🧫 Experiment Protocol
ExploratoryMycoplasma pneumoniae pneumoniaGPR109A, MAPK1, MAPK14, MAPK8, MAPK9GPR109A knockout mouse macrophagesproposed
GPR109A knockout mouse macrophages were used to confirm that the G-protein coupled receptor GPR109A mediates butyric acid's action in QTJD's therapeutic mechanism. This experiment was designed to validate the butyrate-GPR109A-MAPK pathway hypothesis. In GPR109A-/- macrophages, QTJD treatment still suppressed p38 and JNK1/2 MAPK proteins but showed no effect on ERK1/2, providing evidence that ERK1/2 inhibition by QTJD specifically requires GPR109A receptor signaling, while p38 and JNK1/2 inhibition may involve additional GPR109A-independent mechanisms. This experiment confirmed the involvement of the butyrate-GPR109A-MAPK pathway in QTJD's anti-inflammatory effects.
PRIMARY OUTCOME
Differential MAPK pathway responses in GPR109A-/- cells
EXPECTED OUTCOMES
GPR109A-dependent effects on ERK1/2 but not p38/JNK1/2
SUCCESS CRITERIA
Selective loss of ERK1/2 inhibition in GPR109A-/- cells
PROTOCOL
Western blotting analysis of MAPK proteins in GPR109A knockout macrophages treated with QTJD