1.Determination of Five Heavy Metals in Chuanxiong Rhizoma and Gastrodiae Rhizoma by ICP-MS
Dawen XIE ; Yuanpei LIAN ; Wenlin LIANG ; Gang DING ; Yongcheng SUN ; Yanjing LI ; Wei XIAO
World Science and Technology-Modernization of Traditional Chinese Medicine 2015;17(3):740-743
This article is aimed to provide a method for the simultaneous analysis of five heavy metals, including Cu, As, Pb, Cd, and Hg, in Chuanxiong Rhizoma and Gastrodiae Rhizoma through ICP-MS. Five heavy metals were determined by an inductively coupled plasma-mass spectrometry method after microwave-assisted digestion. The linear correlation coefficients were all better than 0.999. The lowest limits of quantification for all target elements were 0.003 0.134 ug·L-1, while the recovery values ranged from 80.04% 118.34%. This method was accurate, convenient, rapid, and highly sensitive, and can be applied to determine five heavy metals, including Cu, As, Pb, Cd, and Hg in Chuanxiong Rhizoma and Gastrodiae Rhizoma.
2. Correlation between intestinal mucosal permeability and prognosis in patients with liver cirrhosis
Xiaoxiao LIAN ; Yuanpei SUN ; Xiaoxia GUO
Chinese Journal of Hepatology 2020;28(1):58-63
Objective:
To investigate the correlation between changes in intestinal mucosal permeability and prognosis of patients with liver cirrhosis.
Methods:
Data of 89 cases with liver cirrhosis who were hospitalized in the Hepatology Department of Shanxi Provincial Hospital of Traditional Chinese Medicine from January 2017 to August 2017 were collected as the liver cirrhosis experimental group, and 40 healthy subjects were randomly selected as the healthy control group. JY-DLT, the Intestinal Mucosal Barrier Biochemical Index Analysis System was used to measure the levels of serum diamine oxidase (DAO), D-lactic acid, and endotoxin (ETX) in two groups to evaluate intestinal mucosal barrier function. Spearman’s rank correlation test was used to evaluate the correlation between liver cirrhosis prognosis and intestinal mucosal permeability. The results of the two groups were compared by Mann-Whitney H test of two independent samples. One-way Anova was used for intergroup comparison. The pairwise comparison between groups was performed using the LSD or SNK test.
Results:
The level of ETX in patients with decompensated cirrhosis was significantly higher than that in the compensated phase, and the difference was statistically significant (
3.Proteomic Research on Characteristic Processing Materials of Porcine Cardiac Blood and Porcine Blood from Menghe Medical School and Their Effect on Anti-microglial Inflammatory Damage of Salviae Miltiorrhizae Radix et Rhizoma
Nan LIU ; Yuanpei LIAN ; Chunyan YIN ; Jiali CAI ; Dijun WANG ; Li ZHU ; Chanming LIU ; Wei HUANG ; Xiaojing YAN
Chinese Journal of Experimental Traditional Medical Formulae 2023;29(18):137-145
ObjectiveProteomics was used to investigate the protein differences between porcine cardiac blood(PCB) and porcine blood(PB) from Menghe medical school and to compare the effects of both on the microglial inflammation of Salviae Miltiorrhizae Radix et Rhizoma(DS). MethodNanoliquid chromatography-quadrupole orbitrap mass spectrometry(nLC-MS/MS) and bioinformatics were utilized to compare the proteomic differences of PCB and PB in simulated gastrointestinal digestion. Furthermore, Western blot was used to verify the contents of some shared proteins and differential proteins identified in PCB and PB. In addition, BV2 neuroinflammation model constructed by corticosterone(CORT) and lipopolysaccharide(LPS) was applied to detect the intervention effects of PCB and PB on the levels of tumor necrosis factor(TNF)-α and interleukin(IL)-6 in BV2 inflammatory cells of DS. ResultA total of 69 common proteins and 68 differential proteins were identified in PCB and PB, among which the common proteins included transferrin(Tf) with brain-targeting effect, and the differential proteins in the two were 41 and 27, respectively. Western blot validation showed that the difference in the content of the same protein Tf between PCB and PB was not statistically significant, while the difference in the contents of the specific proteins of creatine kinase M and heart-type fatty acid binding protein(H-FABP) were statistically significant(P<0.05). Moreover, in vitro experimental studies revealed that compared with the same concentration of DS group, in addition to the 100 mg·L-1 PB-DS group, PCB-DS and PB-DS groups could significantly inhibit the levels of TNF-α and IL-6 in BV2 inflammatory cells(P<0.05, P<0.01), and PCB-DS group had more significant anti-inflammatory effect than PB-DS group with the same concentration(P<0.05, P<0.01). ConclusionBoth of PCB and PB can enhance the inhibitory effect of DS on the release of inflammatory factors, thus playing a neuroprotective role, and PCB promotes DS inhibition more significantly, which may be due to the existence of the two involved in energy metabolism-related differential proteins, which can lay a foundation for revealing the scientific connotation of the processing of PCB-DS and PB-DS.
4.Correlation Analysis Between Anti-cerebral Ischemia Oxidative Damage and Contents of Active Components in Characteristic Processed Products with Porcine Cardiac Blood and Other Processed Products of Salviae Miltiorrhizae Radix et Rhizoma from Menghe Medical School
Zhen ZENG ; Yuanpei LIAN ; Jiali CAI ; Chunyan YIN ; Dijun WANG ; Li ZHU ; Chanming LIU ; Wei HUANG ; Xiaojing YAN
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(15):162-171
ObjectiveTo analyze the correlation between 11 small molecule active components and 1 protein component of characteristic processed products with porcine cardiac blood and other products of Salviae Miltiorrhizae Radix et Rhizoma(SMRR) from Menghe medical school and anti-cerebral ischemic oxidative damage, and to identify its key component markers of characteristic processed products with porcine cardiac blood for anti-cerebral ischemic oxidative damage. MethodHigh performance liquid chromatography(HPLC) was established to simultaneously determine the contents of 11 active ingredients in SMRR and its processed products[processed with porcine cardiac blood, porcine blood, wine and transferrin(Tf) in porcine cardiac blood], and the content of Tf in different processed products of SMRR was detected by enzyme-linked immunosorbent assay(ELISA). Furthermore, A zebrafish ischemic stroke model was constructed to evaluate the effects of different processed products of SMRR on the behavioral trajectory of cerebral ischemic zebrafish, the neuronal damage of transgenic zebrafish Tg(elavl3:eGFP) brain, as well as the levels of malondialdehyde(MDA) and superoxide dismutase(SOD) in the brain tissues. The hippocampal neurons oxygen-glucose deprivation/reoxygenation(OGD/R)-induced ischemia-hypoxia model was constructed to evaluate the effects of different processed products of SMRR on oxidative damage of neuronal cells by taking lactate dehydrogenase(LDH), reactive oxygen species(ROS), MDA and SOD as indexes. Finally, principal component analysis(PCA), partial least squares-discriminant analysis(PLS-DA) and Spearman correlation analysis were used to analyze the 11 small molecule active components and 1 protein component with efficacy indicators, in order to screen the key components of the characteristic processed products with porcine cardiac blood for cerebral ischemic oxidative damage. ResultCompared with the raw products, the contents of water-soluble and fat-soluble components in processed products of SMRR increased to different degrees, while the content of salvianolic acid A decreased. Compared with the wine-processed products, the contents of salvianolic acid B, danshensu, rosmarinic acid and other components in the porcine cardiac blood-processed products, porcine blood-processed products, Tf-processed products were increased, while the content of salvianolic acid A was decreased. ELISA results showed that there was no significant difference in Tf content between the porcine cardiac blood-processed products, porcine blood-processed products, Tf-processed products. Pharmacological results showed that different processed products of SMRR could improve the behavioral deficits, brain neuronal injury and oxidative stress after ischemic stroke in zebrafish, and the effect of the porcine cardiac blood-processed products was most pronounced. PCA results showed that salvianolic acid B, salvianolic acid A, rosmarinic acid, lithospermic acid, danshensu, tanshinone ⅡA, caffeic acid, cryptotanshinone and tanshinone Ⅰ were the main contributing components of SMRR and its processed products. And the results of correlation analysis showed that the contents of cryptotanshinone, rosmarinic acid, caffeic acid, dihydrotanshinone Ⅰ, salvianolic acid B, tanshinone ⅡA and tanshinone Ⅰ were negatively correlated with MDA level in zebrafish brain tissue, while the contents of lithospermic acid, protocatechuic aldehyde, rosmarinic acid, dihydrotanshinone Ⅰ, salvianolic acid B and Tf were positively correlated with SOD level, and the contents of rosmarinic acid, caffeic acid, dihydrotanshinone Ⅰ, salvianolic acid B, tanshinone ⅡA, tanshinone Ⅰ, danshensu, Tf were positively correlated with neuronal fluorescence intensity in the zebrafish brain. And the contents of lithospermic acid, protocatechuic aldehyde, rosmarinic acid, dihydrotanshinone Ⅰ, salvianolic acid B, tanshinone ⅡA and Tf were negatively correlated with LDH, ROS and MDA levels and positively correlated with SOD level. ConclusionThere are differences in the anti-ischemic oxidative damage effects of SMRR and its different processed products, among which the porcine cardiac blood-processed products has the strongest effect on improving oxidative damage, which may be related to the content changes of salvianolic acid B, danshensu, rosmarinic acid and other components. This study can provide a basis for clarifying the quality markers of SMRR processed with porcine cardiac blood for cerebral ischemia and elucidating its processing mechanism.