[Effect of Electro-acupuncture on the Expression of Slit2 and HSPGs in Rats with Focal Cerebral Infarction].

OBJECTIVE: To determine the expression of Slit2 and heparan sulfate proteoglycans (HSPGs) in cortex of rats with focal cerebral infarction and the effect of acupuncture (EA) on the expression of Slit2 and HSPGs. METHODS: 40 male Sprague Dawley (SD) rats were equally randomized into four groups: Control group, model group, non-acupoint EA group, and EA group. Thread-tying method was used in the model group, non-acupoint EA group and EA group to clog arteries and then open up after 1.5 h. Morphology changes of tissues around the infarction area were observed 14 days later by Nissl staining. The expressions of Slit2 and HSPGs in the ischemic brain tissues were detected by indirect immunofluorescence staining (IF) and Western blot (WB). RESULTS: Modified neurologicalseverity scores (mNSS) showed zero in the control group, lower than the score of EA group. The EA group had lower mNSS score than the non-acupoint group. The highest mNSS score appeared in the model group. Paired comparisons showed statistical differences (P < 0.01). Nissl's staining showed that EA group had increased Nissl bodies in alignment; non-acupoint EA group had increased and disordered Nissl bodies; model group had decreased and disordered Nissl bodies with edema in the brain. IF and WB showed that non-acupoint EA group had higher levels of Slit2 and HSPGs than model group (P < 0.05); EA group had higher levels of Slit2 and HSPGs than non-acupoint EA group (P < 0.05); model group had higher levels of Slit2 and HSPGs than control group (P < 0.05). The results of Nissl' s staining, IF and WB were consistent. CONCLUSION: EA can enhance the expressions of Slit2 and HSPGs. This may be one of the mechanisms of EA promoting recovery of neural functions after cerebral infarction.

[Effect of Electroacupuncture on Expression of Cortical srGAP 1 and Cdc 42 in Rats with Focal Cerebral Infarction].

OBJECTIVE: To observe the effect of electroacupuncture (EA) intervention on the neurological function and the expression change of Slit-Robo GTPase-activating protein-1 (srGAP 1) and cell division-cycle 42 (Cdc 42) in the cortex of rats with cerebral ischemic injury (CIRI) , so as to explore the mechanism of EA in the management of cerebral infarction. METHODS: A total of 48 male Sprague Dawley (SD) rats were randomly and equally divided into control, model, non-acupoint EA and EA groups (n = 12/group). The CIRI model was established based on the modified Zea Longa method. EA intervention was applied for 30 min, once a day for 14 days. Modified neurologic severity scores (mNSS) were assessed on day 1,3,7 and 14 after mode- ling. Immunofluorescence assay was used to detect the immunoactivity and distribution of srGAP 1 and Cdc 42 in the cortical ischemic region. Western blot was employed to detect the expression of srGAP 1 and Cdc 42 in the affected cortex. RESULTS: The mNSS displayed that the neurological score in the EA group was significantly lower than that in the model group and non-acupoint EA group at the 7th d and 14th d (P<0. 01). Immunofluorescence results showed that cerebral srGAP 1 and Cdc 42 were ex- pressed mainly in the cytoplasm. The fluorescence intensity of srGAP 1 of the EA group was significantly lower than that of the model group and non-acupoint EA group(P<0. 01). Meanwhile the fluorescence intensity of Cdc 42 of the EA group was markedly higher than that in the model group and non-acupoint EA group(P<0. 01). Western blot assay indicated that the expression level of srGAP 1 in the model group was significantly higher than that of the control group( P<0. 01) ,and that of the EA group was much lower than those of the model group and non-acupoint EA group(P<0. 01). There was no significant difference of srGAP 1 expression levels between the non-acupoint EA group and the model group(P>0. 05). Additionally, the protein expression of Cdc 42 in the model group was slightly higher than that of the control group(P>0. 05), and that of the EA group was significantly higher than those of the model group and non-acupoint EA group(P<0. 01). There was no significant difference of Cdc 42 expression levels between the non-acupoint EA group and the model group(P>0. 05). CONCLUSION: Cerebral infarction induced increase of cerebral srGAP 1 and decrease of Cdc 42 can be reversed by acupoint EA intervention in CIRI rats, which may be responsible for its effect in improving impaired neurological function after cerebral infarction.

[Effect of electroacupuncture of "Neiguan" (PC 6) and "Zusanli" (ST 36) on expression of cerebral cortical slit 2/Robo 1 in the focal cerebral infarction rats ].

OBJECTIVE: To observe the effect of electroacupuncture (EA) intervention on expression of Slit 2 and its transmembrane receptor Robo 1 in the cortex tissue of cerebral infarction rats so as to study its mechanism underlying improvement of cerebral ischemia. METHODS: Ninety male Sprague Dawley rats were randomly divided into control group (n = 10), model group (n = 40) and EA group (n = 40), and the latter two groups were further randomized into four subgroups: 1 d, 3 d, 7 d, and 14 d (n = 10 in each subgroup) according to the cerebral ischemia duration. Cerebral infarction model was established by occlusion of the middle cerebral artery (MCAO). EA (80-100 Hz, 1-3 mA) was applied to bilateral "Neiguan" (PC 6) and "Zusanli" (ST 36) for 30 min, once daily for 1 d, 3 d, 7 d, and 14 d, respectively. The animals' neurological defect was assessed using Zea-Longa scoring. The expression of Slit 2 and Robo 1 proteins in the cerebral cortex on the ischemic side was assayed using immunohistochemistry and Western blotting, respectively. RESULTS: In comparison with the control group, the neurological score was significantly higher in the model group (P < 0.05), and reduced considerably on day 7 and 14 after MCAO in the EA group compared with the model group (P < 0.05). Immunohistochemical results showed that in comparison with the control group, the immunoactivity levels of cerebral Slit 2 and Robo 1 were remarkably upregulated on day 1, 3 and 7 after MCAO in the model group (P < 0.05, P < 0.01), and backed to the control levels on day 14 (P > 0.05). While compared with the model group, the immunoactivity levels of cerebral Slit 2 and Robo 1 were further obviously upregulated on day 1, 3, 7 and 14 after cerebral ischemia in the EA group (P < 0.05, P < 0.01). The results of Western blotting about the expression levels of cerebral Slit 2 and Robo 1 proteins were nearly the same to those of immunohistochemical outcomes in the 4 subgroups apart from that the expression levels of both Slit 2 and Robo 1 proteins were still obviously higher on day 14 after MCAO in the model group (P < 0.01). CONCLUSION: EA intervention can significantly improve cerebral ischemia rats' neurological function and obviously upregulate the expression of cerebral Slit 2 and Robo 1 proteins, which may be one of the mechanisms of EA therapy for relieving cerebral infarction in clinic.