BuShenKangShuai Tablet Alleviates Hepatic Steatosis via Improving Liver Adiponectin Resistance in ApoE-/- Mice

Abstract

BuShenKangShuai tablet (BSKS) is a Chinese herbal compound, which has been used to treat nonalcoholic fatty liver disease and cardiovascular diseases in clinic for over four decades. This study intends to explore whether BSKS administration can alleviates hepatic steatosis via improving liver adiponectin resistance in ApoE^-/- mice. ApoE^-/- mice were fed with western-type diet for 6 weeks and then were administrated with BSKS or atorvastatin for 6 weeks by gavage, and then blood and liver were collected for analysis. The results showed that BSKS attenuated hepatic steatosis, decreased blood lipids, and increased the serum level of adiponectin. We also found that adiponectin resistance in the liver was improved by BSKS, while the expression of TLR4 and NF-κB p65 was inhibited, followed by the suppression of proinflammatory mediators of TNF-α. Our data provided evidence that BSKS was able to alleviate hepatic steatosis in vivo. The underlying mechanism of BSKS was focused on improving liver adiponectin resistance, thereby regulating dyslipidemia and inhibiting inflammatory signaling pathway.

Figure 1

Effect of BuShenKangShuai tablet (BSKS) on the hepatic steatosis in ApoE−/− mice. Representative photomicrographs of Hematoxylin and Eosin staining of liver. The area of liver tissue was measured by Image-Pro Plus 6.0 (magnification, ×400). (a) No significant hepatic steatosis could be found in control group. (b) Hepatocyte nuclear ballooning, hepatocyte apoptosis, Mallory's hyaline, and inflammation foci were the main types of hepatic histopathological changes in model group. (c) The above histopathological changes were significantly improved in BSKS group. (d) Atorvastatin could also significantly alleviate the above histopathological changes. Except for this, the hepatic cells were arranged more closely in atorvastatin group. Scale bars 50 μm.

Figure 2

Effect of BSKS on blood lipids. The serum TG, TC, LDL-C, and HDL-C of mice were detected and compared between the control group and model group, model group and drug treatment groups (n = 10). Data are shown as mean ± S.D and compared by one-way analysis of variance (ANOVA) followed by Fisher's Least Significant Difference (LSD) test for individual comparisons. ^n.s.P > 0.05; ^⋆P < 0.05.

Figure 3

Effect of BSKS on the serum level of adiponectin and the expression of adiponectin and its receptors in the liver tissue. (a) The serum level of adiponectin was detected by ELISA (n = 10). (b) The expressions of adiponectin, AdipoR1, and AdipoR2 in the liver were detected by western blot (n = 3). The results were quantitatively compared between the control group and model group, model group and drug treatment groups. Data are shown as mean ± S.D and compared by ANOVA followed by LSD test for individual comparisons. ^n.s.P > 0.05; ^⋆P < 0.05.

Figure 4

Effect of BSKS on TLR4 mediated signaling pathway, including TLR4, NF-κB p65, and TNF-α, which were detected by immunohistochemical staining. The percentage of immunohistochemical staining area was quantitatively analyzed by using Image-Pro Plus 6.0 (magnification, ×400). The results were quantitatively compared between the control group and model group, model group and drug treatment groups (n = 10). TLR4 mediated signaling pathway was activated in model group. BSKS and atorvastatin could inhibit TLR4 mediated signaling pathway. Data are shown as mean ± S.D and compared by ANOVA followed by LSD test for individual comparisons. ^n.s.P > 0.05; ^⋆P < 0.05. Scale bars 50 μm.

Figure 5

Effect of BSKS on TLR4 mediated signaling pathway, including TLR4, NF-κB p65, and TNF-α. The results were quantitatively compared between the control group and model group, model group and drug treatment groups (n = 3). The western blot results were coincidentally consistent with the immunohistochemical staining results. Data are shown as mean ± S.D and compared by ANOVA followed by LSD test for individual comparisons. ^n.s.P > 0.05; ^⋆P < 0.05.