复方益肾胶囊对大鼠糖尿病肾病的药效学研究

[目的]探讨复方益肾胶囊对糖尿病肾病的疗效。[方法]采用长期高脂肪乳剂饲养(10 mL/kg,7 d)加小剂量链尿佐菌素(STZ)(25 mg/kg,ip 1次)诱导大鼠2型糖尿病动物模型,随机将动物分为6组:正常对照组、糖尿病模型组、土鳖虫水提物组、土鳖虫脂提物、复方益肾胶囊组、阳性药物对照组(依那普利治疗组)。每组大鼠10只,各组均为灌胃给药。实验4周后大鼠尾部静脉取血测血糖,并测定空腹血糖、肾功能(尿微量白蛋白、血肌酐、尿素氮)指标,以及肾脏病理学的改变。[结果]复方益肾胶囊可降低糖尿病肾病大鼠血糖,治疗组较糖尿病模型组及阳性药物对照组血糖降低差异显著(P0.01);复方益肾胶囊治疗后大鼠尿微量白蛋白、血肌酐与模型组比较有统计学意义(P0.01)。[结论]复方益肾胶囊可以降低实验性糖尿病大鼠的高血糖、尿微量白蛋白、血肌酐,能够治疗糖尿病,改善肾功能。     

Tangshenping granule inhibits pyroptosis in a rat model of streptozotocin-induced diabetic nephropathy via the NLRP3/caspase-1/GSDMD pathway

ObjectiveTo explore the inhibitory effect of Tangshenping (TSP) on pyroptosis in a streptozotocin-induced diabetic nephropathy (DN) rat model.MethodsDN was established in Sprague–Dawley rats. Rats were randomly divided into DN (model group), irbesartan, and TSP low-, medium-, and high-dose groups, besides the control group. The 24 h albuminuria content, and serum content of TC, TGs, Scr, IL-1β, UREA, LDLs, and IL-18 were assessed. Hematoxylin & eosin and Mallory staining were performed to examine pathological changes in the kidney. The mRNA and protein expression of NLRP3, caspase 1, and GSDMD in the kidney were also examined.ResultsThe 24 h albuminuria content was obviously lower in the treatment groups compared to the model group (all P < .01). Levels of TC, TGs, Scr, UREA, LDLs, and IL-18 after drug interventions were obviously lower compared to the model group (all P < .05). The serum content of IL-1β in the TSP medium- and high-dose groups were much lower compared to the model group (P = .013 and P = .001, respectively). Through immunohistochemistry and western blotting, we observed that the protein expressions of NLRP3, caspase-1, GSDMD, IL-1β, and IL-18 were lower after drug interventions compared to the model group (all P < .05). Using qPCR, we observed that the mRNA expressions of caspase-1, IL-1β, IL-18, and GSDMD after drug interventions were significantly lower compared to the model group (all P < .05). The mRNA expressions of NLRP3 in the TSP medium- and high-dose groups were both lower compared to the model group (all P < .05).ConclusionTSP downregulated mRNA and protein expressions of NLRP3, caspase-1, and GSDMD. Our findings demonstrate that the beneficial effects of TSP on renal function are at least partly mediated by the inhibition of micro-inflammation and modulation of the expression of pyroptosis-related factors.     

Electro-acupuncture promotes gut motility and alleviates functional constipation by regulating gut microbiota and increasing butyric acid generation in mice

ObjectiveAbnormalities in the gut microbiota and intestinal short-chain fatty acid (SCFA) levels are implicated in the pathogenesis of functional constipation (FC). Electro-acupuncture (EA) has been shown to improve constipation-related symptoms and rebalance the gut microbiota. However, it is currently unknown whether the gut microbiota is a key mechanistic target for EA or how EA promotes gut motility by regulating the gut microbiota and SCFAs. Therefore, we assessed the effects of EA in FC mice and pseudo-germfree (PGF) mice to address these questions.MethodsForty female Kunming mice were randomly separated into a normal control group (n = 8), an FC group (n = 8), an FC + EA group (n = 8), a PGF group (n = 8) and a PGF + EA group (n = 8). The FC group and FC + EA group were treated with diphenoxylate to establish the FC model; the PGF group and PGF + EA group were given an antibiotic cocktail to initiate the PGF model. After maintaining the model for 14 d, mice in the FC + EA and PGF + EA groups received EA stimulation at the ST25 and ST37 acupoints, once a day, 5 times per week, for 2 weeks. Fecal parameters and intestinal transit rate were calculated to assess the efficacy of EA on constipation and gastrointestinal motility. Colonic contents were used to quantify gut microbial diversity using 16S rRNA sequencing, and measure SCFA concentrations using gas chromatography-mass spectrometry.ResultsEA significantly shortened the first black stool defecation time (P < 0.05) and increased the intestinal transit rate (P < 0.01), and fecal pellet number (P < 0.05), wet weight (P < 0.05) and water content (P < 0.01) over 8 h, compared with the FC group, showing that EA promoted gut motility and alleviated constipation. However, EA treatment did not reverse slow-transit colonic motility in PGF mice (P > 0.05), demonstrating that the gut microbiota may play a mechanistic role in the EA treatment of constipation. In addition, EA treatment restored the Firmicutes to Bacteroidetes ratio and significantly increased butyric acid generation in FC mice (P < 0.05), most likely due to the upregulation of Staphylococcaceae microorganisms (P < 0.01).ConclusionEA-mediated resolution of constipation occurs through rebalancing the gut microbiota and promoting butyric acid generation.Please cite this article as: Xu MM, Guo Y, Chen Y, Zhang W, Wang L, Li Y. Electro-acupuncture promotes gut motility and alleviates functional constipation by regulating gut microbiota and increasing butyric acid generation in mice. J Integr Med. 2023; Epub ahead of print.     

当归多糖对糖尿病肾病KK-Ay小鼠肾脏AMPK信号通路及线粒体自噬的影响

目的 研究当归多糖对糖尿病肾病（diabetic nephropathy,DN）KK-Ay小鼠肾脏磷酸腺苷激活的蛋白激酶（AMPactivated protein kinase,AMPK）信号通路及线粒体自噬的影响。方法 SPF级雄性KK-Ay小鼠用高糖高脂饲料喂养,随机分为模型组、厄贝沙坦（25 mg/kg）组和当归多糖高、中、低剂量（400、200、100 mg/kg）组,每组10只;将10只雄性C57BL/6J小鼠作为对照组。给予药物干预4周,观察小鼠一般情况,每周称定体质量并检测血糖;末次给药后,心脏取血并处死小鼠,分离血清检测尿微量白蛋白（urine microalbuminuria,U-ALB）、肌酐（creatinine,SCr）、尿素氮（urea nitrogen,BUN）;采用苏木素-伊红（HE）染色观察肾组织病理变化;采用Western blotting检测肾组织线粒体自噬相关蛋白[微管相关蛋白1轻链3(microtubule-associated protein 1 light chain 3,LC3)、p62、Nix]和线粒体裂变蛋白[线粒体动力相关蛋白1(dy...     

Effect of transcutaneous auricular vagus nerve stimulation on fasting blood glucose and serum insulin concentration in Zucker diabetes fatty rats: 经皮耳迷走神经刺激对Zucker糖尿病大鼠血糖及血清胰岛素浓度的影响

ObjectiveTo observe the influence of transcutaneous auricular vagus nerve stimulation (taVNS) on fasting plasma glucose (FPG) and serum insulin (INS) in Zucker diabetes fatty (ZDF) rats and explore the regulatory effect of taVNS on blood glucose in ZDF rats.MethodsA total of 20 male ZDF rats were randomized into a model group and a taVNS group, 10 rats in each one. Besides, the other 10 Zucker Lean (ZL) rats were selected to be a control group. The rats in the control group were fed with common forage and those in the model group and the taVNS group were fed with high-sugar and high-fat forage. The intervention of electric stimulation was applied in the rats of taVNS group, with 2/15 Hz in frequency, disperse-dense wave and 2 mA in intensity. Each intervention lasted 30 min, once a day, consecutively for 12 weeks. No any intervention was given in the control group and the model group. The body mass and FPG level were recorded once every 2 weeks in the rats. After the experiment, the rats were sacrificed and blood sample was collected. Enzyme linked immunosorbent assay (ELISA) was adopted to determine the level of serum INS in the rats.ResultsAfter the modeling, compared with the control group, the body mass and FPG level were all higher (P < 0.05, P < 0.01) and the concentration of serum INS was lower (P < 0.01) in the rats of the model group. After intervention, compared with the model group, the body mass was lower in week 6 to 10 (P < 0.01, P < 0.05), FPG level was lower in week 8 to 12 (P < 0.01, P < 0.05) and serum INS concentration was higher (P < 0.01) in the rats of the taVNS group.ConclusiontaVNS apparently improves in hyperglycemia in ZDF rats and increases serum insulin concentration in the rats.     

荞麦黄酮对大鼠糖尿病肾病的保护作用

目的:观察荞麦黄酮对大鼠糖尿病肾病的保护作用及其机制。方法:SD大鼠ip四氧嘧啶150 mg·kg-1,每隔7 d注射一次,连续3次,建立大鼠糖尿病肾病模型。将成模大鼠随机分为模型组、荞麦黄酮低、中、高剂量组(50,100,200 mg·kg-1)及厄贝沙坦组(17 mg·kg-1),另设正常组。造模后各组大鼠ig给药1次/d,连续12周。给药前及给药后每隔3周测定大鼠空腹血糖(FBG)及24 h尿蛋白含量,末次给药后2 h处死大鼠,采用生化仪测定血清中血肌酐(SCr),尿素氮(BUN),甘油三酯(TG),总胆固醇(TC)水平。肾组织匀浆测定果糖胺(FTS),糖基化终产物(AGEs),超氧化物歧化酶(SOD)及丙二醛(MDA)水平。透射电镜观察各组大鼠肾组织形态。结果:与正常组比较,模型组大鼠FBG和24 h尿蛋白含量持续升高,血清中SCr,BUN,TG,TC水平及肾组织中FTS,AGEs,MDA含量均明显增高,SOD水平下降(P0.05,P0.01),且肾组织形态明显损伤。与模型组比较,荞麦黄酮各剂量组可不同程度改善上述指标,且中剂量组作用更为明显(P0.05,P0.01)。结论:荞麦黄酮对糖尿病肾病大鼠具有一定保护作用,其机制与其具有降糖、降脂、降低非酶糖基化及氧化应激水平相关。     

Influence of moxibustion on NR2B and PKMζ after neural stem cell transplantation in the rats with vascular dementia: 艾灸对血管性痴呆大鼠神经干细胞移植后海马NR2B和PKMζ的影响

ObjectiveTo observe the influence of moxibustion on learning and memory ability in the rats with vascular dementia (VD) and explore the potential effect mechanism.MethodsA total of 80 rats, screened by Morris water maze, were randomly divided into a sham-operation group, a model group, a neural stem cells (NSCs) group, a NSCs + piracetam group and a NSCs + moxibustion group, 16 rats in each group. After corresponding treatments, Morris water maze and immunofluorescence technique were adopted to evaluate the therapeutic effect respectively.ResultsComparison among groups after modeling: compared with the sham-operation group, the escape latency was longer (P < 0.01) and the times of crossing platform were reduced (P < 0.01) in the rats of the model group. Comparison among groups after treatment: compared with the model group, the escape latency was shortened (P < 0.01) and the times of crossing platform were increased (P < 0.05) in the rats of the NSCs group. Compared with the NSCs group, the escape latency was shorter and the expressions of the hippocampus NR2B/EGFP and PKMζ/EGFP expression level increased (all P < 0.05) in the rats of the NSCs + piracetam group and the NSCs + moxibustion group (all P < 0.05). Compared with the NSCs + piracetam group, the escape latency was shorter and the expressions of the hippocampus NR2B/EGFP and PKMζ/EGFP expression level were higher in the rats of the moxibustion + NSCs group (all P < 0.05)ConclusionMoxibustion improves the spatial learning and memory ability of the VD rats and promotes the reconstruction of neurogenesis and synaptic function, which may be related to the up-regulation of the expressions of hippocampus NR2B and PKMζ expressions.     

Antihyperglycemic and hepatoprotective properties of miracle fruit (Synsepalum dulcificum) compared to aspartame in alloxan-induced diabetic mice

ObjectiveThis study was undertaken to investigate the antihyperglycemic potential of miracle fruit (MF) as well as its hepatic safety as compared to aspartame in alloxan-induced diabetic mice.MethodsMF extracts were prepared and screened for their phytochemical composition using high-performance liquid chromatography (HPLC). Total phenolic, flavonoid and tannin contents and antioxidant potential were also determined. Additionally, MF was evaluated for its sensory attributes. For in vivo work, MF ethanol extract at high (MFH: 500 mg/kg body weight [BW]) and low (MFL: 250 mg/kg BW) doses as well as aspartame were injected intraperitoneally into alloxan-induced diabetic mice. Blood glucose levels were determined following acute and subchronic treatment. At the end of the study, animals were sacrificed, serum was collected for biochemical analysis and liver tissues were obtained for histopathological examination.ResultsMF ethanol extract contained more flavonoids and tannins, and had higher 1,1-diphenyl-1-picrylhydrazyl radical-scavenging activity (79.61%) compared to MF aqueous extract (P < 0.05). HPLC analysis of MF ethanol extract also revealed the presence of 10 antioxidants with quercetin comprising the major polyphenol. Additionally, sensory analysis of MF showed that its intake is effective in masking undesirable sourness. Subchronic administration of MFH proved amelioration of hyperglycemia in mice as compared to aspartame. Moreover, aspartame treatment significantly elevated (P < 0.05) the level of alanine aminotransferase and had destructive effects on the liver histopathology; however, hepatic architecture was restored by low and high doses of MF.ConclusionMF is an effective antihyperglycemic with hepatoprotective properties that can be used as a healthier alternative sweetening agent in place of aspartame for sour beverages.     

附子-干姜对慢性溃疡性结肠炎小鼠肠黏膜微循环障碍的改善作用

目的研究附子-干姜对慢性溃疡性结肠炎(ulcerativecolitis,UC)小鼠肠黏膜微循环障碍的改善作用。方法建立慢性UC小鼠模型,给予附子-干姜水煎液进行干预,记录各组小鼠体质量、大便性状及便血情况,进行疾病活动指数(disease activityindex,DAI)评分;采用激光多普勒血流仪检测各组小鼠肠黏膜血流灌注量、血细胞移动速度和移动血细胞浓度;采用ELISA法检测各组小鼠血清中血小板活化因子(platelet activating factor,PAF)和组织因子(tissue factor,TF)水平;采用苏木素-伊红(HE)染色法观察各组小鼠结肠组织病理改变情况;采用免疫组化法检测各组小鼠肠黏膜毛细血管内皮细胞CD31表达并计算微血管密度(microvessel density,MVD)。结果与对照组比较,模型组小鼠DAI评分显著增加(P0.01),肠黏膜血流灌注量、血细胞移动速度显著下降(P0.01),肠黏膜移动血细胞浓度显著升高(P0.01),血清PAF、TF水平显著升高(P0.01),结肠组织CD31表达减少,肠黏膜微血管密度显著减少(P0.01);与模型组比较,附子-干姜高剂量(6.06 g/kg)组小鼠DAI评分明显降低(P0.01),肠黏膜血流灌注量、血细胞移动速度显著升高(P0.01),肠黏膜移动血细胞浓度显著降低(P0.01),机体凝血因子PAF、TF水平显著降低(P0.01),肠黏膜MVD显著增加(P0.01)。结论附子-干姜可通过改善肠黏膜微循环障碍从而治疗慢性UC。     

左归降糖舒心方对糖尿病心肌病MKR鼠心肌细胞损伤和凋亡的影响＊

目的 观察左归降糖舒心方对糖尿病心肌病（Diabetic cardiomyopathy， DCM）模型鼠心肌细胞损伤和凋亡的影响，并探讨其作用机制。方法 以转基因2型糖尿病小鼠（MKR鼠）为实验对象，通过链脲菌素（streptozotocin， STZ）腹腔注射+持续高脂喂饲建立DCM模型，分为模型组、中药组、阳性药物组，另设C57小鼠为空白对照组。中药组给予左归降糖舒心方14.27 g生药/（kg·天）灌胃，阳性药物组给予二甲双胍250 mg/（kg·天）+依那普利4.5 mg/（kg·天）灌胃，连续给药4周。采用电化学法血糖仪测定空腹血糖（fasting blood glucose， FBG），光镜和透射电镜观察小鼠心肌组织形态学变化，ELISA法检测血清心肌肌钙蛋白I（cardiac troponin I， cTnI）、乳酸脱氢酶（lactic dehydrogenase， LDH）水平，免疫组化法检测I型胶原蛋白（Collagen-I）、III型胶原蛋白（Collagen-III）、Bcl-2相关X蛋白（Bcl-2 associated X protein， Bax）、半胱氨酸天冬氨酸蛋白酶-9（Caspase-9）、B淋巴细胞瘤-2基因（B-cell lymphoma-2， Bcl-2）的蛋白表达，TUNEL法检测心肌细胞凋亡。结果 模型组FBG较空白对照组显著升高，经左归降糖舒心方干预后明显降低（P<0.01）。与模型组比较，中药组和阳性药物组的心肌组织病理形态明显改善，Collagen-I、Collagen-III蛋白表达减弱（P<0.01），血清cTnI、LDH水平明显降低（P<0.01），细胞凋亡阳性表达减弱（P<0.01），Bax、Caspase-9表达减弱（P<0.01），Bcl-2表达增强（P<0.01）。中药组下调Collagen-I和Collagen-III表达、减少心肌细胞凋亡、下调Bax和Caspase-9表达等作用弱于阳性药物组（P<0.01），但上调Bcl-2表达的作用强于阳性药物组（P<0.01）。结论 左归降糖舒心方对糖尿病心肌病MKR鼠的心肌细胞损伤和凋亡具有保护作用，其作用机制可能与降糖、改善心肌纤维化、抑制促凋亡因子Bax和Caspase-9表达、促进抗凋亡因子Bcl-2表达等有关。     

莪术对糖尿病肾病大鼠的保护作用

目的:探讨莪术(Curcuma wenyujin)对糖尿病肾病大鼠的保护作用。方法:Wistar大鼠40只,分为假手术组、模型组和莪术治疗组,利用肾切除和链脲佐菌素(streptozotocin,STZ,45 mg.kg-1 ip)诱导糖尿病大鼠模型并用莪术进行干预(3 g.kg-1.d-1 ig),观察肾脏病理学变化,检测血糖(glucemia,Glu)总胆固醇(total cholesteral,TC)、血肌酐(blood serum creatinine,Scr)、尿微量白蛋白及结缔组织生长因子(connective tissue growth factor,CTGF)在肾脏的表达变化。结果:模型组较假手术组Glu,TC,尿白蛋白排泄率明显增高(P<0.01),CTGF在肾组织中的表达显著增加(P<0.01),肾脏病理变化显著;而莪术治疗组上述指标较模型组显著减少(P<0.01),CTGF在肾组织中的表达显著降低(P<0.01),肾脏病理变化有明显改善。结论:莪术对糖尿病大鼠的肾脏有保护作用,可能与其抑制CTGF在肾组织中的表达有关。     

Analgesic and anti-inflammatory properties of hydroalcoholic extracts of Malva sylvestris,Carum carvi or Medicago sativa,and their combination in a rat model

ObjectiveThe aim of this study was to evaluate the analgesic and anti-inflammatory effects of the hydroalcoholic extracts of Malva sylvestris flowers or Carum carvi and Medicago sativa seeds, alone and in combination, which have been used in traditional Iranian medicine.MethodsMale Wistar rats were divided into 6 treatment groups: distilled water, sodium salicylate (SS), M. sylvestris extract (600 mg/kg), C. carvi extract (600 mg/kg), M. sativa extract (300 mg/kg) and combined extract (including 300 mg/kg M. sylvestris and C. carvi extracts, and 150 mg/kg M. sativa extract). The formalin pain model was used to evaluate the antinociceptive effects of the treatments. For anti-inflammatory effect, acute (one hour after injection) and chronic (during a week after injection) paw inflammation was measured after subcutaneous injection of 2.5% formalin in the hindpaw. Finally, tissue samples from all groups were prepared for histopathological studies.ResultsThe combined extract significantly inhibited the nociception in the acute phase of the formalin test (P < 0.001). In the chronic phase, all the extracts and SS had significant analgesic effect (P < 0.001). Analgesic activity of the combined extract was significantly stronger than SS (P < 0.01). In the acute inflammation model, M. sylvestris, C. carvi and the combined drug had significant inhibitory effects against paw edema (P < 0.05). All extracts, individually and in combination, significantly alleviated chronic paw inflammation (P < 0.01). The combined extract had much more anti-inflammatory activity than SS (P < 0.05). Histopathological results indicated improvement and reduction of inflammatory factors in the treatment groups.ConclusionM. sylvestris, C. carvi and M. sativa have analgesic and anti-inflammatory properties. Potentially, each of these extracts or a mixture of them might be a valuable alternative drug to control pain and inflammation.     

Electroacupuncture at “Zusanli(ST36)” alleviates 5-fluorouracil-induced renal injury in colorectal cancer-bearing mice by exerting effects on oxidative stress,inflammatory responses,and cell apoptosis: 电针“足三里”减轻大肠癌荷瘤小鼠5-FU化疗后肾损伤及其氧化应激、炎症反应和细胞凋亡的研究

ObjectiveTo observe the effect of electroacupuncture at “Zusanli (ST36)” on 5-fluorouracil (5-FU)-induced renal injury in colorectal cancer-bearing mice, and to further investigate its effects on oxidative stress, inflammatory responses, and cell apoptosis in mouse renal tissue.MethodsThirty-five male BALB/c mice were randomly divided into five groups of seven mice each, namely the control, CT26, 5-FU, sham point (SP), and ST36 (which received EA at the “ST36”) groups. With the exception of the control group, each group was subjected to establishment of a subcutaneous implantation tumor model using the murine CT26 colorectal cancer cell line. Once the models were successfully established, the 5-FU, SP, and ST36 groups received 5-FU injection solution intraperitoneally at a dose of 5 mg/mL once every three days over a 21-day period. Mice in the SP and ST36 groups additionally received an EA intervention after each intraperitoneal 5-FU injection. EA were performed on mice of the SP group at bilateral sham acupoints and on mice of the ST36 group at the bilateral “ST36” using the continuous wave mode at a frequency of 2 Hz for a duration of 5 min, intervention was administered once every two days for a duration of 21 days. Samples were collected from the mice at the end of the experiment. The pathological morphology of the renal tissue was observed using hematoxylin and eosin (HE) staining; the contents of creatine (Cre), blood urea nitrogen (BUN) and malondialdehyde (MDA), and superoxide dismutase (SOD) activity were measured using biochemical assays; the expression and nuclear translocation of nuclear factor kappa B p65 subunit (NF-κB p65) were measured by immunofluorescence; the expression levels of tumor necrosis factor alpha (TNF-α), interleukin-6 (IL-6), and interleukin-1β (IL-1β) in serum were measured by ELISA; cell apoptosis in renal tissue was detected using the TUNEL assay; and the expression levels of the anti-B-cell lymphoma/leukemia 2 (Bcl-2), Bcl-2-associated X protein (Bax), cleaved caspase-3, cleaved caspase-9, and cytochrome C (cyt c) in renal tissue were measured by Western blotting.ResultsCompared with the control group, mice of the CT26 group showed a significant increase in serum Cre content (P<0.01), but the difference in BUN content was not statistically significant (P>0.05). HE staining of renal tissue revealed clear structures with normal glomerular and renal tubular morphology. SOD activity was decreased (P<0.01); MDA content was increased, but the increase was not statistically significant (P>0.05). Differences in NF-κB p65 protein expression in the cytoplasm of renal tissue and serum levels of TNF-α, IL-6, and IL-1β were not statistically significant (all P>0.05). Results of immunofluorescent TUNEL staining indicated an absence of significant cell apoptosis. In the renal tissue, Bcl-2 protein expression was slightly increased (P<0.05), and the expression levels of Bax (P<0.01), cleaved caspase-3 (P>0.05), cleaved caspase-9 (P<0.01), and cyt c (P>0.05) were increased. Compared with the CT26 group, mice of the 5-FU group exhibited an increase in Cre (P<0.01) and BUN (P<0.05) content. HE staining of renal tissue revealed the presence of glomerular atrophy and dilated Bowman's capsular spaces. SOD activity was significantly decreased (P<0.01), while the increase of MDA content was not significant (P>0.05). The expression of NF-κB p65 in the nucleus and serum TNF-α, IL-6, and IL-1β levels showed significant increases (P<0.05). The cell apoptosis level was significantly increased. The protein expression of Bcl-2 (P<0.05), Bax (P<0.01), cleaved caspase-9 (P<0.01), and cyt c (P<0.05) was significantly increased. Compared with the 5-FU group, the ST36 group showed decreased serum Cre and BUN levels (both P<0.01). HE staining of renal tissue showed less renal tissue injury and less dilation of renal capsular cavities. SOD activity was significantly higher (P<0.01), while MDA content was lower (P<0.05). Nuclear expression of NF-κB p65 and serum TNF-α (P<0.05), IL-6 (P<0.05), and IL-1β (P<0.01) levels were lower. The cell apoptosis level was decreased relative to the 5-FU group. Bcl-2 protein expression was significantly increased (P<0.01), while the protein expression levels of Bax (P<0.01), cleaved caspase-3 (P<0.01), cleaved caspase-9 (P<0.01), and cyt c (P<0.05) also were reduced.ConclusionEA at “ST36” attenuated 5-FU-induced renal injury in colorectal cancer-bearing mice. The mechanism may be related to the regulation of renal oxidative stress, alleviation of inflammatory responses, and inhibition of cell apoptosis.     

Effects of scalp cluster needling on cognition and oxidative stress in a rat model of schizophrenia☆头穴丛刺对精神分裂症模型大鼠认知功能及氧化应激系统的影响

ObjectiveTo analyze the effects of cluster needling at scalp acupoints (CNSA) on behavioral performance and expression of superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GSH-Px) in the hippocampus of rats with schizophrenia, and therefore, to shed light on the mechanism of action of CNSA in attenuating schizophrenia.MethodsThirty-six Wistar rats were randomly divided into the control, model, risperidone, and CNSA groups (9 rats per group). The schizophrenia model was prepared by injecting 0.1 mg/mL dizocilpine maleate (MK-801) for 14 consecutive days. Subsequently, rats in the risperidone and CNSA groups were subjected to the following therapy for 14 consecutive days: (1) Risperidone group: intragastric administration of risperidone suspension (0.4 mg/kg); (2) CNSA group: the “GV 20″ “Qiánd?ng (前顶GV 21) ” “Shéntíng (神庭GV 24) ” “Xìnhuì (囟会 GV 22) ” “Tōngtiān (通天BL 7) ” “Luòquè (络却BL 8) ” “Qūchā (曲差BL 4) ” and “W?chù (五处 BL 5) ” acupoints were selected for needle positioning. Following 14-day intervention period, the Morris water maze experiment and open field experiment were performed. Finally, hippocampal tissue specimens were collected and SOD, CAT, and GSH-Px expression levels were measured by ELISA.Results(1) Morris water maze experiment: Following the 14-day model construction period, the model, risperidone, and CNSA groups showed a significant increase in escape latency (all P < 0.05) and a significant decrease in the number of platform crossings (all P < 0.05) compared with the control group, indicating successful induction of schizophrenia in the rat model. At the end of the intervention period (28d), the risperidone and CNSA groups showed a significant decrease in escape latency (both P < 0.05), and the CNSA group showed a significant increase in the number of platform crossings (P < 0.05) compared with the model group. (2) Open field experiment: At 14d, the model, risperidone and CNSA groups exhibited a significant decrease in the travelled distance and amount of time spent in the central zone (all P < 0.05) compared with the control group (all P < 0.05). At 28d, the risperidone and CNSA groups showed a significant increase in the travelled distance and percentage of time spent in the central zone (P < 0.05 or P < 0.01). (3) Antioxidant enzyme expression: At 28d, the model group exhibited significant decreases in the hippocampal SOD, CAT, and GSH-Px levels, compared with the control group (P < 0.01 or P < 0.001).ConclusionCNSA enabled the attenuation of cognitive impairment and enhancement of memory and learning abilities in the rat model of schizophrenia, plausibly through inhibition of the expression of oxidative stress factors in the hippocampus.     

Effect of hypoglycemic anti-deafness capsules in diabetic patients with deafness and toxicological assessment in rats

Objective

Through experiment on animals and clinical trials to explore the safety and efficacy of hypoglycemic anti-deafness capsules on diabetic patients with deafness.

Methods

Total 296 patients with non-insulin dependent diabetes mellitus (NIDDM) were randomly divided into two groups. A treatment group of 164 patients (208 ears) was treated with hypoglycemic anti-deafness capsules based on TCM syndrome differentiation. A control group of 132 patients (184 ears) was treated with glibenclamide and conventional drug treatment for deafness. The following were observed: hearing, fasting plasma glucose (FPG), 2 h postprandial plasma glucose (2hPG), 24 h urine glucose (24hUG), improvement of main symptoms, platelet function, and changes in superoxide dismutase (SOD) and lipid peroxide (LPO) levels. In animal studies, Kunming mice, weighing 18-22 g were used. Half of the mice were males and half were females. Wistar rats, weighing 80-120 g were used. Half of the rats were males and half were females. Male Wistar rats, weighing 200-220 g, were also used. Their acute and chronic toxicity was studied.

Results

The hearing improvement was 56.7% in the treatment group and 26.6% in the control group. FPG, 2hPG, and 24hUG were improved significantly (P<0.05, P<0.01, P<0.01, respectively) in the treatment group and 2hPG and 24hUG improved significantly in the control group (P<0.05, P<0.05). The improvement in 2hPG and 24hUG in the treatment group was significantly greater than that in the control group P<0.01). There was no significant difference in FPG between the two groups (P<0.05). Main symptoms in the treatment group were significantly more improved than those in the control group (P<0.05, P<0.01). In the treatment group, platelet adhesion and aggregation, SOD, and LPO were all significantly improved from before treatment (P<0.05, P<0.01). However, in the control group, except LOP (P<0.05), there were no significant differences from before treatment to after (P<0.05). In animal studies, no obvious acute or long-term toxicity was observed from capsule administration.

Conclusion

Through experiment on animals and clinical trials, we can found that hypoglycemic anti-deafness capsules could decrease blood glucose and serum triglycerides of alloxan-induced diabetic rats. This herbal capsule is effective for safely treating diabetic patients with deafness.     

Intervention of the Mahuang Lianqiao Chixiaodou decoction on immune imbalance in atopic dermatitis-like model mice

ObjectiveTo explore the potential mechanism of intervention on the immune imbalance of atopic dermatitis (AD) by studying the effects of Mahuang Lianqiao Chixiaodou decoction (MLCD) on skin damage and inflammation factors in an AD-like mouse model.MethodsNinety-six male BALB/c mice were divided into normal, model, positive control (mometasone furoate), and traditional Chinese medicine treatment (MLCD) groups by a random number table. 2,4-dinitrofluorobenzene was used to induce AD-like mice in all groups except the normal group. The treatment or intervention was administered for seven consecutive days on days 4, 18, 32, and 39. The mRNA relative expressions of interleukin-4 (IL-4), IL-10, interferon-γ (IFN-γ), thymic stromal lymphopoietin (TSLP), and the TSLP receptor (TSLPR) were measured using quantitative real-time polymerase chain reaction, and the serum immunoglobulin E, IL-4, IL-10, and IFN-γ levels were detected using enzyme-linked immunosorbent assay.ResultsCompared with the normal group, the hematoxylin-eosin staining of the skin lesions of the mice in the model group was significantly thickened on days 11, 25, and 39. Compared with the model group, the epidermal thickness of the positive control group was significantly alleviated on day 39 (P < .001), and that of the MLCD group was significantly improved on days 25 and 39 (P < .001). Compared with the four observation time points, MLCD had the best treatment effect on day 39 of the experiment and significantly improved the skin damage performance and relieved pathological lesions. On day 39, compared with the model group, MLCD downregulated the skin mRNA relative expressions of IL-4 (P = .009), TSLP (P = .030), and TSLPR (P < .001), and reduced the mouse serum levels of IL-4 (P = .003). For other serum indicators, no significant difference was observed between the model and MLCD groups.ConclusionMLCD improved AD-like mice skin damage by regulating the Th1/Th2 immune imbalance.     

怀菊花提取物对皮质酮诱导的抑郁模型的保护作用及机制研究

目的 通过皮质酮诱导的小鼠抑郁症模型和肾上腺嗜铬细胞瘤PC-12细胞损伤模型,研究怀菊花提取物的抗抑郁作用。方法 将雄性C57BL/6小鼠随机分为对照组、模型组、氟西汀（5 mg/kg）组和怀菊花提取物低、中、高剂量（1.67、3.33、6.67 g/kg）组,模型组和各给药组连续21 d sc皮质酮（20 mg/kg）,造模同时ig相应药物,给药结束后进行糖水偏好实验、悬尾实验和旷场实验;采用ELISA法检测小鼠血清中皮质酮及神经递质水平;采用Western blotting检测海马组织中环磷腺苷效应元件结合蛋白（cyclic adenosine monophosphate-response element binding protein,CREB）通路关键蛋白表达。皮质酮诱导PC-12细胞建立细胞损伤模型,采用流式细胞术检测怀菊花提取物对细胞凋亡及活性氧（reactive oxygen species,ROS）水平的影响;采用In-Cell Western法检测凋亡相关蛋白及CREB通路关键蛋白表达。结果 怀菊花提取物能够显著改善小鼠的糖水偏好率（P＜0.05、0.01）,增加小鼠的站立次数（P＜0.05、0.01）,减少小鼠悬尾实验的不动时间（P＜0.01）,显著降低血清中皮质酮水平（P＜0.01）,升高血清中5-羟色胺（5-hydroxytryptamine,5-HT）、去甲肾上腺素（norepinephrine,NE）、多巴胺（dopamine,DA）、乙酰胆碱（acetylcholine,Ach）水平（P＜0.05、0.01）,上调海马组织中CREB、磷酸化CREB（phosphorylated CREB,p-CREB）、环磷酸腺苷（cyclic adenosine monophosphate,cAMP）蛋白表达水平（P＜0.05、0.01）。怀菊花提取物显著抑制皮质酮诱导的PC-12细胞凋亡率及细胞内ROS水平（P＜0.01）,抑制凋亡相关蛋白表达（P＜0.01）,上调CREB通路关键蛋白表达（P＜0.01）。结论 怀菊花提取物可能通过调节CREB通路、抑制神经细胞凋亡,从而改善抑郁。