Objective

2

To observe the effect of Buyang Huanwutang on ferroptosis in diabetic kidney disease （DKD） mice and explore the protective effect and mechanism of Buyang Huanwutang on the kidneys of DKD mice.

Method

2

Seventy-three C57BL/6 mice were randomly divided into model group （63 mice） and normal group （10 mice）. After the model group mice were fed with high-sugar and high-fat diets for six weeks， they were injected with streptozotocin （STZ） of 50 mg·kg

-1

intraperitoneally for five days for preparing the diabetes model and then fed with high-sugar and high-fat diets for eight weeks. When the mice showed positive urine protein， the DKD model was successfully prepared. DKD mice were randomly divided into model group （

n

= 10）， rosiglitazone （7.05 × 10

-4

g·kg

-1

） group （

n

= 9）， Buyang Huanwutang low-dose （3.21 g·kg

-1

） group （

n

= 9）， middle-dose （6.41 g·kg

-1

） group （

n

= 10）， and high-dose （12.82 g·kg

-1

） group （

n

= 10） for gavage. The normal group and model group were given the same volume of normal saline once a day for eight weeks. Fasting blood glucose （FBG）， 24 h urinary protein （24 h-UTP）， and renal weight index （KI） were measured after administration. Hematoxylin-eosin （HE） staining， Periodic acid Schiff （PAS） staining， and Masson staining were used to observe the pathological changes in mouse kidneys. Western blot was used to detect the protein expressions of long-chain acyl-CoA synthetase 4 （ACSL4）， member 11 of solute carrier family 7 （SLC7A11）， glutathione peroxidase 4 （GPX4）， ferritin heavy chain （FTH-1）， and transferrin receptor 1 （TFR-1） in mouse kidneys. The activities of glutathione （GSH） and contents of malondialdehyde （MDA） and 4-hydroxynonenal （4-HNE） were measured. The expression level of reactive oxygen species （ROS） in mouse kidneys was detected by fluorescence probe labeling.

Result

2

Compared with the normal group， KI， FBG， and 24 h-UTP in the model group increased significantly， and mesangium in the glomerulus proliferated. The basement membrane thickened， and glycogen particles were deposited around the glomerulus. FTH-1 expression decreased， while TFR-1 and ROS expressions increased. MDA and 4-HNE increased， but GSH activity decreased. ACSL4 expression increased， and SLC7A11 and GPX4 expressions decreased （

P

<

0.01）. Compared with the model group， in Buyang Huanwutang and rosiglitazone groups， KI and 24 h-UTP decreased， and FBG showed a downward trend， but there was no statistical significance. Pathological damage of kidney tissue was improved to different degrees， FTH-1 expression increased， and TFR-1 and ROS expressions decreased. MDA and 4-HNE contents decreased， and GSH activity increased. ACSL4 expression decreased， and SLC7A11 and GPX4 expressions increased （

P

<

0.05，

P

<

0.01）.

Conclusion

2

Buyang Huanwutang can alleviate the pathological damage of kidney tissue in DKD mice， and its mechanism is related to the regulation of ferroptosis.