柴胡桂枝汤对肝纤维化大鼠转化生长因子-β_1表达的影响

目的观察柴胡桂枝汤对肝纤维化大鼠转化生长因子-β1(TGF-β1)表达的影响。方法采用CCl4复合法制备肝纤维化大鼠模型,用免疫组化方法观察柴胡桂枝汤对TGF-β1表达的影响。结果柴胡桂枝汤可抑制模型大鼠肝组织TGF-β1的表达,与模型组及西药对照组相比均有显著性差异(P<0.01和0.05)。结论柴胡桂枝汤可有效防治大鼠肝纤维化,其作用机制可能是通过抑制肝纤维组织TGF-β1的表达实现的。     

Therapeutic effect of tripterine on adjuvant arthritis in rats

AIMS OF THE STUDY: Tripterygium wilfordii Hoog f., a perennial vine, is used in traditional Chinese medicine for treatment of rheumatoid arthritis. This study was to determine whether tripterine, isolated from Tripterygium wilfordii Hoog f., had therapeutic effects on adjuvant arthritis. MATERIALS AND METHODS: Adjuvant arthritis (AA) was induced in rats on day 0. Tripterine 5, 10 and 20 mg kg(-1)day(-1), or prednisone 10 mg kg(-1)day(-1) was given to rats intragastrically from day 19 to day 24. RESULTS: Tripterine significantly inhibited paw swelling and bone destruction in AA rats. Serum level of IgG anti-Mycobacterium tuberculosis antibodies and delayed-type hypersensitivity (DTH) induced by Mycobacterium tuberculosis were also decreased by tripterine. The effects of tripterine were associated with decreased interleukin-1beta (IL-1beta) mRNA expression in ankle joint synovial membrane and tumor necrosis factor-alpha (TNF-alpha) mRNA expression in homogenized paws from adjuvant-induced arthritic rats. CONCLUSIONS: These findings suggested that tripterine had a therapeutic effect on adjuvant arthritis.     

益肾解毒通络方治疗慢性肾炎导致的慢性肾功能衰竭失代偿期30例疗效观察

[目的]观察益肾解毒通络方治疗慢性肾炎导致的慢性肾功能衰竭失代偿期的疗效。[方法]将60例患者随机分为两组,治疗组30例,口服益肾解毒通络方治疗,对照组30例,在常规治疗的基础上联合海昆肾喜胶囊治疗。4周为1个疗程。[结果]2个疗程后,治疗组总有效率为83.33%,对照组为56.67%,两组比较治疗组优于对照组,P<0.05。治疗组治疗后肌酐、尿素氮的水平均明显下降,P<0.01,两组中医症状均有改善,治疗组明显优于对照组,P<0.05。[结论]益肾解毒通络方治疗慢性肾功能衰竭失代偿期,可明显降低患者肌酐及尿素氮水平,延缓肾衰进程。     

蒺藜总皂苷对缺氧复氧心肌细胞NF-κB核移位及其调控下游基因表达的影响

目的观察蒺藜总皂苷(GSTT)对模拟缺血再灌注心肌细胞核因子-κB(NF-κB)核移位及其调控下游基因TNF-α、IL-1β蛋白表达的影响。方法利用原代培养的SD乳鼠心肌细胞建立模拟心肌缺血再灌注模型,分为正常组、模型组和GSTT大、小剂量组。放免法检测TNF-αI、L-1β,免疫组化定性观察NF-κB p65核移位。结果与模型组比较,GSTT大、小剂量组TNF-α、IL-1β含量显著降低(P<0.01或0.05),NF-κB p65核移位明显。结论GSTT对模拟缺血再灌注损伤心肌细胞有保护作用,可减轻炎症损伤,其机制与上游调控枢纽核转录因子NF-κB激活有关。     

HBeAg阳性和阴性慢性乙型肝炎患者病理与细胞因子对比研究

目的探讨HBeAg阳性和HBeAg阴性慢性乙型肝炎(CHB)患者病理分级和分期频数分布情况及与肝纤维化有密切关系的细胞因子α平滑肌肌动蛋白(α-SMA)与转化生长因子β1(TGF-β1)的半定量评分之间的关系。方法随机选取进行肝组织活检的CHB患者118例,HBeAg阳性组57例,HBeAg阴性组61例。所有病例进行肝穿刺活组织检查,并进行炎症分级(G)和纤维化分期(S)诊断,免疫组化标记肝组织中α-SMA及TGF-β1表达,并进行半定量评分。对比2组α-SMA与TGF-β1表达情况。结果 HBeAg阴性组炎症分级G3～4级、纤维化分期S3～4期患者明显多于HBeAg阳性组;TGF-β1表达在1分组HBeAg阳性组频数分布明显高于HBeAg阴性组,而在3分组及4分组中HBeAg阴性组频数分布明显多于HBeAg阳性组;α-SMA在3分组及4分组中HBeAg阴性组频数分布明显高于HBeAg阳性组。结论 HBeAg阴性CHB患者更需要进行早期及时有效的抗病毒治疗,TGF-β1及α-SMA的阳性表达情况可评估对乙型肝炎患者肝纤维化程度。     

Attenuation of diabetic nephropathy by Chaihuang-Yishen granule through anti-inflammatory mechanism in streptozotocin-induced rat model of diabetics

Ethnopharmacological relevance

Traditional Chinese medical herbs have been used in China for a long time to treat different diseases. Based on traditional Chinese medicine (TCM) principle, Chaihuang-Yishen granule (CHYS) was developed and has been employed clinically to treat chronic kidney disease including diabetic nephropathy (DN). The present study was designed to investigate its mechanism of action in treatment of DN.

Materials and methods

Diabetic rats were established by having a right uninephrectomy plus a single intraperitoneal injection of STZ. Rats were divided into four groups of sham, diabetes, diabetes with CHYS and diabetes with fosinopril. CHYS and fosinopril were given to rats by gavage for 20 weeks. Samples from blood, urine and kidney were collected for biochemical, histological, immunohistochemical and molecular analyses.

Results

Rats treated with CHYS showed reduced 24 h urinary protein excretion, decreased serum TC and TG levels, but CHYS treatment did not affect blood glucose level. Glomerular mesangial expansion and tubulointerstitial fibrosis in diabetic rats were significantly alleviated by CHYS treatment. Moreover, CHYS administration markedly reduced mRNA levels of NF-κB p65 and TGF-β1, as well as decreased protein levels of NF-κB p65, MCP-1, TNF-α and TGF-β1 in the kidney of diabetic rats.

Conclusions

CHYS ameliorates renal injury in diabetic rats through reduction of inflammatory cytokines and their intracellular signaling.     

丹参对自发性高血压大鼠炎症因子干预作用的实验研究

目的初步探讨炎症反应在高血压左室重构中的作用,以及中药丹参对自发性高血压大鼠（SHR）左室重构的干预作用与机制。方法选用SHR8周龄40只,随机分为模型对照组、治疗组（丹参＋络活喜）、西药对照组（络活喜）、中药对照组（牛黄降压丸＋络活喜）4组,并以sD大鼠10只作为对照组。疗程20周。观察各组血压及测定心脏重量指数（HWI）;HE染色观察病理学改变;采用放射免疫法检测大鼠血清中白细胞介素-6（IL-6）、肿瘤坏死因子-α（TNF-α）的表达水平。免疫组化法及RT—PCR检测心肌组织内转化生长因子-β1（TGF-β1）蛋白及mRNA水平表达。结果丹参可显著减轻SHR心肌细胞肥大、心肌问质炎性细胞浸润,也有降低心脏重量指数的趋势,但对收缩压无明显改变。SHR血清中TNF-α、IL-6,心肌组织内TGF-β1蛋白及mRNA表达水平均较sD大鼠显著升高（P〈0．01）;治疗组与模型对照组比较可明显降低SHR的TNF-α、IL-6、TGF-β1表达水平（P〈0．01）,优于其他给药组。结论SHR血清中TNF-α、IL-6和心肌组织内TGF-β1的过度表达参与了高血压左室重构的过程。中药丹参具有独立于降压之外的逆转SHR左室重构的作用,其机制可能与降低TNF-α、IL-6以及TGF-β1的表达水平有关。     

甘草甜素对大鼠肾小球硬化早期的防护作用

目的探讨甘草甜素对大鼠肾小球硬化早期的保护作用。方法采用两次经尾iv阿霉素方法建立大鼠肾小球硬化模型。实验随机分为3组,模型组、甘草甜素治疗组和对照组。检测各组大鼠第4、6、8周各项指标的变化,包括24 h尿蛋白定量、血清尿素氮(BUN)、肌酐(SCr)、胆固醇(TC)和白蛋白(Alb)。各组取肾皮质进行光镜等组织病理学检测。应用免疫组织化学方法检测肾组织内转化生长因子β1(TGF-β1)、结缔组织生长因子(CTGF)蛋白的表达。采用标准曲线法进行荧光定量PCR检测肾组织内TGF-β1、CTGF和金属蛋白酶组织抑制物-1(TIMP-1)mRNA的表达。结果甘草甜素治疗组大鼠24 h尿蛋白定量、BUN、SCr、TC和Alb等指标与模型组相比较均有不同程度改善(P0.05)。肾小球硬化程度治疗组明显轻于模型组。治疗组与模型组比较肾组织内TGF-β1和CTGF蛋白表达均有不同程度降低,表达峰值下降(P0.05)。治疗组与模型组比较TGF-β1、CTGF和TIMP-1 mRNA表达均有不同程度降低,表达峰值下降(P0.05)。结论从蛋白和mRNA水平同时证实甘草甜素对TGF-β1、CTGF和TIMP-1的表达有明显的抑制作用,证实甘草甜素对大鼠肾小球硬化有早期保护作用。     

HMCO5, herbal extract, inhibits NF-kappaB expression in lipopolysaccharide treated macrophages and reduces atherosclerotic lesions in cholesterol fed mice

HMCO5 is a herbal extract which comprises of eight different herbs. We studied whether this extract has anti-atherosclerotic effects. In lipopolysaccharide (LPS) stimulated RAW264.7 cells, HMCO5 inhibited NF-kappaB activation as well as iNOS promoter activity, inhibited the secretion of TNF-alpha and IL-1beta, and directly inhibited the intracellular accumulation of reactive oxygen species. ApoE knock-out mice fed a high-fat high-cholesterol diet with HMCO5 for 10 weeks showed a significant reduction in atherosclerotic lesions. A notable finding was the preservation of the smooth muscle cell layer in the media of aorta in the HMCO5 co-treated mice. HMCO5 treated mice did not show significant decrease in serum level of cholesterol. These results suggest that HMCO5 has anti-atherosclerotic effects which in part may be attributable to the inhibition of production of NF-kappaB dependent pro-inflammatory cytokines.     

Anti-atherosclerotic effects of Polygonum aviculare L. ethanol extract in ApoE knock-out mice fed a Western diet mediated via the MAPK pathway

Ethnopharmacological relevance

Polygonum aviculare L. has been used in traditional Korean medicine to treat obesity and symptoms associated with hypertension. The effectiveness or mechanism of Polygonum aviculare L. ethanol extract (PAE) on atherosclerosis disease has not been examined experimentally. This study investigated the protective effect of PAE in atherosclerotic mice.

Materials and methods

ApoE KO mice were fed a Western diet (WD) alone or with PAE or a statin for 12 weeks, followed by analysis of bodyweight, serum lipid levels, and blood pressure. Staining of the aorta and adipose tissue, expression levels of adhesion molecules, and the MAPK pathway were also examined. Cell viability, NF-κB activity, and protein levels of adhesion molecules were assessed in vitro.

Results

ApoE KO mice fed PAE (50 and 100 mg/kg) or statin (10 mg/kg) gained less body weight, and has less adipose tissue and lower serum lipid levels and blood pressures than the WD group. Aorta ICAM-1, VCAM-1, and NF-κB levels were decreased by PAE in a dose-dependent manner, consistent with the in vitro observations. PAE and statin decreased atherosclerotic plaque and adipocyte size versus the WD group. Furthermore, PAE decreased phosphorylation of MAPK pathway components in the aorta of PAE-treated mice, suggesting that PAE's anti-atherosclerotic effects are mediated via a MAPK pathway-dependent mechanism.

Conclusions

PAE may protect against the development of atherosclerotic disease. The beneficial effects are associated with lowering bodyweight, serum lipids, blood pressure, adhesion molecular protein levels, atherosclerotic plaque, and adipocyte size, involving the MAPK pathway.     

虎杖及其有效成分虎杖苷对肾缺血再灌注损伤的保护作用

目的探讨虎杖对肾缺血再灌注损伤大鼠肾脏的保护作用及其机制。方法健康雄性SD大鼠72只随机分为4组,假手术组、缺血再灌注组、虎杖预处理及虎杖苷预处理组,每组18只。再灌注时问分为3个时间点（缺血前10分钟、再灌注后6小时、再灌注后12小时）,每组每个时间点取6只大鼠。采用切除大鼠右侧肾脏,夹闭左侧肾蒂60分钟的肾缺血后再灌注模型。免疫组化法检测肾组织细胞间黏附分子-1（intercellular adhesion molecule-1,ICAM-1）的表达,酶联免疫吸附测定法检测血清中肿瘤坏死因子-α（TNF—α）的含量,并分别进行比较。结果再灌注6小时、12小时后虎杖预处理及虎杖苷预处理组肾组织ICAM-1阳性表达均较缺血再灌注组降低（P〈0．05）,术前10分钟各组间无明显变化。再灌注6小时后虎杖预处理及虎杖苷预处理组血清TNF-α含量明显低于缺血再灌注组（P〈0．05）,再灌注12小时后无明显变化。结论在肾缺血再灌注损伤的过程中,虎杖可能具有抑制肾组织中ICAM-1表达,减少血清中TNF-α的含量,其中虎杖苷起了主要作用。     

Protective and immunomodulatory effect of Gingyo-san in a murine model of acute lung inflammation

To investigate the effects of Gingyo-san (GGS), the traditional Chinese medicinal formula, on the acute lung inflammation induced by LPS in vivo, mice were challenged with intratracheal LPS before treatment with GGS or vehicle. In lung morphology, GGS reduced the infiltration of activated polymorphonuclear neutrophils in the airways, decreased pulmonary edema, reduced nitrosative stress, and improved lung morphology. ELISA or RT-PCR detected the expression of cytokines in BALF and lung tissue. The mechanism of these benefits by treatment with GGS including attenuating expression TNFalpha, IL-1 beta, IL-6, KC, MCP-1, MIP-2, iNOS, and activation of nuclear factor (NF-kappaB and AP-1) in BALF and lung tissue. Particularly, GGS also enhanced the anti-inflammatory cytokine (IL-10) and limited the acute lung inflammation. Therefore, its protection activity against LPS-induced lung inflammatory mediators release might be beneficial in the treatment of endotoxin-associated inflammation.     

The neuroprotective effect of modified “Shengyu” decoction is mediated through an anti-inflammatory mechanism in the rat after traumatic brain injury

Ethnopharmacological relevance

“Shengyu” decoction, a traditional Chinese medicine, has been used to treat diseases with deficit in “qi” and “blood” induced frequently by profound loss of blood or by long sores with heavy pus, in which a potential anti-inflammatory effect is implied. The modified “Shengyu” decoction (MSD) used in the present study was designed on the basis of the “Shengyu” decoction, additional four herbs were added in. Many ingredients in these herbs have been demonstrated to be anti-inflammatory and thus MSD may be used for the treatment of traumatic brain injury (TBI). To evaluate the neuroprotective effect and the underlying mechanisms of MSD on the rat brain after TBI.

Materials and methods

TBI was induced in the right cerebral cortex of male adult rats using Feeney's weight-drop method. The rats were administered a gavage of MSD (0.5, 1.0 or 2.0 ml/200 g) 6 h after TBI. The neurological functions, brain water content, contusion volume, and neuron loss were determined. The levels of TNF-α, IL-1β, IL-6, and IL-10 and the number of GFAP- and Iba1-positive cells in the brain ipsilateral to TBI were also measured. Moreover, the influence of MSD on these variables was observed at the same time.

Results

The neurological deficits, brain water content, and neuron loss were significantly reduced after 1.0 or 2.0 ml/200 g of MSD treatment but not after 0.5 ml/200 g. In addition, treatment with MSD (1.0 ml/200 g) significantly increased the level of IL-10 and reduced the level of TNF-α and IL-1β and the number of GFAP- and Iba1-positive cells after TBI. However, the contusion volume of brain tissue and the expression of IL-6 were not significantly changed.

Conclusion

MSD may be a potential therapeutic for the treatment of TBI because MSD alleviated secondary brain injury induced by TBI. In addition, MSD inhibited the inflammatory response through reducing the expression of inflammatory cytokines and the activation of microglial cells and astrocytes in the brain tissue of rats after TBI. Therefore, a potential anti-inflammatory mechanism of the “Shengyu” decoction was confirmed, which may be one of the main reasons of “Shengyu” decoction used to treat diseases with obvious inflammatory responses.     

Suppression of lipopolysaccharide-induced of inducible nitric oxide synthase and cyclooxygenase-2 by Sanguis Draconis, a dragon's blood resin, in RAW 264.7 cells

Sanguis Draconis (SD) is a kind of dragon's blood resin that is obtained from Daemomorops draco (Palmae). It is used in traditional medicine and has shown anti-inflammatory activity in some diseases. In this study, we examined the effects of Sanguis Dranonis ethanol extract (SDEE) on LPS-induced inflammation using RAW 264.7 cells. Our data indicated that SDEE inhibits LPS-stimulated NO, PGE2, IL-1 beta and TNF-alpha release, and iNOS and COX-2 expression. Furthermore, SDEE suppressed the LPS-induced p65 expression of NF-kappa B, which was associated with the inhibition of I kappa B-alpha degradation. We also found that the expression of HO-1 was significantly increased in RAW 264.7 cells by SDEE. These results suggest among possibilities of anti-inflammation that SDEE inhibits the production of NO and PGE2 by the down-regulation of iNOS and COX-2 gene expression via the suppression of NF-kappaB (p65) activation. SDEE can induce HO-1 over-expression in macrophage cells, which indicates that it may possess antioxidant properties. This result means that SEDD its anti-inflammatory effects in macrophages may be through a novel mechanism that involves the action of HO-1. Thus, SD could provide a potential therapeutic approach for inflammation-associated disorders.     

Antitumor effect of laticifer proteins of Himatanthus drasticus (Mart.) Plumel - Apocynaceae

Ethnopharmacological relevance

Himatanthus drasticus (Mart.) Plumel - Apocynaceae is a medicinal plant popularly known as Janaguba. Its bark and latex have been used by the public for cancer treatment, among other medicinal uses. However, there is almost no scientific research report on its medicinal properties.

Aim of the study

The aim of this study was to investigate the antitumor effects of Himatanthus drasticus latex proteins (HdLP) in experimental models.

Materials and methods

The in vitro cytotoxic activity of the HdLP was determined on cultured tumor cells. HdLP was also tested for its ability to induce lysis of mouse erythrocytes. In vivo antitumor activity was assessed in two experimental models, Sarcoma 180 and Walker 256 carcinosarcoma. Additionally, its effects on the immunological system were also investigated.

Results

HdLP did not show any significant in vitro cytotoxic effect at experimental exposure levels. When intraperitoneally administered, HdLP was active against both in vivo experimental tumors. However, it was inactive by oral administration. The histopathological analysis indicates that the liver and kidney were only weakly affected by HdLP treatment. It was also demonstrated that HdLP acts as an immunomodulatory agent, increasing the production of OVA-specific antibodies. Additionally, it increased relative spleen weight and the incidence of megakaryocyte colonies.

Conclusion

In summary, HdLP has some interesting anticancer activity that could be associated with its immunostimulating properties.