## 3 Results and Discussion
### Anti-Inflammatory Potential
#### 3.4.2 Immunohistochemistryistochemical Analysis  
As one of the eleven members of the cytokine family, IL1 is mainly produced by the majority of immune cells in reaction to other cytokines or microbial stimuli [65]. Since IL-1\(\upbeta\) plays a part in both autoimmune and inflammatory diseases, it is crucial to inhibit its activity in order to lower the inflammatory response. In necrotizing enterocolitis and inflammatory bowel disease, IL-1\(\upbeta\) and other proinflammatory cytokines are increased, causing intestinal inflammation via breaking down the intestinal barrier [66]. This causes inflammation in the intestine. Furthermore, through regulating neuro-immune interactions across neuronal ganglia in the central nervous system, spinal cord, and digestive tract, IL-1\(\upbeta\) is essential for the development and maintenance of neuropathic pain in a range of chronic illnesses [67]. Inflammation is one of the adaptive responses to a variety of traumas, including those caused by physical, chemical, and biological agents [68]. IL-1\(\upbeta\) can cure acute inflammation and activate adaptive anti-tumor responses. Chronic inflammation, however, increases the risk of cancer [69]. Along with phenotypic changes in B and T cells, the production of cytokines through processes mediated by lipotoxicity and macrophage infiltration into adipose tissue, chronic and low-grade inflammation is the foundation of metabolic disorders [70; 71]. Pro-inflammatory cytokines such as TNF-a, IL-8, and IL-2 are signs of intestinal dysbiosis. TH1 cells and macrophages produce the pro-inflammatory cytokines IFN-, IL-2, IL-3, and TNF, which can be blocked by the anti-inflammatory cytokine IL-10 [72]. On the other hand, cytokine IL-1\(\upbeta\) directly affects the SMs and neurons in the stomach [73]. The innate immune system produces the potent pro-inflammatory cytokine IL-1\(\upbeta\). Unlike the usual method, it secretes proteins through one or more unconventional secretion pathways and is manufactured without the need for a signal sequence.  
The immunohistochemical expression of IL-1\(\upbeta\) in smooth muscle preparations (SMPs) incubated with SQ (**5**) was utilized in a semiquantitative manner (Figure 6, Table 2).  
\begin{table}
\begin{tabular}{c c c} \hline \hline  & **Control** & **SQ’s Response** \\ \hline IL-1\(\upbeta\) & ++ & \(+/-\) \\ SM & ++ & \(+/-\) \\ MP & ++ & \(-\) \\ \hline \hline \end{tabular} SM and neurons in the MP were shown to express IL-1\(\upbeta\) on a panel. By measuring the expression of IL-1\(\upbeta\) in tissue slices using immunohistochemical staining, the semiquantitative method for reporting its expression was established. \(+\), moderate; \(+\), low; \(-\), absent.  
\end{table}
Table 2: Expression of IL-1\(\upbeta\) and SQ’s response.  
Any histological changes between the SQ-incubated preparations stained with H-E and the control were noted (Figure 6B). Weak IL-1\(\beta\) expression was observed in the MP in control preparations (Figure 6C). The preparations incubated with SQ (**5**) in the MP showed no expression of IL-1\(\beta\), with only a tiny percentage of SM cells labeled with IL-1\(\beta\) (Figure 6D).  
We found that both SM and MP neurons showed the anti-inflammatory activity of SQ, which we observed as reduced or absent IL-1\(\beta\) expression in preparations. This suggests that the SQ might be able to lessen inflammatory processes in many somatic constituents of cells. For this reason, SQ may be able to reduce gastrointestinal tract inflammation [29]. Therefore, SQ (**5**) will be established as a potential medication to treat inflammatory illnesses through future preclinical experiments.