目的 研究白藜芦醇通过抑制T样受体4/核因子-κB(TLR4/NF-κB)通路对呼吸道合胞病毒(RSV)毛细支气管炎小鼠的保护作用。方法 选取30只小鼠随机分为对照组、RSV组、给药组,建立RSV毛细支气管炎小鼠模型,检测小鼠肺组织中TLR4、NF-κB的变化;利用肺组织HE染色、ELISA法检测白藜芦醇给药前后气道炎症病变、IL-6、TNF-α因子水平,Western Blot法及实时定量PCR法检测TLR4、 NF-κB蛋白及基因表达等相关变化。结果 与对照组相比,RSV组小鼠组肺组织中TLR4、NF-κB水平升高,肺组织切片HE染色显示气道炎症细胞浸润加剧,ELISA检测炎性因子IL-6、TNF-α升高;而给药组处理后,肺组织TLR4、NF-κB的表达下调,病理改变减轻,炎性因子IL-6、TNF-α下降。结论 白藜芦醇可通过抑制TLR4/NF-κB通路抑制炎性因子的释放,从而减轻毛细支气管炎小鼠的气道炎症反应。

Objective To study the protective effect of resveratrol on lung tissue of respiratory syncytial virus(RSV) bronchiolitis mice by regulating TLR4/NF-κB pathway. Methods Thirty mice were randomly divided into control group, RSV group and resveratrol group. The mice models of RSV bronchiolitis were established, and the changes of TLR4 and NF-κB levels in lung tissues of mice were detected. HE staining and ELISA were used to detect airway inflammation, IL-6 and TNF-α levels before and after resveratrol administration. TLR4 and NF-κB protein and gene expressions were detected by Western Blot and real-time quantitative PCR. Results Compared with the control group, the levels of TLR4 and NF-κB in the lung tissues of mice with bronchiolitis were significantly increased. Airway inflammatory cell infiltration was aggravated in HE staining of lung tissue sections, and inflammatory factors IL-6 and TNF-α were significantly increased showing by ELISA. The expressions of TLR4 and NF-κB in resveratrol group were down-regulated after treatment. Conclusions Resveratrol can inhibit the release of inflammatory factors by inhibiting the TLR4/NF-κB pathway, play a protective role in mice with bronchiolitis.

目的 探讨新生儿坏死性小肠结肠炎(NEC)炎症损伤与肠道微生态-LPS-TLR4通路之间的关系。方法 本研究收集2019年3月1日—2021年1月31日在中山市人民医院新生儿监护室确诊为NEC新生儿11例为实验组,随机选取30 例同期在新生儿科病房住院喂养顺利,排除NEC及败血症诊断的新生儿为对照组。采集2组新生儿的粪便标本,进行Real-time PCR表达谱分析2组粪便肠道菌群;取2组外周静脉血检测外周血单核细胞Toll样受体4(TLR4)和血清PCT、CRP、IL-6、SAA等指标,对比2组肠道菌群、外周血单核细胞TLR4和炎症指标水平,通过统计学分析组间差异。结果 本研究结果提示实验组变形菌门占82%(9/11),厚壁菌门占9%(1/11),放线菌门占9%(1/11),对照组变形菌门占20%(6/30),厚壁菌门占73%(22/30),放线菌门占7%(2/30),2组患儿的粪便肠道菌群分布有差异(χ²=11.521,P<0.05);实验组患儿外周血单核细胞TLR4水平高于对照组,组间差异有统计学意义(P<0.001);实验组患儿血清PCT、CRP、IL-6和SAA等炎症指标高于对照组,组间差异有统计学意义(P<0.001)。结论 NEC患儿的肠道菌群以变形菌门为主,伴外周血单核细胞TLR4和外周血炎症指标升高。可见,肠道微生态-LPS-TLR4通路可能与新生儿坏死性小肠结肠炎炎症损伤相关,具体的机制仍需进一步深入研究。

Objective To investigate the relationship between intestinal flora-LPS-TLR4 pathway and the inflammatory injury of neonatal necrotizing enterocolitis (NEC). Methods Eleven neonates with NEC from March, 2019 to January, 2021 were enrolled as the experimental group, and 30 neonates without NEC and septicemia who were admitted in the department of neonatology in the same period were included as the control group. Faecal flora from the two groups were collected and analyzed by Real-time PCR. Toll-like receptor 4 (TLR4) and serum PCT, CRP, IL-6, SAA in peripheral blood were measured. The intestinal flora, the expression of TLR4 in peripheral blood leukocytes and inflammatory markers were compared between two groups. Results It showed that the ratio of Proteobacteria was 82% (9/11), Firmicutes was 9% (1/11), Actinobacteria was 9% (1/11) in the experimental group. In the control group, the ratio of Proteobacteria was 20% (6/30), Firmicutes was 73% (22/30), Actinobacteria was 7% (2/30). There was a significant difference in the distribution of faecal flora between the two groups (χ² = 11.521, P<0.05), and the level of TLR4 in peripheral blood of the experimental group was significantly higher than that of the control group (P＜0.001). The levels of serum PCT, CRP, IL-6 and SAA in the experimental group were significantly higher than those in the control group (P＜0.001). Conclusions The main intestinal flora of neonates with NEC is Proteobacteria, with elevated TLR4 expression and inflammatory markers in peripheral blood. Therefore, the intestinal flora-LPS-TLR4 pathway may be associated with inflammatory injury in neonatal necrotizing enterocolitis.The specific mechanism still needs further study.

目的 探讨血必净注射液对SAP大鼠TLR4信号通路介导肠黏膜屏障功能障碍的影响。方法 24只SD大鼠随机分成空白组(n=8)、对照组(n=8)和治疗组(n=8)。对照组和治疗组用4.5%牛磺胆酸钠溶液胆胰管逆行注射制备SAP模型,空白组采用等量生理盐水逆行注射。治疗组在造模3 h后经鼠尾静脉注射血必净注射液(3 mL/kg)。三组大鼠造模后观察24 h,然后处死取胰腺和小肠组织送病理检查,采用荧光RT-PCR技术检测TLR4和NF-κB表达水平,采用ELSIA法检测血清TNF-α、IL-6、淀粉酶(AMS)及二胺氧化酶(DAO)水平,比较三组大鼠各项指标。结果 对照组和治疗组小肠组织TLR4和NF-κB表达以及血清TNF-α、IL-6、AMS及DAO水平均高于空白组(P>0.05),治疗组小肠组织TLR4和NF-κB表达以及血清TNF-α、IL-6、AMS及DAO水平低于对照组(P＜0.05)。结论 血必净注射液通过干预SAP大鼠TLR4信号通路,降低小肠组织TLR4和NF-κB的表达,减轻小肠组织的炎症反应,对肠黏膜屏障具有一定的保护作用。

Objective To investigate the effect on intestinal mucosal barrier dysfunction (IBF) of Xuebijing injection mediated by Toll-like receptor 4 (TLR4) signal pathway in rats of severe acute pancreatitis (SAP). Methods 24 Sprague Dawley (SD) rats were randomly divided into Sham group (n=8), control group (n=8) and treatment group (n=8). The SAP model was established by retrograde injection of 4.5% sodium taurocholate into the biliopancreatic duct in control group and treatment group, while control group was injected with the same amount of saline. In treatment group, Xuebijing injection (3 mL/kg) was injected via tail vein after 3h of modeling. All rats were monitored and sacrificed after 24 hours of modeling. Samples of pancreas and intestine were collected for pathologic determination. A fluorescent RT-PCR was used to determine the expression of TLR4 and NF-κB of small intestine. The serum levels of TNF-α, IL-6, amylase (AMS) and diamine oxidase (DAO) were measured by using ELISA. All parameters of three groups were compared. Results The expression of TLR4 and NF-κB of small intestine in control group and treatment group were higher than it in control group (P<0.05), as well as the serum levels of TNF-α, IL-6, AMS and DAO (P<0.05). The expression of TLR4 and NF-κB of small intestine in treatment group were lower than it in control group (P<0.05), as well as the serum levels of TNF-α, IL-6, AMS and DAO (P<0.05). Conclusion Xuebijing injection may not only reduce the expression of TLR4 and NF-κB of small intestine, but also alleviate the inflammation reaction of small intestine by interfering with TLR4 signal pathway, which may have a protective effect on intestinal mucosal barrier in SAP rats.

目的 探讨川芎嗪对链脲佐菌素(STZ)诱导2型糖尿病大鼠肾病的治疗作用及机制。方法 SD大鼠50只,随机分为正常组和模型组。除正常组外,其余大鼠均给予高脂-高糖饲料喂养4周,再给予链脲佐菌素(40 mg/kg,ip),72 h后测定空腹血糖,将血糖值高于16.67 mmol/L的大鼠随机分成4个组即模型组,二甲双胍阳性组(250 mg/kg),川芎嗪低、高剂量组(80、160 mg/kg),连续给予相应试药8周。其中正常组和模型组的大鼠均给予同等量蒸馏水灌胃。实验结束时,测定大鼠血糖、尿蛋白、血尿素氮和血肌酐含量;免疫组化法测定大鼠肾组织TLR4和caspase3蛋白表达。光镜下观察肾脏病理学变化。结果 与模型组比较,二甲双胍组和川芎嗪高剂量组给药8周后,大鼠动态空腹血糖均能明显降低(P<0.05),大鼠动态尿蛋白显著性降低(P<0.01,P<0.05); 二甲双胍和高剂量组TLR4和caspase3蛋白表达明显低于模型组(P<0.05);肾脏组织病理性损伤明显减轻。结论 川芎嗪对STZ诱导2型糖尿病大鼠肾病具有保护作用,其机制可能与下调TLR4表达作用有关。

Objective To investigate the therapeutic effects and mechanisms of tetramethylpyrazine (TMP) on streptozocin(STZ)-induced-nephropathy in type 2 diabetic rats. Methods 50 SD rats were randomly divided into normal group(n=10) and model group(n=40). The model rats were fed on high fat and sugar diets for 4 weeks, then given STZ(40 mg/kg,ip). After 72 hours, the fasting blood glucose (FBG) was measured. Rats with high FBG above 16.67mmol/L were randomly divided into four groups: model, metformin(Met, 250 mg/kg)and TMP (80 mg/kg, 160 mg/kg) groups for treating 8 weeks, and both the control and model groups were given equals distilled water by intragastric administration. At the end of the experiment, blood glucose, urine protein, blood urea nitrogen and creatinine were measured. The expression of TLR4 and caspase3 protein in kidney tissue of rats was determined by immunohistochemistry. Pathological changes of kidney were observed under light microscope. Results Compared with the model group, metformin and high dose of TMP administered after 8 weeks, rats can significantly reduce the dynamic fasting blood glucose(P<0.05). Urinary protein excretion of total dynamic decreased significantly (P<0.01, P<0.05); the protein expression of TLR4 and caspase3 in the metformin group and high dose group was significantly lower than that in the model group (P<0.05); kidney tissue pathological damage was significantly reduced. Conclusion TMP has a protective effect on STZ induced nephropathy in type 2 diabetic rats, and its mechanism may be related to the down-regulation of TLR4 expression.