Professor Bi Yunfeng of Jilin Agricultural University, et al.: Protective effect of burdockinogenin on tetraoxamine-induced liver injury in diabetic mice

According to the statistics of clinical data in 2019, the number of people suffering from diabetes mellitus (DM) in China is about 116 million, accounting for 12.
8% of the total population, with the improvement of people's living standards and the increase of unhealthy eating habits, DM patients have shown a trend
of early disease and an increase in the number of patients at a young age.
DM patients' blood sugar is at a high level for a long time, which can cause various complications, such as liver, kidney, blood vessels and heart-related diseases, among which the rate of liver lesions is as high as 21%~78%.

At present, the treatment of patients with DM type liver disease is mainly lifelong medication, which cannot achieve radical cure, and long-term drug use will produce a variety of toxic side effects, resulting in a decrease
in body weight.
Therefore, the search for natural active ingredients with high efficiency and low toxicity to protect DM liver damage has become the focus
of attention.

Gao Fei, Tang Jinxin, Bi Yunfeng* and others from the College of Food Science and Engineering of Jilin Agricultural University studied the protective effect of arctilin glycoside (ATG) on tetraoxamine-induced liver injury in DM mice, aiming to provide scientific basis
for the prevention and treatment of DM-type liver injury and the development of ATG-related health care products.

1.
Analysis of modeling situation

The intraperitoneal dose of tetraoxazine was 180 mg/kg mb, and the second half-dose modeling (90 mg/kg mb) was performed on mice whose first modeling was unsuccessful, with a model success rate of 62%.

After modeling, 6 mice died, and the causes of death were low immunity of mice, discomfort caused by DM and mutual bites of mice, etc.
, which had nothing to do with
gavage ATG.
The symptoms of DM in mice were obvious, manifested as rough and messy body hair, walking on the ground, not moving well, slow response, wet litter, more feces and increased dietary water intake
.

2.
Analysis results of routine indicators of mice

It can be seen from Figure 1 that compared with the control group, the blood glucose concentration of mice in the model group was significantly increased (P<0.
01), all of which exceeded 11.
1 mmol/L, combined with the physiological conditions of 2.
1 mice to prove that the DM mouse model of this experiment was successfully<b10> established.
At 28 d, compared with the model group, the blood glucose concentration of mice in the Met group and the ATG high-dose group decreased significantly (P<0.
01), and the blood glucose concentration decreased by 30.
75% and 26.
12%,<b11> respectively, compared with 0 d.

It can be seen from Table 1 that compared with 0 d, the body mass of mice in each group increased to different degrees at 28 d, and the body mass of mice in the control group, Met group and ATG high-dose group increased significantly (P<0.
01), which increased by 20.
22%, 13.
47% and 11.
03%, respectively.
The growth of body mass of mice in the model group was not significant (P>0.
05).

At 0~7 d, some mice were disturbed by metabolism and negative growth in body weight after injection of tetraoxamine, while different dose groups of Met and ATG could improve the decline of body mass of DM mice
.

As shown in Table 2, compared with the model group, the effect of ATG on liver index was very significant in the high- and medium-dose groups (P<0.
01), while the effect in the low-dose group was not significant (P>0.
05).

In summary, ATG can improve the loss of body weight and liver fat accumulation in DM mice, and the treatment effect of ATG high-dose group is comparable
to that of Met group.

3.
Serum index analysis results

It can be seen from Figure 2 that compared with the control group, ALT activity and AST activity in the model group were significantly increased (P<0.
01), indicating that the liver organs of DM mice were damaged<b10>.
Compared with the model group, the ALT activity and AST activity of the Met group and the ATG high-dose group were significantly reduced (P<0.
01), while the AST activity of the ATG medium-dose group was significantly reduced (P<0.
05), and there was no significant change in ALT activity (P>0.
05).

The above results showed that ATG treatment could improve ALT activity and AST activity
in the serum of DM mice.

The effect of ATG on the mass concentrations of liver inflammatory factors IL-6 and TNF-α in DM mice is shown in Table 3, and compared with the control group, the mass concentrations of IL-6 and TNF-α in the model group were significantly increased (P<0.
01), indicating that inflammatory changes<b10> had appeared in DM mice in this experiment.
Compared with the model group, the mass concentrations of IL-6 and TNF-α in all dose groups of Met and ATG were significantly reduced (P<0.
01, P<0.
05).
<b11> The above results indicate that ATG gavage can reduce the inflammatory response
in the liver of DM mice.

4.
Analysis results of liver homogenization related indicators

As shown in Figure 3, compared with the control group, GSH content, CAT activity and SOD activity in the model group decreased significantly (P<0.
01).
<b10> Compared with the model group, GSH content, CAT activity and SOD activity were increased to different degrees in the ATG high-dose group (P<0.
01, P<0.
05), and the improvement effect on CAT activity was comparable to that in the positive Met group, and CAT activity and SOD activity in the ATG medium-dose group were significantly increased (P<0.
05).
<b11> The above results showed that ATG had an effect on improving hepatic stress in DM mice, and could make GSH content, CAT activity and SOD activity close to normal, thereby improving the level of liver oxidative stress in
DM mice.

5.
Observation results of liver histopathological section

As shown in Figure 4, the liver tissue morphology of the control group was complete and clear, there was no steatosis, the tissue cells were stained with normal red, the cells were uniform, the internal serous fluid was clearly visible, and the middle blue was the nucleus; The liver of mice in the model group showed abnormality, the arrangement was obviously loose and irregular, and the vacuole morphology was displayed in the cells, and there was bleeding, indicating that the fat had been denatured.
High and medium doses of ATG and Met have a certain effect on liver tissue damage, the area of intracellular redness increases, and the cell vacuole and bleeding area decreases.
The morphology of liver slice regions in the ATG low-dose group was basically no different from that in the model group, and there was no significant improvement effect
.

6.
Protein immunoassay results

Compared with the control group, the relative expression of TLR4, MyD88 and NF-κB p65 protein in the model group was significantly increased (P<0.
01), and the ATG high-dose group and Met group had similar therapeutic effects, which could significantly reduce the relative expression of TLR4, MyD88 and NF-κB p65 protein (P<0.
01, P<0.
05); the intervention effect in the dose group of ATG was not significant (P>0.
05), and the relative expression of MyD88 and NF-κB p65 proteins in the ATG low-dose group was significantly reduced (P<0.
01, P<0.
05), but there was no significant reduction in the expression of TLR4 protein (P>0.
05).

The above results showed that ATG had a certain therapeutic effect on liver injury in DM mice, and was closely related
to TLR4, MyD88 and NF-κB p65 signaling pathways.

Conclusion