Effects of Houttuynia Cordata Thumb on Expression of BMP-7 and TGF-β_1 in the Renal Tissues of Diabetic Rats

Objective： To explore the effects of Houttuynia Cordata Thumb （HCT 鱼腥草 Yu Xing Cao） on expression of transforming growth factor-β1 （TGF-β1） and bone morphogenetic protein-7 （BMP-7） in the renal tissues of diabetic rats. Methods： The diabetic rats induced by intravenous injection of streptozotocin（STZ） were randomly divided into a model group, a HCT group and a lotensin group, with normal rats designated as the controls. 8 weeks later, the ratio of kidney weight to body weight, the glomerular area, the excretion of β 2-microglobin （β2-MG） in 24-hr urine, the albumin excretion in 24-hr urine, and creatinine clearance rate （CCR） were investigated. The expression of TGF- β 1, BMP-7 and collagen I in the renal tissues was observed with the immunohistochemical method and by the semi-quantitative assay. Results： The overgrowth of glomerulus, the excretion of β 2-MG in 24-hr urine, the albumin excretion rate in 24-hr urine and CCR in the HCT group significantly reduced （P〈0.05）, and the expression of TGF-β1 and collagen I significantly decreased （P〈0.05）, but BMP-7 significantly increased （P〈0.05） in the HCT group as compared with those in the model group, with no significant difference as compared with the lotensin group （P〉0.05）. Conclusion： HCT has a protective effect on the renal tissues in diabetic rats, which is probably correlated with the decrease of the expression of TGF-β1 and collagen I and with the increase of the expression of BMP-7 in the renal tissues.

Chemical composition and biological activity of Houttuynia cordata

Houttuynia cordata is an important traditional Chinese medicine.It is widely used for clearing heat and detoxification,reducing swelling and treating sores,diuresis and dehumidification.It is also used to treat lung carbuncles,ulcers,hemorrhoids,rectal bleeding,spleen and stomach heat,which are caused by excess heat,heat toxicity,dampness,and disease heat.Modern pharmacological experiments have shown that H.cordata has antibacterial,antiviral,immune enhancing,diuretic and other effects.This article reviews the chemical components and biological activities of H.cordata to provide reference for clinical application.

Study on Underground Stem Tissue Structure and Agronomic Traits of Different Populations of Wild Houttuynia cordata Thumb

[Objectives] To study the underground stem tissue structure and agronomic traits of different populations of wild Houttuynia cordata Thumb. [Methods] 8 different populations of wild Houttuynia cordata Thumb. in southern China were selected,the preliminary study was carried out about underground stem tissue structure( stem wall thickness,underground stem cross section diameter,medullary cavity,large vascular bundle area,small vascular bundle area,and number of vascular bundles),and main agronomic traits of aboveground and underground parts. [Results]There were significant differences in the underground stem tissue structure and agronomic traits of some populations of wild Houttuynia cordata Thumb. [Conclusions] From comparison,it reached the conclusion that the populations of wild Houttuynia cordata Thumb. in Kunming of Yunnan,Hechuan of Chongqing,and Nanning of Guangxi showed better underground stem tissue structure and agronomic traits. The results are expected to provide certain guidance for cultivation,reasonable development,and utilization of wild populations of Houttuynia cordata Thumb.

铅、锌及其交互作用对鱼腥草(Houttuynia cordata)叶绿素含量及抗氧化酶系统的影响

采用营养液培养方法研究铅、锌及其交互作用对鱼腥草叶绿素含量及抗氧化酶系统的影响。实验结果显示,随着Pb浓度的增加,鱼腥草叶绿素含量逐渐降低但无显著变化。Zn在一定浓度下能提高鱼腥草叶绿素含量,而在高浓度Zn胁迫下,叶绿素含量急剧下降。鱼腥草叶片中SOD、POD和CAT3种酶活性都是随着Pb浓度的增加先上升后下降。随着Zn浓度的增加,SOD和CAT也是先上升后下降,POD则是逐渐上升。Pb-Zn交互作用增加了鱼腥草叶绿素含量,对SOD和POD活性具有抑制作用,对CAT活性影响不明显。同时研究结果还表明,单一Pb、Zn对鱼腥草叶绿素含量和抗氧化酶系统的影响大于Pb、Zn二者的共同作用,其中高浓度Zn对鱼腥草的伤害作用最大,而当溶液Pb处理浓度达到400mg/L时,鱼腥草仍能正常生长,说明鱼腥草具有较强的耐Pb能力。

峨眉山不同山峪和海拔高度鱼腥草(Houttuynia cordata Thunb.)居群挥发油成分的变化

以分布于峨眉山不同山峪不同海拔（500-1800m）的17个鱼腥草（Houttuynia cordata Thunb.）居群及生长地土样作为试验材料，按土壤农化分析常规方法对土壤pH值及土壤营养元素进行测定，用水蒸馏法提取植株地上部分的挥发油，并用GCMS对挥发油化学成分进行分离和鉴定，峰面积归一法测定各成分含量。结果显示：峨眉山鱼腥草生长地土壤肥力水平较高，对鱼腥草的生长极为有利。从所有居群中共鉴定出35种化合物，各居群挥发油成分组成及含量存在较大的差异。月桂烯作为四个山峪唯一共有的主要成分在各居群中均能检测到，且在各居群中的平均含量最高。乙酸龙脑酯和trans-β-罗勒烯的含量与海拔高度间分别存在显著和极显著正相关。方差分析及多重比较的结果表明，不同山峪以及海拔高度对各居群挥发油成分和含量有显著影响。通过偏相关分析发现土壤微量元素对鱼腥草挥发油共有成分合成与积累的作用大于大量元素，微量元素中以有效钼的影响最为显著，大量元素中以速效钾的影响最为显著。采用逐步多元回归分析筛选出了对挥发油共有成分含量影响显著的土壤营养元素。

Expression of Exogenous Antimicrobial Peptide and Resistance to Stem Rot Disease of Houttuynia cordata Thunb.

[Objective] This study was to identify the expression of exogenous antimicrobial peptide in transgenic Houttuynia cordata Thunb. plants,and analyze their resistance to stem rot disease. [Methods] SDS-PAGE and Western blot analysis were employed to detect expression of exogenous antimicrobial peptide in transgenic H. cordata plants. Both wild type and transgenic H. cordata plants were inoculated with different concentrations of Rhizoctonia solani spores for detecting their resistance. [Results] The exogenous antimicrobial peptide was detected at translation level. The optimal parameters for detecting the resistance of transgenic H. cordata plants to R. solani was inoculation of spores at a concentration of 3×105 ind./ml and cultured for three days. The results showed that resistance of transgenic H. cordata plants to R. solani was enhanced in comparison with CKs. [Conclusion] Expression of exogenous antimicrobial peptide can enhance the resistance of transgenic H. cordata plants to stem rot disease.

Standard Operating Procedure of Houttuynia cordata Thunb.Plastic Greenhouse Cultivation

This standard operating procedure stipulated the natural condition of pro- ducing area, cultivation technique, disease and pest control, harvest, quality stan- dard, packaging, transportation and storage of H. cordata, in order to provide a ba- sis for the standard cultivation of H. cordata.

Evaluation of antiviral activities of Houttuynia cordata Thunb.extract,quercetin,quercetrin and cinanserin on murine coronavirus and dengue virus infection

Objective:To evaluate the in vitro activities of the ethyl acetate(EA) fraction of Houttuynia cordata(H.cordata) Thunb.(Saururaceae) and three of its constituent flavonoids(quercetin.quercitrin and rutin) against murine coronavirus and dengue virus(DENV).Methods:The antiviral activities of various concentrations of the EA fraction of H.cordata and flavonoids were assessed using virus neutralization tests against mouse hepatitis virus(MHV) and DENV type 2(DENV-2).Cinanserin hydrochloride was also tested against MHV.The EA fraction of H.cordata was tested for acute oral toxicity in C57BL/6 mice.Results:The EA fraction of H.cordata inhibited viral infectivity up to 6 d.Cinanserin hydrochloride was able to inhibit MHV for only 2 d.The 50%inhibitory concentrations(IC_(50)) of the EA fraction of H.cordata added before the viral adsorption stage were 0.98 μg/mL for MHV and 7.50 μg/mL for DENV-2with absence of cytotoxicity.The mice fed with the EA fraction up to 2 000 mg/kg did not induce any signs of acute toxicity,with normal histological features of major organs.Certain flavonoids exhibited comparatively weaker antiviral activity,notably quercetin which could inhibit both MHV and DENV-2.This was followed by quercitrin which could inhibit DENV-2but not MHV,whereas rutin did not exert any inhibitory effect on either virus.When quercetin was combined with quercitrin,enhancement of anti-DENV-2 activity and reduced cytotoxicity were observed.However,the synergistic efficacy of the flavonoid combination was still less than that of the EA fraction.Conclusions:The compounds in H.cordata contribute to the superior antiviral efficacy of the EA fraction which lacked cytotoxicity in vitro and acute toxicity in vim.H.cordata has much potential for the development of antiviral agents against coronavirus and dengue infections.

鱼腥草(Houttuynia cordata)提取物对结肠癌细胞HCT116增殖的抑制作用

运用细胞生存活力测定实验(MTT)研究鱼腥草提取物对人结肠癌细胞HCT116生长的抑制情况.采用倒置显微镜观察不同浓度鱼腥草提取物对结肠癌细胞HCT116形态的影响.实验结果表明,鱼腥草提取物对人结肠癌细胞HCT116的生长增殖有明显的抑制作用,且存在浓度和时间的依赖性.鱼腥草作为天然抗肿瘤化合物的筛选材料具有潜在的价值.

Inhibitory effect of Houttuynia cordata Thunb on LPS-induced retinal microglial activation

AIM: To identify the effect of Houttuynia cordata Thunb(HCT) on lipopolysaccharide(LPS)-induced microglial activation and investigate its possible molecular mechanisms. METHODS: The primary retinal microglial cells were cultured from the retinas of newborn Sprague-Dawley rats and exposed to LPS, and/or HCT with different concentrations. The survival ability of retinal microglia cells was tested by standard MTT method. BrdU cell proliferation assay was used to evaluate the proliferation of retinal microglia. Inflammatory factors in the culture supernatants, including TNF-α, iNOS and IL-1β, were measured using ELISA. Microglia cells’ migration was determined with Transwell migration assay. The total p38-MAPK and phosphorylation of p38-MAPK(p-p38-MAPK) were detected with Western blot.RESULTS: Primary retinal microglia in culture exposed to LPS to induce microglia activation. Pretreatment with HCT significantly inhibited the LPS-induced cell proliferation, but not the cell viability. LPS induced inflammatory reaction in microglia and cell migration. HCT significantly reduced LPS-stimulated release of pro-inflammatory factors and decreased the number of migrating cells substantially in a concentration-dependent manner. Moreover, the protein levels of p-p38 MAPK were identified as the up regulation and co-treatment with HCT obviously inhibited the upregulation of p-p38 MAPK, but had no effect on the levels of total p38-MAPK.CONCLUSION: The data suggest that HCT inhibits LPSinduced retinal microglial activation via suppression of the p-p38-MAPK. HCT may be used for the treatment of ocular diseases characterized by over-activated microglia.

Systematic analysis of components and contents in Houttuynia cordata Thunb

Objective:This study aims to provide the material basis for the molecular mechanism of Yuxingcao(Houttuynia cordata Thunb,HCT)through the method of evidence-based study to summarize the natural constituents isolated from HCT.Methods:We searched CNKI,Wanfang,VIP,Pubmed and relevant conference compilations.The keywords were'Yuxingcao or Houttuynia cordata Thunb'and'components'or'ingredients'or'constituent'or'volatile oil'or'flavonoids'or'terpene'or'content',both in Chinese and English.According to the inclusion and exclusion criteria,the reported compositions and contents have been summarized,and SPSS software was used to draw the boxplot of contents.Results:A total of 603 natural compounds in 11 categories were obtained from pooled articles.In the diverse components,the number of aliphatic compounds(n=259)and terpenoids(n=158)are more than those of flavonoids(n=26),alkaloids(n=42)and aromatics(n=42).While,in the part of volatile oils of HCT,the largest components are aliphatic compounds(mainly distributed on the ground)and terpenoids(mainly distributed in the underground).Although,methyl n-nonylketone is distributed in the whole herb plant,a large proportion is present in the underground parts.As for non-volatiles,the flavonoid content(mainly distributed on the ground)was the highest,among which quercetin and its glycosyl derivatives were the prominent.Conclusion:The results of this study provide a more comprehensive material basis for the further study of HCT.It is also helpful to explain the mechanisms of anti-oxidant,anti-inflammatory,anti-bacterial,anti-viral and anti-cancer effects from the molecular target and molecular network levels.

<i>Houttuynia cordata</i>Thunb: A Review of Phytochemistry and Pharmacology and Quality Control

Houttuynia cordata Thunb is an important medicinal plant widely distributed in East Asia. The collected information is an attempt to cover recent developments in the pharmacology, phytochemistry and quality control of this species. During the past several decades, the medicinally important phyto-constituents have been identified including essential oil, flavonoids and other polyphenols, fatty acids and alkaloids. A survey of the literatures shows H. cordata possesses a variety of pharmacological activities including antiviral, antitumor, antimicrobial, anti-inflammatory, and antioxidative effects. Little attempt has been done to review the techniques used for its quality control. Future efforts should concentrate more on in vitro, in vivo studies and clinical trials in order to confirm traditional wisdom in the light of a rational phytotherapy. The information summarized here is intended to serve as a reference tool to practitioners in the fields of ethnopharmacology and natural products chemistry.

Effects of frying oil and Houttuynia cordata thunb on xenobiotic-metabolizing enzyme system of rodents

AIM: To evaluate the effects of frying oil and Houttuynia cordata Thunb (H. cordata), a vegetable traditionally consumed in Taiwan, on the xenobiotic-metabolizing enzyme system of rodents. METHODS: Forty-eight Sprague-Dawley rats were fed with a diet containing 0%, 2% or 5% H. cordata powder and 15% fresh soybean oil or 24-h oxidized frying oil (OFO) for 28 d respectively. The level of microsomal protein, total cytochrome 450 content (CYP450) and enzyme activities including NADPH reductase, ethoxyresorufin 0-deethylase (EROD), pentoxyresorufin 0-dealkylase (PROD), aniline hydroxylase (ANH), aminopyrine demethylase (AMD), and quinone reductase (QR) were determined. QR represented phase Ⅱ enzymes, the rest of the enzymes tested represented phase Ⅰ enzymes. RESULTS: The oxidized frying oil feeding produced a significant increase in phase Ⅰ and Ⅱ enzyme systems, including the content of CYP450 and microsomal protein, and the activities of NADPH reductase, EROD, PROD, ANH, AMD and QR in rats (P<0.05). In addition, the activities of EROD, ANH and AMD decreased and QR increased after feeding with H. cordata in OFO-fed group (P<0.05). The feeding with 2% H. cordata diet showed the most significant effect. CONCLUSION: The OFO diet induces phases I and II enzyme activity, and the 2% H. cordata diet resulted in a better regulation of the xenobiotic-metabolizing enzyme system.

Phenolics and Antioxidant Activities of Houttuynia cordata Thunb.

[Objectives] To evaluate the phenolic components of Houttuynia cordata Thunb. and their antioxidant activities from different sources. [Methods] Samples of 35 different sources of H. cordata were collected from Guizhou,and polyphenols,total flavonoids,total chlorogenic acid and 7 kinds of flavonoids( xanthol-3-O-glucoside,hyperoside,quercitrin,rutin,quercetin,kaempferol and isorhamnetin),and DPPH free radical scavenging rate of the extracts were determined. Relationship between the contents of phenolics and the DPPH free radical scavenging rate of the extracts were analyzed by SPSS software. [Results] There were some significant differences in polyphenols,total flavonoids,total chlorogenic acid and the seven flavonoids between samples of different habitats,and the contents of polyphenols,total flavonoids and total chlorogenic acid increased linearly with altitude. There was a significant positive correlation between polyphenols,total flavonoids and total chlorogenic acid content and DPPH free radical scavenging rate of extracts. The contents of kaempferol-3-O-glucoside,hyperoside,quercitrin,rutin,quercetin kaempferol and isorhamnetin were positively correlated with the DPPH free radical scavenging rates of the extracts,except for the significant correlation with quercitrin. [Conclusions]The experimental results suggest that the antioxidant capacity of H. cordata is derived from the total contribution of the phenolic components,indicating that these components have overall equivalence of antioxidant capacities. This provides a new research idea for solving the key problems of quality control of traditional Chinese medicine.

Determination of Total Flavonoids and Its Antioxidant Ability in Houttuynia cordata

The total flavonoids of Houttuynia cordata are determined by UV-Vis spectrophotometry. The ex-traction condition of flavonoids from Houttuynia cordata includes: the extraction temperature is 70℃, solid-liquid ratio is 1:20, solvent is 80% ethanol, extraction time is 65 min. We studied the antioxidation of flavonoids in Houttuynia cordata, and results showed that flavonoids had strong reducing capacity, hydroxyl free radical scavenging activity and the superoxide anion radical scavenging ability, thus can provide a basis for the development of natural antioxidants.

Houttuynia cordata Thunb rescues retinal ganglion cells through inhibiting microglia activation in a rat model of retinal ischemia-reperfusion

AIM:To determine whether Houttuynia cordata Thunb(HCT)can increase the survival of the retinal ganglion cells(RGCs)and inhibit microglia activation following retinal ischemia-reperfusion(RIR)injury.METHODS:Rat model of RIR was induced by transient elevation of the intraocular pressure(IOP).HCT was orally administered for 2 d before the performance of retinal RIR model and once a day for the next 14 d.After 14 d of RIR injury,the rats were sacrificed for further analysis.Survival RGCs were stained with haematoxylin and eosin(H&E).Apoptosis of RGCs was detected by TUNEL staining.Retinal function was examined by flash-electroretinography(F-ERG).Retinal microglia were labeled using Iba-1,one specific marker for microglia.The m RNA expression levels of inducible nitric oxide synthase(i NOS),tumor necrosis factor alpha(TNF-α),and interleukin 1 beta(IL-1β)were assessed by quantitative real time reverse transcription polymerase chain reaction(q RT-PCR).RESULTS:Systemic HCT treatment significantly reduced RGCs death by H&E staining and exhibited an anti-apoptotic effect as assessed by TUNEL staining at day 14 after RIR injury.HCT greatly improved the retinal function as examined by F-ERG.The number of activated microglia significantly increased after RIR injury,which was significantly attenuated by HCT treatment.Besides,RIR injury induced a strong upregulation of pro-inflammatory genes TNF-α,i NOS and IL-1βm RNAs at day 14 post injury,which was suppressed by HCT.CONCLUSION:Neuroprotective effects of HCT encourage the survival of RGCs through inhibiting microglia activation due to RIR injury.Together these results support the use of HCT as promising therapy for the ischemic events of the retina diseases.

Network pharmacology-based analysis on three amicoumacin-type isocoumarin compounds from an endophytic bacterium in Houttuynia cordata

Amicoumacin isocoumarin-type compounds are mainly derived from the secondary metabolites of Bacillus bacteria,and have potential biological activities such as antibacterial,anti-tumor,anti-inflammatory,and anti-ulcer.In our previous research,three amicoumacin-type isocoumarin compounds,Paenicoumacin A,Paenicoumacin B,and Paenicoumacin C(B and C are novel),were isolated from an endophytic bacterium in Houttuynia cordata.In this study,network pharmacology and bioinformatics techniques were used to analyze the potential activities of amicoumacin compounds in the treatment of liver cancer.These compounds were found to mainly regulate cellular oxidative stress,cancer process,nucleic acid metabolism,nutritional metabolism and other pathways.The findings provide useful information for subsequent experimental verification.

Primary Research on Protective Effect of Houttuynia cordata Flavonoid and Sophora japonica L. Polysaccharide as Anti-UV Sun-screening Supplement

Houttuynia cordata flavonoid and Sophora japonica L. polysaccharide were investigated to develop anti-UV sun-screening agent through UV wavelength scanning spectrum, antioxidant ability and cell viability after UVB irradiation. The results showed that the extracts had strong UVA and UVB absorption ability and hydroxyl free radical scavenging ability. The S. japonica L. polysaccharide showed strong anti-UVB irradiation ability according to the cell viability experiment. It suggests that the H. cordata flavonoid and S. japonica L. polysaccharide could be combined as a candidate of anti-UV sun-screening supplement and compensate for the shortcomings of chemical sunscreens against UVA ray.

Analysis of Amino Acids in Houttuynia Cordata Thunb. by High Performance Liquid Chromatography-Diode Array Detector (HPLC-DAD)

[Objectives] The biologically active compounds of Houttuynia cordata Thunb. as a medicinal and edible plant are well documented. However, amino acid as an important indicator of the value of H. cordata is little reported. [Methods] Amino acids of the H. cordata were analysed by pre-column derivatization with 2,4-dinitrochlorobenzne(CDNB) coupled to HPLC/DAD. The optimized conditions of the amino acids derivatized by CDNB were followed: 110 ℃ of hydrolysis temperature, 1∶12 of the equivalent proportion between amino acid and CDNB, 80 min of derivatization reaction time and 80 ℃ of derivatization reaction temperature. The method was used to determine the amino acids of H. cordata samples from 11 different geographical sites in Guizhou. [Results] The results suggested that the contents of 17 amino acids were quite variable at different H. cordata samples. [Conclusions] This study will provide a basis for the quality evaluation of H. cordata nutrition.

Establishment of Cytokines Quantitative PCR Methods and Its Application in Anaphylaxis Study of Houttuynia Cordata Injection

We design specific primers, according to the four kinds of important macaque cytokines, based on macaque β-action genes, then extract total RNA in peripheral blood mononuclear cells of macaques, and trascriptase cDNA reversely after establishing real-time quantitative PCR, and test the relative expression amount of several kinds of important cytokines during the hypersensitive process. The testing method of quantitative PCR in this experiment is highly sensitive, all cytokines are up to 102 copies/μL and highly reproducible, the intraassay and interassay coefficients of variation are less than 6%. The method applied to the sensitive reaction experiment on cytokines induced by houttuynia cordata turns out that the cytokines relative expression of houttuynia cordata distillate with low dose group and high dose group is significantly higher than that of saline group (P > 0.01). Meanwhile, significantly increased IL-4, IL-10 and IL-13 and decreased IFN-γ are found in both group A and group B after houttuynia injection (P > 0.05).