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Objective To observe the effect of Shenghui Granule on EC and CA1 regions of scopolamine-induced dementia rats and explore its mechanism based on p38 MAPK signal pathway.Methods 40 SD rats were randomly divided into four groups(blank group,model group,Shenghui granule group,donepezil group),and were treated with scopolamine.Morris water maze and open field test were used to evaluate the cognition and anxiety behavior of rats.The nerve injury of EC and CA1 was observed by HE staining.The activity of neurons in EC and CA1 regions was observed by c-Fos immunofluorescence staining.Western blot was used to detect p38 MAPK pathway related proteins.Results The behavioral experiment found that Shenghui Granule could improve the cognitive impairment and anxiety-like behavior of AD model rats.The results of HE staining showed that Shenghui granules had protective effects on EC and CA1 regions.The results of c-Fos immunofluorescence staining showed that Shenghui granules could increase the activity of neurons in EC and CA1 regions.Western blot results showed that Shenghui Granule could down-regulate the expression of Bax,reduce the levels of phosphorylated p38 and Tau,and increase the expression of Bcl-2.Conclusion Shenghui granule has protective effect on EC and CA1 regions of AD model rats,and may play a therapeutic role through p38 MAPK signal pathway.
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Objective To investigate the effects of Abnormal Phlegmatic Temperament Granules on spatial learning and memory, histopathology morphological change in hippocampus CA1 zone; To discuss its mechanism of action.Methods Three-month-old APP/PS1 transgenic mice were randomly divided into 5 groups: model control group, positive control (donepezil 0.92 mg/kg) group, Abnormal Phlegmatic Temperament Granules high-, medium-, and low-dose groups (3, 2, 1.5 g/kg), 18 mice in each group. Another 18 three-month-old C57BL/ 6J mice were chosen as normal control group. All administration groups received relevant medicine for successive 6 months. Then the changes in learning and memory ability of mice were detected by Morris water maze test; pathomorphism in hippocampus CA1 zone was detected by HE staining method; changes of myelin sheath, microtubule, and microfilament in myelinated nerve of hippocampus CA1 zone were detected by electron microscope. Results Morris water maze test results showed that escape incubation period of APP/PS1 transgenic mice was significantly longer than the normal control group (P<0.01), and the original platform time was significantly shorter than normal control group (P<0.01); compared with model control group, Abnormal Phlegmatic Temperament Granules treatment groups escape latency time was significantly reduced (P<0.05, P<0.01). Space experiments and escape incubation period of Abnormal Phlegmatic Temperament Granules high-, medium-, low-dose groups were significantly shortened (P<0.05, P<0.01), and spatial searching test showed that the times of mice in Abnormal Phlegmatic Temperament Granules high-, medium-, low-dose groups passing through effective area increased (P<0.01). The integrity of HE staining pyramidal cell layer in the hippocampus CA1 zones of Abnormal Phlegmatic Temperament Granules high-, medium-, and low-dose groups was relatively good; cells arranged orderly; distribution was normal. Electron microscopic observation showed that compared with model control group, the hippocampus neurons nuclear had irregular shape; nuclear membrane was clear and complete; chromatin was clear; nucleolus was obvious; cell matrix was uniform; organelles were abundant; mitochondrial cristae was obvious; endoplasmic reticulum and free ribosomes were obvious. Conclusion Abnormal Phlegmatic Temperament Granules can improve spatial learning and memory in APP/PS1 mice, alleviate neuronal ultrastructure damage and ultimately improve cognitive function.
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Objective To investigate the alteration of chaperonine hsp40 and its influence on delayed death of neurons in the CA1 region of hippocampus after transient cerebral ischemia. Method After transient global ischemia for 20 minutes, rat model was made. Following different lengths of reperfusion, all the 28 wistar rats were divided into sham-operation group,4 hour recovery group, 24 hour recovery group and 72 hour recovery group ( n = 7 rats in each group). Immunochemistry and laser scanning confocal microscopy were used to observe the distributional alteration of hsp40 in the neurons. Differential centrifuge and western blot assay were used to analyze the quantitative alteration of hsp40 and its redistribution in the neurons. Results Immunochemistry and laser scanning confocal microscopy showed the progressive reduction of hsp40 occurred at first in the cytosol, then in the nucleus until the death of all the neurons in the CA1 region died. Differential centrifuge and western blot assay showed the level of hsp40 decreased from 1.00 ± 0.21 to 0.23 ± 0.13 ( P ＜ 0.01 ) 24 hours after reperfusion; the quantity of hsp40 in the protein aggregates increased from 1.00±0.18 to 8.61 ± 1.89 (P ＜0.01 =24 h after reperfusion.Conclusions The reduction of hsp40 in the neurons of hippocampus CA1 region is an important role in protein aggregates formation.
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Objective To investigate the serum concentration and expressions of S100β protein in hippocampus CA1 region and the changes of water content in rats with asphyxia following ulinastatin injection after cardiopul-monary resuscitation (CPR). Method One hundred twenty male adult SD rots were randomly divided into 3 groups:sham-operation group, CPR group and ulinastafin group. And each group was further divided into 5 sub-groups (n=8) based on various intervals, 0.5 h,3 h,6 h,12 h and 24 h after tracheotomy in sham-operation group or after ROSC in CPR group and ulinastatin group. Asphyxial cardiac arrest and CPR model of rat was used in CPR group and ulinastatin group in which bolus dose of 100 000 U/kg ulinastatin was injection into arteria carotis. Anaesthesia, tracheotomy and vascular canratlafion without asphyxia and CPR in sham-operation group. Samples from subgroups were taken at different intervals. Brain water content was measured by using wet-dry weight method. Serum S100β protein was measured with enzyme-linked immunosorbent assay ( ELISA). The expres-sion of S100β protein in hippocampus CA1 region was measured by using immunohistochemistry. Data were ana-lyzed by SPSS version 10.0 software. Results Compared with sham-operation group, the brain water content of rats elevated significantly in all CPR subgroups after ROSC (P<0.05 or P<0.01). The brain water content of rats decreased significantly 12 h and 24 h after ROSC in ulinastatin group in comparison with CPR group (P<0.05). The serum S100β protein started to elevated significantly 0.5 h after ROSC in CPR group, and reached the peak 12 h after ROSC (P<0.01).serum S100β decreased 6 h,12 h and 24 h after ROSC in ulinastatin group compared with CPR group (P<0.01).The expression of S100β protein in hippocampus CA1 region remained at a low level in sham-operation group. The expression of S100β protein elevated significantly in all CPR subgroups after ROSC compared with sham-operation group (P<0.05 or P<0.01). Compared with CPR group, the ex-pression of S100β protein decreased after ROSC in ulinastatin group(P<0.05) .However,the expression of S100β protein in hippocampus CA1 region was significantly correlative with brain edema in all subgroups of CPR (r=0.862, P<0.05). Conclusions Ulinastatin can decrease serum S100β protein and the expression of S100β pro-tein in hippocampus CA1 region and lessen the severity of cerebral edema, alleviate the brain isehemic injury in rats after cardiopulmonary resuscitation.