清补通络方改善2型糖尿病小鼠认知障碍的机制研究OA

Objective To investigate the improving effect of the Qingbu Tongluo Formula on cognitive impairment in type 2 diabetic mice and to explore its mechanism of action in reducing amyloid-beta(Aβ)deposition.Methods Seventy C57BL/6J were randomly divided into a normal control group(NC,n=10)and a modeling group(n=60).Mice in the model group were fasted overnight and then injected with STZ(50 mg/kg)intraperitoneally for 3 consecutive days to establish a type 2 diabetes model.Fifty successfully modeled mice were selected to maintain on the high-fat diet and then were subjected to induce cognitive impairment.After the successful modeling,fifty mice were divided into the following five groups:model group(MG),low-dose Qingbu Tongluo Formula group(Q-L group),medium-dose Qingbu Tongluo Formula group(Q-M group),high-dose Qingbu Tongluo Formula group(Q-H group),and donepezil hydrochloride group(DN group),with 10 mice in each group.NC group received standard chow and was gavaged with normal saline;MG group received high fat diet and was gavaged with normal saline;Q-L,Q-M,and Q-M groups were gavaged with Qingbu Tongluo Formula at doses of 15.66,31.32,and 62.64 g/kg respectively;DN group was gavaged with donepezil hydrochloride at dose of 0.65 mg/kg.All administrations were performed once daily and continued for 8 consecutive weeks.The binding affinity between Qingbu Tongluo Formula and transforming growth factor-beta 1(TGF-β1)was analyzed using a surface plasmon resonance(SPR)-based biomolecular interaction analysis system(Biacore T200).After 8 weeks of intervention,the Morris water maze test was conducted.The mRNA expression level of TGF-β1 in brain tissue was detected via RT-qPCR.Pathological changes in the brain tissue were observed through HE staining.The levels of Aβ1-40 and Aβ1-42 in serum and brain tissue were measured by ELISA.The expression of TGF-β1,IDE,and Aβ protein in the hippocampal tissue were examined using immunohistochemistry.Results Biacore analysis demonstrated a strong binding affinity between the Qingbu Tongluo Formula and TGF-β1.Compared with the model group,mice in the drug intervention groups exhibited shortened escape latency,a decreased number of platform crossings,and reduced time spent in the target quadrant.The morphology of brain tissue was found to be more intact.TGF-β1 mRNA expression was reduced,levels of Aβ1-42 and Aβ1-40 were significantly decreased,protein expression of TGF-β1 and Aβ was downregulated,and IDE expression was upregulated(P<0.05).Conclusion The Qingbu Tongluo Formula can improve cognitive impairment in type 2 diabetic mice by regulating TGF-β1 expression,which leads to reduced Aβ deposition.