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Cellular signal transduction includes many cell signal
pathways, an area having received considerable research interest. The
cell endoplasmic reticulum ER Ca2+ pool attempts to maintain an
ion concentration balance of cellular calcium, which is an extremely
important organelle. Among the calcium, an ion is cellular ER to
proceed with the protein translation, protein translocation, folding and
cellular ER translation protein to confirm ER Ca2+ pool plays a
significant role. However, these functions should mainly cause the
implementation of many resident ER Ca2+binding proteins.The
existence of many Ca2+-binding proteins in the ER is widely
documented. Additionally, although passive in producing ER inside the
high concentration, these proteins have an important physiological
function. Additionally, the protein kinase C accepts some physiological
functions after stimulation. Protein kinase C belongs to serine-
threonine kinase about the message transduction The cell-related
hormone or growth factor is used to proceed to the next step for
activation of phospholipase C, thus producing DAG. Finally, protein
kinase C is activated.
However, in addition to having many isoforms, PKC can
differentiate between conventional PKCs (a, b1, b2, and g), novel PKCs
(d, e,and), atypical PKCs (and/) . Previous research focused on
ER and how to regulate the calcium concentration of the intracellular
pathway. . Therefore, these PKC isoforms activate to some degree
correlation. However, PKC cannot clearly identify the different types of
cell lines that inhibit ER Ca2+ sequestering activity. PKC has also not
been investigated with respect to the intracellular Ca2+ pool.
Still, ER Ca2+ pool has not been investigated with
respect to PKC isoforms in cell signal transduction, the functions of
PKC and DNA transcription or translation as well as various
intracellular pathways. Moreover, conventional cell culture methods
can not thoroughly understand the cellular pathway of PKC, making
the cellular apoptosis mechanism unclear as well.
The inability to thoroughly understand protein kinase
C and the cell signal pathway will negatively impact the physiological
characteristics of PKC with respect to cellular life and death.
Based on the above, we should attempt to
understand what role macrophage secretion cytokine plays
in the human immune system.
To do so, NO can be examined, as found
between different macrophage cells, RAW 264.7 and J774.
An identical stimulator and inhibitor can then be inserted
into RAW 264.7 and J774 murine macrophage cells,
allowing RAW264.7 and J774 cells not only to produce the
cytokine but also to induce apoptosis physiology.
Moreover, NO content and cell apoptosis can be examined.
Furthermore, RAW 264.7 and J774 cells can be treated
with the LPS in different glucose concentration media.
As anticipated, analysis results can indicate
that NO and other cytokines can locate many signal
pathways of the macrophage. Additionally, the NO content
and protein kinase C of cellular signal regulation can
identify the RAW 264.7 or J774 cell morphology in different
glucose concentration media.
Results in this study can demonstrate that, in
addition to possibly inducing the cell apoptosis pathway,
NO can promote the human immune system to achieve an
appropriate balance between cytokines.
Commonly found in the RAW 264.7 cell,
endoplasmic reticulum (ER) calcium pool plays a
significant role in regulating the concentration of cellular
calcium ion. Additionally, , ER calcium pool can facilitate
protein translation, protein transfer, and protein
embellishment. According to recent investigations,
elevated intracellular Ca2+ concentration, [Ca2+]i, can
initiate apoptosis; in addition, [Ca2+]i increases before
genome fragmentation and cell death. As well known, as a
major intracellular reservoir of Ca2+ in nonmuscular cells,
endoplasmic reticulum ER is essential for many cellular
functions, including protein processing within ER.
However, while previous studies investigated how murine
macrophage cell line regulates the signal pathway of ER calcium pool,
exactly how the signal pathways of cellular TNF-, NF-B, and MAPK
regulates the concentration of cellular calcium ion remains unclear.
For instance, cell ER pool investigations have not
identified the signal pathways of TNF-, NF-B, and MAPK within an
accuracy of 80, thus making it impossible to determine how RAW
264.7 cell regulates the signal pathway.
The inability to thoroughly understand the signal
pathways of intracellular TNF-, NF-B, and MAPK makes it
impossible to determine what role ER calicium pool and induced
cytokine play in the RAW 264.7 cell.
Based on the above, we should analyze TNF- and NF-B of
the RAW 264.7 cell by using cell culture methods, providing insight into
how the cell signal pathway and immunity regulation of nitro oxide in
cellular endoplasmic reticulum (ER) calcium poolare related. TNF-
,NF-B and MAPK can also be determined to be closely associated
with the signal pathways of both human diseases such as cancer.
To do so, the RAW 264.7 cell can be analyzed using
Griess reagent (1% Sulfanilamide, 0.1% NED) and chemiluminescence.
The PKC protein can then be analyzed using western blot analysis.
Next, NF-B and MAPK can be analyzed using primary and secondary
antibodies. Additionally, the mouse fibroblasts can be used by adding
L929 to serial dilutions of the conditioned media at 5104 cells per well
(in 96-well plates), followed by treatment with 1 g/ml actinomycin D.
Moreover, after 24 h of treatment, the viability of cells can be measured
by MTT assay. Finally, a standard curve can be defined using TNF-.
As anticipated, analysis results can indicate
that the TNF-,NF-B and MAPK can be found in the RAW
264.7 cell of the cellular endoplasmic reticulum (ER)
calcium pool signal pathway. Additionally, this pathway can
be understood with respect to elucidating the
characteristics of cancer.
Results of this study can provide further insight
into not only the signal pathways of intracellular TNF-, NF-
B, and MAPK, but also the role in which ER calicium pool
and induced cytokine play in the RAW 264.7 cell.
Further details can be found at