1.Danggui Shaoyaosan Regulates Nrf2/SLC7A11/GPX4 Signaling Pathway to Inhibit Ferroptosis in Rat Model of Non-alcoholic Fatty Liver Disease
Xinqiao CHU ; Yaning BIAO ; Ying GU ; Meng LI ; Tiantong JIANG ; Yuan DING ; Xiaping TAO ; Shaoli WANG ; Ziheng WEI ; Zhen LIU ; Yixin ZHANG
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(16):35-42
ObjectiveTo investigate the effect of Danggui Shaoyaosan on ferroptosis in the rat model of non-alcoholic fatty liver disease (NAFLD) and explore the underlying mechanism based on the nuclear factor E2-related factor 2 (Nrf2)/solute carrier family 7 member 11 (SLC7A11)/glutathione peroxidase 4 (GPX4) signaling pathway. MethodsThe sixty SD rats were randomly grouped as follows: control, model, Yishanfu (0.144 g·kg-1), and low-, medium-, and high-dose (2.44, 4.88, and 9.76 g·kg-1, respectively) Danggui Shaoyaosan. A high-fat diet was used to establish the rat model of NAFLD. After 12 weeks of modeling, rats were treated with corresponding agents for 4 weeks. Then, the body weight and liver weight were measured, and the liver index was calculated. At the same time, serum and liver samples were collected. The levels or activities of total cholesterol (TC), triglycerides (TG), alanine aminotransferase (ALT), aspartate aminotransferase (AST), and Fe2+ in the serum and TC, TG, free fatty acids (FFA), malondialdehyde (MDA), superoxide dismutase (SOD), glutathione peroxidase (GPX), and Fe2+ in the liver were measured. Hematoxylin-eosin staining and oil red O staining were employed to observe the pathological changes in the liver. Immunofluorescence was used to assess the reactive oxygen species (ROS) content in the liver. Mitochondrial morphology was observed by transmission electron microscopy. The protein levels of Nrf2, SLC7A11, GPX4, transferrin receptor 1 (TFR1), and divalent metal transporter 1 (DMT1) in the liver were determined by Western blot. ResultsCompared with the control group, the model group showed increases in the body weight, liver weight, liver index, levels or activities of TC, TG, ALT, AST, and Fe2+ in the serum, levels of TC, TG, FFA, MDA, Fe2+, and ROS in the liver, and protein levels of TFR1 and DMT1 in the liver (P<0.01), and decreases in the activities of SOD, GPX and the protein levels of Nrf2, SLC7A11, and GPX4 in the liver (P<0.05, P<0.01). Meanwhile, the liver tissue in the model group presented steatosis, iron deposition, mitochondrial shrinkage, and blurred or swollen mitochondrial cristae. Compared with the model group, all doses of Danggui Shaoyaosan reduced the body weight, liver weight, liver index, levels or activities of TC, TG, ALT, AST, and Fe2+ in the serum, levels of TC, TG, FFA, MDA, Fe2+, and ROS in the liver, and protein levels of TFR1 and DMT1 in the liver (P<0.01), while increasing the activities of SOD and GPX and the protein levels of Nrf2, SLC7A11, and GPX4 in the liver (P<0.01). Furthermore, Danggui Shaoyaosan alleviated steatosis, iron deposition, and mitochondrial damage in the liver. ConclusionDanggui Shaoyaosan may inhibit lipid peroxidation and ferroptosis by activating the Nrf2/SLC7A11/GPX4 signaling pathway to treat NAFLD.
2.Traditional superior disciplines empower the development of potential or new disciplines
Zuoqi DING ; Zhen TANG ; Jiewen LUO ; Jinnan SUN
Journal of China Pharmaceutical University 2025;56(3):397-404
The scientific evaluation of the construction and development of university disciplines is an important part and a key challenge in China’s “Double First-Class” initiative. Based on the ESI and InCites databases, this study conducted a year-long observation and analysis of the disciplines at universities in Jiangsu Province under the “Double First-Class” framework. The research focused on the development trends of potential and emerging disciplines, while exploring the influence of traditional predominant disciplines on the growth of these emerging fields and the pathways through which such influence might occur. The study aims to provide some reference and insights for the development of disciplines in the universities in Jiangsu. The findings reveal that, overall, the discipline development maturity of the universities is high, with a positive growth trend and significant development potential in certain fields, which are strongly supported by the traditional predominant disciplines. However, an imbalance in the development across various discipline categories has also been observed. Looking ahead, universities are encouraged to capitalize on their unique academic strengths to foster collaborative development, emphasize the growth of social science disciplines, promote interdisciplinary integration, and cultivate high-level talent for society. This approach will better facilitate the establishment of high-level universities and contribute to the realization of the goal of building a strong educational nation.
3.Analysis of information labeling in instructions of antitussive and expectorant drugs in 34 children’s medical institutions
Zhen DING ; Hai LIANG ; Wenyi RUAN ; Heping CAI
China Pharmacy 2024;35(5):607-611
OBJECTIVE To understand the information labeling of children’s medication in the instructions of antitussive and expectorant drugs commonly used in children’s medical institutions, analyze the existing problems, and propose relevant suggestions. METHODS The instructions for 100 antitussive and expectorant drugs commonly used in 20 tertiary children’s hospitals (centers) and 14 maternal and child health hospitals (centers) with regional representativeness in China were collected, and the information labeling of children’s medication in the instructions was investigated and analyzed. RESULTS There were only 7 kinds of antitussive and expectorant drugs for children, and the others were non-specific drugs for children. Among antitussive and expectorant drugs, tablets accounted for 18.00%, injections for 18.00%, and capsules for 4.00%. Among 100 antitussive and expectorant drugs, 72 (72.00%) labeled the usage and dosage for children, 63 (63.00%) labeled the medication items for children, 59 (59.00%) indicated the information of children in the precautions, and pharmacokinetic parameters for children were absent. Compared with imported antitussive and expectorant drugs or the drugs manufactured by joint ventures, there were many missing labeling of medication information for children with Chinese drugs. Among 63 kinds of drug instructions labeled with medication items for children, various instructions information had little guiding significance. CONCLUSIONS There is a lack of labeling in the instructions of antitussive and expectorant drugs for children, and the proportion of special drugs for children remains low. Pediatric drug information in the instructions has little guiding significance for pediatric medication. Relevant departments should further promote the completeness of pediatric medication information in the instructions of antitussive and expectorant drugs to ensure the rational use of children’s medication.
4.Mechanism of Zhishi Xiebai Guizhitang in Treating AS Based on Regulation of Cholesterol Metabolism in Foam Cells by TRPA1
Zhanzhan HE ; Zhen YANG ; Xuguang TAO ; Xiangyun CHEN ; Wei DING ; Ce CHU ; Yulu YUAN ; Yuxin ZHANG ; Yongqi XU ; Peizhang ZHAO ; Hongxia ZHAO ; Wenlai WANG
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(10):1-10
ObjectiveTo explore the effect and mechanism of Zhishi Xiebai Guizhitang on the progression of atherosclerosis (AS) mice based on the regulation of cholesterol metabolism in foam cells by transient receptor potential channel ankyrin 1 (TRPA1). MethodThe AS model was established on apolipoprotein E knockout (ApoE-/-) mice with a high-fat diet. The mice were randomly divided into low-dose, middle-dose, and high-dose groups of Zhishi Xiebai Guizhitang (2.97, 5.94, 11.88 g·kg-1) and simvastatin group (0.002 g·kg-1), and the drug was administered along with a high-fat diet. C57BL/6J mice were fed an ordinary diet as a normal group. After the above process, the aorta and serum of mice were taken. The pathological changes of the aortic root were observed by hematoxylin-eosin (HE) staining. The lipid plaques in the aorta were observed by gross oil redness. Serum levels of total cholesterol (TC), triglyceride (TG), low density lipoprotein cholesterol (LDL-C), and high density lipoprotein cholesterol (HDL-C) were detected, and the levels of interleukin-1β (IL-1β) and interleukin-18 (IL-18) were detected by enzyme-linked immunosorbent assay (ELISA). Western blot and immunohistochemical method were used to analyze the expression of TRPA1, ATP-binding cassette transporter A1 (ABCA1), ATP-binding cassette transporter G1 (ABCG1), and mannose receptor (CD206). ResultFrom the perspective of drug efficacy, compared with the normal group, pathological changes such as plaque, a large number of foam cells, and cholesterol crystals appeared in the aorta of the model group, and the serum levels of TC, LDL-C, IL-1β, and IL-18 were significantly increased (P<0.01). The HDL-C level was significantly decreased (P<0.01), and the CD206 level in aortic tissue was significantly decreased (P<0.01). Compared with the model group, the lipid deposition in the aorta was alleviated in all drug administration groups. In addition, except for the high-dose group of Zhishi Xiebai Guizhitang, all drug administration groups could significantly decrease the levels of TC and LDL-C (P<0.01). In terms of inflammation, except for the middle-dose group of Zhishi Xiebai Guizhitang, the levels of IL-1β and IL-18 were significantly decreased in all drug administration groups (P<0.05). Moreover, Zhishi Xiebai Guizhitang could also up-regulate the levels of CD206, and the difference was significant in the middle-dose and high-dose groups (P<0.05). From the perspective of mechanism, the expression levels of TRPA1, ABCA1, and ABCG1 in the aorta in the model group were lower than those in the normal group (P<0.05). Compared with the model group, all drug administration groups significantly increased the expression of TRPA1 in the aorta (P<0.05), and the expressions of ABCA1 and ABCG1 were increased. The differences in the middle-dose and high-dose groups and the simvastatin group were significant (P<0.05), which was basically consistent with the trend of immunohistochemical results. ConclusionZhishi Xiebai Guizhitang can effectively reduce blood lipid and inflammation levels and inhibit the formation of aortic plaque. The mechanism may be explained as follows: the expressions of ABCA1 and ABCG1 downstream are increased through TRPA1, which promotes cholesterol outflow in foam cells, thereby regulating cholesterol metabolism, intervening in inflammation level to a certain extent, and finally treating AS.
5.Efficacy of Lianhua Qingyou Decoction Combined with Acupuncture in the Treatment of Helicobacter Pylori-Infected Chronic Atrophic Gastritis and Its Effect on Gastrointestinal Function
Ding-Ming CHEN ; Zhen LIU ; Zhi-Hua TIAN
Journal of Guangzhou University of Traditional Chinese Medicine 2024;41(4):828-834
Objective To investigate the clinical efficacy of Lianhua Qingyou Decoction combined with acupuncture in the treatment of Helicobacter pylori(Hp)-infected chronic atrophic gastritis(CAG),and to observe the effect on gastrointestinal function.Methods Ninety-eight patients with Hp-infected CAG of heat stagnation in the liver and stomach type were randomly divided into a study group and a control group,with 49 patients in each group.The control group was treated with standard anti-Hp quadruple therapy,and the study group was treated with Lianhua Qingyou Decoction combined with acupuncture on the basis of treatment for the control group.The treatment course for the two groups covered 12 weeks.Before and after the treatment,the two groups were observed in the scores of pathohistological changes in the gastric mucosal atrophy,intestinal epithelial hyperplasia,inflammatory response,activity and Hp infection,gastrointestinal function indicators of serum gastrin,motilin,vasoactive intestinal peptide(VIP),and somatostatin,and the levels of pepsinogens of PGⅠand PGⅡ.After treatment,the clinical efficacy and Hp negative-conversion rate in the two groups were compared.Results(1)After 12 weeks of treatment,the total effective rate in the study group was 95.92%(47/49)and that in the control group was 73.47%(36/49),and the intergroup comparison(by chi-square test)showed that the therapeutic efficacy of the study group was significantly superior to that of the control group(P<0.01).(2)After treatment,the scores of pathohistological changes in the gastric mucosal atrophy,intestinal epithelial hyperplasia,inflammatory response,activity and Hp infection in the two groups were decreased compared with those before treatment(P<0.05),and the decrease in the study group was significantly superior to that in the control group(P<0.01).(3)After treatment,the serum levels of gastrointestinal function indicators of gastrin,motilin and somatostatin in the two groups were all higher than those before treatment(P<0.05),and the serum VIP level was lower than that before treatment(P<0.05).The increase in the serum gastrin,motilin and somatostatin levels and the decrease in the serum VIP level of the study group were significantly superior to those of the control group(P<0.01).(4)After treatment,the serum pepsinogen levels of PGⅠ and PGⅡ in the two groups were higher than those before treatment(P<0.05),and the increase in the study group was significantly superior to that in the control group(P<0.01).(5)The Hp negative-conversion rate of the study group was 95.92%(47/49),which was significantly higher than that of the control group(79.59%,39/49)and the difference was statistically significant(χ2 = 6.078,P = 0.014).Conclusion For the treatment of patients with Hp-infected CAG of heat stagnation in the liver and stomach type,Lianhua Qingyou Decoction combined with acupuncture can effectively enhance the clinical efficacy and Hp negative-conversion rate,improve the pathohistological scores and gastrointestinal function,and regulate the serum PGⅠand PGⅡlevels.
6.Study on the Evolution Principles of Traditional Chinese Medicine syndromes and fNIRS Cerebral Hemodynamic Characteristics in Patients with Alzheimer's Disease
Mengxue ZHANG ; Yanjie QU ; Qian LI ; Chao GU ; Limin ZHANG ; Minrui DING ; Tong ZHANG ; Rongrong ZHEN ; Hongmei AN
Journal of Traditional Chinese Medicine 2024;65(6):600-608
ObjectiveTo explore the evolution principles of symptoms including deficiency, phlegm and blood stasis, and of the functional near-infrared spectroscopy (fNIRS) cerebral hemodynamic characteristics at various stages in patients of Alzheimer's disease. MethodsA total of 497 patients with complaint of memory loss were included, and were divided into subjective cognitive decline (SCD) group (198 participants), mild cognitive impairment (MCI) group (228 participants) and dementia (AD) group (71 participants). Neuropsychological evaluation, traditional Chinese medicine (TCM) syndrome investigation, and fNIRS data collection of prefrontal cortex were performed in each group. Descriptive statistics were used to analyze the distribution of TCM syndromes and the difference of TCM syndrome scores in each group; logistic regression was used to analyze the influence of TCM syndromes on the incidence of the patients; association rules were used to analyze the TCM syndromes of the patients; the hemodynamic characteristics of fNIRS in the prefrontal cortex of each group were compared. ResultsKidney essence deficiency syndrome was the dominant syndrome in all stages of AD. There were statistically significant differences in the distribution frequency of kidney essence deficiency, phlegm turbidity obstructing orifices, blood stasis obstructing collaterals, qi and blood deficiency, heat toxin in the interior, and fu-organ stagnation and turbidity retention syndromes among the three groups (P<0.01), and the scores of kidney essence deficiency syndrome among the three groups were statistically significant (P<0.01). Logistic regression analysis showed that kidney essence deficiency, and qi and blood deficiency syndromes were the main risk factors for the SCD group (P<0.05), phlegm turbidity obstructing orifices syndrome was the main risk factor for the MCI group (P<0.05), and heat toxin in the interior, and fu-organ stagnation and turbidity retention syndromes were the main risk factors for the AD group (P<0.05). The association rule analysis showed that the combination of kidney essence deficiency plus phlegm turbidity obstructing orifices had the highest support (33.33%) in the SCD group, and the combination of kidney essence deficiency plus blood stasis obstructing collaterals had the highest support (32.90% and 52.13%) in both the MCI and AD group. The prefrontal fNIRS results showed that the mean ∆HbO2 concentration in the left dorsolateral prefrontal cortex (LDLPFC) decreased sequentially among the three groups (P<0.05), and the mean ∆HbO2 concentration in the LDLPFC was negatively correlated with the MoCA score among the three groups (r = -0.142, P<0.05). Further analysis showed that the mean ∆HbO2 concentration in the LDLPFC of patients with kidney essence deficiency syndrome were statistically significant differences among the three groups (P<0.05). ConclusionKidney deficiency is the basis of the pathogenesis of AD, and the key brain area damaged is the LDLPFC. Turbid pathogens such as phlegm and blood stasis are the pathological factors that aggravate the disease, and the syndromes of AD show the evolution law of deficiency and excess as “kidney deficiency→phlegm turbidity→blood stasis→turbid toxin”. The changes in prefrontal hemodynamics based on fNIRS are consistent with the changes in the characteristics of symptoms, which can be used to assess the degree of cognitive impairment in AD patients.
7.Treatment of Paroxysmal Sympathetic Hyperactivity by the Method of “Returning Fire to Its Origin”
Yingchun XU ; Yi GUO ; Jing DING ; Wanyu LIU ; Zhen TIAN ; Jiangying WU ; Xiaozhe WU
Journal of Traditional Chinese Medicine 2024;65(5):537-540
This paper summarized the clinical experience of using the method of “returning fire to its origin” for treatment of paroxysmal sympathetic hyperactivity (PSH). According to the causes and clinical characteristics of PSH, the author believes that the deficiency of kidney qi, and the loss of yin and yang are the basis of the pathogenesis of PSH. Fright causes qi to be chaotic as the triggering mechanism of PSH. The key mechanism of PSH is that the deficiency yang with upper manifestation, and the fire does not return to its origin. The treatment should be nourishing yin and astringing yang, by taking modified Yinhuo Decoction (引火汤) internally, and receiving warm moxibustion as the first choice externally with selected acupoints Guanyuan (CV 4), Mingmen (GV 4), and bilateral Yongquan (KI 1); For prevention, attention should be paid to take care of stomach qi, support healthy qi, and cultivate original qi.
8.Linggui Zhugantang Treats Chronic Bronchitis in Rats via PLA2-TRPV1/TRPA1 Pathway
Wei DING ; Wenlai WANG ; Zhenhong LIU ; Xiangyun CHEN ; Zhanzhan HE ; Ce CHU ; Yulu YUAN ; Yongqi XU ; Yuxin ZHANG ; Peizhang ZHAO ; Zhen YANG ; Hongxia ZHAO
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(14):1-9
ObjectiveTo study the effect and mechanism of Linggui Zhugantang in treating chronic bronchitis (CB) induced by exposure to cigarette smoke combined with tracheal instillation of lipopolysaccharide (LPS). MethodSixty SPF-grade SD rats were randomly divided into normal, model, dexamethasone (1 mg·kg-1), and high-, medium-, and low-dose (30.06, 15.03, 7.515 g·kg-1, respectively) Linggui Zhugantang groups by the body weight stratification method, with 10 rats in each group. Each group was administrated with 200 μL LPS (1 g·L-1) by tracheal instillation on days 1 and 14, respectively, while the normal group was administrated with an equal volume of normal saline. Except the normal group, the other groups were exposed to cigarette smoke on days 2-13 and 15-30 (10 cigarettes/time/30 min, twice/day) for the modeling of CB. The rats were administrated with corresponding drugs by gavage for 30 consecutive days from day 2 of modeling, and the mental status, behavior, and body weights of the rats were observed and measured. The wet/dry mass ratio (W/D) of the left lung was measured 30 days after modeling. Hematoxylin-eosin staining was employed to observe the pathological changes in the lung and bronchial tissues. The bronchial mucus secretion and goblet cell proliferation were observed by Alcian blue-periodic acid Schiff (AB-PAS) staining. The levels of mucin 5AC (MUC5AC), interleukin (IL)-13, IL-6, and tumor necrosis factor (TNF)-α in the serum were determined by enzyme-linked immunosorbent assay. The expression of phospholipase A2 (PLA2), transient receptor potential vanilloid receptor 1 (TRPV1), and transient receptor potential ankyrin 1 (TRPA1) in the lung tissue was quantitatively analyzed by immunohistochemistry and Western blot. ResultCompared with the normal group, the model group showcased abnormal mental status and behaviors, bloody secretion in the nose and mouth, the mortality rate of 40%, decreased body weight, severe lung bronchial structure damage, a large number of inflammatory mediators and inflammatory cell infiltration in the tube wall, hyperemia, edema, and fibroplasia, massive proliferation of goblet cells, excessive secretion and accumulation of mucus, stenosis and deformation of the lumen, and aggravation of pulmonary edema (P<0.01). In addition, the model group had higher levels of MUC5AC, IL-13, IL-6, and TNF-α in the serum and higher expression of PLA2 in the lung tissue than the normal group (P<0.01). Compared with the model group, the medication groups showed normal mental status and behaviors, reduced mortality rate, stable weight gain, reduced lung and bronchial injuries, decreased goblet cell proliferation and mucus secretion, and alleviated pulmonary edema (P<0.01). Furthermore, Linggui Zhugantang lowered the levels of MUC5AC, IL-13, IL-6, and TNF-α in the serum and down-regulated the protein levels of PLA2, TRPV1, and TRPA1 in the lung tissue (P<0.01). ConclusionLinggui Zhugantang can reduce the pulmonary inflammation and airway mucus hypersecretion in the rat model of chronic bronchitis. It may exert the effects of reducing inflammation and resolving phlegm by regulating the PLA2-TRPV1/TRPA1 pathway.
9.Effect of Qingfei Paidu Decoction on Acute Lung Injury Model Mice Based on TRPV1/TRPA1 Heat-sensitive Channel
Yulu YUAN ; Zhanzhan HE ; Ce CHU ; Xuguang TAO ; Zhen YANG ; Xiangyun CHEN ; Wei DING ; Yongqi XU ; Yuxin ZHANG ; Peizhang ZHAO ; Wanping CHEN ; Hongxia ZHAO ; Wenlai WANG
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(24):95-102
ObjectiveTo investigate the mechanism and effect of Qingfei Paidu decoction on transient receptor potential vanilloid-1/Transient receptor potential ankyrin1 (TRPV1/TRPA1) based on heat-sensitive channel and inflammatory response. MethodAccording to body weight, 80 8-week-old C57BL/6 mice were randomly divided into the normal group, model group, dexamethasone group (5 mg·kg-1), and low-dose, medium-dose, and high-dose groups of Qingfei Paidu decoction (14.865, 29.73, 59.46 g·kg-1), with 12 mice in each group. In addition to the normal group, the other groups were administered 20 μL (1×10-3 g·kg-1) to each mouse by airway infusion to establish the acute lung injury (ALI) model. In the administration group, the drug was given 1 h after modeling and again after an interval of 24 h. The lung tissue was taken 36 h after modeling. Double lung wet/dry weight ratio(W/D), hematoxylin-eosin (HE) staining, enzyme-linked immunosorbent assay (ELISA), and Western blot were used to observe and detect the pathological changes of lung tissue, expression levels of inflammatory cytokine tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6), and expressions of TRPV1 and TRPA1 proteins in heat-sensitive channel, nuclear factor kappa-B (NF-κB), inhibitor of NF-κB (IκBα) in inflammatory pathway, and phosphorylated proteins. The phosphorylated protein/total protein ratio was calculated. ResultCompared with that in the normal group, the lung tissue of mice in the model group was seriously damaged, and pulmonary capillary permeability increased. Alveolar capillary congestion and dilation destroyed the complete structure of the alveolar, and the alveolar wall thickened. A large number of inflammatory cells and red blood cells were infiltrated, and pulmonary edema was significantly aggravated. The expressions of TNF-α, IL-6, TRPV1, TRPA1, phosphorylated NF-κB p65/NF-κB p65, and phosphorylated IκBα/IκBα were significantly increased (P<0.01), and the whole lung W/D was significantly increased (P<0.01). Compared with the model group, the dexamethasone group and low-dose, medium-dose, and high-dose groups of Qingfei Paidu decoction could significantly improve pulmonary edema. TNF-α, IL-6, TRPV1, TRPA1, lung tissue NF-κB p65, and IκBα phosphorylated protein/total protein ratio decreased significantly (P<0.05, P<0.01). The whole lung W/D also decreased significantly (P<0.05, P<0.01). ConclusionQingfei Paidu decoction has anti-inflammatory and protective effects on LPS-ALI mice, which can effectively reduce inflammation, induce diuresis, and alleviate edema. Its mechanism may be related to the regulation of the expression of TRPA1 and TRPV1 and the inhibition of the activation of the NF-κB pathway.
10.Liuwei Dihuang Pills-elicited inhibition of MMP-2/MMP-9 via RAGE on tight junction protein of Aβ1-40-injured bEnd.3 cells
Rui DING ; Yong YUAN ; Ya-Quan JIA ; Ai-She GAO ; Zhen-Qiang ZHANG ; Jun-Ying SONG
Chinese Traditional Patent Medicine 2024;46(2):424-430
AIM To investigate the protective effects and the mechanism of the Liuwei Dihuang Pills on mouse brain microvascular endothelial(bEnd.3)cells damaged by β-Amyloid protein1-40(Aβ1-40).METHODS CCK8 method was used to detect the effects of Aβ1-40 and medicated serum of Liuwei Dihuang Pills(MSLDP)on cell activity,and to screen the appropriate concentration.bEnd.3 cells of the control group,the Aβ1-40 group,the MSLDP+Aβ1-40 group and the MSLDP group had their low density lipoprotein-associated protein 1(LRP1),receptor for advanced glycation end products(RAGE),matrix metalloproteinase-2(MMP-2),MMP-9,scaffold protein zonule protein-1(ZO-1)detected by Western blot.bEnd.3 cells assigned into the control group,the Aβ1-40 group,the FPS-ZM1(RAGE inhibitor)+Aβ1-40 group and the FPS-ZM1+Aβ1-40+MSLDP group had their expressions of RAGE,MMP-9,MMP-2 and ZO-1 detected by Western blot as well.RESULTS The cell activity of bEnd.3,was dose-dependently decreased by Aβ1-40(P<0.01),but was protected by MSLDP(P<0.05,P<0.01).And 10 μmol/L Aβ1-40 and 10%MSLDP were selected for subsequent experiments.Compared with the control group,the Aβ1-40 group displayed increased protein expressions of RAGE,MMP-2 and MMP-9(P<0.01),decreased protein expressions of LRP1,ZO-1 and BDNF(P<0.05,P<0.01),and decreased fluorescence intensities of LRP1 and ZO-1(P<0.01).Compared with the Aβ1-40 group,the MSLDP group shared decreased expressions of RAGE,MMP-2,MMP-9 proteins and RAGE fluorescence intensity(P<0.05,P<0.01),and increased expressions of LRP1,ZO-1 and BDNF proteins,and the fluorescence intensity of LRP1 and ZO-1(P<0.05,P<0.01);the Aβ1-40+FPS-ZM1 group displayed decreased protein expressions of MMP-2,MMP9 and RAGE(P<0.05,P<0.01),and increased ZO-1 protein expression(P<0.05);and the Aβ1-40+FPS-ZM1+ MSLDP group displayed an even more decreased protein expressions of MMP-2,MMP9 and RAGE(P<0.01),increased ZO-1 protein expression(P<0.01)due to the the combination use of FPS-ZM1 and MSLDP.CONCLUSION Liuwei Dihuang Pills can protect the tight junction of bEnd.3 injured by Aβ1-40 and neurovascular units from Alzheimer's disease by alleviating the dysfunction of the blood-brain barrier via RAGE-mediated MMP-2/MMP-9 pathway inhibition.

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