Analysis of Patent Application Status of Scutellaria Barbata Industrial Chain Based on Patent Data

Objective To analyze patent application status of Scutellaria Barbata industrial chain and provide some suggestions for its patent application and product development.Methods Patent data were collected through IncoPat patent analysis system.Meanwhile,the patent analysis method combined with text mining method was adopted to analyze the situation and development trend of patent application in China’s Scutellaria Barbata industrial chain by using pie chart,bubble chart,trend chart and other visual charts to display the results.Results and Conclusion The patent application of Scutellaria Barbata in China mainly experienced three stages:Slow development,rapid development,and recession period.The number of patents is large,but the authorization rate is low.Individuals and enterprises are the main applicants for patent applications.Product development is involved in the whole industrial chain,but it basically focuses on its efficacy in downstream drugs,health food and other aspects.Therefore,government should enhance the awareness of patent protection,encourage collaborative innovation in industry-university-research to promote the combination of basic research and market application.Besides,it should provide theoretical support to tackle the problem of short board products,which can promote the transformation of scientific and technological achievements and contribute to the upgrading of Scutellaria Barbata industrial chain.

Scutellaria barbata attenuates diabetic retinopathy by preventing retinal inflammation and the decreased expression of tight junction protein

AIM： To observe the attenuation of ethanol extract of Herba Scutellaria barbata （SE） against diabetic retinopathy （DR） and its engaged mechanism. METHODS： C57BL/6J mice were intraperitoneally injected with streptozotocin （STZ, 55 mg/kg） for 5 consecutive days to induce diabetes, The diabetic mice were orally given with SE （100, 200 mg/kg） for lmo at lmo after STZ injection. Blood-retinal barrier （BRB） breakdown was detected by using Evans blue permeation assay. Real-time polymerase chain reaction （RT-PCR）, Western blot and immunofiuorescence staining were used to detect mRNA and protein expression. Enzyme-linked immunosorbent assay （ELISA） was used to detect serum contents of tumor necrosis factor-e （TNF-a） and interleukin （IL）-II. RESULTS： SE （100, 200 mg/kg） reversed the breakdown of BRB in STZ-induced diabetic mice. The decreased expression of retinal claudin-1 and claudin-19, which are both tight junction （T J） proteins, was reversed by SE. SE decreased the increased serum contents and retinal mRNA expression of TNF-a and IL-113. SE also decreased the increased retinal expression of intercellular cell adhesion molecule-1 （ICAM-1）. SE reduced the increased phosphorylation of nuclear factor kappa B （NFKB） p65 and its subsequent nuclear translocation in retinas from STZ- induced diabetic mice. Results of Western blot and retinal immunofluorescence staining of ionized calcium-binding adapter molecule 1 （Ibal） demonstrated that SE abrogated the activation of microglia cells in STZ-induced diabetic mice. CONCLUSION： SE attenuates the development of DR by inhibiting retinal inflammation and restoring the decreased expression of TJ proteins including claudin-1 and claudin-19.

Attenuation of Scutellaria barbata flavonoids on cortical cytoplasm apoptotic factors disorders induced by complex Aβ_(25-35) in rats

OBJECTIVE To investigate the effect of SBF on cortical cytoplasm apoptotic factors disturbances induced by complex Aβ25-35in rats.METHODS The cerebral injuries model was established by rats received intracere-broventricular injection of RHTGF-β1,Aβ25-35and Al Cl3and then accepted SBF treatment.All the rats were sacrificed by decapitation for indicators detection at last the drug treatment.Western blotting method was for caspase-3 protein expression and RT-PCR method detected cytochrome c,apoptotic protease activating factor-1(Apaf-1),caspase-9 m RNA expression in cortical cytoplasm.RESULTS The protein expression of caspase-3in cortical cytoplasm of rats was assayed by Western blotting.The results indicated that compared with the sham group,the caspase-3 protein expression of cortical cytoplasm in Aβgroup was significantly increased(P<0.01).However,the increased expression can be obviously reversed by SBF at doses of 35,70 and 140mg·kg-1,as compared with model group(P<0.01).The Cyt-C,Apaf-1 and caspase-9 m RNA expressions in cortical cytoplasm of rats were determined by RT-PCR.The results indicated that compared with the sham group,the Cyt-C Apaf-1 and caspase-9 m RNA relative expressions of cortical cytoplasm in Aβgroup was significantly increased(P<0.01).However,these increased expressions can be differently reversed by SBF at doses of 35,70 and 140 mg·kg-1,as compared with model group(P<0.01).CONCLUSION SBF can definitely improve rats′cortical cytoplasm apoptotic factors disorders induced by complex Aβ25-35,which maybe benefit for treatment of degenerative disease.

Study on Infrared Fingerprint of Sculellaria brbata

[Objective] This study was conducted to establish the characteristic finger- prints of herbal materials in the Weinan section of the Yellow River Wetland. [Method] Scutellaria barbata materials were collected from the Weinan section of the Yellow River Wetland. Fingerprint analysis was performed using their mid-IR spectra, and an IR fingerprint analysis method of S. barbata was established.[Result] The four collected S. barbata materials had basically the same IR finger- print characteristics, [Conclusion] S. barbata was identified by IR fingerprint method, and FTIR could serve as a rapid and scientific detection means for macro-control of S. barbata materials.

Chemical profiling of Scutellaria barbata by ultra high performance liquid chromatography coupled with hybrid quadrupole-orbitrap mass spectrometry

An ultra high performance liquid chromatography coupled with hybrid quadrupole-orbitrap mass spectrometry （UHPLC-quadrupole-orbitrap MS） method was developed to systematically analyze chemical constituents of Scutellaria barbata D. Don. The 70% methanol extract was separated on an Agilent Eclipse Plus Cl8 column （1.8μm, 150 mm×2.1 mm）, and eluted with a gradient of methanol-acetonitrile-water containing 0.1% formic acid at a flow rate of 0.25 mL/min. Constituents were identified by HRMS in the negative ion mode using both full scan and two-stage threshold-triggered mass modes. A total of 56 compounds, including 20 free flavonoids, 20 flavonoid O-glycosides, 2 flavonoid C-glycosides, 4 phenylethanoid glycosides, and 10 phenolic compounds were identified by analyzing their high resolution mass spectral data. Among them, 8 flavonoids and 2 phenylethanoid glycosides were unambiguously identified by comparing with reference standards. This study demonstrates that hybrid quadrupole-orbitrap mass spectrometry is a powerful tool in structural identification of unknown compounds in comolicated herbal extracts.

Two New neo-Clerodane Diterpenoids from Scutellaria barbata

Two new neo-clerodane diterpenoids, 6,7-dibenzoyloxybarbatin C （1, named barbatin D） and 6-（2-acetoxy-3-methylbutanoloxy）-7-（2-carbonyl-3-methylbutanoyloxy） barbatin C （2, named barbatin E） were isolated from the whole plant of Scutellaria barbata D. Don. Their structures were elucidated by spectroscopic methods including extensive 1 D and 2D NMR analyses. In vitro, compounds 1-2 showed cytotoxic activities against three human cancer lines, namely, HONE-1 nasopharyngeal, KB oral epidermoid carcinoma, and HT29 colorectal carcinoma cells, and with IC50 values in the range of 3.5-6.7μM.

Scutellaria barbata D.Don Inhibits Colorectal Cancer Growth via Suppression of Wnt/β-Catenin Signaling Pathway

Objective： To investigate the effect of the ethanol extract of Scutellaria barbata D. Don（EESB） on colorectal cancer（CRC） growth and Wnt/β-catenin signaling pathway in vivo and in vitro. Methods： In vivoexperiment, CRC xenograft mouse model was constructed with injection of HT-29 cells. Following xenograft implantation, twenty mice were randomly divided into EESB-treated group（n=10） and control group（n=10） by a random number table, and were given with intra-gastric administration of 2 g/kg EESB or saline, 5 days a week for 16 days, respectively. At the end of experiment, tumors were removed and weighed by electronic scales. The proliferation biomarker Ki-67 of tumor was evaluated by immunohistochemistry（IHC） assay. In vitro study, HT-29 cells were treated with 0, 0.5, 1.5, 2.5 mg/m L EESB for 24 h. At the end of the treatment, the viability and survival of HT-29 cells were determined by methylthiazolyldiphenyl-tetrazolium bromide（MTT） assay and colony formation assay, respectively. The m RNA expression of c-Myc, Survivin and adenomatous polyposis coli（APC） was examined by reverse transcription-polymerase chain reaction（RT-PCR） both in tumor tissues of CRC xenograft mice and HT-29 cells. Protein expression of c-Myc, Survivin, APC, and β-catenin as well as β-catenin phosphorylation level were evaluated by IHC assay or Western blotting. Results： EESB significantly reduced tumor weight in CRC xenografts mice, compared with the control group（P〈0.05）. IHC assay showed that EESB significantly inhibited protein expression of Ki-67 in tumor tissues（P〈0.05）. MTT assay showed that EESB significantly reduced HT-29 cell viability in a dose-dependent manner（P〈0.05）. Colony formation assay showed that EESB dose-dependently decreased the survival of HT-29 cells（P〈0.05）. In addition, RT-PCR assay showed that EESB decreased the m RNA expression of c-Myc and Survivin and increased APC expression, both in tumor tissues of CRC xenograft mice and HT-29 cells（P〈0.05）. IHC assay or Western blotting showed that EESB decreased protein expression of β-catenin, c-Myc and Survivin, as well as increased APC expression and β-catenin phosphorylation in tumor tissues or HT-29 cells（P〈0.05）. Conclusions： EESB significantly reduced tumor growth in CRC xenografts mice, and inhibited the viability and survival of HT-29 cells. EESB could suppress the activation of the Wnt/β-catenin pathway, which might be one of the mechanisms whereby Scutellaria barbata D. Don exerts its anticancer activity.

The genomes of medicinal skullcaps reveal the polyphyletic origins of clerodane diterpene biosynthesis in the family Lamiaceae

The presence of anticancer clerodane diterpenoids is a chemotaxonomic marker for the traditional Chinese medicinal plant Scutellaria barbata,although the molecular mechanisms behind clerodane biosynthesis are unknown.Here,we report a high-quality assembly of the 414.98 Mb genome of S.barbata into 13 pseudochromosomes.Using phylogenomic and biochemical data,we mapped the plastidial metabolism of kaurene(gibberellins),abietane,and clerodane diterpenes in three species of the family Lamiaceae(Scutellaria barbata,Scutellaria baicalensis,and Salvia splendens),facilitating the identification of genes involved in the biosynthesis of the clerodanes,kolavenol,and isokolavenol.We show that clerodane biosynthesis evolved through recruitment and neofunctionalization of genes from gibberellin and abietane metabolism.Despite the assumed monophyletic origin of clerodane biosynthesis,which is widespread in species of the Lamiaceae,our data show distinct evolutionary lineages and suggest polyphyletic origins of clerodane biosynthesis in the family Lamiaceae.Our study not only provides significant insights into the evolution of clerodane biosynthetic pathways in the mint family,Lamiaceae,but also will facilitate the production of anticancer clerodanes through future metabolic engineering efforts.