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ObjectiveTo compare the effects of total alkaloids， matrine， and sophoridine extracted from Sophora alopecuroides on the activity of pheochromocytoma cells （PC12 cells）. MethodThe effect of S. alopecuroides total alkaloids， matrine， and sophoridine at concentrations of 2， 1， 0.5， 0.25， 0.125， and 0.062 5 g·L-1 on the proliferation of PC12 cells was evaluated using the 3-（4，5-dimethylthiazol-2-yl）-2，5-diphenyl tetrazolium bromide （MTT） assay. The lactate dehydrogenase （LDH） leakage rate was measured by enzyme-linked immunosorbent assay （ELISA）. Flow cytometry was used to assess cell apoptosis rate， cell cycle distribution， and intracellular Ca2+ levels. Real-time quantitative polymerase chain reaction （Real-time PCR） was performed to determine the mRNA transcription levels of B-cell lymphoma-2 （Bcl-2） and Bcl-2-associated X protein （Bax）. Protein expression levels of apoptosis-related proteins Caspase-3， Caspase-8， Bcl-2， and Bax were detected by Western blot. ResultCompared to the control group， S. alopecuroides total alkaloids， matrine， and sophoridine inhibited the proliferation of PC12 cells， increased LDH leakage rate， enhanced intracellular Ca2+ fluorescence intensity， and induced cell apoptosis in concentration-dependent manner （P<0.05， P<0.01）. Among them， S. alopecuroides total alkaloids had the strongest inhibitory effect on cell proliferation and induction of apoptosis in PC12 cells （P<0.01）. After treatment with S. alopecuroides total alkaloids， matrine， and sophoridine， the cell cycle progression of PC12 cells was arrested at G1/G0 in the S. alopecuroides total alkaloids group， and at G1/S in the matrine and sophoridine groups. The expression levels of Bax mRNA were significantly increased （P<0.05， P<0.01）， while the expression levels of Bcl-2 mRNA were significantly decreased （P<0.05， P<0.01）. All treatments significantly downregulated the expression of the anti-apoptotic protein Bcl-2 （P<0.05， P<0.01） and upregulated the expression of the pro-apoptotic proteins Bax， Caspase-3， and Caspase-8 （P<0.05， P<0.01）， with the most significant protein expression changes observed in the S. alopecuroides total alkaloids group. ConclusionAmong the S. alopecuroides total alkaloids， matrine， and sophoridine， S. alopecuroides total alkaloids exhibit the strongest inhibitory effect on the activity of PC12 cells， and its mechanism of action may be related to the inhibition of anti-apoptotic protein expression， upregulation of pro-apoptotic protein expression， and activation of the mitochondrial Caspase-8 apoptotic pathway.
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Objective To evaluate the effect of total alkaloids of Sophora alopecuroides L on the nervous behavior and the expression of neurotransmitters in rats. Methods 48 male SD rats were randomly divided into 4 groups:blank group,total alkaloids of Sophora alopecuroides L treatment with 4 mg·kg-1 ·d-1 ( low dose group) ,8 mg·kg-1 ·d-1( medium dose group) and 16 mg·kg-1 ·d-1( high dose group) groups. After successive intragastric administration for 30 days,the locomotor activity was applied to test the nervous behavior and emotional state of rats in each group. After behavioral tests were finished,the contents of trypto-phan (Trp),5-hydroxytryptophan (5-HTP),5-serotonin (5-HT),5-hydroxyindoleacetic acid (5-HIAA), norepinephrine (NE),epinephrine (E) and dopamine (DA) were detected by ELISA in serum and brain. Results In the experiment of locomotor activity,compared with blank group ((95.33±12.75) times),the numbers of horizontal movement of Sophora alopecuroides L in medium and high dose group ( ( 61. 64 ± 5.91),(64.62±5.79)times both P0.05). Correlation analysis indicated that the degree of au-tonomic activity in rats with the content of 5-HT,5-HIAA and DA in serum was negatively correlated (P<0.05, P<0.01) ,the degree of emotional stress and the content of 5-HT,5-HIAA in brain was negatively cor-related (P<0.05, P<0.01) . Conclusion The total alkaloids of Sophora alopecuroides L can reduce the ac-tivity of rats and increase the degree of emotional stress. And the mechanism may be correlated with the in-creasing level of 5-HT and 5-HIAA in serum and brain.
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OBJECTIVE:To study the effects of butin in Vernohia anthelmintica(VW)on proliferation of human immortal ke-ratinocyte cell strain HaCaT and cell secretory factors,and explore the mechanism of butin in VW in the treatment of vitiligo. METHODS:MTT method was used to determine the survival rate of HaCaT cells cultured by 0 (blank control),0.1,0.5,1.0, 5.0,10.0 μg/mL of butin for 48 h. Enzyme-linked immunosorbent assay was used to determine the contents of cell secretory factors as endothelin 1 (ET-1),ET-3,melanocyte stimulating hormone (MSH),stem cell factor (SCF),basic fibroblast growth factor (bFGF)in culture medium after HaCaT cells were cultured by 0.5,1.0,5.0 μg/mL of butin for 48 h. RESULTS:Compared with blank control,cell survival rate was increased to varying degrees after cultured by 0.1-5.0μg/mL of butin for 48 h,while decreased after cultured by 10.0 μg/mL of butin. Contents of ET-1,SCF,bFGF in culture medium were significantly increased after cultured by 0.5,1.0,5.0μg/mL of butin for 48 h(P<0.01);and contents of ET-3,MSH in culture medium were significantly increased af-ter cultured by 1.0,5.0 μg/mL of butin for 48 h(P<0.01). CONCLUSIONS:Butin can promote the proliferation of HaCaT cells, the mechanism may be associated with promoting the secretion of cell secretory factors of ET-1,ET-3,MSH,SCF,bFGF.
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OBJECTIVE:To study the effects of butin in Vernohia anthelmintica(VW)on proliferation of human immortal ke-ratinocyte cell strain HaCaT and cell secretory factors,and explore the mechanism of butin in VW in the treatment of vitiligo. METHODS:MTT method was used to determine the survival rate of HaCaT cells cultured by 0 (blank control),0.1,0.5,1.0, 5.0,10.0 μg/mL of butin for 48 h. Enzyme-linked immunosorbent assay was used to determine the contents of cell secretory factors as endothelin 1 (ET-1),ET-3,melanocyte stimulating hormone (MSH),stem cell factor (SCF),basic fibroblast growth factor (bFGF)in culture medium after HaCaT cells were cultured by 0.5,1.0,5.0 μg/mL of butin for 48 h. RESULTS:Compared with blank control,cell survival rate was increased to varying degrees after cultured by 0.1-5.0μg/mL of butin for 48 h,while decreased after cultured by 10.0 μg/mL of butin. Contents of ET-1,SCF,bFGF in culture medium were significantly increased after cultured by 0.5,1.0,5.0μg/mL of butin for 48 h(P<0.01);and contents of ET-3,MSH in culture medium were significantly increased af-ter cultured by 1.0,5.0 μg/mL of butin for 48 h(P<0.01). CONCLUSIONS:Butin can promote the proliferation of HaCaT cells, the mechanism may be associated with promoting the secretion of cell secretory factors of ET-1,ET-3,MSH,SCF,bFGF.
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OBJECTIVE:To investigate penetrative effects of a penetration enhancer and multiple penetration enhancers combi-nation with different proportions on Euodia rutaecarpa superfines cataplasm,so as to optimize enhancer and their concentrations. METHODS:Modified Franz diffusion cell was employed with isolated mice abdominal skin as barrier. HPLC method was adopted to detect the effects of a penetration enhancer (propanediol, azone, oleic acid), multiple penetration enhancers (propanedi-ol-azone,propanediol-oleic acid,propanediol-azone-oleic acid),their proportion and amount on accumulative permeation quantity (Qn) of evodiamine and rutaecarpin in Euodia rutaecarpa superfines cataplasm. RESULTS:The penetrative effect of a penetration enhancer propanediol was significantly better than that of other one penetration enhancers and multiple penetration enhancers;the higher proportion of propanediol in multiple penetration enhancer system was,the better penetrative effects of evodiamine and rutae-carpin had. Using 3%,5 % and 7 % propanediol as enhancer,Q36 h of evodiamine were 11.290,14.332 and 13.537 μg/cm2,and those of rutaecarpin were 11.965,14.856 and 13.901 μg/cm2. CONCLUSIONS:The penetrative effect of 5% propanediol is the best,and can be used as enhancer for E. rutaecarpa superfines cataplasm.
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objective To explore the effect of application of cluster-based strategies on preventing the complication of fibrosclerosis of patients with systemic scleroderma.Methods A total of 59 patients with systemic scleroderma were randomly assigned to the experimental group (34 cases) and the control group (25 cases) during the period of hospitalization in our department.The experimental group took the cluster-based strategies after admission,including a gymnastics,two kinds of respiratory breathing,three kinds of habits,four kinds of protection to prevent the complication of fibrosclerosis.The control group received the conventional care procedures of dermatology.The skin stiffness,joint function,Raynaud's phenomenon and swallowing function were evaluated when patients were admitted and before discharge in two groups.The length of stay between the two groups was also compared.Results The skin stiffness,joint function,Raynaud's phenomenon and swallowing function scored (14.74±1.33),(7.06±0.69),(0.88±0.33),(0.54±0.21) in the experimental group,significantly better than those of the control group,(24.08±1.12),(13.48±0.77),(1.24±0.60),(0.96±0.34),P ＜ 0.01.The length of stay in the experimental group and the control group was (23.65±5.45),(32.00±6.56) days,t=-5.340,P ＜ 0.01.Conclusions Application of cluster-based strategies can be effective against the fibrosclerosis damage of patients with systemic scleroderma and reduce the occurrence of complications.
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Objective To study the relationship of T, B and NK lymphocytes with the pathogenesis of chronic urticaria. Methods Flow cytometry was applied to assess the proportion of T, B and NK lymphocyte subgroups in the peripheral blood of 51 patients with chronic urticaria and 30 sex and age-matched human controls. The CD4:CD8 ratio was calculated. Moreover, the symptoms, disease course and response to antihistamines of these patients were evaluated by one physician. Results The percentage of CD8+ T and NK cells, CD4:CD8 ratio were (27.20±8.22)%, (21.20±10.84)% and 1.48±0.62, respectively, in these patients,(29.9±3.74)%, (17.5±3.56)%, 1.24±0.27, respectively, in the controls; the differences were significant between the two groups (all P＜0.05). Decreased levels of CD3+ T cells, CD8+ T cells and B cells were noted in patients resistant to antihistamines compared with those responsive to antihistamines[(61.81±11.70)% vs (75.74±2.36)%, (24.00±7.79)% vs (34.22±9.30)%, (10.78±2.07)% vs (15.25±4.10)%, P＜0.05, 0.01, 0.05, respectively)], while the CD4:CD8 ratio and percentage of NK cells were increased in antihistamine-resistant patients compared to those in antihistamine-sensitive patients [1.67±0.76 vs 1.17±0.41, (28.61±12.62)% vs (12.78±6.02)%, both P＜0.01 ]. In these patients with chronic urticaria, the percentages of CD3+ T and CD8+ T cells were negatively correlated with the symptom scores (R = -0.31, -0.28, respectively, both P＜0.05 ), while the percentage of B cells was positively correlated with the symptom scores and disease course (R = 0.53, 0.55, respectively, both P＜0.01 ). Conclusions There is an abnormality in the proportion of T, B and NK lymphocyte subgroups in patients with chronic urticaria,which indicates that humoral immunity may be involved in the pathogenesis of chronic urticaria and the mechanism for responsiveness to antihistamine.
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Objective To study the effect of RNA interference on the expression of CTGF in skin fibroblasts of systemic sclerosis(SS). Methods Four CTGF specific siRNAs and a negative control siRNA were designed and then synthesized by in vitro transcription. siRNAs labeled with carboxyfluorescein-6-succimidyl ester (FAM) were transiently transfected into SS skin fibroblasts. Forty-eight hours after the fibroblasts were treated with siRNAs, the mRNA and protein expression of CTGF was detected by semiquantitative RT-PCR and Western blot analysis, respectively. Results The mRNA and protein expression of CTGF in fibroblasts was significantly down-regulated by 4 and 3 CTGF specific siRNAs (both P