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1.
Article in Chinese | WPRIM | ID: wpr-1031876

ABSTRACT

ObjectiveTo observe the regulatory effect of Huangwu Ganfu ointment on transient receptor potential anchor protein 1 (TRPV1) receptor expression, macrophage polarization, and p38 mitogen-activated protein kinase (p38 MAPK) signaling pathway in synovial tissue of knee osteoarthritis (KOA) rats with yang deficiency and cold coagulation syndrome and explore the mechanism of relieving peripheral inflammatory hyperalgesia of KOA. MethodForty-eight male SD rats were randomly divided into blank group, model group, high-dose, middle-dose, and low-dose groups of Huangwu Ganfu ointment (9.3, 4.65, 2.325 g·kg-1), and celecoxib group (20.82 mg·kg-1). The KOA rat model of yang deficiency and cold coagulation syndrome was established through climate box and swimming for two weeks combined with an injection of sodium iodoacetate (MIA) in the articular cavity. After continuous administration for four weeks, the general condition of rats in each group was observed, and the pain withdrawal threshold (PWT) and joint diameter induced by mechanical stimulation were recorded. The expression levels of interleukin-1β (IL-1β), tumor necrosis factor-α (TNF-α), nerve growth factor (NGF), and calcitonin gene-related peptide (CGRP) inflammatory factor were detected by enzyme-linked immunosorbent assay (ELISA), and the histopathological changes of synovial tissue of the knee joint were observed by hematoxylin-eosin (HE) staining. Western blot was used to detect the protein expression of TRPV1, p38 MAPK, and p-p38 MAPK in synovial tissue of the knee joint, and immunofluorescence (IF) was used to evaluate the polarization of M1/M2 macrophages. ResultCompared with that in the blank group, the overall mental state of the model group was worse, and the autonomous activity was decreased. The body mass was lower, and the joint diameter was increased. The X-ray showed that the osteophyte at the edge of the joint proliferated, and the articular surface was obviously rough. The articular cavity was significantly narrowed, and the PWT was significantly decreased (P<0.01). The contents of IL-1β, TNF-α, CGRP, and NGF in serum and synovium Krenn score increased significantly (P<0.01). The protein expression of TRPV1 and p-p38 MAPK/p38 MAPK increased significantly (P<0.01), and the proportion of M1 macrophages and M1/M2 increased (P<0.01), while the proportion of M2 macrophages decreased (P<0.01). Compared with model group, the body mass in the low, middle, and high dose groups of Huangwu Ganfu ointment increased to different degrees (P<0.05, P<0.01). The diameter of the knee joint in the high dose group of Huangwu Ganfu ointment and celecoxib group decreased (P<0.01). The recovery of PWT in the high and middle dose groups of Huangwu Ganfu ointment groups was more obvious (P<0.05). The contents of IL-1β and CGRP in the serum of rats in each administration group were significantly decreased (P<0.01), and the content of serum TNF-α in the celecoxib group and high dose group of Huangwu Ganfu ointment decreased significantly (P<0.05). The content of serum NGF in the middle dose group of Huangwu Ganfu ointment decreased significantly (P<0.05), and the synovium Krenn score decreased in the high dose group of Huangwu Ganfu ointment (P<0.05). In addition, the protein expression of TRPV1 and p-p38 MAPK/p38 MAPK in synovial tissue decreased significantly in all groups of Huangwu Ganfu ointment (P<0.01). The proportion of M1 macrophages in synovial tissue in the celecoxib group and all groups of Huangwu Ganfu ointment decreased (P<0.01), and the proportion of M2 macrophages in the high dose group of Huangwu Ganfu ointment increased (P<0.05). The M1/M2 in the middle and high dose groups of Huangwu Ganfu ointment decreased (P<0.05). ConclusionHuangwu Ganfu ointment can mediate the polarization of macrophages to reduce the inflammatory reaction of KOA, alleviate the release of inflammatory pain mediators, and lower the protein expression of TRPV1. The mechanism may be related to the p38 MAPK signaling pathway, so as to improve the peripheral hyperalgesia of KOA.

2.
Herald of Medicine ; (12): 1791-1795, 2023.
Article in Chinese | WPRIM | ID: wpr-1023651

ABSTRACT

Objective To develop an accurate,rapid and sensitive flow cytometry method for the determination of anti-mesenchymal stem cell antibody in cynomolgus monkey serum.Methods After the solutions of mesenchymal stem cell were centrifuged and washed,and the suspension was taken,positive controls or actual samples were added and incubated with mesenchymal stem cell,then were incubated with protein L-PE solution.After the removal of the free protein L-PE,the mean fluorescence intensity of the PE was detected by flow cytometry.Results The method sensitivity is 115.54 ng·mL-1,far higher than the non-clinical research recommended sensitivity of 250-500 ng·mL-1.The precision of intra-assay and inter-assay were less than 20%.Assay cut points,low positive control concentration determination,sensitivity,precision and stability were validated in this study.Conclusion The method is proved to be sensitive,specific,rapid and suitable for the determination of anti-mesenchymal stem cell antibody in monkey serum and immunogenicity study.

3.
Article in Chinese | WPRIM | ID: wpr-883503

ABSTRACT

Alismatis Rhizoma(AR)is widely used in Chinese medicine,and its major bioactive components,tri-terpenes,reportedly possess various pharmacological activities.Therefore,it is very important to study the metabolism of triterpenes in vivo.However,the metabolism of AR triterpene extract has not been comprehensively elucidated due to its complex chemical components and metabolic pathways.In this study,an ultra-performance liquid chromatography quadrupole time-of-flight mass spectrometry method,which was based on the characteristic ions from an established database of known triterpenes,was used to analyze the major metabolites in rats following the oral administration of Alismatis Rhizoma extracts(ARE).As a result,a total of 233 constituents,with 85 prototype compounds and 148 metabo-lites,were identified for the first time.Hydrogenation,oxidation,sulfate and glucuronidation conjugation were the major metabolic pathways for triterpenes in AR.In addition,the mutual in vivo transformation of known ARE triterpenes was discovered and confirmed for the first time.Those results provide comprehensive insights into the metabolism of AR in vivo,which will be useful for future studies on its pharmacodynamics and pharmacokinetics.Moreover,this established strategy may be useful in meta-bolic studies of similar compounds.

4.
Progress in Modern Biomedicine ; (24): 4285-4288, 2017.
Article in Chinese | WPRIM | ID: wpr-615361

ABSTRACT

Objective:To explore the effect of laparoscopic minimally invasive on anorectal dynamics and serum carcino-embryonic antigen (CEA) and carbohydrate antigen 724 (CA724) in patients with rectal cancer.Methods:38 cases of rectal cancer patients were selected as the research group from March 2014 to March 2016 and 40 cases of non neoplastic intestinal polyps as control group,Preoperative anorectal dynamics index (anal resting pressure (ARP),rectal resting pressure (RRP)and anal maximal contraction pressure (MSP),rectal maximum tolerated volume (MTV) of two groups were compared,Preoperative and 3 d,1,2 weeks after operation of serum CEA and CA724 levels were detected in the research group and the clinical curative effect was observed.Results:There was no significant difference of ARP,RRP,MSP,MTV before operation in research group compared with the control group(P>0.05),and was significantly reduced 2,4 weeks after operation(P<0.05),then gradually restored,and basically recovered to the preoperative level 12 weeks after operation.There was no significant difference of serum CEA and CA724 levels 3 d after operation compared with preoperative in research group (P>0.05),and were significantly lower 1,2 weeks after operation compared with preoperative (P<0.05).The clinical effective rate was 65.8%.Conclusion:Efficacy of laparoscopic minimally invasive in treatment of rectal cancer is effective,can effectively reduce the levels of tumor markers,although there are some influence on anorectal dynamics,but it can be returned to normal in the short term.

5.
Article in Chinese | WPRIM | ID: wpr-570228

ABSTRACT

Objective The research is to study the temperature rising situation in the thermochemotherapy process of the domestic dog through the hepatic artery, and the bearing ability of heart, liver, and kidney during the process, as well as the biochemical and histological change of the liver after the procedure. Methods A thermostated container of water bath and an infusion pump, both designed domestically, were adopted. Ten domestic dogs were divided into 2 groups of A and B with 5 dogs respectively. Through hepatic artery infusion for group A, glucose solution (GS) was administered at 43?C for 30 minutes, with the flowing rate of 30ml/min, in group B 5% GS at 45?C for 30 minutes, with the flowing rate of 40ml/min added with chemotherapy drug MMC(0.2mg/kg) was administered to 3 dogs and 5% GS with the same flowing rate added with CDDP(1mg/kg) into other 2 dogs. Results The temperature of anus and liver was 0.3~0.5?C and 0.5~1.0?C higher than before the procedure respectively, there was no significant rate changes of heart and respiration during and before the procedure. Four dogs were radiographied after 15 minutes' infusion, showing different degrees of hepatic artery dilation accompanied with relevant increase of infusion. Six dogs were radiographied after 30 minutes' infusion, 5 of them showed arterial contraction astescal on accompanied with relevant decrease of infusion. There appeared slight or moderate increase of aminopherase after the procedure, but in actually returned to normal within a week. Pathologically, there appeared no acute effect on heart, lung and kidney other than slight and reversible damage to hepatic tissue. Conclusions Healthy dogs can endure the chemotherapy treatment of infusion through hepatic arterial fistula at 43?C 45?C for 30 minutes.

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