活体成像技术观察胶质瘤荷瘤鼠中过表达SASH1基因的作用*

张思铭1**,马 1,胡樱子2,尤正晨3,陈 3,巫荣华4,杨 3,刘 4

(南通大学1杏林学院医学部;2医学院口腔系,江苏226001

3南通大学附属医院神经外科;4南通大学神经再生重点实验室)

[ ] 目的:采用小动物活体成像技术,观察过表达SASH1对荷胶质瘤裸鼠的抗肿瘤生长作用。方法:将稳定表达GFP绿色荧光蛋白及荧光素酶报告基因(luciferase)的胶质瘤SHG-44细胞接种于裸鼠皮下,待成瘤后,将ADV4-SASH1慢病毒注射入裸鼠肿瘤组织中,对照组注射相同剂量的ADV4-NC空载体对照病毒;1周后,腹腔注射底物荧光素(luciferin),用小动物活体成像仪采集荧光值,分析肿瘤生长情况。结果:过表达SASH1的裸鼠中有70%7/10)肿瘤减小;对照组中有71.4%5/7)裸鼠肿瘤增加,且有30%3/10)裸鼠死亡。过表达SASH1 1周后肿瘤大小减小至90%,对照组肿瘤增大至166%。结论:本实验表明活体成像技术可以实时观察荷瘤鼠异位接种的胶质瘤体的生长情况,初步结果表明在胶质瘤体中过表达SASH1基因可以抑制肿瘤的生长。

[关键词] SASH1;病毒载体;胶质瘤荷瘤鼠;小动物活体成像

[中图分类号] Q334+.13 [文献标志码] A

In vivo bioluminescent analysis on investigating effects of overexpressing SASH1 gene on glioma growthof the nude mice

ZHANG Siming1, MA Chao1, HU Yingzi2, YOU Zhengchen3, CHEN Han3, WU Ronghua4, YANG Liu3, LIU Mei4

(1Division of Medicine, Xinglin College; 2Departmet of Stomatology, Medical College; 3Departmant of Neurosurgery, Affiliated Hospital of Nantong University; 4Key Laboratory of Neuroregeneration, Nantong University, Jiangsu 226001)

[Abstract] Objective: To investigate theeffect of SASH1 gene overexpression on glioma by using bioluminescent imaging of animal in vivo imaging system. Methods: Glioma bearing mice model was established by inoculatingsubcutaneously SHG-44 cells with stably expressing GFP-Luci gene. After neoplasm emergence, ADV4-SASH1 were injected into tumors, while control mice were treated with ADV4-NC virus. After 1 week, luciferin was injected and the bioluminescent values were collected by animals in vivo imaging system, and the growth of the tumor was analyzed. Results: 70% of the tumors growth of the nude mice with an injection of ADV4-SASH1 decreased, while in the nude mice with an injection of the control virus, 50% of the tumors growth increased, and 30% of the nude mice died. Conclusion: In this study, it is shown that in vivo imaging technique could trace the growth of inoculated glioma in the nude mice. The preliminary results showed that overexpression of SASH1 could inhibit the glioma growth in vivo.

[Key words] SASH1; virus vector; glioma bearing the nude mice; animal in vivo image system

健肠Ⅰ号方联合“治未病”健康管理防治腹泻型肠易激综合征*

1**,余利华2,季 1,朱时林1,田祖成1,梅 1,代海峰1,葛乃建3

1海安县中医院消化科,江苏2266002无锡市中西医结合医院消化科;

3第二军医大学东方肝胆外科医院微创介入中心)

[ ] 目的:探讨“健肠Ⅰ号”方联合“治未病”健康管理治疗腹泻型肠易激综合征的效果,观察患者血清脑肠肽水平变化。方法:120例腹泻型肠易激综合征患者随机分为观察1组、观察2组和对照组各40例,对照组予以匹维溴铵治疗,观察1组予以健肠Ⅰ号方治疗,观察2组在健肠Ⅰ号方基础上给予“治未病”健康管理,观察3组有效率、消化道症状评分、血清脑肠肽水平变化及不良反应。结果:总有效率观察2组为97.5%,观察1组为85.0%,均高于对照组的65.0%,差异均有统计学意义(P0.05);治疗4周后,3组消化道症状各项评分较治疗前降低,差异均有统计学意义(P0.05),观察2组、观察1组与对照组比较,症状评分降低更明显,差异均有统计学意义(P0.05);治疗10周后,对照组、观察1组消化道症状评分又升高,与治疗前相比差异均无统计学意义(P0.05),而观察2组治疗10周后消化道症状评分无明显升高,与治疗前相比差异仍有统计学意义(P0.05)。治疗4周后,3组脑肠肽水平均显著下降,与治疗前相比差异均有统计学意义(P0.05),观察2组、观察1组与对照组比较,脑肠肽水平降低更明显,差异均有统计学意义(P0.05);治疗10周后,对照组、观察1组脑肠肽水平又升高,与治疗前相比差异均无统计学意义(P0.05),而观察2组治疗10周后脑肠肽水平无明显升高,与治疗前相比差异仍有统计学意义(P0.05)。观察1组、观察2组不良反应发生率均为2.5%,低于对照组的20.0%,差异有统计学意义(P0.05)。结论:健肠Ⅰ号方治疗腹泻型肠易激综合征的效果优于单用匹维溴铵,联合“治未病”健康管理则疗效更佳,且能有效预防复发,值得临床推广应用。

[关键词] 肠易激综合征;健肠Ⅰ号;治未病;脑肠轴

[中图分类号] R574.4 [文献标志码] A

The clinical effect of the“Jianchang NO.1”recipe combined with health management mode in the prevention and treatment of IBS-D

GE Fei1, YU Lihua2, JI Yu1, ZHU Shilin1, TIAN Zucheng1, MEI Li1, DAI Haifeng1,GE Naijian3

(1Department of Gastroenterology, Hai’an County Hospital of Traditiona Chinese Medicine, Jiangsu 226600;

2Department of Gastroenterology, Wuxi Hospital of Integrated Traditional Chinese and Western Medicine;

3Minimally Invasive Interventional Center, Eastern Hepatobiliary Surgery Hospital, Second Military Medical University)

[Abstract] Objective:To study the clinical effect of “Jianchang NO.1”recipe combined with health management mode of “preventive treatment of disease”in the prevention and treatment of diarrhea-predominant irritable bowel syndrome. Methods:120 outpatients with diarrhea-predominant irritable bowel syndrome were randomly divided into the observation group1, observation group2 and the control group with 40 cases in each group. The control group was orally treated with pinaverium bromide,the observation group1 was given“Jianchang NO.1”recipe treatment,observation group2 was given“Jianchang NO.1”recipe combined with health management mode of “preventive treatment of disease”. Then the efficiency,gastrointestinal symptoms scores,changes of brain gut peptide levels and adverse reactions in these three groups were observed. Results:The total effective rate of the observation group2(97.5%) and group1(85.0%) were significantly higher than that of the control group(65.0%)(P<0.05); the gastrointestinal symptom score of the three groups were significantly lowered after 4 weeks of treatment(P<0.05), and the observation group2 was most obviously lowered. However, after ten weeks’follow-up observation, the gastrointestinal symptom scores of the control group and the observation group1 both rose much higher(P>0.05), comparing with the scores at the prior treatment; Also, there were significant changes in digestive symptom score in the observation group2, that compared with the score of the prior treatment(P<0.05). The levels of the gut-brain peptide in these three groups were significantly decreased after treatment(P<0.05), and the observation group2 decreased most significantly(P<0.05). The follow-up resultsat the 10th week after treatment:The brain gut peptide levels of the control group and observation group1 were not significantly higher than the levels of the prior treatment (P>0.05), but the brain-gut peptide levels of the observation group2 didn’t significantly change when compared with the levels after the prior treatment (P<0.05). The incidence of the adverse reactionsin the two observation groups(2.5%) was significantly lower than that of the control group(20.0%)(P<0.05). Conclusions:The curative effects of the “Jianchang NO.1”recipe were much better than that of pinaverium bromide in the treatment of IBS, and the clinical effects were much better when it was combined with health management mode of “preventive treatment of the disease”. This combination therapy could also prevent the recurrence of IBS effectively, and was worthy of clinical application.

[Key words] irritable bowel syndrome; Jianchang NO.1 recipe; preventive treatment of disease; brain gut axis

RhoGDI1TNF-α诱导内皮细胞损伤中的作用研究*

秦峥幸**,王小丽,周楚绮,张 香,任 颖,姚文娟

(南通大学药学院药理系,江苏 226001

[ ] 目的:阐明Rho特异鸟苷酸解离抑制因子1RhoGDI1)和Rho/ROCK信号通路在肿瘤坏死因子α(TNF-α)诱导内皮细胞损伤中的作用机制。方法:采用siRNA技术干扰人脐静脉内皮细胞RhoGDI1表达,实验分为正常对照组,10μg·L-1 TNF-α处理组和RhoGDI1干扰组。MTT法检测细胞存活率,TUNEL法检测细胞凋亡,Western blot检测RhoGDI1RhoAROCK表达及活化。结果:TNF-α处理和RhoGDI1干扰均显著降低内皮细胞存活率,提高内皮细胞的凋亡。TNF-α处理后RhoGDI1表达显著下降,TNF-α处理和RhoGDI1干扰均可显著提高RhoA表达以及ROCK活化。结论:TNF-α可能通过抑制RhoGDI1促进Rho/ROCK信号通路的活化,最终引起内皮细胞凋亡。

[关键词] 内皮损伤;肿瘤坏死因子α;Rho/ROCK信号通路;Rho特异鸟苷酸解离抑制因子1

[中图分类号] Q25 [文献标志码] A

The role of RhoGDI1 in endothelial cell injury induced by TNF-α

QIN Zhengxing, WANG Xiaoli, ZHOU Chuqi, ZHANG Xiang, REN Ying, YAO Wenjuan

(Department of Pharmacology, School of Pharmacy, Nantong University, Jiangsu 226001)

[Abstract] Objective: To elucidate the mechanism of Rho-specific guanine nucleotide dissociation inhibitor 1 (RhoGDI1) and Rho/ROCK signaling pathway in endothelial cell injury induced by tumor necrosis factor-α(TNF-α). Methods: RhoGDI1 expression was interfered in human umbilical vein endothelial cells by siRNA technique. The experiment was divided into 3 groups: the normal control group, the 10μg·L-1 TNF-α treatment group, and the RhoGDI1 interference group. MTT method was used to detect cell viability. TUNEL assay was used to detect cell apoptosis. The expression and activation of RhoGDI1, RhoA and ROCK were detected by Western blot analysis. Results:TNF-α treatment or RhoGDI1 suppression could significantly decrease the survival rate of endothelial cells and increase the endothelial apoptosis. Western blot showed that TNF-α treatment significantly decrease the expression of RhoGDI1. Both TNF-α treatment and RhoGDI1 suppression could significantly enhance the expression of RhoA and activation of ROCK. Conclusion:TNF-α may promote the activation of Rho/ROCK signaling pathway by inhibiting RhoGDI1, and finally induce apoptosis in endothelial cells.

[Key words] endothelial dysfunction; tumor necrosis factor-α; Rho/ROCK signaling pathway; Rho-specific guanine nucleotide dissociation inhibitor 1