2.Effects of sera containing Kang xian ling on c-Met and its downstream MAPK signal molecules in HK-2 cells treated with TGF-β_1
Haiying LU ; Kejian LIU ; Yue ZHANG ; Liqun HE ; Yumin LIU ; Jing LI ; Zhifen HAN ; Huifang PANG
Chinese Journal of Pathophysiology 2010;26(1):154-157
AIM: To observe the effects of sera containing Kang xian ling (a traditional Chinese medicine) on c-Met and its downstream MAPK signal molecules in HK-2 cells treated with transforming growth factor-β_1(TGF-β_1), so as to further explore the effective mechanism of anti-renal fibrosis of the traditional Chinese medicine. METHODS: Rat sera containing herbs were collected after rats were intragastrically administered with the herbs. HK-2 cells activated by TGF-β_1 were incubated with different heat-inactivated sera. The proliferation and expression of MAPK and the phosphorylation of MAPK in HK-2 cells were detected by Alarm blue and immunoblotting. RESULTS: No significant difference of cell proliferation between any two groups was observed. C-Met and the phosphorylation of its downstream molecules were also observed after incubated with different sera. CONCLUSION: The receptor c-Met of hepatocyte growth factor, and the phosphorylation of its downstream MAPK molecules can be regulated by the sera containing Kang xian ling.
3.Simultaneous Determination of 4 Components in Regan Saibisitan Granules by RP-HPLC
Xin ZHAI ; Kejian PANG ; Hui TANG ; Xinyu LIU ; Xiaoya SUN ; Yaqiao WANG
China Pharmacy 2017;28(21):2963-2966
OBJECTIVE:To establish the method for simultaneous determination of gallic acid,rosmarinic acid,liquiritin and ammonium glycyrrhetate in Regan saibisitan granules. METHODS:RP-HPLC method was adopted. The determination was per-formed on Waters RP-C18 column with mobile phase consisted of acetonitrile-0.2% phosphoric acid (gradient elution) at the flow rate of 1.0 mL/min. The detection wavelengths were 210 nm(gallic acid,rosmarinic acid and liquiritin),230 nm(ammonium glyc-yrrhetate). The column temperature was 28 ℃,and sample size was 20 μL. RESULTS:The linear ranges were 0.2744-7.546 μg for gallic acid(r=0.9998),0.1870-5.143 μg for rosmarinic acid(r=0.9996),0.1300-3.575 μg for liquiritin(r=0.9999)and 0.2540-6.985μg for ammonium glycyrrhetate(r=0.9998),respectively. The LOQ were 2.67,1.36,1.09 and 2.11 ng,respective-ly. The LOD were 1.03,0.62,0.87 and 0.91 ng,respectively. RSDs of precision,stability and repeatability tests were all less than 2.0%. The average recoveries were 97.26%-101.00%(RSD=1.1%,n=9),97.66%-101.80%(RSD=1.3%,n=9),97.45%-101.70%(RSD=1.4%,n=9),97.74%-101.70%(RSD=1.4%,n=9). CONCLUSIONS:The method is simple,accurate and reproducible, and can be applied for simultaneous determination of gallic acid,rosmarinic acid,liquiritin and ammonium glycyrrhetate in Regan saibisitan granules.
4.Effect of Kang Xianling decoction on TGF-?_1-Smad pathway in a unilateral ureteral obstruction rat model
Yumin LIU ; Yue ZHANG ; Liqun HE ; Kejian LIU ; Jing LI ; Haiying LU ; Zhifen HAN ; Huifang PANG ; Min LIN
Chinese Journal of Pathophysiology 2000;0(12):-
AIM: To study the effect of Kang Xianling decoction,comprised of dahuang,danshen,taoren,niuxi and danggui,on TGF-?1-Smad pathway in unilateral ureteral obstruction rat model.METHODS: Eighteen male SD rats were divided into 3 groups,sham group,model group and model group treated with Kang Xianling decoction randomly.Renal interstitial fibrosis model was established in rats by unilateral ureteral obstruction(UUO).After treatment for additional 14 d,parameters of hydroxyproline in obstructed kidney from 3 groups were analyzed.Rats were sacrificed and the pathological statuses of their kidneys were checked by HE staining and electron microscopy.Transforming growth factor-?1(TGF-?1) mRNA in kidney tissue was determined by RT-PCR.TGF-?1 receptor Ⅰ(T?RⅠ),TGF-?1 receptorⅡ(T?RⅡ),phosphorylated Smad2 and Smad2 protein were determined by Western blotting.RESULTS: Parameters of hydroxyproline in animals of model group were significantly increased than those in sham operation group(P
5.Study on quality standard of Onosma echioides
Qianqian WANG ; Kejian PANG ; Wei GAO ; Ke ZHANG ; Hanying CHEN ; Bo ZHANG
China Pharmacy 2022;33(19):2321-2325
OBJECTIVE To establish the quality standard of Onosma echioides . METHODS Sixteen batches of O. echioides from different sources were collected to observe their appearance ,microscopic powder identification ,and TLC identification ;the impurity,moisture,total ash and acid -insoluble ash of the medicinal materials were examined . The total pigment content of hydroxynaphthoquinone in O. echioides was determined by UV -visible spectrophotometry ;the contents of alkannin and β,β′- dimethylacrylalkannin in O. echioides were determined by HPLC . RESULTS Medicinal material and powder of O. echioides were purplish red ;non-glandular single cells ,embolized cells ,parenchyma cells ,reticulate ducts could be seen microscopically . TLC results showed that the color and change of the spots in the chromatogram of test sample were consistent with that of the control . The contents of moisture ,total ash and acid insoluble ashshall not exceed 13.0%,18.0%,6.0%. The total pigment content of hydroxynaphthoquinone should not be less than 0.80%. The content of alkannin was recommended to be no less than 0.06 mg/g. The content of β,β′-dimethylacrylalkannin was recommended to be no less than 0.60 mg/g. CONCLUSIONS The established standard can provide reference for the scientific evaluation of the quality of the medicinal materials of O. echioides.
6.Improvement of Quality Standard for Mongolian Medicine Juniperus rigida
Xiaohong YUAN ; Kejian PANG ; Hui TANG ; Hong ZHANG ; Hui GUAN ; Feng WEI ; Ping TANG
China Pharmacy 2021;32(16):1949-1955
OBJECTIVE:To provide scientifi c evidence for improving the quality standard of Mongolian medicine Juniperus rigida. METHODS :Totally 10 batches of J. rigida from different places were taken as samples to observe their characters and identify them by microscope ;TLC method was adopted to qualitatively identify isoquercitrin ,quercitrin,amentoflavone, podocarpusflavone A and hinokiflavone ;the contents of total ash ,acid-insoluble ash ,ethanol-soluble extract and heavy metals were determined by related method stated in 2020 edition of Chinese Pharmacopeia (part Ⅳ). The contents of above 5 components in samples were determined by HPLC. RESULTS :The powder of J. rigida was green or yellowish green ,polygonal tracheids , closely arranged in longitudinal with unequal stomatal ;epidermal cells were nearly rectangular ;sclerenchyma cells were quasi rectangular and the wall beadedly thickening. Results of TLC showed that the spots of the same color were found in the corresponding positions of chromatogram for test sample and substance control. The contents of total ash ,acid-insoluble ash and ethanol-soluble extract in 10 batches of samples were 7.37%-11.18%,0.75%-2.98%,16.55%-26.42%,respectively;average contents were 8.51%,1.27%,22.35%. The contents of lead ,arsenic,cadmium,mercury and copper were 2.00-5.44,0.65-1.65, 0.044-0.100,0.034-0.160,4.59-6.79 mg/kg,respectively;average conte nts were 3.73,0.97,0.078,0.061,5.23 mg/kg. The linear ranges of isoquercitrin ,quercitrin,amentoflavone,podocarpus- flavone A and hinokiflavone were 4.98-20.02,49.99-199.96, 19.94-99.96,9.99-40.00,20.20-159.98 μg/mL(all r>0.999 7); com RSDs of precision ,repeatability and stability (24 h) tests were all less than 3.00%(n=6);the average recoveries were 话:0993-2057878。E-mail:Tanghuishz@qq.com 100.62%-102.96%,RSDs were 1.21%-1.88%(n=6). Average contents of the above-mentioned 5 compounds in 10 batches of samples were 0.089-0.379,1.379-4.250,1.077-2.026,0.162-0.423, 0.016 9-0.117 0 mg/g,respectively. CONCLUSIONS :The qualitative and quantitative analysis methods of Mongolian medicine J. rigida are established. It is preliminarily proposed that the total ash content shall not exceed 10.22%,the acid-insoluble ash content shall not exceed 1.53%,ethanol-soluble extract content shall not be less than 17.88%,heavy metal lead should not exceed 5 mg/kg,arsenic should not exceed 2 mg/kg,cadmium should not exceed 0.3 mg/kg,mercury should not exceed 0.2 mg/kg,copper should not exceed 20 mg/kg.
7.Improvement of Quality Standard for Mongolian Medicine Artemisia sacrorum
Xiaohong YUAN ; Kejian PANG ; Hui TANG ; Hong ZHANG ; Feng WEI ; Lele LIU ; Guozhen JIANG
China Pharmacy 2021;32(5):536-541
OBJECTIVE:To improve the quality standard of M ongolian med icine Artemisia sacrorum ,and to provide scientific basis for comprehensive quality evaluation. METHODS :The appearance and microscopic characteristics of A. sacrorum were identified;scopoletin,chlorogenic acid ,caffeic acid ,scopoletin and 3,5-dicaffeoylquinic acid were identified quantitatively by TLC;the contents of above 5 components were determined by HPLC. The water content ,total ash and extract were examined. RESULTS:The stem of A. sacrorum was cylindrical ,and its surface was purple or purple-brown or cyan-brown ;the leaves were ovate or oblong-ovate ,fragrant;the flowers were yellow ,head-shaped,subglobose or hemispherical. The powder was green or yellow-green,its pollen grain had three germination ;the parenchymal cell clusters with sharp edges and numerous threaded ducts , occasionally having marginal pitted ducts ;its wood fibers were in bundles mostly. Results of TLC showed that the spots of the same color were found in the corresponding positions of chromatogram for 5 substance control and samples. The linear range of scopoletin, chlorogenic acid , caffeic acid , scopolactone and 3,5-dicaffeoylquinic acid were 85.60-428.00, 10.16-101.60, 10.20-102.00,40.84-408.40 and 40.80-408.00 μg/mL(all r>0.999 0). RSDs of precision ,stability,repeatability tests were all less than 3.00%(n=6). The average recoveries were 103.07%,99.66%,98.37%,97.78%,98.40%(all RSDs <3.00%,n=6). The contents of the above-mentioned 5 compounds in 10 batches of samples were 0.36%-1.23%,0.09%-0.51%,0.04%-0.13%, 0.61% -1.13% ,0.12% -1.11% ,respectively;the average com contents of water ,total ash and water soluble extract were 6.25%,5.86%,26.50%,respectively. CONCLU SIONS:O the basis of the original quality standard of A. sacrorum , microscopic identification,TLC identification ,content determination and examination items of water ,total ash and extract are added. The method shows good precision ,accuracy and stability ,which can provide reference for more scientific and standardized evaluation of the quality of this medicinal material.