, 0.672, 1.342 g/kg, i.p.) was injected intraperitoneally 1 h before being administered with dasatinib (70 mg/kg for 3 d, i.p.). (a, b) Representative western blots and the quantitative analysis of the ratio of ROCK-1 and P-MLC. Data are expressed as the mean ± SD, n = 3. ^###P < 0.001 vs. control group; ^∗∗∗P < 0.001 vs. model group.