Toll樣受體在先天免疫和細(xì)胞損傷中的作用.pdf

1、SECTION-Ⅰ ROLES OF TLR2 IN OPIOIDS MEDIATED APOPTOSIS AND DEPENDENCE Part 1st：Toll-Like Receptor 2 Is Required for Opioids-Induced NeuronAlApoptosis Running title:TLR2-Mediated Apoptosis Promoted by Opioids Toll-like

2、 receptor 2 (TLR2), a key immune receptor in the TLR family, is widely expressed in varioussystems, including the immune and nervous systems and plays a criticAlrole in controlling innate andadaptive immune responses. We

3、 previously reported that opioids inhibit cell growth and trigger apoptosis.However, the underlying mechanism by which TLR2 mediates apoptosis in response to opioids is not yetknown. Here we show that chronic morphine tr

4、eatment in primary neurons dramatically increased theexpression of TLR2 at both the messenger RNA and protein levels. In addition, TLR2 deficiencysignificantly inhibited chronic morphine-induced apoptosis in primary neur

5、ons. Activation of caspase-3after morphine treatment is impaired in TLR2 deficient primary neurons. Moreover, morphine treatmentfailed to induce an increased level of phosphorylated glycogen synthase kinase 3 beta (GSK3β

6、) in TLR2deficient primary neurons, suggesting an involvement of GSK3βin morphine-mediated TLR2 signaling.These results thus demonstrate that opioids prime neurons to undergo apoptosis by inducing TLR2expression. Our dat

7、a suggest that inhibition of TLR2 is capable of preventing opioids-induced damage toneurons. Part 2nd：PivotAlRole of Toll-Like Receptor 2 in Opioid Dependence Running title:TLR2-Mediated Opioid Dependence Opioids are

8、powerful pain relievers, but also potent inducers of dependence and tolerance. Chronicmorphine administration (via subcutaneous pellet) induces morphine dependence in the nucleusaccumbens, a key dependence region in the

9、brain, yet the cellular mechanisms are mostly unknown.Toll-like receptor 2 (TLR2) plays an essentiAlfunction in controlling innate and inflammatory responses.Using a knockout mouse lacking TLR2, we assessed the contribut

10、ion of TLR2 to the development ofmorphine dependence and microglia activation. We report here that mice deficient in TLR2 inhibitmorphine-induced the levels of microglia activation and proinflammatory cytokines. Moreover

11、, in TLR2knockout mice the main symptoms of morphine withdrawAlwere significantly attenuated. Our datademonstrate that TLR2 is criticAlfor opioid dependence and is a factor in response to innate immuneresponse. Part 3rd

12、：ROLES OF TLR2 IN R6A MEDIATED APOPTOSIS Synthetic Resveratrol Aliphatic Acid Inhibits TLR2-Mediated Apoptosis and An Involvement of Akt/GSK3βPathway Running title:Resveratrol Aliphatic Acid Inhibits TLR2-Mediated Apopt

13、osis As resveratrol derivatives, resveratrol aliphatic acids were synthesized in our laboratory. Previously, wereported the improved pharmaceuticAlproperties of the compounds compared to resveratrol, includingbetter sol

14、ubility in water and much tighter binding with human serum albumin. Here, we investigate therole of resveratrol aliphatic acids in Toll-like receptor 2 (TLR2)-mediated apoptosis. We showed thatresveratrol aliphatic acid

15、(R6A) significantly inhibits the expression of TLR2. In addition,overexpression of TLR2 in HEK293 cells caused a significant decrease in apoptosis after R6A treatment.Moreover, inhibition of TLR2 by R6A decreases serum d

16、eprivation-reduced the levels of phosphorylatedAkt and phosphorylated glycogen synthase kinase 3β(GSK3β). Our study thus demonstrates that theresveratrol aliphatic acid inhibits cell apoptosis through TLR2 by the involve

17、ment of Akt/GSK3βpathway. SECTION-Ⅱ ROLE OF B-ARRESTIN 2 IN TLR4 MEDIATED APOPTOSIS AND CHRONIC STRESS Part 1st：β-Arrestin 2 Regulates TLR4-Mediated Apoptotic Signaling through GSK-3β Running title:β-Arrestin 2 Regula

18、tes TLR4-Mediated Apoptosis TLR4 (Toll-like receptor-4), a key member of the TLRs family, has been well characterized by itsfunction in induction of inflammatory products of innate immunity. However, the involvement of

19、TLR4in a variety of apoptotic events with an unknown mechanism recently interests great research focus. Ourinvestigation found that TLR4 promoted apoptotic signaling through affecting glycogen synthasekinase-3β(GSK-3β) p

20、athway in the serum deprivation (SD)-induced apoptotic paradigm. SD inducesGSK-3βactivation in a pathway that leads to subsequent cell apoptosis. Intriguingly, this apoptoticcascade is amplified in presence of TLR4 where

21、as greatly attenuated by β-arrestin 2, another criticalmolecule implicated in TLR4 mediated immune responses. Our data suggest the association of β-arrestin2 with GSK-3βcontributes to the stabilization of phospho-GSK-3β,

22、 an inactive form of GSK-3β. Itbecomes a criticAldeterminant for the attenuation of TLR4-initiated apoptosis by β-arrestin 2. Takentogether, we demonstrate that the TLR4 possesses the capability of accelerating GSK-3βact

23、ivationthereby deteriorating SD-induced apoptosis; β-arrestin 2 represents an inhibitory effect onTLR4-mediated apoptotic cascade, through controlling the homeostasis of activation and inactivation ofGSK-3β. Part 2nd：β-

24、Arrestin 2-Mediated Immune Suppression Induced By Chronic Stress Running title:Stress-Induced Lymphocyte Reduction viaβ-Arrestin 2 Stress, either physicAlor psychological, can modulate immune function. However, the mec

25、hanismsassociated with stress-induced immune suppression remains to be elucidated. β-arrestin 2 serves asadaptors, scaffolds, and/or signAltransducers. The role of β-arrestin 2 in stress-induced immunesuppression is not

26、known yet. Here, we demonstrate that β-arrestin 2 deficiency in mice increases thesensitivity of chronic stress-induced lymphocyte reduction. Interestingly, the stress-induced suppressionof T help 1(Th1) cytokine and inc

27、reased production of Th2 cytokine was greatly increased in β-arrestin2 deficient mice compared with wild type mice. Moreover, inhibition of PI3K in β-arrestin 2-deficientmice exerts an additive effect on stress-induced l

28、ymphocyte reduction. Our study thus demonstrates thatβ-arrestin 2 plays an important role in stress-induced immune suppression. SECTION-Ⅲ CriticAlRole of Toll-like receptor 9 in Morphine and Mycobacterium Tuberculosis-

29、Induced Apoptosis in Mice Running title：TLR9-Mediated Apoptosis in TB Infected Mice Background：Although it is established that opioid and Mycobacterium tuberculosis are both publichealth problems, the mechanisms by whi

30、ch they affect lung functions remain elusive.Methodology/PrincipAlFindings：We report here that mice subjected to chronic morphine administrationand M. tuberculosis infection exhibited significant apoptosis in the lung in

31、 wild type mice as demonstratedby the terminAldeoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick end labeling assay.Morphine and M. tuberculosis significantly induced the expression of Toll-like recepto

32、r 9 (TLR9), a keymediator of innate immunity and inflammation. Interestingly, deficiency in TLR9 significantly inhibitedthe morphine and M. tuberculosis induced apoptosis in the lung. In addition, chronic morphine treatm

33、entand M. tuberculosis infection enhanced the levels of cytokines (TNF-， IL-1β, and IL-6) in wild type mice,but not in TLR9 knockout (KO) mice. The bacteriAlload was much lower in TLR9 KO mice comparedwith that in wild t

34、ype mice following morphine and M. tuberculosis treatment. Morphine alone did notalter the bacteriAlload in either wild type or TLR9 KO mice. Moreover, administration of morphine and M.tuberculosis decreased the levels o

35、f phosphorylation of Akt and GSK3βin the wild type mice, but not inTLR9 KO mice, suggesting an involvement of Akt/GSK3βin morphine and M. tuberculosis-mediatedTLR9 signaling. Furthermore, administration of morphine and M

36、. tuberculosis caused a dramatic decreasein Bcl-2 level but increase in Bax level in wild type mice, but not in TLR9 KO mice, indicating a role ofBcl-2 family in TLR9-mediated apoptosis in the lung following morphine and

37、 M. tuberculosisadministration. Conclusions/Significance：These data reveAla role for TLR9 in the immune response toopioids during M. tuberculosis infection. SECTION-Ⅳ Glycogen Synthase Kinase-3 and p38 MAPK are Require

38、d for Opioid-Induced Microglia Apoptosis Running title：Morphine Mediates Microglia Apoptosis via GSK3βand p38 Opioids have been widely applied in clinics as one of the most potent pain relievers for centuries, but thei

39、rabuse has deleterious physiologicAleffects beyond addiction. We previously reported that opioids inhibitcell growth and trigger apoptosis in lymphocytes. However, the underlying mechanism by whichmicroglia apoptosis in

40、response to opioids is not yet known. In this study, we show that morphine inducesmicroglia apoptosis and caspase-3 activation in an opioid-receptor dependent manner. Morphinedecreased the levels of microglia phosphoryla

41、ted Akt (p-Akt) and p-GSK-3β(glycogen synthase kinase 3beta) in an opioid receptor dependent manner. More interestingly, GSK-3βinhibitor SB216763significantly increased morphine-induced apoptosis in both BV-2 microglia a

42、nd mouse primarymicrogliAlcells. Moreover, co-treatment of microglia with SB216763 and morphine led to a significantsynergistic effect on the level of phospho-p38 mitogen-activated protein kinase (MAPK). In addition,inhi

43、bition of p38 MAPK by its specific inhibitor SB203580significantly inhibited morphine-inducedapoptosis and caspase-3 activation. Taken together, our data clearly demonstrates that morphine inducesapoptosis in microgliAlc