## 4 Discussion  
The pathogenesis of chronic inflammatory diseases such as type 2 diabetes, cancer, rheumatoid arthritis, neurological, and cardiovascular and inflammatory bowel diseases are linked with inflammation and oxidative stress [24]. Interaction of multiple phytochemicals in medicinal plants and herbs provide synergistic effects on anti-inflammatory and antioxidant activity [25]. A large number of studies have determined the pharmacological activities of SGE and its chemical constituents, such as anti-viral, anti-tumor, anti-bacterial, antioxidant, anti-inflammatory, hepatoprotective, and neuroprotective activities [26]. Processing is a traditional herbal method to achieve clinical benefits by improving therapeutic efficacy and also by lowering toxicity [27]. The alteration in therapeutic and toxic effects induced by processing is suggested to be related with structural transformation such as hydrolysis, oxidation, decomposition, isomerization, and other chemical reactions [28]. Comparative analysis of the major flavonoids, baicalin, baicalin, and wogon contained in  
Figure 4: Effect of PSGE against LPS-induced intracellular ROS levels, KEAP1/NRF2 signaling antioxidant gene expression in RAW 264.7 cells. (**A**) Intracellular ROS levels were assayed using oxidation of H2 DCFDA fluorescence probe. RAW 264.7 cells were pre-treated with indicated concentration of PSGE or DXM for 2 h followed by LPS for 24 h.(**B**) Protein expression and quantitative analysis of (**C**) KEAP1 relative to \(\beta\)-actin, (**D**) Western blot analysis of nuclear and cytosolic fractions using NRF2 antibody, quantitative analysis of (**E**) cytosolic NRF2 relative to \(\beta\)-actin, (**F**) nuclear NRF2 relative to Lamin A/C. RAW 264.7 cells were pre-treated with indicated concentration of the PSGE or DXM for 2 h followed by LPS for 30 min. The levels of (**G**) _Hmox1_, (**H**) _Nq01_ and (**I**) _Txn1_ mRNA expression were analyzed by RT-qPCR. RAW 264.7 cells were pre-treated with indicated concentration of the PSGE or DXM for 2 h followed by LPS for 2 h. (**J**) Changes in ROS levels by SnPPIX. RAW 264.7 cells were pre-treated with indicated concentration of SnPPIX (HO-1 inhibitor) for 1 h and then DXM or PSGE for 2 h followed by LPS for 24 h. Lamin A/C and HSP90 were used as cytosolic and nuclear markers. DXM as positive control. Data shown represent mean \(\pm\) SEM (\(n\) = 3). (# \(p\) \(<\) 0.01 and ### \(p\) \(<\) 0.001 vs. control); * _p_\(<\) 0.05, **_p_\(<\) 0.01, and ***_p_\(<\) 0.001 vs. LPS). SGE and PSGE suggested that bacialin content was higher in SGE than PSGE. Interestingly, bacialein and wogonin contents were higher in PSGE than SGE. A report on the change in chemical components of SG after processing showed that stir-frying with alcohol and stir-frying till charred decreased bacialin but increased bacialein and wogonin contents [29]. Earlier studies suggested that the high temperature inhibited glucuronidase activity and prevented hydrolysis of bacialin or wogonin in SG [30,31]. However, understanding the diverse factors influencing glucuronidase activity remains unclear, since it has been reported that bacialin, bacialein, and wogonin contents are increased by heat processing [21,32]. NO is considered as an inflammatory mediator and regulator. In response to inflammatory stimuli, iNOS synthesize NO, inducing proinflammatory and destructive effects that lead to pathological conditions [33]. PSGE pretreatment suppressed NO increase after LPS treatment, which was considerably more potent than DXM treatment (Figure 2B). PGE\({}_{2}\) is recognized as a bioactive lipid that mediates proinflammatory and immunomodulatory effects. In response to inflammatory stimuli, COX-2 and mPGES-1 mediate concomitant generation of PGE\({}_{2}\). The elevated PGE\({}_{2}\) levels can also be inactivated by the catabolic enzyme, 15-PGDH [34]. Our data showed that PSGE strongly suppressed PGE\({}_{2}\) production to a greater extent than DXM, while COX-2 and mPGES-1 expression remained unchanged (Figure 2C,D,F,G).