Effect of Wenyang Zhuyu Decoction on Expressions of Wnt3a and β-catenin after Acute Cerebral Ischemic Injury in Rats with Kidney-Yang Deficiency Syndrome

HE Hongdun; WEI Liang; SUN Shijie; LI Tian; HU Jiajing; ZHU Meizhen

doi:10.12102/j.issn.1672-1349.2024.14.012

Abstract：Objective:To observe the effect of Wenyang Zhuyu decoction on the expression of wingless MMTV integration site family member 3A(Wnt3a)and β-catenin in the ischemic brain tissue of rats with kidney-Yang deficiency syndrome after acute cerebral ischemia injury,and to explore the mechanism of Wenyang Zhuyu decoction in preventing and treating cerebral ischemic injury in rats with kidney-Yang deficiency syndrome.Methods:One hundred and seventy-six male SD rats were divided into Blank group(44 rats)and kidney-Yang deficiency group(132 rats).After the successful establishment of kidney-Yang deficiency syndrome model,the rats were randomly divided into Sham operation group(Sham group,44 rats),cerebral ischemia model group(MCAO group,44 rats)and Wenyang Zhuyu decoction group(WYZY group,44 rats).WYZY group was gavaged with Wenyang Zhuyu decoction,MCAO group and Sham group were gavaged with equal volume of distilled water.One day after operation,neurobehavioral scores of rats in each group were collected by Zea Longa score criteria.Materials were collected on 3 day,7 day,14 day and 21 day after operation,and 11 rats were selected at each time.The morphological changes of nerve cells were observed by hematoxylin-eosin(HE)staining.The contents of cyclic adenosine monophosphate(cAMP)and cyclic guanosine monophosphate(cGMP)in the plasma were detected by enzyme-linked immunoadsorbent assay(ELISA),and the expressions of Wnt3a and β-catenin in ischemic cerebral tissue were detected by Western Blot.Real-time fluorescence quantitative polymerase chain reaction(RT-PCR)was used to detect the mRNA expressions of Wnt3a and β-catenin mRNA.Results:Compared with the Blank group,cAMP content in the WSYX group decreased(P＜0.05),and cGMP content increased(P＜0.05).The results of HE staining showed that the MCAO group showed disordered cell arrangement,necrosis,disappearance of cell membrane and contraction of nuclei,and showed obvious ischemic injury.The WYZY group showed disordered cell arrangement and partial cell necrosis in the early stage.With the extension of time,the cell damage was alleviated,the cell arrangement was gradually dense,the necrotic cells reduced,and the ischemic state improved significantly.Compared with MCAO group and Sham group,cAMP content increased and cGMP content decreased in the WYZY group(P＜0.05).Compared with MCAO group,the neuroethological scores of WYZY group on 1 day,3 day,7 day,14 day and 21 day were decreased(P＜0.05).Compared with Blank group and Sham group,the expressions of Wnt3a and β-catenin in ischemic brain tissue of MCAO group and WYZY group increased on 3 day,7 day,14 day and 21 day(P＜0.05).Compared with MCAO group,the expressions of Wnt3a and β-catenin in the WYZY group significantly increased(P＜0.05),and Wnt3a and β-catenin in the WYZY group reached their peak value on 3 day,7 day,respectively.Compared with Blank group and Sham group,the mRNA expressions of Wnt3a and β-catenin in MCAO group and WYZY group were increased on 1 day,3 day,7 day,14 day and 21 day(P＜0.05).Compared with MCAO group,the mRNA expressions of Wnt3a and β-catenin in the WYZY group were increased(P＜0.05),and the mRNA expression of Wnt3a reached its peak value on 7 day and the mRNA expression of β-catenin reached its peak value on 14 day.Conclusion:Wenyang Zhuyu decoction could improve the levels of cAMP,cGMP and the pathological state in rats with kidney-Yang deficiency.Wenyang Zhuyu decoction might relieve acute cerebral ischemia and reduce neurological function injury in rats with kidney-Yang deficiency by up-regulating the expressions of Wnt3a and β-catenin.