Material Basis Of Huangpu Tongqiao Capsule And Its Effect On EGFR-PLC? Signal Pathway In AD Rats

Objective: In this study,the Chinese medicine compound Huangpu Tongqiao Capsule was taken as the research object to clarify the material basis of Huangpu Tongqiao Capsule in vitro.At the same time,the neuroprotective effects of Huangpu Tongqiao Capsules on AD animal models and their effects on the EGFR-PLC? signal pathway were explored,with a view to providing a certain reference for the modernization and internationalization of Huangpu Tongqiao Capsules,and to provide a more reference for Huangpu Tongqiao Capsules to treat AD As a solid experimental foundation,Huangpu Tongqiao Capsules can play a maximum role and efficacy in the clinical treatment of AD.Method: 1.In vitro material basis of Huangpu Tongqiao Capsule An ultra high performance liquid chromatography(UPLC)method was developed for the simultaneous determination of Stilbene glycoside,Ferulic acid,Ginsenoside Rg1,Aloe-emodin,?-asarone,Emodin,Chrysophanol in Huangpu Tongqiao Capsule.The main active ingredients in Huangpu Tongqiao Capsule were initially identified.2.Effects of Huangpu Tongqiao capsule on EGFR-PLC? signal pathway of hippocampus in rats with Alzheimer's disease Clean-grade healthy adult Sprague-Dawley(SD)rats were randomly divided into: sham operation group(Sham),model group(Model),Huangpu Tongqiao capsule group(HPTQC),nimodipine group(NMP)4 groups,There are 10 animals in each group.Intraperitoneal injection of D-galactose combined with bilateral hippocampal injection of A?25-35 was used to create the model.The sham operation group was injected with normal saline.After 28 days of continuous administration,Morris water maze test was used to detect the learning and memory ability of rats in each group;HE staining was used to detect hippocampal neuron damage in each group;immunofluorescence method was used to detect EGFR and p in hippocampal CA1 region of each group of rats.-tau expression;Western blot was used to detect hippocampal EGFR,PLC?,IP3 R,and p-tau protein expression in each group;RT-PCR was used to detect hippocampal EGFR,PLC?,IP3 R,and tau m RNA content in each group.Results: 1.Seven components in Huangpu Tongqiao Capsules were detected: Stilbene glycoside,Ferulic acid,Ginsenoside Rg1,Aloe-emodin,?-asarone,Emodin,Chrysophanol.The main effects of Huangpu Tongqiao Capsules were initially identified ingredient.2.Pharmacodynamics of Huangpu Tongqiao Capsule in AD Rats(1)Water maze results: When compared with the sham operation group,the escape latency of the model group rats was markedly prolonged(P <0.01),and the number of times to cross the platform was markedly reduced(P <0.01);When compared with the model group,the escape latency of the rats in Huangpu Tongqiao Capsule group was markedly shortened(P <0.05),and the number of crossing platforms was markedly increased(P <0.05).(2)HE staining results: In the sham operation group,the neurons in hippocampal CA1 area had more layers,arranged tightly and neatly,uniformly stained,deeply stained nucleoli,and intact morphology.In the model group,the neuron cell morphology of hippocampal CA1 area obviously changed,the number of layers was reduced,the arrangement was loose and uneven,the nucleoli contraction was accompanied by obvious rupture,and dead neurons were more common.The neurons in the hippocampal CA1 region of the Huangpu Tongqiao Capsule group were relatively evenly distributed and arranged more closely.3.Effect of Huangpu Tongqiao Capsule on EGFR-PLC? Signaling Pathway in AD Rats(1)Immunofluorescence results:When compared with the sham operation group,the fluorescence intensity of EGFR and p-tau in the hippocampal CA1 region of the model group was markedly enhanced(P <0.01);When compared with the model group,the fluorescence intensity of EGFR and p-tau in the hippocampal CA1 region of the Huangpu Tongqiao Capsule group rats were markedly reduced(P <0.01).(2)Western Blot results: When compared with the sham operation group,the expressions of EGFR,PLC?,IP3 R,and p-tau protein in the hippocampus of the model group were markedly increased(P <0.01);When compared with the model group,the expressions of EGFR,PLC?,IP3 R,and p-tau protein in the hippocampus of the Huangpu Tongqiao Capsule group were markedly reduced(P <0.05).(3)RT-PCR results: When compared with the sham operation group,the m RNA expression of EGFR,PLC?,IP3 R,and tau in the hippocampus of the model group markedly increased(P <0.01);When compared with the model group,the m RNA expression of EGFR,PLC?,IP3 R,and tau in the hippocampus of the Huangpu Tongqiao Capsule group rats was markedly reduced(P <0.05).Conclusion: 1.The main components of Huangpu Tongqiao Capsule was clarified,mainly Stilbene glycoside,Ferulic acid,Ginsenoside Rg1,Aloe-emodin,?-asarone,Emodin,Chrysophanol.2.Huangpu Tongqiao Capsule can improve the learning and memory ability of AD rats and has neuroprotective effect.3.The mechanism of Huangpu Tongqiao Capsule on AD animal model may be related to the regulation of EGFR-PLC? signal pathway.