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OBJECTIVE To explore the antidepressant effect and potential mechanism of icariside Ⅱ （ICSⅡ） based on the GABAergic nervous system. METHODS The male Kunming mice were randomly divided into a control group （group C， 10 mice） and a modeling group （50 mice）. The depression model was induced by chronic unpredictable mild stress （CUMS） method in the modeling group. After 21 days of stimulation， the rats of modeling group were randomly divided into depression model group （NS group）， positive control group [ECS group， oxalate escitalopram 15 mg/（kg·d）] and ICSⅡ low-dose， medium-dose and high-dose groups [ICSⅡ-L group， ICSⅡ-M group， ICSⅡ-H group； ICSⅡ 10， 20， 30 mg/（kg·d）]， with 10 mice in each group. Administration groups were given relevant medicine intragastrically， once a day， for 14 consecutive days. The sugar water preference rate， total exercise distance， immobility time in tail suspension and forced swimming experiments were detected in each group. The morphology of neurons and Nissl bodies in the hippocampal CA3 region were observed； the contents of γ-aminobutyric acid （GABA） and glutamic acid （Glu）， GABA/Glu ratio， and the expressions of GABAergic nervous system-related proteins （GABA A receptor α1， GABA B receptor 1， vesicular GABA transporter， glutamate decarboxylase 67， GABA membranal transporter 3） were detected in hippocampus. RESULTS Compared with group C， the sugar water preference rate and the total exercise distance significantly reduced in NS group， while the values of immobility time in the tail suspension test and forced swimming test were significantly prolonged （P＜0.05）. The morphology of neurons in the CA3 area of the hippocampus was irregular and the Nissl bodies were reduced， with a significant decrease in the number of structurally intact neurons （P＜0.05）； the content of Glu was significantly increased， while the content of GABA， GABA/Glu ratio， and the expressions of GABAergic nervous system-related proteins were significantly decreased （P＜0.05）. Compared with NS group， depression behavior in each administration group was improved， and the above indexes were mostly reversed （P＜0.05）. CONCLUSIONS ICSⅡ can improve depression behavior of depression model mice. The mechanisms may be associated with regulating the balance of GABA and Glu， increasing the synthesis， transport and release of GABA， and regulating the expressions of GABA-related receptors， so as to improve GABAergic nervous system.
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Objective:To investigate the protective effect and mechanism of Icaritin Ⅱ (ICAⅡ) on bladder in radiation cystitis model.Methods:A total of 18 10-week-old male SD rats were selected from July 2021 to March 2022 and divided into control group, model group and treatment group by random number table method, with 6 cases in each group. Model group and treatment group were given a single dose of 20 Gy X-ray irradiation in the pelvic area. 24 h after irradiation, the treatment group was given ICAⅡ 4.5 mg/(kg·d) gavage, while the control group and model group were given the same volume of solvent (10% anhydrous ethanol, 20% isopropyl alcohol, 30% polyethylene glycol and 40% deionized water) gavage for 4 consecutive weeks. Drug eluting for 1 week. The bladder volume and leakage point pressure of the three groups of rats were measured by multi-conducting physiological apparatus, and the bladder function was evaluated. HE staining, Masson staining, ELISA, TUNEL staining and western blotting were performed on the bladder samples of the three groups of rats. The pathological changes (thickness of bladder mucosa, ratio of smooth muscle to collagen fiber), oxidative stress level (superoxide dismutase SOD, malondialdehyde), apoptosis rate, protein levels of inflammatory factors (IL-6, NF-kB) and anti-oxidative stress signaling pathway factors (Nrf2, HO-1) of the three groups of rats were compared.Results:After 4 weeks of modeling, in the model group, the bladder volume [(1.01±0.12)ml vs. (1.58±0.21)ml, P=0.001], the bladder leakage point pressure [(38.79±4.12) cmH 2O (1 cmH2O=0.098 kPa)vs.(60.59±3.81) cmH 2O, P=0.001], the ratio of smooth muscle of bladder wall to collagen fiber [1.78±0.17 vs.3.15±0.57, P=0.001], SOD[(6.31±0.73) U/mg vs.(14.67±1.04) U/mg, P=0.001] were lower than the control group, and the differences were statistically significant. In the model group, the thickness of bladder mucosa [(47.33±1.78)μm vs.(20.83±2.33)μm, =, P=0.001], malondialdehyde [(1.01±0.13) nmol/mg vs.(0.49±0.03) nmol/mg, P=0.001], IL-6 (0.87±0.11 vs. 0.33±0.10, P=0.001), NF-kB (0.71±0.14 vs. 0.29±0.07, P=0.001), apoptosis rate [(11.60±3.04)% vs. (3.91±1.40)%, P=0.007] was higher than the control group, and the differences were statistically significant. In the treatment group, the bladder volume [(1.27±0.13)ml, P=0.030], bladder leakage point pressure [(47.83±2.50)cmH 2O, P=0.004], smooth muscle to collagen fiber ratio (2.78±0.68, P=0.015), SOD[(10.48±0.85) U/mg, Compared with model group, bladder mucosa thickness [(31.94±3.20)μm, P=0.001], malondialdehyde [(0.64±0.09) nmol/mg, P=0.001], IL-6 (0.69±0.11, P=0.035), NF-kB (0.45±0.06, P=0.002) and apoptosis rate [(6.05±0.60)%, P=0.030] were lower than those in model group. The protein expression level of Nrf2 in model group (0.73±0.08 vs. 0.58±0.11, P=0.023) was higher than that in control group, but there was no significant difference in the protein expression level of downstream antioxidant factor HO-1 (0.50±0.14 vs. 0.35±0.06, P=0.060). Nrf2 protein expression level (0.88±0.03, P=0.027) and HO-1 expression level (0.68±0.07, P=0.026) in treatment group were higher than those in model group. Conclusion:ICAⅡ can reduce radiation cystitis injury, and its mechanism may be related to anti-oxidative stress and reducing inflammation.
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OBJECTIVE：To study the improvement effects of icariside Ⅱ（ICS Ⅱ）on neurological function of focal cerebral ischemia model rats by regulating miR- 141-3p/Notch/nuclear factor erythroid- 2-related factor 2（Nrf2）axis（miR-141-3p/Notch/ Nrf2）. METHODS ：The rats were divided into sham operation group ，model group ，nimodipine group （20 mg/kg）and ICS Ⅱ low-dose，medium-dose and high-dose groups （4，8，16 mg/kg），with 20 rats in each group. Twenty-four hours after establishing focal cerebral ischemia model ，model rats were given re levant medicine or normal saline intragastrically ，twice a day ，for consecutive 3 d. The neurological deficit of rats in each group was scored ；the volume of cerebral infarction was measured by 2，3， 5-triphenyltetrazolium chloride （TTC）staining；water content of cerebral tissue and the permeability of blood-brain barrier were measured；HE staining was performed to observe the pathological change of cerebral tissue of rats ；the expression of miR- 141-3p in cerebral tissue of rats was measured by qRT-PCR ；the protein expression of Notch and Nrf 2 in cerebral tissue of rats were measured by Western blotting assay. RESULTS ：Compared with sham operation group ，the neurological deficit score ，expression of Notch-1 and Nrf 2 in model group were significantly lowered （P＜0.05）；infarction volume ，brain water content ，the permeability of blood-brain barrier and the expression of miR- 141-3p in cerebral tissue were increased significantly （P＜0.05）；the distribution of cortical cells was disordered ，and inflammatory infiltration and necrosis were observed in a large number of nerve cells. Compared with model group ，the neurological deficit score ，the protein expression of Notch- 1 and Nrf 2 in cerebral tissue were significantly increased in ICS Ⅱgroups（P＜0.05）；infarction volume ，brain water content ，the permeability of blood-brain barrier and the expression of miR- 141-3p in cerebral tissue were decreased significantly （P＜0.05）；the arrangement of cortical cells was regular，and the inflammatory infiltration and necrosis of nerve cells were decreased significantly. CONCLUSIONS ：ICS Ⅱ can promote the recovery of neurological function in focal cerebral ischemic model rats ，which may be related to down-regulation of miR-141-3p and activation of Notch/Nrf 2 axis.
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Aim To investigate the effects of IcarisideⅡ ( ICS Ⅱ) on myocardial fibrosis in spontaneously hypertensive rat( SHR) . Methods Twenty male SHR rats were randomly divided into the model group (group SHR) , ICS Ⅱ low ( ICS Ⅱ-L) , middle ( ICSⅡ-M) and high ( ICS Ⅱ-H) group, and male WKY rats were set as control group ( group WKY) . ICS Ⅱ-L, ICSⅡ-M and ICSⅡ-H groups were intragastrically administered with ICS Ⅱ for 12 weeks. After that the blood pressure was measured in rats. Then, the rats were sacrificed and the left ventricles were separated in order to calculate the left ventricular mass index. Mas-son staining was used to detect the occurrence of inter-stitial fibrosis in cardiac tissues. Real time PCR was used to observe the gene expression of MMP-2, MMP-9 and TIMP-1 in the left ventricle in SHRs. The protein expression levels of MMP-2, MMP-9, TIMP-1, Colla- gen Ⅰ and Collagen Ⅲ were measured by Western blot. Results Compared with SHR group, the myo-cardial fibrosis was reduced after ICS Ⅱ (8, 16 mg· kg-1) treatment. The blood pressure and left ventricu-lar mass index decreased(P<0.05). The expressions of MMP-2, MMP-9, CollagenⅠand CollagenⅢwere down-regulated in left ventricular tissues( P <0.05 ) , while the expression of TIMP-1 was up-regulated( P<0.05) . Conclusion Icariside Ⅱ ameliorates myocar-dial fibrosis in SHR, and the mechanisms might be re-lated to the decrease of blood pressure and down-regu-lation of MMP-2, MMP-9 expression and up-regulation of TIMP-1 expression.
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Aim To observe the effect of icariside Ⅱ(ICS Ⅱ)on cardiomyocyte apoptosis in spontaneously hypertensive rats (SHR)and to explore its possible mechanism. Methods Thirty male 13-week-old SHRs were randomly divided into model group,ICS Ⅱ low, medium,high and positive drug group (n = 6),ho-mologous male Wistar-Kyoto rats as control group (n =6). After a week of adaptive feeding,ICS Ⅱ low,me-dium and high dose groups were given ICS Ⅱ 4,8,16 mg · kg - 1 (ig,qd),and the positive drug group was given losartan 20 mg·kg - 1 . At the same time,the WKY and SHR group were given equal volume double distilled water. After 12 weeks of administration,the blood pressure was measured in rats. Then,the rats were sacrificed and the left ventricles were separated in order to calculate the left ventricular mass index. HE staining was used to observe the pathological changes of the left ventricle,and the apoptosis of the left ventricu-lar myocardium was detected by TUNEL staining. The expressions of Bcl-2 and Bax mRNA in left ventricle were detected by real time RT-PCR,and Bcl-2,Bax and cleaved-caspase-3 protein expressions were detec-ted by Western blot. Results Compared with WKY group,the blood pressure and left ventricular mass in-dex increased in SHR group (P < 0. 05),and the my-ocardial cell arrangement was disordered and the cell hypertrophy and apoptosis were obvious,accompanied by rupture of filament ;the level of Bax mRNA was up-regulated (P < 0. 05),and Bcl-2 mRNA was down-regulated (P < 0. 05 );the expressions of Bax and cleaved-caspase-3 protein were up-regulated (P <0. 05),and the level of Bcl-2 protein was down-regu-lated (P < 0. 05 ),and the ratio of Bax / Bcl-2 in-creased (P < 0. 05). Compared with SHR group,the blood pressure and left ventricular mass index de-creased in ICS Ⅱ middle,high group and the positive drug group (P < 0. 05);moreover,the arrangement of myocardial cells became more orderly,the cell hyper-trophy and the apoptosis of myocardial cells were im-proved;the level of Bax mRNA was down-regulated and Bcl-2 mRNA was up-regulated (P < 0. 05);the expression of Bax and cleaved-caspase3 protein were down-regulated and the level of Bcl-2 protein was up-regulated (P < 0. 05 );the ratio of Bax / Bcl-2 de-creased (P < 0. 05). Conclusions ICS Ⅱ can im-prove the left ventricular cardiomyocytes apoptosis in SHR,and its mechanism is related to the decrease of blood pressure and the inhibition of mitochondrial ap-optosis pathway.
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Objective To observe the effect of IcarisideⅡ (ICSⅡ) on spatial learning and memory impairments and axonal regeneration induced by chronic cerebral hypoperfusion (CCH) in rats.Methods 90 male SD rats were randomly divided into normal group,sham operation group,CCH group and ICS Ⅱ low,middle and high-dose treatment groups.The chronic cerebral hypoperfusion model was established by permanent bilateral common carotid artery occlusion.Then these rats in ICS Ⅱ low,middle and high-dose treatment groups were given ICS Ⅱ4,8 and 16 mg/(kg · d) by gavage on the 1st day after modeling.There were 5 rats in every group at each observing time(4,8 and 12 week).Morris water maze experiment was utilized to assess the escape latency and the target quadrant residence time while HE and immunohistochemistry analysis were applied to test the morphology change and expressions of GAP-43,MAP-2 and Nogo-A in hippocampal CA 1.Results Compared with those of sham operation groups at 4,8 and 12 week respectively,the escape latency in CCH group were significantly prolonged(40.02±4.95) s,(42.29±5.75) s,(53.68±6.14) s vs (26.43±2.68) s,(26.84±2.06) s,(31.53±4.12) s,P＜0.05;the target quadrant residence time were significantly reduced(28.53±2.40) s,(28.02±4.28) s,(22.60±4.03) s vs (33.34±2.89) s,(33.31 ±4.14) s,(31.63±2.20)s,P＜0.05);the expressions of GAP-43 and Nogo-A were increased with that of MAP-2 reduced(P＜0.05).Meanwhile,the neuropathological changes with more denatured neurons and less normal neurons were found in hippocampal CA1.However,compared with those of CCH group,the escape latency of ICS Ⅱ middle and high-dose groups (30.58±3.03) s,(29.19±4.23) s,(38.77±5.80) s;(28.90±2.98) s,(26.91 ±6.63) s,(36.51 ±3.98) s) were shortened (P＜0.05);the target quadrant residence time (32.54± 3.41) s,(32.69±3.47) s,(28.27±3.57) s;(32.69±3.54) s,(33.20±4.29) s,(28.07±4.04) s) were increased (P＜ 0.05);the expression of Nogo-A was decreased while those of GAP-43 and MAP-2 were conversely increased (P＜0.05).Moreover,few denatured neurons were observed in hippocampal CA1.But there were no differences for those indexs between CCH group and ICS Ⅱ low-dose treatment groups (P＞0.05).Compared with those in 8 week and 4 week,the escape latency and the target quadrant residence time were prolonged and reduced with the expression of Nogo-A increased in all groups except normal group and sham operation group(P＜0.05),the expressions of GAP-43 and MAP-2 were decreased in CCH group and ICS Ⅱ low-dose treatment group(P＜0.05),but there were no significant differences in ICS Ⅱ middle and high-dose treatment groups at 12 week(P＞0.05).However,there were no statistical significance of all indexes between 8 week and 4 week(P＞0.05).Conclusion ICS Ⅱ can improve the spatial learning and memory in chronic cerebral hypoperfusion rats,which may be achieved by neuroprotective effects and reducing the expression of Nogo-A consequently promotes the regeneration of axons.
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To explore the effect of icariside ? on osteoprotegerin(OPG) expression in osteoblasts, primary osteoblasts were cultured with different concentration of icariside ? and 17?estradiol. As a major metabolite of icarrin in vivo, icariside ? exerted a better effect on OPG expression which plays a key role in osteoporosis. 20 ng/dl icariside ? also exerted better effect on OPG secretion than that of positive control group. Consequently, icariside ?could be a candidate for osteoporosis treatment.