BACKGROUND: Retinal ganglion cell (RGC) death caused by acute ocular hypertension is an important characteristic of acute glaucoma. Receptor-interacting protein kinase 3 (RIPK3) that mediates necroptosis is a potential therapeutic target for RGC death. However, the current understanding of the targeting agents and mechanisms of RIPK3 in the treatment of glaucoma remains limited. Notably, artificial intelligence (AI) technologies have significantly advanced drug discovery. This study aimed to discover RIPK3 inhibitor with AI assistance. METHODS: An acute ocular hypertension model was used to simulate pathological ocular hypertension in vivo . We employed a series of AI methods, including large language and graph neural network models, to identify the target compounds of RIPK3. Subsequently, these target candidates were validated using molecular simulations (molecular docking, absorption, distribution, metabolism, excretion, and toxicity [ADMET] prediction, and molecular dynamics simulations) and biological experiments (Western blotting and fluorescence staining) in vitro and in vivo . RESULTS: AI-driven drug screening techniques have the potential to greatly accelerate drug development. A compound called HG9-91-01, identified using AI methods, exerted neuroprotective effects in acute glaucoma. Our research indicates that all five candidates recommended by AI were able to protect the morphological integrity of RGC cells when exposed to hypoxia and glucose deficiency, and HG9-91-01 showed a higher cell survival rate compared to the other candidates. Furthermore, HG9-91-01 was found to protect the retinal structure and reduce the loss of retinal layers in an acute glaucoma model. It was also observed that the neuroprotective effects of HG9-91-01 were highly correlated with the inhibition of PANoptosis (apoptosis, pyroptosis, and necroptosis). Finally, we found that HG9-91-01 can regulate key proteins related to PANoptosis, indicating that this compound exerts neuroprotective effects in the retina by inhibiting the expression of proteins related to apoptosis, pyroptosis, and necroptosis. CONCLUSION AI-enabled drug discovery revealed that HG9-91-01 could serve as a potential treatment for acute glaucoma.
Animals ; Glaucoma/metabolism* ; Neuroprotective Agents/pharmacology* ; Mice ; Receptor-Interacting Protein Serine-Threonine Kinases/metabolism* ; Artificial Intelligence ; Retinal Ganglion Cells/metabolism* ; Disease Models, Animal ; Molecular Docking Simulation ; Mice, Inbred C57BL ; Male

Aim Tostudytheanalgesiceffectofoxyso-phoridine (OSR)on GABA transporter-1 (GAT-1 )mR-NA expression and its influence on GAT-1 expression inmice.Methods Formalintestwasusedtodetectthe analgesic effect of OSR(iv).Immunohistochemis-try was taken to inspect the expression of GAT-1 in cerebral cortex and thalamus in mouse brain. The quantitative real-time PCR method was used to inspect the influence of OSR on GAT-1 mRNA expression of braininmice.Results OSR(500,250,125mg· kg-1 ,iv ) could significantly increase the foot-licking latency.OSR(500 mg·kg-1,ip)could significantly decrease the number of GAT-1 immuopositive cells incerebral cortex and thalamus in mouse brain,and re-duce GAT-1 mRNA expression in brain(P<0. 01,P<0.05)intheformalintest.Conclusion OSRhasa significant analgesic effect,and its analgesic mecha-nism is related to the GAT-1 expression in mouse brain.

OBJECTIVETo study the effect of GABA transporter (GAT-1) on the analgesic action of oxysophoridine (OSR) in the central nervous system of mice.

METHODHot plate test was used to observe and analyze the effect of gamma aminobutyric acid and the inhibitor of GAT-1 (NO-711) on the analgesic action of oxysophoridine. Real time RT-PCR was used to investigate the influence of OSR on the expression of GAT-1 mRNA induced by formalin in spinal cord and brain of mice.

RESULTBoth GABA (2.0 mg x kg(-1), icv) and NO-711(0.125 mg x kg(-1), icv) enhanced the analgesic action of OSR (32.0 mg x kg(-1), iv) in the hot plate test, and the latencies was markedly increased (P < 0.05, P < 0.01). OSR (500.0 mg x kg(-1), iv) significantly inhibited the expression of GAT-1 mRNA induced by formalin (P < 0.05).

CONCLUSIONGAT-1 was involved in the analgesia effect of OSR and the down-regulation of GAT-1 mRNA enhanced the analgesic effect.

Alkaloids ; pharmacology ; Analgesics ; pharmacology ; Animals ; Brain ; drug effects ; metabolism ; Down-Regulation ; drug effects ; Female ; GABA Plasma Membrane Transport Proteins ; genetics ; metabolism ; Gene Expression Regulation ; drug effects ; Male ; Mice ; RNA, Messenger ; analysis ; Spinal Cord ; drug effects ; metabolism

.This study is to investigate the analgesic effect produced by intrathecal injection (ith) of oxysophoridine (OSR) and the mechanism of GABAA receptor. Warm water tail-flick test was used to detect the analgesic effect of OSR (12.5, 6.25, and 3.13 mg.kg-1 ith) and to observe the influence of GABA (gamma aminobutyric acid) agonist or antagonist on the analgesic effect of OSR in mice. Immunohistochemistry method were used to detect the influence of OSR (12.5 mg.kg-1, ith) on the GABAARalpha1 protein expression in spinal cord. The results obtained covers that OSR (12.5 and 6.25 mg.kg-, ith) alleviates pain significantly with the warm water tail-flick test (P<0.05, P<0.01), the rate of pain threshold increases by 68.45%; GABA and muscimol (MUS) produces analgesic synergism together with the OSR, picrotoxin (PTX) and bicuculline (BIC) antagonize the analgesic effect of OSR; OSR (12.5 mg.kg-1, ith) significantly increase the positive number of GABAARalpha1 nerve cell in spinal cord (P<0.01) and significantly decrease the average grey levels (P<0.01). In conclusion, OSR intrathecal injection has significant analgesic effect. And GABAA receptor in spinal cord is involved in the analgesic mechanism.

Aim To study the analgesic action of oxysophoridine and its effect on the expression of protein kinase C?(PKC?) in dorsal horn of spinal cord(it should be expressed as in dorsal horn of the spinal),cerebral cortex and thalamus of the mice.Methods Hot plate test was used to observe and analyze the analgesic strength and action position of OSR through iv and icv approaches,immunohistochemistry(SABC) was taken to inspect the expression of PKC? in dorsal horn of spinal cord(it should be expressed as in dorsal horn of the spinal),cerebral cortex and thalamus of the mice after administrating OSR.Results The foot-licking latencies of mice were prolonged both iv OSR(500、250、125 mg?kg-1)and icv OSR(100,50,25 mg?kg-1)in the hot plate test(P