Effect of Jinmaitong (筋脉通) serum on the proliferation of rat Schwann cells cultured in high glucose medium

Objective:To investigate the effect of Jinmaitong(筋脉通,JMT)serum on the proliferation of rat Schwann cells(SCs)primarily cultured in high glucose medium.Method:SOs were primarily cultured in Dulbecco's minmum essential medium(DMEM control),50 mmol/L glucose medium(50 mmol/L Glu),75 mmol/L glucose medium(75 mmol/L Glu),as well as 50 mmol/L glucose medium,with different concentrations of JMT serum(undiluted,1:2 diluted and 1:8 diluted)and Neurotropin(Ntp), respectively.The proliferation of SCs under different conditions was detected by MTT.Result:SCs grew exuberantly in DMEM within 24-72 h,but slowed down at 96 h.The proliferation of SCs was inhibited in 50 mmol/L Glu and 75 mmol/L Glu after cultures of 48,72 and 96 h,which showed that both were significantly different compared to the control group(P<0.01).The inhibition was more significant in 75 mmol/L Glu than in 50 mmol/L Glu(P<0.05).Spearman's rho analysis revealed that the proliferation of SCs had a negative correlation with the concentration of glucose(r=-0.471,P<0.01).Excluding the time factor,partial correlation showed similar results(r=-0.679,P<0.01).After 48 h,the proliferation of SCs increased significantly in JMT1:2 and Ntp compared with 50 mmol/L Glu(control 0.437±0.019,50 mmol/ L Glu 0.367±0.035,JMT1:2 0.426±0.024,Ntp 0.422±0.013;P<0.01),and there were no statistically significant differences among the JMT groups,the Ntp group and the control group(P>0.05).Conclusions: The proliferation of SCs was inhibited in high glucose medium,and the inhibition was reduced by different concentrations of JMT serum,especially at JMT1:2.     

针灸脑缺血再灌注损伤大鼠百会及足三里穴位对热休克蛋白70及肿瘤坏死因子-α表达的作用

Objective:To explore the effects of acupuncture on the peripheral serum expression of heat shock protein 70(HSP70)and tumor necrosis factor a(TNF-α)in rats with cerebral ischemia-reperfusion injury(CIRI).Methods:In total,152 Sprague-Dawley(SD)rats were randomly divided into an operated group and a non-operated group according to a random digits table.The operated group included a sham-operated group,a model group and an acupuncture group,whereas the non-operated group consisted of a normal group.Except for the normal group,each group was further divided into 12,24,48,72,96,and 144 h time points according to different reperfusion times.Eight rats were assigned in each operated group and in the normal group.The rat model of CIRI was established by the thread occlusion method in the model and acupuncture groups.The acupuncture group was treated with electroacupuncture at Baihui(DU20)and Zusanli(ST36)for the required time after successful operation.Blood was sampled to detect the HSP70 and TNF-αcontent by enzyme linked immunosorbent assay.Results:The expression of HSP70 protein in the peripheral serum of the experimental groups was higher than that in the normal control group.The peak time in both the model and the sham-operated groups was 12 h,and the peak time in the acupuncture group was 24 h.The expression in the acupuncture group declined to a lower level at 72 h and was lower than that in the model and sham-operated groups(P0.05).The peak time for the expression of TNF-αprotein in the peripheral serum of both the model and the acupuncture groups was 24 h,but the expression in the acupuncture group was lower than the model group.Additionally,the expression of TNF-αin all experimental groups was higher than the normal group(P0.05).Conclusions:Acupuncture at DU20 and ST36 in rats attenuated CIRI,which was associated with a reduction in the expression of HSP70 and TNF-α.These results provide clues to acupunctural neuroprotective properties.Acupuncture at DU20 and ST36 in rats after CIRI can adjust the expression of HSP70 and TNF-αin the peripheral serum,which might be one of the mechanisms of acupuncture's attenuation of CIRI.     

Anti-fibrotic Effects and Mechanism of Shengmai Injection(生脉注射液)on Human Hepatic Stellate Cells LX-2

Objective: To investigate the effects of Shengmai Injection(生脉注射液, SMI) on the proliferation, apoptosis and N-myc downstream-regulated gene 2(NDRG2, a tumour suppressor gene) expression in varying densities of human hepatic stellate cells LX-2. Methods: LX-2 cells were cultured in vitro. Then, cells were plated in 96-well plates at an approximate density of 2.5×10~4 cells/mL and cultured for 48, 72, 96 or 120 h followed by the application of different concentrations of SMI(0.6, 1.2, 2.4, 4.8 or 6 μL/mL). Cell proliferation was measured after an additional 24 or 48 h using the 3(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide(MTT) assay. The effects of SMI on different cell growth states(cultured for 48, 72, 96, or 120 h) were observed by light microscopy at 24 h after treatment. When the cells reached 80% confluence, apoptosis was detected by flow cytometry after 24 h. Lastly, LX-2 cells were treated with different concentrations of SMI and extracted with protein lysis buffer. The levels of NDRG2 were measured by Western blot. Results: When the LX-2 cells grew for 48, 72, 96 and 120 h, 4.8 and 6 μL/m L of SMI significantly inhibited cell proliferation at 24 and 48 h after treatment(P0.05). And 2.4 μL/mL of SMI also inhibited cell proliferation at 24 h after treatment when cell growth for 48 h(P0.05) and at 48 h after treatment when cell growth for 72, 96 and 120 h(P0.05). The NDRG2 expression level in the LX-2 cell was significantly increased when treated with SMI at concentrations of 1.2, 2.4, 4.8 or 6 μL/mL(P0.05). Conclusions: The inhibitory effects of SMI on the proliferation of LX-2 cells were related to not only concentration dependent but also cell density. In addition, SMI(2.4, 4.8 and 6 μL/mL) could accelerate apoptosis in LX-2 cells, and the mechanism might be associated with NDRG2 over-expression.     

Inhibitory Effect of Curcumin on Proliferation of Human Pterygium Fibroblasts

In order to investigate the effect of curcumin on proliferation and apoptosis of human pterygium fibroblasts (HPF) in culture and search for a new method to prevent the recurrence after pterygium surgery, HPF was incubated with 0-160 μmol/L curcumin for 24-96 h. The MTT method was used to assay the biologic activities of curcumin at different time points and different doses. The expression of proliferating cell nuclear antigen (PCNA) in each group was detected by immunohistochemistry. The cell cycle distribution was detected by flow cytometry (FCM). Admini- stration of 20-80 μmol/L curcumin for 24-72 h could significantly inhibit HPF proliferation in a dose- and time-dependent manner (P<0.05). After treatment with curcumin at different concentrations of 20, 40, 80 and 160 μmol/L for 24 h, FCM revealed there was a significant sub-G1 peak at each concentration. The number of HPF in G0/G1 phase was increased, while in S phase, it was decreased (P<0.05). At the concentration of 20-80 μmol/L, curcumin, in a dose-dependent manner (P<0.05), could inhibit the expression of PCNA in HPF. It was suggesterd that curcumin could significantly in- hibit the proliferation of HPF, make HPF arrest in G0/G1 phase and induce the apoptosis of HPF in a dose- and time-dependent manner.     

Rabbit annulus fibrosus cell apoptosis induced by mechanical overload via a mitochondrial apoptotic pathway

In order to investigate the apoptotic pathway of rabbit annulus fibrosus(AF) cells induced by mechanical overload,an experimental air-pressure model was established in this study to pressurize the rabbit AF cells in vitro.Cells were randomly divided into five groups in which the cells were exposed to a continuous pressure of 1.1 MPa for different lengths of time(0,5,12,24 and 36 h).The cell proliferation and apoptosis were detected by cell counting kit-8(CCK-8) assay and flow cytometry;the alterations in mi...     

The Effect of Ligustrazine on Peritoneal Transport in Peritoneal dialysis

In order to investigate the effect of ligustrazine (Lig)i.p.on peritoneal permeability in peritoneal dialysis and its side effects,creatinine was given intravenously and continuously to maintain the high plasma creatinine level.All the rabbits were divided into three groups:normal control group (goup A),group B treated with 0.12% Lig and group C treated with 0.24% Lig.The peritoneal dialysis of all rabbits lasted 2h.The plasma and dialysate levels of glucose,protein and creatining were observed immediate,30min,60min,90min,120min after dialysis.Creastinine dialysate/plasma ratio (D/P),protein D/P ratio,grucose D/Do at different time points after dialysis and creatinine mass transfer area coefficient (MTAC) at 120min were calculated.The structures of peritoneum were observed under optical microscope and electron microscope after continuously intraperitoneal injection of Lig for 14 days.The results showed that the 90-min and 12-min creatinine D/P ratios in the group C were higher than in the group A.The 120-min creatinine MATC in the group C was higher than in the group A.The rabbits treated with Lig did not show significant structure changes of peritoneum and signs of peritoneal irritation.It was suggested that Lig could increase mass transfer ability of peritoneum without significant side effects.     

Testosterone Secretion in Response to Human Chorionic Gonadotropin （hCG） in Eugonadal and Hypogonadal Men

Two injections of hCG in dose of 2000 IU were administered in an interval of 96 hours, and venous blood samples were drawn at 0, 1, 2, 3, 4, 6, 24, 48, 72 and 96 hr for testosterone determination. A biphasic curve of testosterone release was foand in normal adult men (n=4), patients with Klinefelter syndrome (n=10) and patients with hypogonadism due to pituitary tumor (n=8), respeetively, but not in prepubertal boys (n=4) and patients with IHH(n=9). Only after the second loading of hCG the first peak of testosterone secretion emerged in the latter two groups. The second peak values after the second injection of hCG were signfieantly greater than those after the first injection in all groups. Whereas the maximal increments of the second peak were much lower in normal adult men and patients with Klinefelter syndrome than those in other 3 groups.It was suggested that (1) the first peak of testosterone secretion was depending upon the previous exposure to high concentration of hCG or LH; (2) repeated administration of hCG had a setf-priming effect on testosterone release; and (3) the desensitization of Leydig cells existed after a single injection of hCG and its removal was incomplete after an intermission of 96 hours.     

染料木素和大豆黄素对海马神经细胞H19-7增殖和神经营养因子表达的影响

Objective To determine the effects of genistein and daidzein on the proliferation and survival of the hippocampal neural cells and underlying mechanism. Methods H19-7/IGF-IR neural cell line was cultured in phenol red free DMEM absented of serum for 72h. Genistein, daidzein or 17β-estradiol was added to the culture at various concentrations. Their proliferation and protective effects on the neuronal cells were determined by BrdU and MTT assay respectively. The effect of phytoestrogens on cell cycle regulation was determined using flow cytometry. The effects of the soy isoflavones on brain-derived neurotrophic factor (BDNF) expression were determined by ELISA and RT-PCR respectively. Results It was observed that, with 72h of treatment, 20nM and 200 nM 17B-estradiol significantly promoted the neuronal cell proliferation at 33% and 36% ;20nM and 200nM genistein significantly promoted the neuronal cell proliferation at 15% and 13% ; 200nM daidzein significantly promoted the neuronal cell proliferation at 11% compared to the control (P＜0.05). Genistein and daidzein induced an significantly increase in the S phase arrest at (17.64 ± 0.43) % and (19.48 ± 1.01) % compared to the control (P ＜ 0. 05). Moreover, genistein and daidzein significantly increased the expression of mature BDNF and BDNF mRNA level (P＜0.05). The effect of genistein and daidzein on hippocampal neuronal cell proliferation was blocked by K252a, selective inhibitor of tyrosine kinase receptors. Conclusion Genistein and daidzein improved hippocampal neuronal cell proliferation and viability in vitro. The effects might be mediated by increasing in BDNF expression.     

Changtong oral liquid for prevention of postoperative intestinal adhesion: an experimental study]

OBJECTIVE: To observe the effect Changtong oral liquid (CTOL), a traditional Chinese drug preparation, on the prevention of postoperative intestinal adhesion. METHODS: Fifty-four male SD rats were randomly divided into 6 equal groups, namely the normal control, model control, Simo decoction (SMD, another traditional Chinese drug preparation), and CTOL groups, and CTOL group included three subgroups treated at low, moderate and high doses. Except for those in the normal control group, all the other rats were subjected to operation with Ellis' method for establishing intestinal adhesion models. After the operation, the rats in the normal control and model groups received intragastric administration of distilled water (10 ml/kg), SMD group received SMD administration in similar manner at the dose of 10 ml/kg, and the three CTOL groups had CTOL at the doses of 4.3, 8.6 and 17.2 g/kg, respectively. On day 7 after surgery, blood sample were taken from all the rats to determine the white blood cell (WBC) count and fibrinogen (FIB) concentrations, and the intestinal adhesion was graded. RESULTS AND CONCLUSION: CTOL evidently reduced the severity of postoperative adhesion and decreased WBC count and plasma FIB level, suggesting the efficacy of CTOL in inhibiting the formation of postoperative intestinal adhesion.     

常通口服液预防术后肠粘连的实验研究

目的 观察中药常通口服液用于术后腹腔肠粘连的效果。方法 选取54只SD雄性大鼠随机分为6组(n=9):正常对照组、模型对照组、四磨汤组、常通口服液低、中、高剂量组。除正常对照组外,其余大鼠均按Ellis法制备成肠粘连模型。正常及模型对照组予以蒸馏水灌胃(10ml/kg);四磨汤组以10ml/kg灌胃给药;常通口服液低、中、高剂量组分别按4.3、8.6、17.2g/kg灌胃给药。各组于术后第7天取血,测定白细胞(WBC)计数及纤维蛋白原(FIB)浓度;同时进行粘连程度肉眼分级。结果 常通口服液能明显减轻肠粘连程度,降低大鼠血浆中WBC计数及FIB浓度。结论 常通口服液能够预防腹腔术后肠粘连形成。     

常通口服液对术后肠粘连大鼠血清细胞因子的影响

目的观察常通口服液对术后粘连大鼠血液中肿瘤坏死因子α(TNF-α)、转化生长因子β(TGF-β1)、白细胞介素-1β、4、6、10(IL-1β、IL-4、IL-6、IL-10)水平的影响。方法选用SD雄性大鼠，随机分为6组：正常对照组、模型对照组、四磨汤组及常通口服液低、中、高剂量组。除正常对照组外，其余大鼠均按Ellis法制备成肠粘连模型。正常及模型对照组予以蒸馏水灌胃(10ml／kg)；四磨汤组以10ml／kg灌胃给药；常通口服液低、中、高剂量组分别按4.3、8．6、17.2g/kg灌胃给药。各组于术后第7天取血，酶联免疫吸附法测定细胞因子水平，同时进行粘连程度肉眼分级。结果常通口服液能显著减轻模型大鼠的肠粘连程度，降低术后肠粘连大鼠血清中致炎细胞因子TNF-α、IL-1β、TGF-β1、IL-6水平，但对抗炎细胞因子IL-4、IL-10水平无影响。结论本实验初步筛选出有意义的术后粘连疗效评价指标，为术后粘连临床疗效评价和新药研发建立客观、准确、可靠的临床疗效评价指标提供了实验依据。     

常通口服液对术后肠粘连大鼠血清细胞因子的影响

目的 观察常通口服液对术后粘连大鼠血液中肿瘤坏死因子α(TN F-α)、转化生长因子β₁(TG F-β1)、白细胞介素-1β、4、6、10(IL-1β、IL-4、IL-6、IL-10)水平的影响。方法 选用SD 雄性大鼠,随机分为6 组:正常对照组、模型对照组、四磨汤组及常通口服液低、中、高剂量组。除正常对照组外,其余大鼠均按Ellis法制备成肠粘连模型。正常及模型对照组予以蒸馏水灌胃(10 ml/kg);四磨汤组以10 ml/kg 灌胃给药;常通口服液低、中、高剂量组分别按4.3、8.6 、17.2 g/kg 灌胃给药。各组于术后第7 天取血,酶联免疫吸附法测定细胞因子水平,同时进行粘连程度肉眼分级。结果 常通口服液能显著减轻模型大鼠的肠粘连程度,降低术后肠粘连大鼠血清中致炎细胞因子TN F-α、IL-1β、TG F-β₁、IL-6 水平,但对抗炎细胞因子IL-4、IL-10 水平无影响。结论 本实验初步筛选出有意义的术后粘连疗效评价指标,为术后粘连临床疗效评价和新药研发建立客观、准确、可靠的临床疗效评价指标提供了实验依据。     

大鼠腹膜纤维化形成过程中TGF-β/Smads2/3信号蛋白的变化及意义

目的 探讨转化生长因子β(TFGF～β)/Smads信号蛋白在4.25%的葡萄糖腹膜透析液与脂多糖(LPS)诱导的大鼠腹膜纤维化形成过程中的动态变化及意义.方法 32只SD雄性大鼠(160～170g),随机分为正常对照组(n=8),大鼠不做任何处理;模型组(n=24),每日给予腹腔注射4.25%的葡萄糖腹膜透析液(10...     

肺纤维化形成过程中大鼠成纤维细胞对肺泡Ⅱ型上皮细胞的影响

目的:研究在平阳霉素诱导的大鼠肺纤维化形成过程中肺成纤维细胞对肺泡Ⅱ型上皮细胞的影响。方法:先将大鼠分成平阳霉素组和生理盐水组,平阳霉素组气管内给药复制大鼠肺纤维化动物模型(平阳霉素3mg/kg),生理盐水组气管内注射等量生理盐水,分别于7,28 d处死平阳霉素组大鼠10只,生理盐水组大鼠4只,其中平阳霉素组及生理盐水组各取3只提取肺成纤维细胞,平阳霉素组6只提取大鼠原代Ⅱ型肺泡上皮细胞,分别进行培养。待两种细胞均已贴壁后,再进行如下分组即:①实验组:用平阳霉素组肺成纤维细胞上清液加入平阳霉素组肺泡Ⅱ型上皮细胞并培养48 h;②对照组:用生理盐水组肺成纤维细胞上清液加入平阳霉素组肺泡Ⅱ型上皮细胞并培养48 h。用半定量RT-PCR法检测实验组和对照组中肺泡Ⅱ型上皮细胞SP-A、TGF-β及HGF mRNA表达的变化。结果:①SP-A mRNA和HGF mRNA的表达实验组低于对照组(P相似文献   

卫氏并殖吸虫两种途径感染实验鼠的结果比较

以经口或经腹途径,分别用卫氏并殖吸虫囊蚴感染小鼠和大鼠各一组,33～45天后剖查,平均获虫率小鼠经口感染组为18.0%,经腹感染组为69.45%;大鼠则分别为48.79%和53.15%,均以经腹感染的方式获虫率显著为高。实验鼠体内童虫的分布不因这两种感染方式而明显不同。小鼠组共检获472条童虫,85.59%分布在肌肉中。大鼠肌肉中的童虫更多,在检获的2553条虫子中占98.63%。     

Effects of all-trans retinoic acid on the expression of TGF-beta 1 and COL-I in rat model of peritoneal dialysis

OBJECTIVE: To investigate the effects of all-trans retinoic acid (ATRA) on the expression of transforming growth factor-beta 1 (TGF-beta 1) and collagen I (COL-I )in rat model of peritoneal dialysis, which may relate to the prevention peritoneal fibrosis. METHODS: Peritoneal dialysis model was established in rats, and then the rats were given ATRA 2 mg/kg (small dose group) or 5 mg/kg (large dose group) by the way of intraperitoneal injection once a day. All the rats were sacrificed on day 28. TGF-beta 1 and COL-I protein expression of peritoneum were measured by immunohistochemistry. TGF-beta 1 mRNA expression were examined with real time polymerase chain reaction (RT-PCR). RESULTS: Masson stain showed that the peritoneum thickness was significantly increased in the rats model, and collagen deposition was evident in the thickened submesothelial compact zone. With the treatment of ATRA, either in small or large dose, pathological changes were significantly lessened. The expression of TGF-beta 1 and COL-I of peritoneum was increased significantly in the rats model, but the levels in the two ATRA treated groups were lower than those of the untreated group. CONCLUSION: ATRA could decrease the experession of TGF-beta 1 and COL-I in peritoneum and delay the progression of peritoneal fibrosis.     

增生性瘢痕成纤维细胞P物质表达的研究

目的研究P物质（substance P，SP）在人增生性瘢痕体外培养的成纤维细胞的表达情况及其与正常皮肤之间的差异，探讨增生性瘢痕组织SP表达增高的原因。方法以酶消化法体外培养成人增生性瘢痕以及正常皮肤的成纤维细胞，将SP（终浓度为10^-7mol／L）加入培养液中刺激成纤维细胞，分别于刺激前和刺激后1、3、6、12、24h采用RT—PCR检测细胞内SP基因的表达情况，ELISA检测培养液中SP浓度。正常皮肤对照组及瘢痕对照组除不应用SP外，余处理均相同。结果未受SP刺激时，人增生性瘢痕及正常皮肤体外培养的成纤维细胞均能分泌一定浓度的SP，两者之间差异有统计学意义（P〈0．01）。SP刺激后，正常皮肤体外培养的成纤维细胞SP基因和蛋白表达均在1～3h内达到高峰，随后迅速下降，24h后仍高于其正常水平；而增生性瘢痕体外培养的成纤维细胞SP基因和蛋白表达在3～6h内达到高峰，随后缓慢下降，24h后仍高于其正常水平。两种成纤维细胞SP的分泌差异有统计学意义（P〈0．05）。结论外源性SP可诱使成纤维细胞内SP的分泌增加，而增生性瘢痕与正常皮肤成纤维细胞SP的分泌情况有明显差异。     

丹参注射液减轻腹透液致大鼠腹膜结构和功能损伤

目的：研究丹参注射液对腹透液（peritoneal dialysis solution,PDS）致大鼠腹膜组织结构和功能损伤的影响及与水通道蛋白1（aquaporin-1,AQP-1）和紧密连接相关蛋白1（zonula occluden-1,ZO-1）的关系。方法：56只SD大鼠分为正常对照组、1.5%PDS组、4.25%PDS组、1.5%PDS＋1%丹参组、4.25%PDS＋1%丹参组、1.5%PDS＋2%丹参组和4.25%PDS＋2%丹参组。腹膜透析液大鼠腹腔注射8周后,检测腹膜透析超滤量、腹膜对物质的清除率和腹透液漏出蛋白含量;光镜和电镜下观察腹膜结构变化;免疫印迹法检测腹膜AQP-1蛋白表达,免疫组织化学法检测腹膜间皮细胞ZO-1和AQP-1蛋白表达。结果：与正常对照组比较,1.5%PDS组和4.25%PDS组大鼠腹膜结构和功能发生改变,包括腹膜病理的改变明显,超滤量减少（P〈0.05）,对肌酐和葡萄糖清除能力下降（P〈0.01）,AQP-1蛋白表达增加（P〈0.05）;1.5%PDS＋2%丹参组和4.25%PDS＋2%丹参组大鼠腹膜结构和功能较单纯腹透液组改善,表现为病理改变缓解,腹膜超滤量以及肌酐和葡萄糖清除率提高。与1.5%PDS组比较,1.5%PDS＋2%丹参组腹膜AQP-1蛋白表达量减少（P〈0.05）。结论：丹参注射液可能通过下调AQP-1蛋白表达拮抗腹透液对腹膜组织结构和功能的损伤。