Research progress of anti-pulmonary fibrosis drugs targeting autotaxin-lysophosphatidic acid axis / 药学学报

Pulmonary fibrosis is an interstitial lung disease characterized by inflammatory injury and tissue structure destruction. Currently, there is a lack of effective therapeutic drugs for pulmonary fibrosis, and the mechanism is still unknown. Therefore, it is urgent to seek new targets for effective drugs. In pulmonary fibrosis, the level of autotaxin (ATX) in bronchoalveolar lavage fluid increases and stimulates the production of lysophosphatidic acid (LPA). The involvement of LPA receptors in activating a variety of G-protein-mediated signal transduction pathways leads to a range of related physiological effects, including pro-inflammatory signaling in epithelial cells, activation of transforming growth factor signaling, and stimulation of fibroblast accumulation. LPA receptor antagonists and ATX inhibitors have been concerned as new targets for pulmonary fiber therapy, and currently related drugs have entered clinical trials. In this paper, the pathophysiological effects of LPA and ATX in pulmonary fibrosis disease and related drug development progress were reviewed to provide reference information of new drug development for pulmonary fibrosis based on the ATX-LPA axis.

Molecular Mechanism and Potential Clinical Value of Lysophosphatidic Acid in Vascular Endothelial Cells / 中国生物化学与分子生物学报

The vascular system constitutes the largest surface of the human body. Vascular endothelial cells are a layer of flat epithelial cells covering the inner wall of blood vessels and have a variety of biolog-

Downmodulation of lysophosphatidic acid by Berberine loaded folate-conjugated glycol chitosan nanoparticles (BFGCN) to mitigate Rheumatoid arthritis (RA) & Cardio-vascular disease(CVD): Current knowledge and future perspectives

The perils of cardiovascular diseases (CVD) are enhanced by systemic chronic inflammation in autoimmune disorders like Rheumatoid arthritis (RA), in which the patients generally exhibit a high inflammatory burden, dyslipidemia causing 50-60% of RA patients susceptible to CVD dependent mortality. Lysophosphatidic acid (LPA) is a polar, pleiotropic lipid molecule that is water soluble and present in the synovial fluid that can be exploited as an effective biomarker for lipid-signalling. Current research on alternative medicine has recognized various new molecular targets of Berberine (BBR) and established novel signals in support of the efficacy and therapeutic potential of BBR to fight CVD. Therefore, BBR, an alkaloid with poor aqueous solubility could be foreseen as a therapeutic strategy for the reduction of inflammation induced lipidemia by targeting the macrophages and modulating their functions. Hence, a novel BBR loaded folate-conjugated glycol chitosan nanoparticles (BFGCN) could be hypothesized as a three-pronged approach to target activated macrophages, fibroblasts of synovial fluid for downmodulation of LPA. The greatest challenge is the heterogeneity, complexity and interdependence of RA and CVD. Investigation of prognostic and predictive biomarkers is urgently required. Therefore, an improved understanding of the pathogenesis of RA would facilitate identifying an improved targeted treatment and management of RA patients.

Significance of lysophosphatidic acid receptor 6 in the large-cell transformation of mycosis fungoides and its effect on the proliferation and apoptosis of cutaneous T-cell lymphoma cells / 中华皮肤科杂志

Objective:To determine lysophosphatidic acid receptor 6 (LPAR6) expression in patients with mycosis fungoides (MF) , a variant of cutaneous T-cell lymphoma (CTCL) , and to investigate its role and mechanism of action in the development and prognosis of CTCL.Methods:A total of 110 patients with confirmed MF were collected from Department of Dermatology, Peking University First Hospital from 2011 to 2020, including 24 with large-cell transformation (LCT) and 25 with non-large cell transformation (NLCT) in the discovery cohort, and 24 with LCT and 37 with NLCT in the validation cohort. RNA sequencing and RT-PCR were conducted to determine the LPAR6 expression in patients in the discovery cohort and validation cohort respectively. LPAR6 expression was compared between patients with LCT and those with NLCT, and its effect on the prognosis of patients was evaluated. Two LPAR6-overexpressing CTCL cell lines MyLa and Sz4 were constructed to evaluate the effect of LPAR6 overexpression on proliferative activity of MyLa and Sz4 cells, with the cells normally expressing LPAR6 as the control group; after the treatment with LPAR6-related ligand lysophosphatidic acid (LPA) , 2S-OMPT, adenosine triphosphate (ATP) or adenosine (ADO) , the effects of LPAR6 activation on the proliferative activity and apoptosis of LPAR6-overexpressing MyLa and Sz4 cells were evaluated by the MTS method and flow cytometry respectively. Log-rank test was used for prognostic analysis, and t test or Mann-Whitney U test was used for comparisons between two groups. Results:As RNA sequencing showed, LPAR6 was one of the significantly underexpressed genes in the LCT group in the discovery cohort; in the validation cohort, LPAR6 expression (median[ Q1, Q3]) was significantly lower in the LCT group (204.90[81.90, 512.70]) than in the NLCT group (809.40[417.50, 1 829.20], U= 242.00, P= 0.002) ; in the two cohorts, the underexpression of LPAR6 was significantly associated with increased risk of poor prognosis (both P < 0.01) . Cell proliferation assay showed no significant difference in the proliferative activity of MyLa or Sz4 cells between the LPAR6 overexpression group and control group at 0, 24, 48 and 72 hours during the experiment (all P > 0.05) ; 48 hours after activation of LPAR6 by LPA, 2S-OMPT, ATP and ADO in MyLa cells, the LPAR6 overexpression group showed significantly decreased cellular proliferative activity (1.38 ± 0.01, 1.04 ± 0.01, 1.09 ± 0.03, 1.23 ± 0.01, respectively) compared the control group (1.73 ± 0.04, 1.23 ± 0.01, 1.24 ± 0.01, 1.42 ± 0.03, t= 30.33, 18.38, 4.78, 5.75, respectively, all P < 0.05) , but significantly increased cell apoptosis rate (17.93% ± 0.88%, 17.75% ± 0.35%, 23.97% ± 0.57%, 31.44% ± 0.34%, respectively) compared the control group (3.98% ± 0.03%, 7.81% ± 0.59%, 11.95% ± 0.85%, 12.02% ± 0.48%, t= 15.93, 14.49, 11.74, 33.01, respectively, all P < 0.05) ; 48 hours after activation of LPAR6 by 2S-OMPT and ADO in Sz4 cells, compared with the control group, the LPAR6 overexpression group also showed significantly decreased cellular proliferative activity (2S-OMPT: 1.29 ± 0.04 vs. 1.48 ± 0.01; ADO: 1.27 ± 0.01 vs. 1.51 ± 0.02; both P < 0.05) , but significantly increased cell apoptosis rate (2S-OMPT: 41.70% ± 0.70% vs. 29.35% ± 0.55%; ADO: 37.05% ± 0.15% vs. 24.60% ± 1.00%; both P < 0.05) . Conclusions:LPAR6 was underexpressed in the patients with LCT, and its underexpression was significantly associated with increased risk of poor prognosis. In vitro activation of LPAR6 could inhibit the proliferation of CTCL cells and promote their apoptosis, suggesting that the decrease of LPAR6 expression may be one of the important mechanisms underlying disease progression in patients with LCT.

Effects of lysophosphatidic acid on ischemia/reperfusion injury and TRPV1 expression in isolated mouse heart / 中国药理学通报

Aim To investigate the effects of lysophosphatidic acid on ischemia / reperfusion injury(IRI)and TRPV1 expression in isolated mouse heart.Methods The IRI model of isolated mouse heart was established by Langendorff device.The hearts in sham group were continuously perfused for 100 min.The hearts in IR group were stabilized for 10 min followed by no perfusion for 30 min and reperfusion for 60 min.Exogenous LPA was added in the K-H solution during IR in IR+LPA group while HA130, an LPA synthesis inhibitor, was intraperitoneally injected before IR in IR+HA130 group.The infarct volume was measured by TTC staining, the determination of LPA and LDH levels in coronary effluent and LPA concentration in serum was measured by ELISA method.Finally, the expression levels of pTRPV1/TRPV1 and Bcl-2/Bax in myocardial tissues were determined by Western blot.Results Compared with sham group, IR caused evident myocardial infarction and increased the levels of LDH and LPA in coronary effluent.The increase of LPA was linearly correlated with myocardial infarction volume.In addition, the protein levels of pTRPV1 and TRPV1 in myocardium increased, while the ratio of Bcl-2/Bax decreased.The myocardial injury in IR+LPA group was aggravated.In contrast, myocardial IRI was reversed in IR+HA130 group.Conclusions Myocardial ischemia-reperfusion induces the release of LPA, which aggravates myocardial post-ischemic injury, while the inhibition of LPA release exerts cardioprotective effects.The underlying mechanisms might be related to the regulation on cardiac TRPV1 expression and apoptotic signals.

LPA / 中国生物化学与分子生物学报

Lysophosphatidic acid(LPA)is a small bioactive phospholipid that mediates various cellular processes such as proliferation, survival, and migration. In particular, LPA signaling has been shown to affect the development of diverse tissues. Our previous work demonstrated that LPA could promote primary neonatal rat cardiomyocytes proliferation. However, the role of LPA and its receptor in postnatal heart de-velopment is unknown. By using databases for biological information and RT-qPCR, we analyzed the ex-pression of six LPA receptors (LPA

Lysophosphatidic Acid Down-regulated SKOV3 Cells of Ovarian Cancer in Doxorubicin Hydrochloride-induced Apoptosis / 中国生物化学与分子生物学报

Lysophosphatidic acid (LPA), a bioactive lipid medium, plays an important role in the development and progression of ovarian cancer. Doxorubicin hydrochloride (DOX) is a first-line drug in the ovarian cancer clinical therapy, while the effect and molecular mechanism of LPA in the ovarian cancer with DOX treatment is still unclear. This study intended to explore the effect and molecular mechanism of LPA in ovarian cancer treated with DOX. SKOV3 and OVCAR-3 cells of human ovarian cancer and Chinese hamster ovary cells were treated with control, LPA (lOp-mol/L), DOX (2jjLmol/L) and LPA (10jJLmol/L) + DOX (2p,mol/L) respectively for 24 hours. The morphological changes of SKOV3 cells were observed under optical microscope and transmission electron microscope. Results showed that LPA reduced cell death and the degree of chromatin aggregation in SKOV3 cells treated with DOX; RT-qPCR showed that LPA treatment could down-regulate the mRNA levels of caspase-3 in DOX-treated SKOV3 cells (P<0. 05); Western blot showed that LPA treatment could reduce caspase-3 and cleaved caspase-3 levels treated with DOX in SKOV3, OVCAR-3 and CHO cells (P<0. 05); Flow cytometry using Annexin V/PI double staining showed that LPA could down-regulate apoptosis in SKOV3 cells treated with DOX (P<0. 05); DCFH-DA method was used to detect intracellular levels of reactive oxygen species (ROS) in SKOV3 cells. It was found that LPA reduced the intracellular ROS level treated with DOX (P<0. 05). Our preliminarily study showed the effect of LPA in the apoptosis of ovarian cancer treated with DOX, which may provide a reference for the drug therapy of ovarian cancer targeting LPA.

Research progress on the biological regulatory function of lysophosphatidic acid in bone tissue cells / 华西口腔医学杂志

Lysophosphatidic acid (LPA) is a small phospholipid that is present in all eukaryotic tissues and blood plasma. As an extracellular signaling molecule, LPA mediates many cellular functions by binding to six known G protein-coupled receptors and activating their downstream signaling pathways. These functions indicate that LPA may play important roles in many biological processes that include organismal development, wound healing, and carcinogenesis. Recently, many studies have found that LPA has various biological effects in different kinds of bone cells. These findings suggest that LPA is a potent regulator of bone development and remodeling and holds promising application potential in bone tissue engineering. Here, we review the recent progress on the biological regulatory function of LPA in bone tissue cells.

Expression and clinical significance of lysophosphatidic acid receptor 5 protein in breast cancer / 中国基层医药

Objective To investigate the expression and clinical significance of lysophosphatidic acid receptor 5 (LPA5) protein in breast cancer tissues.Methods The specimens of breast cancer tissues and adjacent tissues of 80 breast cancer patients from January 2015 to December 2016 in the First People's Hospital of Yongkang were selected in the study.The expression of LPA5 in breast cancer tissues and adjacent tissues was determined by immunohisto-chemistry.The clinical case characteristics of patients were collected.Results The positive expression rate of LPA5 in breast cancer tissues (72.5%) was higher than that in adjacent tissues (8.75%) (χ2 ＝67.394,P＜0.05).The positive rate of LPA5 in clinical stage Ⅲ-Ⅳ(91.3%) was higher than that in stage Ⅰ-Ⅱ(64.9%)(χ2 ＝5.725, P＜0.05).The positive rate of LPA5 in patients with lymph node metastasis (94.7%) was higher than that in patients without lymph node metastasis (52.4%)(χ2 ＝17.951,P＜0.05).The positive rate of LPA5 in patients with intravascular tumor thrombus ( 92.3%) was higher than that in patients without intravascular tumor thrombus (63.3%)(χ2 ＝7.580,P＜0.05).The expression of LPA5 in breast cancer tissues was not related to age and tumor diameter (P＞0.05).There were no significant differences in the positive expression rates of LPA 5 in the breast cancer tissues among luminal type ,human epidermal growth factor receptor 2 (Her2) overexpressing type and triple negative type (P＞0.05).The disease-free survival rate(86.3%) and overall survival rate (87.5%) of patients with negative expression of LPA5 in breast cancer tissues were higher than those in breast cancer tissues with positive expression (65.0%,66.3%)(χ2 ＝4.517,4.328,all P＜0.05).Conclusion LPA5 expression is up-regulated in breast cancer tissues,and LPA5 may be involved in the development and progression of breast cancer ,which is closely related to the prognosis of breast cancer.

Expression and clinical significance of lysophosphatidic acid receptor 5 protein in breast cancer / 中国基层医药

Objective@#To investigate the expression and clinical significance of lysophosphatidic acid receptor 5 (LPA5) protein in breast cancer tissues.@*Methods@#The specimens of breast cancer tissues and adjacent tissues of 80 breast cancer patients from January 2015 to December 2016 in the First People's Hospital of Yongkang were selected in the study.The expression of LPA5 in breast cancer tissues and adjacent tissues was determined by immunohistochemistry.The clinical case characteristics of patients were collected.@*Results@#The positive expression rate of LPA5 in breast cancer tissues(72.5%) was higher than that in adjacent tissues(8.75%)(χ2=67.394, P<0.05). The positive rate of LPA5 in clinical stage Ⅲ-Ⅳ(91.3%) was higher than that in stage Ⅰ-Ⅱ(64.9%)(χ2=5.725, P<0.05). The positive rate of LPA5 in patients with lymph node metastasis(94.7%) was higher than that in patients without lymph node metastasis(52.4%)(χ2=17.951, P<0.05). The positive rate of LPA5 in patients with intravascular tumor thrombus(92.3%) was higher than that in patients without intravascular tumor thrombus (63.3%)(χ2=7.580, P<0.05). The expression of LPA5 in breast cancer tissues was not related to age and tumor diameter (P>0.05). There were no significant differences in the positive expression rates of LPA5 in the breast cancer tissues among luminal type, human epidermal growth factor receptor 2 (Her2) overexpressing type and triple negative type (P>0.05). The disease-free survival rate(86.3%) and overall survival rate(87.5%) of patients with negative expression of LPA5 in breast cancer tissues were higher than those in breast cancer tissues with positive expression (65.0%, 66.3%)(χ2=4.517, 4.328, all P<0.05).@*Conclusion@#LPA5 expression is up-regulated in breast cancer tissues, and LPA5 may be involved in the development and progression of breast cancer, which is closely related to the prognosis of breast cancer.

Effect and mechanism of YAP in hepatic ischemia-reperfusion injury / 器官移植

Objective To explore the effect and mechanism of Yes-associated protein (YAP) in hepatic ischemia-reperfusion injury (IRI) of mice. Methods Forty male C57BL/6 mice were randomly divided into the sham operation group (Sham group), lysophosphatidic acid (LPA) + Sham group, IRI group and LPA+IRI group, 10 mice in each group. Liver tissue and serum samples were collected at 6 h after ischemia-reperfusion. The levels of serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) were detected. Histopathological changes and macrophage infiltration of liver tissues were detected by hematoxylin-eosin (HE) staining and immunohistochemical staining. The protein expression level of YAP was detected by Western blot. The messenger ribonucleic acid (mRNA) expression levels of inflammatory cytokines including tumor necrosis factor (TNF)-α, inducible nitric oxide synthase (iNOS), interleukin (IL)-1 and IL-6 were quantitatively measured by reverse transcription polymerase chain reaction (RT-PCR). Results Western blot results demonstrated that the protein expression level of YAP in the LPA+IRI group was significantly up-regulated than that in the IRI group. Compared with the Sham group, the ALT and AST were significantly higher in the IRI group (both P < 0.05). The serum levels of ALT and AST in the LPA+IRI group were significantly lower than those in the IRI group (both P < 0.05). HE staining revealed that the morphology of hepatocytes was normal in the Sham group and LPA + Sham group. Pathological changes, such as liver congestion, liver cell swelling and structural abnormalities of hepatic lobule, occurred in the LPA+IRI group and IRI group. Compared with the IRI group, pathological changes were alleviated in the LPA+IRI group. RT-PCR indicated that the mRNA expression levels of TNF-α, iNOS, IL-1 and IL-6 in the LPA+IRI group were lower than those in the IRI group (all P < 0.05). Immunohistochemical demonstrated that LPA partially inhibited macrophage infiltration in ischemic tissues after IRI. Conclusions YAP can significantly mitigate hepatic IRI. The mechanism is associated with the regulation of macrophage recruitment and activation.

Progress of autotaxin in tumors / 肿瘤研究与临床

Autotaxin (ATX) as a member of nucleotide pyrophosphatase/phosphodiesterase (ENPP) family, possesses the activity of lysophospholipase D, and plays a key role in lipid signaling pathway.Increased ATX expression has been detected in a large variety of cancers,implying its important functions in tumor origination and development, especially tumor metastasis. Moreover, ATX has been shown to contribute to drug resistance and angiogenesis. ATX emerges as an attractive target in tumor biology due to its multiple activities in tumor biology.This review summarizes the current progress of ATX in tumors.

Lysophosphatidic acid enhances breast cancer cells-mediated osteoclastogenesis

Lysophosphatidic acid (LPA) is known to play a critical role in breast cancer metastasis to bone. In this study, we tried to investigate any role of LPA in the regulation of osteoclastogenic cytokines from breast cancer cells and the possibility of these secretory factors in affecting osteoclastogenesis. Effect of secreted cytokines on osteoclastogenesis was analyzed by treating conditioned media from LPA-stimulated breast cancer cells to differentiating osteoclasts. Result demonstrated that IL-8 and IL-11 expression were upregulated in LPA-treated MDA-MB-231 cells. IL-8 was induced in both MDA-MB-231 and MDA-MB-468, however, IL-11 was induced only in MDA-MB-231, suggesting differential LPARs participation in the expression of these cytokines. Expression of IL-8 but not IL-11 was suppressed by inhibitors of PI3K, NFkB, ROCK and PKC pathways. In the case of PKC activation, it was observed that PKCδ and PKCμ might regulate LPA-induced expression of IL-11 and IL-8, respectively, by using specific PKC subtype inhibitors. Finally, conditioned Medium from LPA-stimulated breast cancer cells induced osteoclastogenesis. In conclusion, LPA induced the expression of osteolytic cytokines (IL-8 and IL-11) in breast cancer cells by involving different LPA receptors. Enhanced expression of IL-8 by LPA may be via ROCK, PKCu, PI3K, and NFkB signaling pathways, while enhanced expression of IL-11 might involve PKCδ signaling pathway. LPA has the ability to enhance breast cancer cells-mediated osteoclastogenesis by inducing the secretion of cytokines such as IL-8 and IL-11.

Design,synthesis and preliminary anti-radiation activityof novel LPA2 receptor agonists / 国际药学研究杂志

Objective To design and synthesize novel G protein-coupled lysophosphatidic acid 2(LPA2)receptor agonists with anti-radiation activity. Methods Nine new LPA2 receptor agonists(Ⅰ2-Ⅰ5 andⅡ1-Ⅱ5)were designed and synthesized using DBIBB as the lead compound. The anti-radiation activity was assayed by the MTS method using the human umbilical vein endothelial cells(HUVEC)irradiated with 8.0 Gy 60Coγray. Results and Conclusion Nine target compounds notreported in the literature were synthesized and their structures were confirmed by 1H NMR and MS. The results showed thatⅠ4 andⅡ1 have obvious anti-radiation ac-tivity,indicating that this kind of compounds is worth further studying.

Research progress in regulation roles of G protein-coupled receptors in hepatocellular carcinoma / 中国药理学与毒理学杂志

G protein- coupled receptors (GPCRs), also known as seven- transmembrane domain receptors, constitute the largest superfamily of cell surface receptors. By coupling to heterotrimeric G proteins, arrestins and other signaling molecules, GPCRs modulate diverse signal transduction pathways under physiological and pathological conditions. Recent studies have revealed crucial roles of GPCRs in tumorigenesis and development of cancer metastasis. This review summarizes roles of GPCRs, particularly the roles of those coupled to chemokines, prostaglandin, lysophosphatidic acid, endothelin, catecholamine and angiotensin in proliferation, invasion, metastasis and angiogenesis of hepatoma cells and development of hepatocellular carcinoma. The potential of GPCRs- based therapeutics being used for hepatocellular carcinoma is also highlighted.

Effect of hepatitis B virus X protein on autotaxin expression and its significance / 实用医学杂志

Objective To explore the effect of hepatitis B virus (HBV) X protein (HBx) on autotaxin (ATX) expression and its significance. Methods The recombinant eukaryotic expression vector of HBx ,pcD-NA3.1(+)-HBx,and the recombinant luciferase reporter gene vector of ATX promoter,pGL3-ATX,were con-structed and used to co-transfect HepG2 cells to examine the effect of HBx on the activity of ATX promoter. The sta-ble cell expressing HBx,HepG2.HBx,was constructed,and Western blot(WB)was used to detect the effect of HBx on ATX expression. Results The luciferase activity of pcDNA3.1(+)-HBx and pGL3-ATX group was 1.47 times as that of the empty vector cDNA3.1(+)and pGL3-ATX group(P<0.000). WB detection showed that the expression of ATX protein was increased in HepG2.HBx cells,and 1.75 times as that of HepG2 cells(P<0.05). Conclusion HBx can activate ATX promoter and up-regulate ATX expression ,thus suggests that HBV infection might enhance ATX/LPA signaling.

Effects of ovarian induction with Raloxifene versus Clomiphene Citrate on endometrial receptivity in mouse endometrium / 实用医学杂志

Objective To investigate the effects of ovarian induction with raloxifene(RAL)versus clomi-phene citrate(CC)on the endometrial receptivity in mouse endometrium during perimplantation period. Methods 48 female Kun-ming mouse were randomly divided into four groups in equal number:RAL 240 mg group,RAL 180 mg group,CC group,natural conception group(NC),all treated with ovulation induction after drug administration.Successfully mated female mouse were killed,and uterus samples were collected for HE stain-ing and immunohistochemistry. Results HE staining showed that the endometrial morphology in the RAL 180 mg group and RAL 240 mg group and NC group were better than that of CC group.The expressions of COX-2 and LP-AR3 in the RAL 180 mg group and RAL 240 mg group were similar to NC group,without significant difference among the three groups(P > 0.05). But in the CC group,it was statistically significantly lower than other three groups(P<0.05),indicating ovarian induction with RAL did not decrease the expressions of COX-2 and LPAR3 in endometrium. Conclusion The mechanism of ovulation induction with RAL is similar to CC,but RAL has fewer adverse effects on the endometrial receptivity compared with CC.

Role of Rho-kinase in remote ischemicpostconditioning against myocardial ischemia/reperfusion injury / 中国药理学通报

Aim To explore the role of Rho-kinase in remote ischemic postcondi-tioning and its possible mechanism.Methods Thirty male Sprague-Dawley rats were divided into five groups(n=6): sham group(Sham), ischemia/reperfusion group(I/R), remote ischemic postconditioning group(RIPostC), I/R with Rho-kinase inhibitor fasudil group(I/R+Fas) and RIPostC with Rho-kinase activator lysophosphatidic acid group(RIPostC+LPA).Throughout the whole process of experiment, mean arterial pressure(MAP), heart rate(HR) and Ⅱ lead electrocardiogram were continuously monitored.At the end of the reperfusion, plasma creatine kinase(CK) and lactate dehydrogenase(LDH) were measured.Myocardial histopathologic changes were observed by hematoxylin and eosin(HE) staining.Infarct size was measured using 2,3,5-triphenyltetrazolium chloride(TTC) staining.The expressions of phospho-myosin light chain(p-MLC) were detected with Western blot analysis.Results Compared with Sham group, the MAP and HR of other groups decreased, while the amplitude of ST segment increased.Compared with I/R group, MAP and HR increased, the amplitude of ST segment decreased, plasma CK and LDH activity decreased, myocardial pathological morphology and infarct size were improved significantly, infiltration of inflammatory cells was reduced, and the expression of p-MLC decreased in RIPostC and I/R+Fas group.Compared with RIPostC group, RIPostC+LPA group attenuated the effects of RIPostC, and the recovery of the above indicators were inhibited.Conclusion Rho-kinase signaling pathway might mediate remote ischemia postconditioning against myocardial ischemia/reperfusion injury.

Research on LPA and Hippo-YAP signaling pathway in promoting invasion and metastasis of TNBC cell MDA-MB-231 / 医学研究生学报

Objective Triple negative breast cancer(TNBC), a special breast cancer subtype, is lack of effective target therapy.The article aimed to investigate the role of lysophosphatidic acid (LPA) and Hippo Yes-associated protein (Hippo-YAP) signaling pathway in TNBC invasion and metastasis and the mechanisms.Methods The specific small interfering RNA (siRNA) of YAP was synthetized in vitro, and was transfected into MDA-MB-231 cells using liposome transfection.The experiment was divided into YAP-siRNA group, positive control group and blank control group.Each group is provided with 2 parallel holes.Evaluation was made on the effects of each group on Hippo-YAP, the mechanisms and regulation on upstream and downstream molecules of Hippo-YAP pathway.Results In experiment group, YAP content, the capacity of invasion and metastasis after transfection ([0.035±0.005], [2.200±1.000], [3.500±0.800]) significantly decreased compared with positive control group([0.343±0.012], [27.600±5.100], [22.300±5.000]) and blank control group([0.384±0.017], [26.500±4.800], [22.350±6.000]) (P<0.05).YAP expression levels at 60 min, 120 min, and 240 min in experiment group significantly decreased compared with positive control group and blank control group (P<0.05).YAP relative expression levels of 10, 20, 50 μmol/Lwere significantly lower than those of positive control group and blank control group (P<0.05).After respective interference of C3 transferase and Y27623, significant difference was found in the pYAP mRNA contents of experiment group([0.255±0.052], [0.326±0.017]), blank control group([0.048±0.032], [0.534±0.017]) and positive control group([0.052±0.021], [0.528±0.024])(P<0.05).The expression levels of YAP mNA and AREG mNA significantly increased in experiment group([0.176±0.032], [0.263±0.008]) compared with blank control group([0.043±0.013], [0.263±0.008]) and positive control group([0.049±0.025], [0.057±0.043])(P<0.05).Conclusion LPA induces breast cancer invasion and metastasis, which is YAP-dependent, time-dependent and concentration-dependent.LPA-Hippo-YAP singaling pathway may be one of the mechanisms promoting delayed metastasis of TNBC.

Gintonin, an exogenous ginseng-derived LPA receptor ligand, promotes corneal wound healing

Ginseng gintonin is an exogenous ligand of lysophosphatidic acid (LPA) receptors. Accumulating evidence shows LPA helps in rapid recovery of corneal damage. The aim of this study was to evaluate the therapeutic efficacy of gintonin in a rabbit model of corneal damage. We investigated the signal transduction pathway of gintonin in human corneal epithelium (HCE) cells to elucidate the underlying molecular mechanism. We next evaluated the therapeutic effects of gintonin, using a rabbit model of corneal damage, by undertaking histochemical analysis. Treatment of gintonin to HCE cells induced transient increases of [Ca²⁺](i) in concentration-dependent and reversible manners. Gintonin-mediated mobilization of [Ca²⁺](i) was attenuated by LPA1/3 receptor antagonist Ki16425, phospholipase C inhibitor U73122, inositol 1,4,5-triphosphate receptor antagonist 2-APB, and intracellular Ca²⁺ chelator BAPTA-AM. Gintonin facilitated in vitro wound healing in a concentration-dependent manner. When applied as an eye-drop to rabbits with corneal damage, gintonin rapidly promoted recovery. Histochemical analysis showed gintonin decreased corneal apoptosis and increased corneal cell proliferation. We demonstrated that LPA receptor activation by gintonin is linked to in vitro and in vivo therapeutic effects against corneal damage. Gintonin can be applied as a clinical agent for the rapid healing of corneal damage.