Role and mechanism of action of Yinchenhao Decoction in inhibiting ferroptosis of hepatocytes in mice with autoimmune hepatitis / 临床肝胆病杂志

ObjectiveTo investigate the role and mechanism of action of Yinchenhao Decoction in inhibiting ferroptosis of hepatocytes in mice with autoimmune hepatitis. MethodsA total of 18 specific pathogen-free female C57BL/6 mice were selected and divided into normal group， model group， and treatment group using a random number table， with 6 mice in each group. The mice in the model group and the treatment group were injected with concanavalin A （Con A） via the caudal vein to establish a mouse model of autoimmune hepatitis， and those in the normal group were injected with normal saline. The mice in the treatment group were given prophylactic treatment with Yinchenhao Decoction （4.68 g crude drug/kg） by gavage at 14 days before modeling， and Con A was injected after the last gavage. The levels of alanine aminotransferase （ALT）， aspartate aminotransferase （AST）， interferon gamma （IFN-γ）， tumor necrosis factor-α （TNF-α）， iron ion， glutathione （GSH）， reactive oxygen species （ROS）， adenosine triphosphate （ATP）， and malondialdehyde （MDA） were measured； liver index and spleen index were calculated； the expression levels of GPX4 and SLC7A11 were measured； liver histopathological changes were compared between groups. A one-way analysis of variance was used for comparison of normally distributed continuous data between three groups， and the least significant difference t-test was used for further comparison between two groups. ResultsCompared with the normal group， the model group had significant increases in liver index， spleen index， ALT， AST， IFN-γ， TNF-α， iron ion， ROS and MDA （all P<0.05） and significant reductions in the content of GSH and ATP and the protein expression levels of GPX4 and SLC7A11 （all P<0.05）. Compared with the model group， the treatment group had significant reductions in liver index， spleen index， ALT， AST， IFN-γ， TNF-α， iron ion， ROS and MDA （all P<0.05） and significant increases in the content of GSH and ATP and the protein expression levels of GPX4 and SLC7A11 （all P<0.05）. HE staining showed that compared with the normal group， the model group showed massive hepatocyte degeneration and necrosis and inflammatory cell aggregation at the portal area， and compared with the model group， the treatment group had alleviation of liver necrosis and inflammatory infiltration. ConclusionLiver injury induced by Con A may be associated with ferroptosis. Yinchenhao Decoction can increase the protein expression levels of SLC7A11 and GPX4 protein and thus inhibit ferroptosis of hepatocytes induced by Con A.

Effect of Yinchenhao decoction on renal oxidative stress injury in rats with obstructive jaundice and its mechanism of action based on the nuclear factor erythroid 2-related factor 2 signaling pathway / 临床肝胆病杂志

Objective To investigate the effect of Yinchenhao decoction on renal oxidative stress injury in rats with obstructive jaundice and its association with the regulation of the expression of nuclear factor erythroid 2-related factor 2 (Nrf2) and nuclear translocation. Methods A total of 32 male Sprague-Dawley rats were randomly divided into sham-operation group (S group), model group (O group), low-dose Yinchenhao decoction group (LY group), and high-dose Yinchenhao decoction group (HY group), with 8 rats in each group. For the rats in the S group, the upper common bile duct was isolated without ligation, and for those in the other groups, double ligation of the middle and upper 1/3 of the common bile duct was performed to establish a model of obstructive jaundice. After 7 days, the rats in the LY group and the HY group were given Yinchenhao decoction by gavage at a dose of 6.3 and 18.9 mL/kg, respectively, while those in the S and O groups were given an equal volume of distilled water by gavage every day for 7 consecutive days, and the rats were treated on day 14. ELISA was used to measure the serum levels of total bilirubin (TBil), direct bilirubin (DBil), alanine aminotransferase (ALT), gamma-glutamyl transpeptidase (GGT), blood urea nitrogen (BUN), and creatinine (Cr); spectrophotometry was used to measure the activity of the oxidative stress factors superoxide dismutase (SOD) and malondialdehyde (MDA) in renal tissue; quantitative real- time PCR and Western blotting were used to measure the mRNA and protein expression levels of Nrf2, Kelch-like ECH-associated protein 1 (Keap1), and NAD(P)H quinone dehydrogenase 1 (NQO1) in renal tissue; immunohistochemistry was used to measure observe the nuclear translocation of Nrf2 protein in renal tissue. A one-way analysis of variance was used for comparison of continuous data between multiple groups, and the least significant difference t -test was used for further pairwise comparison within groups. Results Compared with the S group, the O group had significant increases in the levels of TBil, DBil, ALT、GGT, BUN, and Cr, a significant reduction in the activity of SOD, and a significant increase in the level of MDA (all P 0.05). Compared with the S group, the O group had a significant reduction in the positive rate of Nrf2 in cell nucleus in renal tissue ( P < 0.05), and compared with the O group, the LY group and the HY group had a significant increase in the positive rate of Nrf2 in cell nucleus ( P < 0.05). Conclusion Yinchenhao decoction can effectively alleviate renal injury caused by obstructive jaundice, possibly by upregulating the protein expression of Nrf2 in renal tissue and regulating the nuclear translocation of Nrf2 protein, so as to mediate the protein expression of downstream NQO1, regulate oxidative stress response caused by obstructive jaundice, and thereby alleviate renal injury in rats.

Professor Liu Guangxian's experience in treatment of cholangiocarcinoma by Yinchenhao decoction combined with Xiaoshi Fanshi powder / 中国中西医结合急救杂志

Professor Liu Guangxian used traditional Chinese medicine (TCM) for the treatment of patient with cholangiocarcinoma after surgery achieved unique effect. On the one hand, the classical prescriptions Yinchenhao decoction and Xiaoshi Fanshi powder combined with processed pangolin scales and cremastrae pseudobulbus were used to promote diuresis and clear jaundice, eliminate phlegm and resolve static blood, and soften hard mass and remove stagnation, in order to eliminate the pathogens; on the other hand, Sijunzi decoction was used as the main prescription to invigorate the spleen and benefit qi to strengthen vital qi, in order to protect the spleen and stomach and strengthen the body resistance. After more than 3 months of treatment for a patient with cholangiocarcinoma, the patient had significant improvement in symptoms, signs, imaging findings, and laboratory results.

Correlation analysis of "compound-target-pathway" of Yinchenhao decoction for cirrhosis based on computed prediction / 中国中药杂志

Chinese medicine prescription is the main form and means to treat diseases in traditional Chinese medicine (TCM). However, the disjointing situation is generally present between "systemic" and "local", "macro" and "micro", " process" and "activity evaluation" in the study of TCM at present. An urgent task for the modernization of TCM is to establish new strategies and methods which can reflect the overall characteristics of TCM. The introduction of integrative pharmacology provided a feasible approach to solve the problem of the fragmentation of TCM. Internet-based computation platform method was adopted in this study to explore the active molecular mechanism of Yinchenhao decoction in the treatment of cirrhosis. Based on the analysis of the functional integration of Internet-based Computation Platform V1.0 version software, the "core components-key target- main pathway" multidimensional network of Yinchenhao decoction in treatment of cirrhosis disease was constructed to explore the potential molecular mechanism of Yinchenhao decoction in treatment of cirrhosis from multiple perspectives. The molecular mechanism analysis of Yinchenhao decoction showed that Yinchenhao decoction can achieve the therapeutic effect on cirrhosis and the mechanism might be associated with oxidative phosphorylation, energy metabolism, circulatory system, glycerophospholipid metabolis, lipid metabolism and other pathways. Yinchenhao decoction in treatment of cirrhosis may be associated with energy metabolism and lipid metabolism.

Fingerprint of serum containing drug of Yinchenhao decoction / 中国药学杂志

OBJECTIVE: To establish the fingerprint of the serum containing drug of Yinchenhao Decoction (YCHD), and analyze the drug-originated constituents in serum. METHODS: An HPLC method was set up on a Symmetry C18(4.6 mm × 250 mm, 5 μm) column with a Security Guard Cartridges C18 (4.0 mm × 3.0 mm) guard column by gradient elution of acetonitrile-0.1% phosphoric acid in water at the flow rate of 1.0 mL · min-1. The column temperature was maintained at 30℃. The detection was carried out at 238 and 440 nm. SD rats were taken as serum donors. Ten batches of serum containing drugs were prepared after ig administration of YCHD. The fingerprints of these serum samples were determined, the common modes were established, and the similarities between fingerprints were evaluated. Drug-originated constituents in blood were distinguished by comparing the fingerprint of serum containing YCHD with the one of control serum. The sources of drug-originated constituents were analyzed by comparing the fingerprint of serum containing YCHD with the ones of serum containing individual herbs of YCHD. Drug-originated constituents in blood were identified by comparing the retention time and UV spectra of peaks in fingerprint of serum containing YCHD with the ones of YCHD and reference substances, respectively. RESULTS: The similarities between the fingerprints of ten batches of serum containing YCHD and common models at 238 and 440 nm were greater than 0.904 and 0.903, respectively. Twenty common peaks of drug-originated constituents were identified in the fingerprints of serum containing drug of YCHD at 238 and 440 nm. Out of them, six were the original compounds, the other 14 were metabolites. Three constituents out of six original form ones were identified as geniposidic acid and genipo-side (iridoids originated from GF), and rhein (anthraquinone from RRR), and the other three were identified as an anthraquinone from RRR and two crocetin derivatives from FG. Out of 14 metabolites, one was identified as the metabolite of iridoid, four were from anthraquinone, and nine were from crocetin derivatives. CONCLUSION: The established method is accurate, reliable and can be used for the analysis of drug-originated constituents in serum containing drug of YCHD.

Contents Determination and Comparison of Active Ingredients in the Yinchenhao Decoction and Its Dispens-ing Granule by HPLC / 中国药房

OBJECTIVE:To establish a method for the contents determination of emodin,rhein,aloe-emodin,chrysophanol, physcion,geniposide,chlorogenic acid in Yinchenhao decoction and its dispensing granule,and to compare the difference among the above-mentioned ingredients. METHODS:HPLC was adopted to determine the contents of emodin, rhein, aloe-emodin, chrysophanol and physcion:the column was Ecosil C18 with mobile phase of acetonitrile-0.5% phosphoric acid(gradient elution)at a flow rate of 1 ml/min,the detection wavelength was 254 nm,column temperature was 35 ℃,and injection volume was 20 μl. When determining the content of geniposide by HPLC,the column was Ecosil C18 with mobile phase of acetonitrile-water(13∶87, V/V)at a flow rate of 1 ml/min,the detection wavelength was 238 nm,column temperature was 35 ℃,and injection volume was 20 μl. When determining the content of chlorogenic acid by HPLC,the column was Ecosil C18 with mobile phase of acetonitrile-0.5% phosphoric acid (10∶90,V/V) at a flow rate of 1 ml/min,the detection wavelength was 327 nm,column temperature was 35 ℃,and injection volume was 20 μl. RESULTS:The linear range was 2-20 μg/ml for emodin(r=0.996 5),5.2-52.0 μg/ml for rhein(r=0.998 5),2.6-26.0 μg/ml for aloe-emodin(r=0.999 9),1.0-10.4 μg/ml for chrysophanol(r=0.999 9),1.0-10.0 μg/ml for physcion(r=0.999 8),20-200 μg/ml for geniposide(r=0.999 9),20-200 μg/ml for chlorogenic acid(r=0.999 9);RSDs of precision,stability and reproducibility tests were lower than 3%;recoveries were 91.1%-96.9%(RSD=2.0%,n=6)for emodin、93.9%-96.1%(RSD=0.8%,n=6)for rhein、90.9%-93.4%(RSD=1.2%,n=6)for aloe-emodin、88.5%-92.7%(RSD=1.8%,n=6) for chrysophanol、82.1% -87.9%(RSD=2.5% ,n=6)for physcion,100.4% -102.0%(RSD=0.7% ,n=6)for geniposide、101.1%-102.2%(RSD=0.4%,n=6) for chlorogenic acid. CONCLUSIONS:The method is simple,stable and reproducible,and can be used for the simultaneous contents determination of active ingredients in Yinchenhao decoction and its dispensing granule. The contents of active ingredients in Yinchenhao dispensing granule are obviously higher than those in its decoction.

Rational Daily Administration Times of Yinchenhao Decoction in Rats with Jaundice Based on PD/PK / 中草药·英文版

Objective To study the rational daily administration times of Yinchenhao Decoction (YCHD) when it was used to treat experimental jaundice in rats based on pharmacodynamics/pharmacokinetics model.MethodsRats were modeled by 4％ 1-naphthylisothiocyanate (75 mg/kg) for 48 h,then YCHD was drenched with doses of 0.324 g/kg (extract,calculated with the clinical dosage) once,0.162 g/kg twice,and 0.108 g/kg thrice a day,respectively.The total bile and the flow rate of bile were observed after the first administration; Blood samples collected from the orbital sinus at different intervals were used to investigate the levels of liver enzymes (ALT and AST) and bilimbins (TBIL and DBIL),and determine the concentration of 6,7-dimethoxycoumarin (DME) in the plasma using UPLC at the same time,then we obtained the time-effect and time-dose curves.The rational daily administration times of YCHD when treating experimental jaundice were determined based on the comprehensive analysis of time-effect and time-concentration relationships.ResultsWithin 10 h the total bile of rats which were administered once daily (G1) was I.65 and 1.33 times higher than that of twice and thrice (G2 and G3) a day,respectively,and the four biochemical indexes (TBIL,ALT,DBIL,and AST) of Gl decreased faster than those of G2 and G3 (P ＜ 0.05).On the other hand,the blood drug level of DME when administrated once daily could maintain at a higher level for a longer time,and its Cmax and AUC0→t were higher than those of G2 and G3,which might be the main reason why its effect was the most significant.ConclusionIt is more appropriate to administrate once daily when YCHD is used to treat jaundice.

Inhibitory effects of Yinchenhao Decoction on fatty deposition and TNF-? secretion in HepG2 cells induced by free fatty acid / 中华中医药杂志

Objective:To study the effects of effective herbs in Yinchenhao Decoction(YD)on fatty deposition and TNF-?secretion of hepatic lipo-toxicity model in vitro.Methods:TG and TNF-?as the inspection target,apply the steatosis with TNF-?secretion lipo-toxicity model in vitro of HepG2 induced by long-chain FFA and uniform design U7(76)form for YD,which had 3 herbs,to compound 6 kinds of different prescriptions,in order to look for the effective herbs or optimized group in YD.Results:The Chinese medicine Yinchen and its optimized group-Yinchen and Zhizi combination can remarkably reduce the TG and TNF-?content.Conclusion:The Chinese medicine Yinchen and its optimized group-Yinchen and Zhizi combination were effective herbs in YD for inhibiting fatty deposition and TNF-?secretion of hepatic lipo-toxicity model in vitro.

Erythrocyte immunity study of intervening 84 cases of ABO maternofetal blood group incompatibility by Yinchenhao Decoction / 中华中医药杂志

Objective: Through treatment based on differentiation of symptoms and signs and classification of defi nited patients of maternofetal blood group incompatibility, detecting anti-A or Anti-B antibody and erythrocyte immune complex chaplet rate(RICR) and erythrocyte receptor chaplet rate(RC3bR) , etc, to observe the therapeutic effect and mechanism of prescriptions mainly with Yinchenhao Decoction. Methods: Patients definited maternofetal blood group incompatibility were divided into control group and observation group according to random principle in 1:3 ratio. Then, observation group was divided into moist heat group(SRG), moist heat and defi ciency of spleen qi group (SR+PQXG) and moist heat and defi ciency of kidney qi group(SR+SQXG) according to syndrome differentiation. Corresponding prescriptions were used, every 30 doses 1 course. Before and after treatment, the anti-A or anti-B antibody and RICR, RC3bR were detected in peripheral blood. Results: In Yinchenhao Decoction group with 21 cases, the anti-A or anti anti-B antibody decreased, including 5 case (1:64) and 4 case (1:32). In group of syndrome differentiation with 63 cases, the anti-A or anti anti-B antibody decreased, including 33 case(1:64) and 20 case (1:32). Before and after treatment, RICR of control group and observation group were(19.43?5.29), (19.57?6.50)and (20.59? 7.96), (23.77?5.12), respectively. RC3bR were(11.62?4.16), (12.42?6.60)and(11.93?4.10), (13.96?6.40)respectively. There was signifi cant difference of before and after treatment in obserbation group(P0.05). Conclusion: In cutting down anti-A and anti-B antibody, and increasing RICR and RC3bR fi eld, the observation groups were better than control group.

Reference value of biochemical indicator and imaging medicine in jaundice-relieving effect of Yinchenhao Decoction / 中华中医药杂志

We can discriminate the cause and classi cation of jaundice by biochemical indicator and imaging medicine.It suggests that Yinchenhao decoction can get a remarkable curative effect on the recessive jaundice and hepatocellular jaundice.But for severe obstructive jaundice,Yinchenhao decoction has no good jaundice-relieving effect.

Clinical Observation on Oral Use of Herbal Medicine Combined with Ultrasonic Wave and Herbal Medicine Mask for Acne Simplex / 广州中医药大学学报

[Objective] To observe the therapeutic effect of oral use of heat-clearing, damp-removing and toxicity-clearing herbal medicine combined with ultrasonic wave and herbal medicine mask treatment in treating acne simplex (AS). [Methods] Two hundred and ten cases of AS were randomized into groups A and B. Group A ( n=113) was treated with modified Yinchenhao Decoction and Qingfei Decoction for oral use and added with ultrasonic wave and mask prepared with herbal medicine including Borneolum Syntheticum, Flos Lonicerae, Radix Scutellariae, Radix Salviae Miltiorrhizae, Fructus Forsythiae; group B ( n=97) was treated with metronidazole 0.2 g, tid, and vitamin Bg 20mg, tid, as well as erythromycin ointment for external application. After a 4-week treatment, the therapeutic effect was compared. [Results] The total effective rale was 80.6% in group A and 41.3% in group B, and the markedly effective rate was 50.5% in group A and 28.9% in group B, the difference being significant (P

Enrichment process of scoparone from Yinchenhao Decoction with macroporous resin / 中草药

Object To study the optimal conditions and parameters for the enrichment process of scoparone in Yinchenhao Decoction (YCHD) with macroporous resin. Methods With the enrichment degree of scoparone in YCHD as an index, the optimal conditions of the enrichment process were investigated. Results The optimal conditions were that the aqueous extract of YCHD was taken into the macroporous resin column, and kept for 30 min, then the resin was washed by five times of distilled water to get rid of impurity and three — four times of 70% ethanol to elute scoparone. Conclusion The process is feasible to enrich effective ingredient scoparone from YCHD.

Research on the pharmacokinetic parameters of 6,7-dimethoxycoumarin after oral administration of Yinchenhao and its decoction / 中成药

Objective: To study the plasma concentrations and pharmacokinetic parameters of 6,7 dimethoxycoumarin after oral administration of Herba Artemisiae Scopariae decoction and Yinchenhao decoction. Methods: A HPLC UV method was developed to determine 6,7 dimethoxycoumarin in rat plasma. The drug was extracted with acetonitrile from plasma and separated on a Kromasil ODS (250mm?4.6mm,5?m)column with methanol 1% solution of acetic acid THF (30∶63∶7) as mobile phase, UV detecter was set at 340nm. The data obtained were analyzed with Topfit program. Results: The liner calibration curves were obtained in the concentration range of 0.026～2.5 ?g?mL -1 . With non compartmental analysis, the main pharmacokinetic parameters after oral administration of Artemisiae Scopariae decoction and Yinchenhao decoction were as follows: T 1/2 was 1.30 and 1.75h, AUC 0→t was 1215.00 and 2527.85ng?h?mL -1 , AUC 0→∝ was 1325.90 and 2612.58ng?h?mL -1 respectively. Conclusion: This method can be used to study the pharmacokinetics of 6,7 dimethoxycoumarin derived from different traditional Chinese medicines containing this component.