Jianpi Gushen Huayu decoction ameliorated diabetic nephropathy through modulating metabolites in kidney,and inhibiting TLR4/NF-κB/NLRP3 and JNK/P38 pathways

BACKGROUND Jianpi Gushen Huayu Decoction(JPGS)has been used to clinically treat diabetic nephropathy(DN)for many years.However,the protective mechanism of JPGS in treating DN remains unclear.AIM To evaluate the therapeutic effects and the possible mechanism of JPGS on DN.METHODS We first evaluated the therapeutic potential of JPGS on a DN mouse model.We then investigated the effect of JPGS on the renal metabolite levels of DN mice using non-targeted metabolomics.Furthermore,we examined the effects of JPGS on c-Jun N-terminal kinase(JNK)/P38-mediated apoptosis and the inflammatory responses mediated by toll-like receptor 4(TLR4)/nuclear factor-kappa B(NF-κB)/NOD-like receptor family pyrin domain containing 3(NLRP3).RESULTS The ameliorative effects of JPGS on DN mice included the alleviation of renal injury and the control of inflammation and oxidative stress.Untargeted metabolomic analysis revealed that JPGS altered the metabolites of the kidneys in DN mice.A total of 51 differential metabolites were screened.Pathway analysis results indicated that nine pathways significantly changed between the control and model groups,while six pathways significantly altered between the model and JPGS groups.Pathways related to cysteine and methionine metabolism;alanine,tryptophan metabolism;aspartate and glutamate metabolism;and riboflavin metabolism were identified as the key pathways through which JPGS affects DN.Further experimental validation showed that JPGS treatment reduced the expression of TLR4/NF-κB/NLRP3 pathways and JNK/P38 pathway-mediated apoptosis related factors.CONCLUSION JPGS could markedly treat mice with streptozotocin(STZ)-induced DN,which is possibly related to the regulation of several metabolic pathways found in kidneys.Furthermore,JPGS could improve kidney inflammatory responses and ameliorate kidney injuries in DN mice via the TLR4/NF-κB/NLRP3 pathway and inhibit JNK/P38 pathwaymediated apoptosis in DN mice.

Ercao Qinggan decoction regulates apoptosis of hepatocytes in mice with acute liver failure via the TLR4-mediated PI3K/Akt/GSK3βsignal pathway

Background:To clarify the inhibitory effect of Ercao Qinggan decoction(EQD)on acute liver failure(ALF)and its related mechanisms.Methods:HL-7702 hepatocytes were pretreated with TLR4 inhibitor CLI-095,glycogen synthase kinase 3β(GSK3β)inhibitor LiCl and different doses of EQD for 2 hours,and lipopolysaccharide(LPS)(10μg/mL)for 24 hours.Cell apoptosis,TNF-αand IL-6 and GSK3βwere detected by flow cytometry,immunofluorescence,quantitative polymerase chain reaction.After mice were gavaged with different concentrations of EQD for 12 days,ALF mouse models were established intraperitoneal injection of D-Gal/LPS.After 24 hours,the mice were euthanized and the liver tissue was stained with hematoxylin and eosin.Liver cell apoptosis,the serum levels of aspartate aminotransferase,alanine aminotransferase,TNF-αand IL-βwere detected by terminal transferase-mediated dUTP nick end-labelling,enzyme linked immunosorbent assay,quantitative polymerase chain reaction,and Western blotting,respectively.These methods were also used to test the mRNA expression of Bax,Bcl-2 and the protein expression of GSK3β,p-Akt/Akt in livers.Results:CLI-095,LiCl,and EQD significantly inhibited apoptosis induced by LPS,the mRNA expression of IL-6,TNF-αand the nuclear translocation of GSK3βin HL-7702 hepatocytes.EQD dose-dependently inhibited hepatocyte apoptosis,the serum concentration of aspartate aminotransferase and ALT,the expression of TNF-αand IL-β,the ratio of p-GSK3β/GSK3β,p-Akt/Akt in alanine aminotransferase mice.Conclusion:EQD can inhibit hepatocyte apoptosis in ALF mice through regulating TLR4/PI3K/Akt/GSK3βsignaling pathway.

Mechanism of Sanshi decoction inhibits macrophage pyroptosis by inhibiting BRD4/NF-κB/NLRP3 pathway in the treatment of gouty arthritis

Objective:To observe the effect of Sanshi decoction on BRD4/NF-κB/NLRP3 pathwaymediated macrophage pyroptosis,so as to elucidate the molecular mechanism of Sanshi decoction in the treatment of gouty arthritis.Methods:THP-1 was induced into macrophages with foboside and the divided into the control group,model group,low-dose,medium-dose,high-dose group of Sanshi decoction,and BRD4 inhibitor group.Except for the control group,the remaining groups were induced with monosodium urate crystals to construct a gouty arthritis cell model.The activity of macrophages was detected by CCK8,the level of macrophage pyroptosis was detected by flow cytometry,the activity of LDH,the content of IL-1β and IL-18 were detected by enzyme-linked immunosorbent assay,and the expression of related proteins in the BRD4/NF-κB/NLRP3 pathway was detected by Western blot.Results:Compared with the control group,macrophage activity was decreased in the model group,and the level of pyroptosis,LDH activity,contents of IL-1β and IL-18,expression levels of BRD4,p-NF-kB p65,NLRP3,Caspase-1 p20,and IL-1β protein were significantly up-regulated,the differences were statistically significant(P<0.05 and P<0.01).Compared with the model group,macrophage activity was up-regulated in the Sanshi Decoction,and the level of pyroptosis,LDH activity,IL-1β and IL-18 contents,expression levels of BRD4,p-NF-kB p65,NLRP3,Caspase-1 p20,and IL-1β protein were significantly decreased with statistically significant differences(P<0.05 and P<0.01).Conclusion:Sanshi decoction inhibits macrophage pyroptosis by inhibiting BRD4/NF-κB/NLRP3 pathway activation,thus improving the inflammation level of gouty arthritis.

Mechanic study of Qushi Kaiyu decoction on non-alcoholic fatty liver disease model rats based on the inhibition of TLR4/NF-ĸB pathway

Background:The objective of this research was to examine the impact of the Chinese herbal formula Qushi Kaiyu(QSKY)on rats with non-alcoholic fatty liver disease(NAFLD)and its inhibitory effect on the TLR4/NF-ĸB pathway.Methods:NAFLD model rats was constructed through high-fat diet.Meanwhile,rats were treated with QSKY(6.4 g/kg)by gavage.The therapeutic effect of QSKY on NAFLD was assessed by testing body weight change,the liver index,lipid concentrations in blood,and antioxidant and inflammatory levels;assessing liver function;and performing pathological staining including hematoxylin-eosin and Oil Red O.The protein levels of key factors in the TLR4/NF-ĸB pathway(TLR4,MyD88,p65 and IKB)in rat liver tissue were determined using western blotting in order to explore the mechanism responsible for the therapeutic effects of QSKY in rats with NAFLD.Results:QSKY significantly reduced the liver index and body weight value;reduced triglyceride,cholesterol,alanine aminotransferase,and aspartate aminotransferase levels in NAFLD rats;improved the pathological changes,such as ballooning degeneration,fat accumulation,necrosis,and inflammation;elevated GSH-Px and superoxide dismutase activities and lowered malondialdehyde levels,indicating that QSKY enhanced the antioxidant capacity;and reduced inflammatory cytokine(IL-6,IL-1β,and TNF-α)levels.Western blotting results showed that QSKY significantly reduced TLR4,MyD88,and decreased the phosphorylation of IKB and p65 protein levels in the livers of rats with NAFLD.Conclusions:QSKY showed therapeutic effects on NAFLD and can alleviate oxidative stress and inflammation.This mechanism may be related to an improvement in TLR4/NF-ĸB pathway.

加减参芪地黄汤治疗慢性肾脏病3-4期的临床研究

目的探讨加减参芪地黄汤在慢性肾病3-4期患者临床治疗中的作用。方法将2019年1月至2023年5月在苏州市中医医院住院及门诊治疗的慢性肾病3-4期患者60例,按随机数字表分组,各30例。对照组在常规治疗的基础上辅以肾衰宁片口服,治疗组在常规治疗的基础上给予加减参芪地黄汤中药治疗。检测两组患者肌酐、尿素氮、尿酸等指标,比较两组患者中医证候积分及临床疗效。结果治疗组总有效率为86.67%,显著高于对照组(77.33%)(P<0.05)。治疗组治疗后血Scr、BUN、UA显著低于对照组,差异具有统计学意义(P<0.05)。治疗组倦怠乏力、食少纳呆、腰膝酸软、肢体困重等证候积分与对照组比较均有统计学差异(P<0.01或P<0.05)。结论加减参芪地黄汤对慢性肾病3-4期患者疗效良好,可降低血肌酐、尿素氮、尿酸,改善临床症状,延缓慢性肾脏病进展。

化痰通遂汤联合督脉三针对脑卒中后吞咽障碍患者脂质过氧化及血清NPAS4、PARK7的影响

目的探讨化痰通遂汤联合督脉三针对脑卒中后吞咽障碍对患者脂质过氧化及血清NPAS4、PARK7的影响。方法研究将前瞻性选取2020年3月—2022年4月在医院诊疗的86例脑卒中后吞咽障碍患者为受试对象,根据数字表法将其分成试验组与对照组,各43例,对照组予以化痰通遂汤治疗,试验组予以化痰通遂汤治疗的同时采用督脉三针治疗,密切观察并对比两组研究对象的疗效,治疗前后的氧化应激和脂质过氧化指标,血清NPAS4、PARK7水平,NIHSS评分、FMA评分、SSA评分及SIS评分。结果应用化痰通遂汤联合督脉三针治疗后的试验组疗效明显高于单纯应用化痰通遂汤治疗的对照组(P<0.05);治疗后两组患者的SOD、iso-PGs指标较治疗前均上升(P<0.05),且试验组SOD指标高于对照组(P<0.05),但试验组iso-PGs指标较治疗前无明显差异(P>0.05),且试验组低于对照组(P<0.05),MDA指标治疗较治疗前显著下降(P<0.05),且试验组低于对照组(P<0.05);治疗前两组的NIHSS评分、SSA评分、FMA评分及SIS评分均无显著性差异(P>0.05),治疗后试验组患者的FMA评分及SIS评分均显著高于对照组(P<0.05),而NIHSS评分、SSA评分显著低于对照组(P<0.05);治疗前两组血清NPAS4、PARK7水平较治疗前均无显著性差异(P>0.05),且试验组患者血清NPAS4、PARK7水平均显著低于对照组(P<0.05)。结论应用化痰通遂汤联合督脉三针治疗脑卒中后吞咽障碍,效果极佳,联用能够改善氧化应激以及脂质过氧化指标,降低血清NPAS4、PARK7水平,提高患者生存水平,安全可靠,临床应用前景较为宽阔。

补阳还五汤增强GPX4表达抑制铁死亡减轻脑缺血再灌注损伤

目的探讨补阳还五汤减轻脑缺血再灌注损伤是否通过增强GPX4的表达抑制铁死亡的机制。方法选取健康雄性SD大鼠48只,随机分为4组:假手术组(Sham组)、模型组(MCAO/R组)、补阳还五汤组(BYHWD组)、阳性对照药组(DFO组)。所有实验动物均建MCAO/R模型(假手术组仅剥离)。BYHWD组给予补阳还五汤灌胃处理,DFO组给予DFO腹腔注射处理,其余组予以生理盐水灌胃处理。再灌注72 h进行指标评定及取材。神经功能评分检测神经功能;TTC染色测定脑梗死面积;尼氏染色观察脑组织形态结构;生化试剂检测血清GSH、丙二醛(MDA)、脑组织铁含量;免疫荧光和Western blot检测谷胱甘肽过氧化物酶4(GPX4)的蛋白表达。结果与假手术组相比,模型组大鼠神经功能损伤明显,脑梗死体积增多,脑组织病理学损伤明显,尼氏体数量减少,脑组织铁含量、血清MDA含量明显增加,血清GSH含量、脑组织GPX4的蛋白表达显著降低(P<0.05);与模型组相比,补阳还五汤组大鼠神经功能明显好转,脑梗死体积降低,脑组织病理学损伤减轻,脑组织铁含量、血清MDA含量降低,血清GSH含量,脑组织GPX4的蛋白表达显著增加(P<0.05)。结论补阳还五汤抑制脑缺血再灌注损伤可能通过增强GPX4的表达抑制铁死亡而发挥作用。

清解化攻方调控NLRP3/TLR4/NF-κB信号通路对重症急性胰腺炎小鼠模型胰腺组织的保护作用

目的观察清解化攻方对重症急性胰腺炎(SAP)小鼠模型的治疗作用,探索清解化攻方抗炎症反应的作用机制。方法将36只C57BL/6J雄性小鼠随机分成空白组,模型组,清解化攻方低、中、高剂量组,西药组(乌司他丁),每组6只,除空白组小鼠,余各组小鼠采用逆行胰胆管注射5%牛黄胆酸钠建立SAP模型,清解化攻方低、中、高剂量组在造模后分别予以清解化攻方1、2、4 g/kg灌胃,西药组在造模后予以腹腔注射乌司他丁(5×10^(4) U/kg),共干预7 d。采用苏木素-伊红染色观察胰腺组织病理改变;酶联免疫吸附测定法(ELISA)检测小鼠α-淀粉酶、脂肪酶、IL-1β、IL-6、IL-8、IL-18和TNF-α水平;RTqPCR检测胰腺组织NOD样受体蛋白3(NLRP3)、Toll样受体4(TLR4)、核因子-κB(NF-κB)mRNA表达水平;免疫组化检测胰腺组织NLRP3、TLR4、NF-κB的阳性表达率;Western Blot技术检测NLRP3、TLR4、NF-κB、IL-1β、IL-6蛋白的表达水平。计量资料多组间比较采用方差分析,进一步两两比较采用LSD-t检验。结果与空白组相比,模型组小鼠胰腺组织结构弥漫性破坏、胰腺小叶间隔局灶性扩张、腺泡萎缩和大量炎症细胞浸润,α-淀粉酶、脂肪酶、IL-1β、IL-6、IL-8、IL-18和TNF-α含量明显升高(P值均<0.05),NLRP3、TLR4、NF-κB mRNA表达水平及阳性表达率均明显上升(P值均<0.05),NLRP3、TLR4、NF-κB、IL-1β、IL-6蛋白表达均明显上调(P值均<0.05)。与模型组相比,清解化攻方各剂量组和西药组可见小鼠胰腺组织结构稍紧密、完整,胰腺腺泡细胞排列有序,伴少量炎症细胞浸润和胰腺小叶出血灶,α-淀粉酶、脂肪酶、IL-1β、IL-6、IL-8、IL-18和TNF-α含量明显下降(P值均<0.05),NLRP3、TLR4、NF-κB mRNA表达水平及阳性表达率均明显降低(P值均<0.05),NLRP3、TLR4、NF-κB、IL-1β、IL-6蛋白表达水平均明显减弱(P值均<0.05)。结论清解化攻方可能通过抑制NLRP3/TLR4/NF-κB信号通路相关蛋白的激活,减少炎症介质的释放,防止炎症级联反应增强,进而对SAP小鼠胰腺组织发挥保护作用。

Effect of Buyanghuanwu Decoction on PI3K/AKT Signaling Pathway and Aquaporin AQP4 in Cerebral Hemorrhage Rats

[Objectives] To explore the effect of Buyanghuanwu decoction on PI3K/AKT signaling pathway and aquaporin AQP4 in cerebral hemorrhage rats and clarify the mechanism to provide clear direction and target for cerebral hemorrhage treatment caused by cerebral edema.[Methods]SD rats were randomly divided into six groups: model group,sham operation group,Buyanghuanwu decoction low,medium and high dose groups,and Ginkgo biloba group. Model group,Buyanghuanwu decoction group,G. biloba group were prepared to be intracerebral hemorrhage rat models by referring to Rosenberg law. While the expression of " polarity" of aquaporin AQP4 was detected by immunofluorescence labeling method,the Evans blue( Evans Blue,EB) content of brain tissue was determined by Spectrophotometry. In addition,the water content of brain tissue was detected by wet and dry weight method. [Results] When compared to the model group,the Buyang Huanwu decoction group,G. biloba group of PI3K and AKT proteins expression increased significantly( P < 0. 05) and AQP4 in Astrocyte end feet membrane concentrated expression significantly increased( P < 0. 05),EB content and water content of brain tissue significantly reduced( P <0. 05).[Conclusions]The protective mechanisms of Buyanghuanwu decoction on cerebral hemorrhage can work might because it can activate PI3K/AKT signaling pathway,regulate AQP4 " polar" expression,and reduce the permeability of the blood brain barrier and cerebral edema.

Effect of Maxingganshi decoction on the expression of STAT4 and STAT6 in lung tissues of rats with chronic obstructive pulmonary disease

Objective:To investigate the effect of Maxingganshi decoction on the expression of STAT4 and STAT6 in lung tissues of rats with chronic obstructive pulmonary disease (COPD).Methods:A total of 30 COPD Wistar rat models with phlegm-heat depression and lung depression were established, with half male and half female. The 30 model rats were fed in different cages, weighed and labeled. They were randomly divided into three groups: Maxingganshi decoction group, roxithromycin tablets control group, model control group. At the same time, 10 normal rats were selected as a blank control group. The white blood cell count and other cell count levels in the alveolar lavage fluid of the rats in the four groups, as well as the STAT4 and STAT6 protein levels in the lung tissues, were observed and compared.Results: After treatment, the white cell counts in Maxingganshi decoction group were significantly higher than that of the model control group (P<0.05);lymph, neutral particles and eosinophil levels were significantly lower than those of the model control group (P<0.05). Compared with roxithromycin tablets control group, white blood cell count and other classification level of cell count in Maxingganshi decoction group were not significantly different (P>0.05). After treatment, STAT4 protein levels in roxithromycin tablets control group and Maxingganshi decoction group were lower than that in the model control group (P<0.05), and STAT6 protein levels in roxithromycin tablets control group and Maxingganshi decoction group were higher than that of the model control group (P<0.05), suggesting the two means of intervention in this study could inhabit the STAT4 protein expression in lung tissue of COPD rats and promote effect on STAT6 protein. In addition, the level of STAT4 and STAT6 in the Maxingganshi decoction group was not significantly different from that in the roxithromycin tablets control group (P>0.05), suggesting that the effect of Maxingganshi decoction was similar to that of roxithromycin tablets.Conclusions: The action mechanism of Maxingganshi decoction group treating COPD may be through the STAT4 and STAT6 protein expression level to impose an effect, and thus interfere with IL-12 / STAT4 and IL-4 / STAT6 these two signaling pathways of Th1 cells and Th2 cells in the body of the gene expression, inhibiting the Th1 polarization and adjusting the imbalance of Thl/Th2 cells, so as to lower inflammatory response mediated by T cells and various kinds of pathological damage.

Effects of Yigan Tiaozhi Decoction on Serum NO, Endotoxin and RBP4 in Patients with Nonalcoholic Fatty Liver Disease

OBJECTIVE: To explore the effects of Yigan Tiaozhi Decoction on serum nitric oxide(NO), endotoxin and retinol binding protein 4(RBP4) in patients with nonalcoholic fatty liver disease(NAFLD), and to provide evidence for clinical treatment of such patients. METHODS: A total of 92 patients with NAFLD admitted to Army General Hospital 263 Clinic from March 2013 to March 2017 were selected, and all patients were divided into the control group(n = 46) and the observation group(n = 46) according to the random number table method. The patients in the control group were given conventional treatment, and the patients in the observation group were treated with Yigan Tiaozhi Decoction on the basis of the control group. Liver function, blood lipid levels, serum NO, endotoxin, RBP4 levels and clinical efficacy were observed and compared between the 2 groups before and after treatment. RESULTS: Before treatment, there was no significant difference in serum alanine aminotransferase, glutamyltranspeptidase, aspartate aminotransferase, total cholesterol and triglyceride levels between the 2 groups(P > 0.05); after treatment, the serum levels of alanine aminotransferase, glutamyltranspeptidase, aspartate aminotransferase, total cholesterol and triglyceride in the observation group were lower than those in the control group, and the difference was statistically significant(P < 0.05); before treatment, there was no significant difference in serum NO, endotoxin and RBP4 levels between the 2 groups(P > 0.05); after treatment, the serum NO, endotoxin and RBP4 levels in the observation group were lower than those in the control group, and the difference was statistically significant(P < 0.05); the total effective rate of treatment in the observation group(89.13%) was significantly higher than that in the control group(76.09%), and the difference was statistically significant(P < 0.05). CONCLUSION: Yigan Tiaozhi Decoction is effective in NAFLD, which can reduce serum NO, endotoxin and RBP4 levels, improve liver function and reduce blood lipid levels.

葛根芩连汤对湿热下注型溃疡性结肠炎患者TLR4/MyD88信号通路的影响

目的:分析葛根芩连汤对湿热下注型溃疡性结肠炎(UC)患者Toll样受体4(TLR4)/髓样分化因子88(MyD88)信号通路相关蛋白和下游炎性因子水平的影响。方法:选择100例唐山市中医医院2020年5月—2023年5月收治的UC患者为研究对象,随机分为观察组和对照组,每组50例。对照组进行常规灌肠配合美沙拉嗪肠溶片治疗;观察组在对照组治疗基础上加用葛根芩连汤内服。比较两组治疗效果、治疗前后中医证候积分变化、TLR4、MyD88水平、炎症因子水平以及不良反应发生情况。结果:观察组总有效率高于对照组(P<0.05);治疗后两组中医证候积分下降,且观察组低于对照组(P<0.05);治疗后两组TLR4、MyD88水平下降,且观察组TLR4、MyD88水平低于对照组(P<0.05);治疗后两组炎症因子水平均降低,且观察组炎症因子水平低于对照组(P<0.05);两组不良反应发生率比较,差异无统计学意义(P>0.05)。结论:葛根芩连汤治疗湿热下注型UC患者的疗效显著,可明显改善临床相应症状,调节TLR4/MyD88信号通路相关蛋白和下游炎性因子水平,降低炎症反应,且安全性较高,值得临床推广。

息喘汤联合治疗慢性阻塞性肺疾病稳定期患者效果及对CXCL13、TLR4、细胞因子影响

目的探讨息喘汤联合治疗慢性阻塞性肺疾病(COPD)稳定期患者效果及对趋化因子CXC配体13(CXCL13)、Toll样受体4(TLR4)、细胞因子影响。方法选取2020年6月—2022年6月收治的COPD稳定期128例,根据治疗方法不同将其分为观察组和对照组2组各64例。观察组给予息喘汤联合茚达特罗格隆溴铵治疗,对照组给予茚达特罗格隆溴铵治疗,疗程均为12周。比较2组治疗后临床效果,治疗前后肺功能、COPD评估测试(CAT)评分、6 min步行距离(6MWT)和血清CXCL13、TLR4、细胞因子,以及治疗期间不良反应发生情况。结果治疗后,观察组总有效率(93.75%,60/64)高于对照组(76.56%,49/64)(P<0.05)。治疗后,2组第1秒用力呼气容积(FEV1)、最大呼气流量(PEF)、FEV1/用力肺活量(FVC)和6MWT高于或长于治疗前,CAT评分和血清CXCL13、TLR4、肿瘤坏死因子-α(TNF-α)、白细胞介素-8(IL-8)、白细胞介素-33(IL-33)低于治疗前;且观察组FEV1、PEF、FEV1/FVC和6MWT高于或长于对照组,CAT评分和血清CXCL13、TLR4、TNF-α、IL-8、IL-33低于对照组(P<0.05)。治疗期间,2组不良反应发生率比较差异无统计学意义(P>0.05)。结论息喘汤联合治疗COPD稳定期患者效果良好,可下调血清CXCL13和TLR4表达,降低血清细胞因子TNF-α、IL-8和IL-33水平。

基于TLR-4/NF-κB信号通路及水通道蛋白1表达探讨大陷胸汤干预重症急性胰腺炎早期肺损伤的效应机制

目的:探讨大陷胸汤对重症急性胰腺炎(SAP)早期肺损伤的影响及作用机制。方法:将60只大鼠随机分为实验组、模型组和对照组,每组20只。实验组予大陷胸汤灌胃,对照组及模型组予蒸馏水灌胃,在连续灌胃7 d后,模型组与实验组以胰胆管注射牛磺胆酸钠的方式制备SAP模型,于造模后12、24、48、72 h各时间点分别处死5只大鼠取材,观察大鼠一般情况、胸腹水量、肺湿干重比、胰腺组织及肺组织病理学改变、肺组织白细胞介素(IL)-1及Toll样受体4(TLR-4)水平、肺组织核因子(NF)-κB及水通道蛋白1(AQP1)表达水平。结果:与对照组相比,模型组大鼠一般情况较差,胰腺炎症反应以及肺损伤程度较重,胸腹水量、肺湿干重比、肺组织NF-κB表达水平、IL-1及TLR-4水平升高(P<0.05),肺组织AQP1表达水平下降。与模型组相比,实验组大鼠一般情况较好,肺损伤程度较轻,胸腹水量、肺湿干重比、肺组织NF-κB表达水平、IL-1及TLR-4水平下降(P<0.05),肺组织AQP1表达水平升高,造模后72 h后,胰腺组织坏死程度与同期模型组相比明显好转。结论:大陷胸汤可在一定程度上减轻SAP大鼠早期肺损伤,其机制可能与抑制TLR-4/NF-κB炎症信号通路,增强AQP1表达有关。

苍附导痰汤对肥胖型PCOS-IR模型大鼠卵巢TLR4/NF-κB p65信号通路的影响

目的 探讨苍附导痰汤对肥胖型PCOS-IR (polycystic ovarian syndrome-insulin resistance, PCOS-IR)模型大鼠卵巢Toll受体4(TLR4)/核转录因子κB p65(NF-κB p65)信号通路的调控作用。方法 48只♀大鼠随机分为正常组8只和模型组40只。来曲唑(1 mg·kg^(-1))联合高脂饮食建立肥胖型PCOS-IR大鼠模型,快速革兰染色法观察动情周期,挑选24只模型大鼠随机分为:模型组、阳性药(二甲双胍135 mg·kg^(-1))组、苍附导痰汤高、低剂量(57.96、14.49 g·kg^(-1))组,每组各6只,药物干预21 d。观察动情周期、卵巢和子宫指数变化;血液生化仪测定空腹血糖(FBG)和血脂(甘油三酯TG和总胆固醇TC)变化;酶联免疫吸附(ELISA)法测定空腹胰岛素(FINS)水平;免疫组化法和荧光定量PCR法检测卵巢中TLR4和NF-κB p65蛋白及基因的表达。结果 与正常组比较,模型组大鼠动情周期紊乱,卵巢多囊性改变明显,FBG、TG、TC含量和FINS、HOMA-IR水平上调,卵巢中TLR4和NF-κB p65蛋白及mRNA表达均增加(P<0.05);与模型组比较,苍附导痰汤高剂量组大鼠排卵周期得到改善,卵巢多囊性改变减轻,上述指标出现明显逆转(P<0.05)。结论 苍附导痰汤能有效改善肥胖型PCOS-IR大鼠卵巢排卵和糖脂代谢功能,作用机制可能与调控卵巢TLR4/NF-κB p65信号通路有关。

活肾通络方联合依那普利对慢性肾脏病患者纤维化标志物、人附睾蛋白4水平的影响

目的:研究活肾通络方联合依那普利对2~4期慢性肾脏病(CKD)患者肾功能、纤维化标志物、人附睾蛋白4(HE4)水平的影响。方法:采用随机对照研究将78例2~4期CKD患者随机分为两组,各39例。对照组予依那普利,观察组予活肾通络方联合依那普利。两组治疗12周。疗程结束后比较两组疗效、主症和次症积分、透明质酸(HA)、Ⅲ型前胶原(PC-Ⅲ)、Ⅳ型胶原(C-Ⅳ)、层粘连蛋白(LN)、尿素(UR)、肌酐清除率(Ccr)、血肌酐(Scr)、β2-微球蛋白(β_(2)-MG)、HE4、高敏-C反应蛋白(hs-CRP)、白细胞介素-6(IL-6)水平、中医症候积分。结果:治疗后观察组总有效率92.31%高于对照组的74.36%(P<0.05);两组治疗前主症和次症积分、HA、PC-Ⅲ、C-Ⅳ、LN、UR、Ccr、Scr、β_(2)-MG、HE4、hs-CRP、IL-6水平比较,差异无统计学意义(P>0.05);两组主症、次症积分、HA、PC-Ⅲ、C-Ⅳ、LN、UR、Scr、β_(2)-MG、HE4、hs-CRP、IL-6水平下降,Ccr水平增加,观察组主症、次症积分、HA、PC-Ⅲ、C-Ⅳ、LN、UR、Scr、β_(2)-MG、HE4水平低于对照组,Ccr水平高于对照组(P<0.05)。结论:活肾通络方联合依那普利可显著延缓2~4期CKD患者肾纤维化进程,保护肾功能,降低HE4水平。

四逆汤对溃疡性结肠炎大鼠TLR4/NF-κB信号通路相关因子及肠道菌群的影响

目的 探讨四逆汤对溃疡性结肠炎大鼠Toll样受体4(Toll-like receptor 4,TLR4)/核因子-κB(nuclear factor-κB,NF-κB)信号通路相关因子及肠道菌群的影响。方法 选取健康雄性SD大鼠28只,常规饲料喂养,自由进食进水,实验开始前适应性饲养7 d。按照随机数字表法将28只大鼠分为正常组、模型组、四逆汤组、美沙拉嗪组,各7只。正常组大鼠注入氯化钠溶液,模型组、四逆汤组、美沙拉嗪组大鼠通过结肠内注入三硝基苯磺酸(trinitrobenzenesulfonic acid, TNBS)建立溃疡性结肠炎模型。造模完成后,四逆汤组大鼠采用四逆汤3.8 g/kg灌胃,美沙拉嗪组采用美沙拉嗪10 g/kg灌胃,1次/d。在给药前1 d及给药7 d后秤测量大鼠的体质量、疾病指数活动评分(disease activity index, DAI)评分,检测大鼠给药7 d后的TLR4、NF-κB、肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)、白细胞介素-10(interleukin-10,IL-10)、白细胞介素-16(interleukin-16,IL-16)、黏蛋白2(mucin 2,MUC2)、三叶因子3(trefoil factor 3,TFF3)表达及肠道菌群。结果 给药前,正常组大鼠体质量显著高于模型组、四逆汤组、美沙拉嗪组(P<0.05);给药7 d后,四逆汤组、美沙拉嗪组大鼠体质量显著升高(P<0.05),正常组、四逆汤组、美沙拉嗪组大鼠体质量显著高于模型组(P<0.05)。给药前,正常组大鼠DAI评分显著低于模型组、四逆汤组、美沙拉嗪组(P<0.05);给药7 d后,四逆汤组、美沙拉嗪组大鼠DAI评分显著降低(P<0.05),模型组、四逆汤组、美沙拉嗪组大鼠DAI评分显著高于正常组(P<0.05),四逆汤组、美沙拉嗪组大鼠DAI评分显著低于模型组(P<0.05)。给药7 d后,正常组、四逆汤组、美沙拉嗪组大鼠TLR4、NF-κB表达均显著低于模型组(P<0.05)。给药7 d后,正常组、四逆汤组、美沙拉嗪组大鼠TNF-α、IL-10、IL-16水平均显著低于模型组(P<0.05)。给药7 d后,正常组、四逆汤组、美沙拉嗪组大鼠肠球菌、肠杆菌显著低于模型组,双歧杆菌、乳酸杆菌显著高于模型组(P<0.05)。给药7 d后,正常组、四逆汤组、美沙拉嗪组大鼠TFF3、MUC2表达显著高于模型组(P<0.05)。结论 四逆汤可用于溃疡性结肠炎的治疗,对于TLR4/NF-κB信号通路相关因子及肠道菌群均有一定调节作用。

补肾止颤方对帕金森病大鼠Caspase-4/11表达的影响

目的探讨补肾止颤方对帕金森病(Parkinson′s disease,PD)大鼠Caspase-4/11表达的影响。方法采用蛋白酶抑制因子注射法建立慢性PD大鼠模型,将造模成功的大鼠分为模型组、多巴丝肼组(2 g·kg^(-1)·d^(-1)),补肾止颤方低、中、高剂量组(15、30、60 g·kg^(-1)·d^(-1)),假手术组采用等量的70%乙醇溶液注射。假手术组、模型组给予10 mL·kg^(-1)·d^(-1)生理盐水;其余各组给予相应药物;连续灌胃3周。观察大鼠的一般情况,于灌药前1周及灌药后第3周进行悬挂实验并评分。采用HE染色、透射电镜观察中脑黑质区神经元病理改变,qPCR技术检测中脑黑质区α-syn、TH、IL-1β、GSDMD、Caspase-4/11的mRNA表达变化。结果补肾止颤方能增加悬挂实验时间及评分,减轻神经元病理损伤,明显抑制α-syn、IL-1β、GSDMD、Caspase-4/11表达,增加TH表达。结论补肾止颤方可有效改善PD模型大鼠的行为障碍、减轻神经元病理损伤,可能通过下调Caspase-4/11,抑制GSDMD蛋白的表达,减少α-syn、IL-1β分泌,从而减轻细胞焦亡,达到神经元保护作用。

桃红四物汤对创伤性股骨头坏死大鼠组织形态学及VEGFR2、Dll4表达的影响

目的 通过观察桃红四物汤对创伤性股骨头坏死大鼠血管内皮细胞生长因子受体2(vascular endothelial growth factor receptor 2, VEGFR2)、Delta样配体4(Delta-like 4, Dll4)蛋白表达的影响,探究其防治创伤性股骨头坏死的作用机制。方法 将44只大鼠采用随机数字表法分为正常组(10只)和造模组(34只)。造模组行创伤性股骨头坏死造模;8周后从正常组和造模组分别随机取2只进行对比检测造模是否成功。将造模成功的剩余32只大鼠按随机数字表法分为模型组及桃红四物汤低、中、高剂量组,每组8只;加上正常组剩余大鼠8只,共计5组。正常组和模型组予以生理盐水灌胃;桃红四物汤低、中、高剂量组分别予以9、18、36 g/kg桃红四物汤灌胃。持续4周后采集股骨头标本,采用micro-CT观察股骨头组织形态学,光镜下检测空骨陷窝率,免疫组织化学法测定VEGFR2和Dll4的蛋白表达水平。结果 与正常组对比,其余4组大鼠股骨头均有不同程度的骨质破坏、软骨面塌陷及空骨陷窝率升高(P<0.05);模型组和桃红四物汤低剂量组VEGFR2蛋白表达均下降(P<0.05);桃红四物汤中、高剂量组Dll4蛋白表达明显增加(P<0.05)。与模型组对比,桃红四物汤中、高剂量组大鼠股骨头软骨形态改善,组织空骨陷窝率下降(P<0.05);桃红四物汤低、中、高剂量组VEGFR2蛋白表达均升高(P<0.05);桃红四物汤中、高剂量组Dll4蛋白表达升高(P<0.05)。结论 桃红四物汤能有效降低创伤性股骨头坏死骨组织空骨陷窝、改善组织形态,可能与增加VEGFR2、Dll4蛋白的表达、激活VEGF/Notch信号通路有关。

Taohong Siwu decoction(桃红四物汤)ameliorates atherosclerosis in rats possibly through toll-like receptor 4/myeloid differentiation primary response protein 88/nuclear factor-κB signal pathway

OBJECTIVE:To investigate the effect of Taohong Siwu decoction(桃红四物汤,TSD)on atherosclerosis in rats as well as investigate the underlying mechanism based on molecular docking.METHODS:Sixty healthy male Sprague-Dawley rats were randomly divided into 6 groups with 10 rats in each group:control group,model group,atorvastatin group(AT,2.0 mg/kg),and TSD groups(20,10,5 g/kg)after 7 d of acclimation.The model of atherosclerosis was successfully established except the control group by high fat diet(HFD)and vitamin D2.Biochemical analyzers were used to detect the levels of triglyceride(TG),total cholestero(TC),low density lipoprotein-cholesterol(LDLC)and high density lipid-cholesterol(HDL-C)in blood lipid.The levels of tumor necrosis factor-α(TNF-α),interleukin-6(IL-6)and interleukin-1β(IL-1β)were determined by enzyme-linked immunosorbent assay.Sudan IV staining and Hematoxylin and eosin staining(HE staining)were performed to observe the pathological changes in aortic tissue.Molecular docking technology was used to predict the best matching between the main components of TSD and the target proteins.The expression of target proteins was further detected by quantitative real time polymerase chain reaction(q RTPCR)and Western blot analysis.RESULTS:The results showed that TSD restricted atherosclerosis development and decreased the inflammatory cytokines in plasma.Molecular docking results predicted that the main components of TSD showed a strong binding ability with toll-like receptor(TLR4),myeloid differentiation primary response protein 88(My D88),and nuclear factor kappa-B(NF-κB).The results of q RT-PCR and Western blot analysis showed that the m RNA and protein expressions of TLR4,My D88 and NF-κB p65 in the aorta were reduced in atorvastatin group and TSD group.CONCLUSIONS:TSD can ameliorate atherosclerosis in rats,and the underlying mechanism is supposed be related to the suppression of inflammatory response by regulating TLR4/My D88/NF-κB signal pathway.