Jatrorrhizine Hydrochloride alleviates tert-butyl hydroperoxide-induced endothelial cell injury through its anti-inflammatory activity and PPAR-γ activation.

The aim of this study was to investigate whether Jatrorrhizine hydrochloride (JAH) can attenuate oxidative damage of endothelial cells by regulating mitochondrial function and inflammatory response. It was found that JAH inhibited tert-butyl hydroperoxide (t-BHP)-induced oxidative damage in mouse brain endothelial cells (MBECs) by increasing cell viability and inhibiting cell apoptosis. Moreover, JAH significantly inhibited the production of reactive oxygen species (ROS) and lipid peroxidation. It enhanced mitochondrial membrane potential (MMP) and maintained ATP synthesis. In addition, JAH regulated the expressions of inflammatory cytokines and increased the activation of endothelial nitric oxide synthase (eNOS). The protective effect of JAH was related to the protein expression of peroxisome proliferator-activated receptor-γ (PPAR-γ) gene. In conclusion, these results suggest that JAH may have therapeutic potential for ischemic stroke associated with endothelial dysfunction through its antioxidant and anti-inflammatory properties.

Jatrorrhizine Hydrochloride Suppresses RANKL-Induced Osteoclastogenesis and Protects against Wear Particle-Induced Osteolysis.

Wear particle-induced aseptic prosthetic loosening is a major complication associated with total joint arthroplasty (TJA). A growing body of evidence suggests that receptor activator of nuclear factor κ-B ligand (RANKL)-stimulated osteoclastogenesis and bone resorption are responsible for peri-implant loosening. Thus, agents which attenuate excessive osteoclast differentiation and function have been considered to offer therapeutic potential for prolonging the life of TJA implants. Jatrorrhizine hydrochloride (JH), a major protoberberine alkaloid isolated from the traditional Chinese herb Coptis chinensis, has been reported to have antimicrobial, antitumor, and antihypercholesterolemic and neuroprotective activities. However, its effects on osteoclast biology remain unknown. Here, we found that JH inhibited RANKL-induced osteoclast formation and bone resorption in vitro and exerted protection against titanium (Ti) particle-induced osteolysis in vivo. Biochemical analysis demonstrated that JH suppressed RANKL-induced activation of MAPKs (p38 and ERK) which down-regulated the production of NFATc1 and NFATc1-regulated osteoclastic marker genes, such as TRAP, CTR and CTSK. Collectively, our findings suggest that JH may be a promising anti-osteoclastogenesis agent for treating periprosthetic osteolysis or other osteoclast-related osteolytic diseases.

Jatrorrhizine Hydrochloride Suppresses Proliferation, Migration, and Secretion of Synoviocytes In Vitro and Ameliorates Rat Models of Rheumatoid Arthritis In Vivo.

Jatrorrhizine hydrochloride (JH), an active component isolated from the traditional Chinese herb Coptis chinensis, has been reported to have antimicrobial, antitumor, antihypercholesterolemic, and neuroprotective activities. However, its antirheumatoid arthritis (RA) property remains unknown. In this study, a collagen-induced arthritis (CIA) rat model was used to evaluate the therapeutic effects of JH on RA by using arthritis score, radiological evaluation, and histopathological assessment. The in vitro effects of JH on proliferation, migration, and production of inflammatory mediators in RA-derived fibroblast-like synoviocyte MH7A cells were determined by the EdU incorporation assay, wound healing assay, real-time PCR, and ELISA, respectively. The in vivo studies showed that JH treatment significantly prevented the progression and development of RA in CIA rats through anti-inflammation and suppressing bone destruction. The in vitro studies revealed that JH could effectively attenuate the destructive phenotypes of MH7A cells, including inhibiting proliferation, migration, and production of inflammatory mediators. Further mechanistic analysis demonstrated that JH suppressed tumor necrosis factor alpha (TNFα)-stimulated activations of nuclear factor of kappaB (NF-κB) and mitogen-activated protein kinases (MAPKs) (ERK and p38) leading to the downregulation of proinflammatory cytokines, which might be beneficial to the antiproliferative and antimigratory activities of FLS cells. Collectively, our results demonstrated that JH has a great potential to be developed into a novel therapeutic agent for treating RA.

Simultaneous Determination of 7 Components in Siwei Jianghuang Decoction Powder by HPLC / 中国药房

OBJECTIVE:To establish a method for simultaneous determination of gallic acid,magnoflorine,ellagic acid,jatrorrhizine hydrochloride,palmatine hydrochloride,berberine hydrochloride and curcumin in Siwei jianghuang decoction powder.METHODS:HPLC method was adopted.The determination was performed on Capcell Pak C18-MG Ⅱ column with mobile phase consisted of acetonitrile-0.1％ phosphoric acid (gradient elution) at the flow rate of 0.8 mL/min.The detection wavelengths were 270 nm (0-60 min,gallic acid,magnoflorine,ellagic acid,jatrorrhizine hydrochloride,palmatine hydrochloride,berberine hydrochloride) and 428 nm (60-70 min,curcumin).The column temperature was set at 30 ℃,and sample size was 10 μL.RESULTS:The linear ranges of gallic acid,magnoflorine,ellagic acid,jatrorrhizine hydrochloride,palmatine hydrochloride,berberine hydrochloride and curcumin were 0.249 6-1.497 6,0.284 0-1.704 0,0.075 6-0.453 6,0.015 9-0.095 9,0.023 6-0.141 6,0.098 2-0.589 0 and 0.060 4-0.362 4 μtg (r≥0.999 8).The limits of detection were 6.24,4.73,7.56,2.36,3.20,6.54,6.04 ng,and the limits of quantitation were 17.47,16.08,20.86,7.31,10.24,19.62,19.32 ng,respectively.RSDs of precision,stability (12 h),reproducibility tests were lower than 2.0％ (n=6).The recoveries were 95.45％-103.47％ (RSD=0.86％-1.98％,n=9).CONCLUSIONS:Established method is simple,accurate,reliable and suitable for simultaneous determination of 7 components such as gallic acid in Siwei jianghuang decoction powder.

Simultaneous determination of six constituents in Qingre Zhili Pills by HPLC / 中成药

AIM To develop an HPLC method for the simultaneous content determination of berberine hydrochloride,jatrorrhizine hydrochloride,palmatine hydrochloride,kaempferol,luteolin and apigenin in Qingre Zhili Pills (Berberidis Radix and Portulacae Herba).METHODS The analysis of methanol extract of this drug was performed on a 35 ℃ thermostatic Agilent TC-C18 column (4.6 mm × 250 mm,5 μm),with the mobile phase comprising of acetonitrile-0.02％ potassium dihydrogen phosphate flowing at 0.9 mL/min in a gradient elution manner,and the detection wavelengths were set at 265 nm and 360 nm.RESULTS Six constituents showed good linear relationships within their own ranges (r≥0.999 3),whose average recoveries were 97.16％-99.90％ with the RSDs of 0.62％-1.48％.CONCLUSION This simple method can be used for the rapid quality control of Qingre Zhili Pills.

Simultaneous determination of six constituents in Jiaotai Pills by HPLC / 中成药

AIM To establish an HPLC method for the simultaneous content determination of six constituents in Jiaotai Pills (Coptidis Rhizoma and Cinnamomi Cortex).METHODS The analysis of 30％ methanol of this drug was performed on a 30 ℃ Agilent ZORBAX SB-C1s column (4.6 mm ×250 mm,5 μm),with the mobile phase comprising of acetonitrile-KH2PO4flowing at 0.8 mL/min in a gradient elution manner,and the detection wavelength was set at 276 nm.RESULTS Epiberberine,jatrorrhizine hydrochloride,coptisine hydrochloride,palmatine chloride,berberine hydrochloride and cinnamaldehyde showed good linear relationships within the ranges of 0.64-41.24 μg/mL (R2 =0.999 9),0.65-43.76 μg/mL (R2 =1.000 0),0.82-52.65 μg/mL (R2 =0.999 9),0.79-50.70 μg/mL (R2 =0.999 9),3.08-197.20 μg/mL (R2=0.999 8) and 0.65-41.65 μg/mL (R2 =0.999 9),whose average recoveries were 98.06％,102.76％,99.27％,99.75％,96.74％ and 101.33％ with the RSDs of 0.56％,0.54％,0.39％,0.55％,0.48％ and 2.14％,respectively.CONCLUSION This accurate,sensitive,stable and reproducible method can be used for the quality control of Jiaotai Pills.

[Determination of six alkaloids in six types of Coptidis Rhizoma pieces by RP-HPLC and spectrum-effect relationships with anti-diabetes pharmacodynamics data].

To establish a method for determining the contents of six alkaloids (jatrorrhizine hydrochloride, columbamine hydrochloride, epiberberine hydrochloride, coptisine hydrochloride, palmatine hydrochloride, berberine hydrochloride) in six types of Coptidis Rhizoma pieces (crude pieces, ginger juice stir-fried pieces, vinegar stir-fried pieces, wine steamed pieces, wine stir-fried pieces, evodiae juice stir-fried pieces) by RP-HPLC, and explore the relationship with the curative effect of traditional Chinese medicine (TCM) and pharmacodynamics results. The chromatographic column was Welch XtimateTM C18 (4.6 mm×250 mm, 5 µm), with 0.1% triethylamine solution (adjust pH at 10 with ammonium bicarbonate and ammonia) as mobile phase A and acetonitrile as mobile phase B for gradient elution (0-15 min, 10%-25%B; 15-25 min, 25%-30%B; 25-40 min, 30%-45%B) at a rate of 1.0 mL•min⁻¹. The column temperature was set at 30 ℃, and the wavelength was set at 270 nm. The six alkaloids showed a good linear relationship within the range of 0.85-16.96 mg•L⁻¹ (r=0.999 7), 1.25-24.96 mg•L⁻¹ (r=0.999 9), 2.05-40.96 mg•L⁻¹ (r=0.999 9), 3.65-72.96 mg•L⁻¹ (r=0.999 9), 2.88-57.60 mg•L⁻¹ (r=0.999 8), and 13.25-264.96 mg•L⁻¹ (r=0.999 6) respectively. The average recoveries (n=9) of the six alkaloids were 102.4% (RSD 1.2%), 101.8% (RSD 1.3%), 100.3% (RSD 1.8%), 100.7%(RSD 1.8%), 101.2% (RSD 1.5%) and 97.90% (RSD 2.0%) respectively, and their average contents were 3.55, 4.49, 9.12, 19.17, 15.69, 62.56 mg•g⁻¹, respectively. This determination method was accurate and repeatable, which could be used for the content determination in six types of Coptidis Rhizoma pieces. Data analysis on contents determination and preliminary pharmacodynamics results was conducted by using principal component analysis (PCA) and hierarchical clustering analysis (HCA). The analysis results showed that three types of Coptidis Rhizoma pieces (wine steamed pieces, wine stir-fried pieces, and evodiae juice stir-fried pieces) had significant differences with crude pieces, and the wine steamed Coptidis Rhizoma pieces showed most difference with crude pieces especially, mainly related to triglyceride (TG) and fasting blood glucose levels (FBG) in serum. In addition, columbamine hydrochloride was most affected among the six alkaloids. Those three types of Coptidis Rhizoma pieces (wine steamed pieces, wine stir-fried pieces, and evodiae juice stir-fried pieces), had more advantages for "anti-diabetes" in TCM clinical application, especially in the treatment of diabetic hyperlipidemia.

Determination of six alkaloids in crude and processed Coptidis Rhizoma with quantitative analysis of multi-components by single marker / 中草药

Objective: To establish a quantitative analysis of multi-components by single marker (QAMS) for the simultaneous determination of six alkaloids in crude and processed Coptidis Rhizoma. Methods: An HPLC method was established to determine the six alkaloids (jatrorrhizine hydrochloride, columbamine hydrochloride, epiberberine hydrochloride, coptisine hydrochloride, palmatine hydrochloride, and berberine hydrochloride) by the external standard method (ESM). With this HPLC method, the berberine hydrochloride was used as the internal standard (IS) to determine five relative correction factors (RCFs) of the five other alkaloids, and their contents in all samples were calculated by their RCFs at the same time. Compared with the content results determined by the ESM and QAMS, the feasibility and accuracy of QAMS method were verified. Results: Within a certain range, the RCFs of jatrorrhizine hydrochloride, columbamine hydrochloride, epiberberine hydrochloride, coptisine hydrochloride, and palmatine hydrochloride to berberine hydrochloride were 1.131, 0.999, 1.011, 1.076, and 1.025, respectively, with the good repeatability in different experimental conditions. There was no significant difference between the QAMS method and ESM method. Conclusion: The QAMS method is feasible and accurate for the simultaneous determination of the six alkaloids in crude and processed Coptidis Rhizoma.

Comparison on content differences of four bioactive compounds in different Wujiwan decoction by HPCE internal standard method / 中草药

Objective: To study the content difference of berberine hydrochloride, palmatine hydrochloride, jatrorrhizine hydrochloride, and paeoniflorin in Wujiwan traditional decoction, granule prescription, and compound granule using HPCE internal standard method. Methods: Capillary zone electrophoresis (CZE) method was used for a complete separation of the components, which was achieved with 50 mmol/L borax buffer-methanol (2:1), constant voltage of 25 kV, pressure injection 5 kPa × 15 s, electrolyle sealing 5 kPa × 10 s, and column temperature of 15°C. Results: The capillary electrophoresis method is simple and accurate. The contents of the three alkaloids in the granule prescription were obviously higher than those in traditional decoction and compound granule. Conclusion: This method is instructive for clinical application and quality control of Wujiwan prescription and compound granule.

Determination of alkaloid from combination of Rhizoma Coptidis and Fructus Evodiae by TLCS / 中成药

AIM: To investigate the changes of alkaloid in Rhizorrm Coptidis and Fructus Evodiae before and after combination. METHODS: The content of berberine hydrochloride,palmatine hydrochloride,jatrorrhizine hydrochloride in Rhizoma Coptidis and Fructus Evodiae before and after combination were determined by TLCS.RESULTS : After combination of Rhizoma Coptidis and Fructus Evodiae,contents of berberine hydrochioride,palmatine hydrochloride,jatrorrhizine hydrochloride significantly reduced,among which the reduction of berberine hydrochloride was more obvious.CONCLUSION: In decoction,Fructus Evodiae appears to restrict the alkaloid from Rhizoma Coptidis in order to lower the side effect of Rhizoma Coptidis.

Simultaneous Determination of5Compositions in Coptis Chinensis Detoxication Capsule by HPLC / 中国药房

OBEJETIVE:To determinate simultaneously the contents of berberine hydrochloride,palmatine hydrochloride,jatrorrhizine hydrochloride,baicalin and jasminoidin in coptis chinensis detoxication capsule by HPLC.METHODS:3different UV wavelengths were adopted simultaneous in the determination in which Diamonsil C 18 was taken as the chromatographic column and water-methanol-0.05%H 3 PO 4 (gradient elution)was taken as mobile phase,the column temperature was set at35℃and the flow rate was1.0ml/min,the detection wavelength were345nm,280nm and238nm respectively and the sample size was10?l.RESULTS:They took good linear relationships when the respective sample size of berberine hydrochloride,pal-matine hydrochloride,jatrorrhizine hydrochloride,baicalin and jasminoidin were at0.105?g～1.680?g(r=0.9999),0.045?g～ 0.720?g(r=0.9998),0.065?g～1.040?g(r=0.9997),0.190?g～3.040?g(r=0.9999)and0.145?g～2.320?g(r=0.9999);Their respective average recovery rates were99.11%,98.19%,97.21%,98.52%and99.22%.CONCLUSION:This method is fast,reproducible,sensitive,and which can provide a more reasonable and reliable quality control for coptis chinensis detoxi-cation capsule.

Comparison of the Main Components between Traditional Decoction and Dispensing Granule Decoction of Huanglian Jiedu Decoction / 中药新药与临床药理

Objective To compare the contents of berberine,palmatine,jatrorrhizine,baicalin and geniposide between traditional decoction and dispensing granule decoction of Huanglian Jiedu Decoction.Methods A HPLC method was carried out on Diamonsil C18 column(250 ? 4.6 mm,5 ?m).The mixture of H2O(A),CH3OH(B)and 0.05 % H3PO4 solution(C)was used as the mobile phase for gradient elution.The column temperature was set up at 35 ℃ and the flow rate was l mL/min.The detection wavelength was 345nm for berberine,palmtine and jatrorrhizine,280 nm for baicalin,and 238 nm for geniposide.Results This method can be used to determine 5 components in Huanglian Jiedu Decoction at the same time and the method was rapid,sensitive and accurate.Conclusion The HPLC chromatograms of traditional sliced decoction of Huanglian Jiedu Decoction are similar to the dispensing granule decoction.The contents of 5 main components in dispensing granule decoction are higher than those in traditional sliced decoction.

Determination of Three Kinds of Alkaloids in Yiqi Zhixue Granules by Ion Pair RP-HPLC Method / 中药新药与临床药理

Objective To develop a method for the determination of berberine hydrochloride,palmatine hydrochloride and jatrorrhizine hydrochloride in Yiqi Zhixue Granules. Methods The samples were extracted by ultrasonication with methanol for 30 min,and an ion pair RP-HPLC method was applied to determine three kinds of alkaloids.The column of C18 with temperature at 30 ℃ was used to separate the target components,the mobile phase consisted of the mixed solution of 0.01 mol/L sodium-heptanesulfonate solution and equal volume of 0.01 mol/L potassium dihydrogen phosphate solution(pH being adjusted at 3.1 with phosphoric acid) combined with acetonitrile(70 ∶ 30),the flow rate was 1.0 mL/min,and the detection wavelength was at 345 nm. Results Three kinds of alkaloids were separated perfectly,the average recoveries(n=6)were 100.95 %(RSD=2.10 % ) for berberine chloride,102.14 %(RSD=2.29 % ) for palmatine chloride,and 100.71 %(RSD=2.65 % ) for jatrorrhizine chloride. Conclusion The developed method is demonstrated to be simple,specific and accurate,which can be used to determine the contents of berberine chloride,palmatine chloride and jatrorrhizine chloride in Yiqi Zhixue Granula,and to control the quality of Folium Mahoniae in Yiqi Zhixue Granules.

Determination of alkaloid from combination of Rhizoma Coptidis and Fructus Evodiae by TLCS / 中成药

AIM:To investigate the changes of alkaloid in Rhizoma Coptidis and Fructus Evodiae before and after combination.METHODS:The content of berberine hydrochloride,palmatine hydrochloride,jatrorrhizine hydrochloride in Rhizoma Coptidis and Fructus Evodiae before and after combination were determined by TLCS.RESULTS:After combination of Rhizoma Coptidis and Fructus Evodiae,contents of berberine hydrochloride,palmatine hydrochloride,jatrorrhizine hydrochloride significantly reduced,among which the reduction of berberine hydrochloride was more obvious.CONCLUSION:In decoction,Fructus Evodiae appears to restrict the alkaloid from Rhizoma Coptidis in order to lower the side effect of Rhizoma Coptidis.

Simultaneously determination of multi-target ingredients in Zuojin Pill by RP-HPLC / 中成药

AIM:To simultaneously determine jatrorrhizine hydrochloride,palmatine hydrochloride,berberine hydrochloride、 evodiamine and rutaecarpine in Zuojin Pill(Rhizoma Coptidis,Fructus Evodiae).METHODS:A RP-HPLC method was established with gradient elution and an agilent C_ 18 column(250 mm?4.6 mm,5 ?m)was used.The mobile phase A was acetonitrile and the mobile phase B was water with 0.3% phosphoric acid-triethylamine.The detective wavelength was at 225 nm.RESULTS:Jatrorrhizine hydrochloride had a good linearity in the range of 0.013-0.065 ?g.The average recovery was 103.1% with RSD of 1.1%.Palmatine hydrochloride had a good linearity in the range of 0.013-0.102 ?g.The average recovery was 99.8% with RSD of 1.9%.Berberine hydrochloride had a good linearity in the range of 0.063-0.317 ?g.The average recovery was 99.8% with RSD of 1.4%.Evodiamine had a good linearity in the range of 0.001-0.007 ?g.The averoge recovery was 100.5% with RSD of 1.5%.Rutaecarpine had a good linearity in the range of 0.002-0.010 ?g.The average recovery was 98.9% with RSD of 1.9%.CONCLUSION:This method is simple,accurate and reproducible.It can be used to determine jatrorrhizine hydrochloride、 palmatine hydrochloride、 berberine hydrochloride、 evodiamine and rutaecarpine together with the same chromatogram condition.