| ObjectivesTo observe the effects of Congrong Shujing Granule granules on the cognitive function of rotenone-induced Parkinson’s disease model rats and the relationship between hippocampal neurotrophic factor MANF,PI3 K / PTEN / Akt-JNK pathway and apoptosis-related factors,and explore its effect on PD Possible mechanism of protective function of model rats’ cognitive function.MethodsParkinson’s disease model rats with cognitive dysfunction were established by subcutaneously injecting rotenone sunflower oil emulsion.The successful modeled rats were randomly divided into model group and Congrong Shujing Granule granules group,and solvent group(injecting an equal volume of sunflower oil)and normal group.Congrong Shujing Granule granule group was administered by intragastric administration,and the other groups were administered intragastrically with the same amount of normal saline for 14 days.After the model was completed and drug intervention,rats were observed in behavior(hanging experiment,square bridge experiment,Morris water maze test,),HE morphology and number of cells in the hippocampal CA1 region of each group were observed,expression of TH was detected by immunohistochemistry,expression of MANF,PI3 K / PTEN / Akt-JNK signaling pathway-related proteins,apoptosis-related factors Bcl-2,Bax in hippocampa of rats was detected by WB.Results1.1 Suspension experiment and square bridge experiment: After 14 days of rotenone model establishment,compared with the normal group,the suspension time of the rats in the model group and the Congrong Shujing granule group was shortened(P <0.01),and the stay time of the square bridge was shortened(P <0.01).);After 7days and 14 days of administration of Congrongshujing granules,compared with the model group,rats in the Congrong Shujing granule group had longer hanging time(P <0.05,P <0.01)and longer square bridge staying time(P <0.01,P <0.01).1.2 Morris water maze test:1.2.1 Positioning and sailing experiment: After 14 days of rotenone model establishment,compared with the normal group,the escape latency time of the model group and the Congrong Shujing granule group was prolonged(P <0.01).After the 7 days and 14 days administration of Congrong Shujing granules,the escape latency time of the model group was longer than that of the normal group(P <0.01);compared with the model group,the escape latency time of the Congrong Shujing granule group was shortened(P<0.01).1.2.2 Space exploration experiment: After 14 days of rotenone model establishment,compared with the normal group,the number of space exploration cross-platforms and the time spent in the original platform quadrant of space exploration decreased in the model group and Congrong Shujing granule group(P <0.01).After the 7 days and 14 days administration of Congrong Shujing granule,compared with the normal group,the number of space exploration cross-platforms in the model group and the stay time of the original platform quadrant in the space exploration were reduced(P <0.01);compared with the model group,The number of cross-platforms in space exploration and the stay time in the quadrant of the original platform of the space exploration group increased in the Congrong Shujing granule group(P <0.01).2 HE staining results: After 14 days of Congrong Shujing granules administration,the number of neurons in the hippocampal CA1 region of normal rats was relatively large,arranged neatly and tightly,with regular morphology,and even distribution.No obvious neuron loss.Compared with the normal group,the number of neurons in the hippocampal CA1 region of the model group was reduced,the arrangement was irregular,loose,irregular shape,neuron loss is significant.Compared with the model group,the number ofneurons in the hippocampal CA1 region of the Congrong Shujing granule group was relatively increased.The arrangement is slightly neat and tight,the shape is slightly regular,and the loss of neurons is relatively reduced.3 Immunohistochemical results: After 14 days of Congrong Shujing granules administration,compared with the normal group,TH positive cells in the model group were significantly reduced(P <0.01);after 14 days of administration of Congrong Shujing Granule granules,compared with the model group,the TH-positive cells in the Congrong Shujing granule group increased significantly(P <0.01).4 Changes of related protein in rats each group4.1 MANF: After 14 days of Congrong Shujing granules administration,compared with the normal group,the expression level of MANF protein in the model group was reduced(P <0.01).Compared with the model group,the expression level of MANF protein in the Congrong Shujing granule group was increased(P <0.05).4.2 PI3 K / PTEN / Akt signaling pathway-related proteins: After 14 days of Congrong Shujing granules administration,compared with the normal group,the expression of PI3 K and Akt proteins was reduced in the model group(P <0.01),and the expression of PTEN protein was increased(P <0.01).Compared with the model group,the PI3 K and Akt protein phosphorylation expressions increased in the Congrong Shujing granule group(P<0.05),while the expressions of PTEN protein was decreased(P <0.05).4.3 Related proteins on the JNK pathway: After 14 days of Congrong Shujing granules administration,compared with the normal group,the expression of JNK proteins in the model group increased(P <0.01).Compared with the model group,the expression of phosphorylated JNK protein in the Congrong Shujing granule group was reduced(P<0.05).4.4 Apoptosis-related factor protein: After 14 days of Congrong Shujing granules administration,compared with the normal group,the expression of Bcl-2 protein in the model group decreased(P <0.01),and the expression of Bax protein increased(P <0.05).Compared with the model group,the expression of Bcl-2 protein increased and the expression of Bax protein decreased(P <0.01).Compared with the normal group,the ratio of Bcl-2 / Bax was decreased in the model group(P <0.01),and compared with the model group,the ratio of Bcl-2 / Bax was increased in the Congrong Shujing group(P <0.01).Conclusions1 Congrong Shujing granules can improve the behavior and cognitive dysfunction of PD model rats;2 Congrong Shujing granules can protect the morphology and number of cells in hippocampal CA1 region,increase the content of TH in the substantia nigra of PD model rats,inhibit neuronal apoptosis;3 Congrong Shujing granules can promote the expression of MANF,PI3 K / Akt pathway related proteins and Bcl-2 protein in PD model rats,and inhibit the activity of PTEN,JNK signaling pathways and Bax protein;4 Congrong Shujing granules may promote the expression of MANF and inhibit the activity of PTEN,increase the activity of PI3 K / Akt pathway,inhibit the activity of JNK signaling pathway,and exert neuroprotective effect on PD cognitive dysfunction. |
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