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\def\RunningAuthor{Ravi Teja P D and Kottai Muthu }
\firstPage{294}
\articleType{original Article}
\receivedDate{25.07.2019}
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\journalVolume{2020, 11}
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\title{Evaluation of the hepatoprotective activity of various extracts of \textit{Dyschoriste littoralis} nees\textit{\space }on paracetamol-induced hepatotoxicity rats}
\author{Ravi~Teja~P~D\textsuperscript{1,2},  
            Balakrishnan~K\textsuperscript{2}, 
            Kottai~Muthu~A\textsuperscript{*}\textsuperscript{1}~\\[5pt]{\textsuperscript{1}Department of Pharmacy, Annamalai University, Annamalainagar-608 002, Tamilnadu, India}~\\{\textsuperscript{2}Seshachala College of Pharmacy, Puttur, Chittoor, Andra Pradesh, India}}

\begin{abstract}
The current investigation was to examine the hepatoprotective potential of \textit{D.littoralis} Nees. (family Acanthaceae)  On paracetamol-induced hepatotoxicity in rats. The aerial parts of \textit{D.littoralis} Nees. The powder was concentrated with various solvents (PE, EA, and methanol) through Soxhlet concentrates and different crude concentrates utilized for hepatoprotective activity.  Hepatotoxicity was induced by paracetamol (2g/kg b.wt.) on the 5\ensuremath{^{th}} day of the investigational period and given orally. Paracetamol-induced rats to exhibit elevated activities of liver enzymes such as SGOT, SGPT, ALP, gamma-glutamyltranspeptidase (GGT), creatinine, urea, total bilirubin, total cholesterol \& triglyc in serum. Furthermore, Oral administration of the ethyl acetate concentrates of \textit{D.littoralis} \textit{\space }(200 mg/ kgb.wt.) given rats were significant reduction the level of SGOT, SGPT, ALP, gamma-glutamyltranspeptidase (GGT) creatinine, urea, total bilirubin, total cholesterol \& triglycerides when compared to other concentrates.  They also significantly elevated the concentration of complete protein and albumin when compared to other concentrates. Thus, results suggested that ethyl acetate concentrates of \textit{D.littoralis } could afford better hepatoprotective activity against paracetamol-induced hepatotoxicity rats.
\end{abstract}\def\keywordstitle{Keywords}

\begin{keywords}D.littoralis,\newline Paracetamol,\newline hepatic markers,\newline acute toxicity,\newline hepatotoxicity
\end{keywords}
\twocolumn[ \maketitle {\printKwdAbsBox}]
\makeatletter{\raggedright\textsuperscript{*}Corresponding Author\par Name:\ Kottai~Muthu~A~\\ Phone:\ +919443171712~\\  Email:\ akottaimuthu@gmail.com
        \par}\vspace*{-11pt}\hrulefill\par{\fontsize{12}{14}\selectfont ISSN: 0975-7538}\par% 
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\section{Introduction}
Synthetic drug-induced hepatic toxicity is a substantial clinical issue in the worldwide. Most of the drug-induced hepatic toxicity and acute liver failures occur due to either unplanned high doses of paracetamol\unskip~\citep{634166:14854697}. Paracetamol acts as an antipyretic and analgesic agent used in the clinical field. During beneficial treatments,paracetamol is metabolized by glucuronidation or sulfation by the cytochrome p450 structure into the reactive metabolite NAPQI. Under normal conditions, NAPQI is rapidly changed to harmless GSH. While a higher amount of paracetamol, elevated concentration of NAPQ1 reacts with hepatic proteins and leads to liver injury \unskip~\citep{634166:14854711,634166:14854701}. Paracetamol-induced liver damage can be considered in animal models, and most mechanisms are translatable to humans \unskip~\citep{634166:14854711}. Paracetamol-induced liver toxicity has been a most important problem for many years, and various strategies have been investigated, together with the utilizing the natural bioactive compounds with hepatoprotective activity \unskip~\citep{634166:14854691,634166:14854689}.

Dyschoriste littoralis Nees. (family Acanthaceae) Leaves were used for the treatment of wounds \textit{Dyschoriste} genus were generally known as snake herb. \textit{D.littoralis } used as a treatment of pain, fever, and inflammation. \textit{D.littoralis}is was used for the treatment of severe coughs along with administrated ginger. \textit{D.littoralis}is leaves were used for asthma in the form of cigarettes\unskip~\citep{634166:14854688} and D.littoralisis has used the treatment of diarrhea and dysentery \unskip~\citep{634166:14854695}. \textit{D.littoralis}is have various activities like anti-microbial \unskip~\citep{634166:14854695,634166:14854696} and wound healing.\unskip~\citep{634166:14854710}. Still, no literature available on the hepatoprotective and antioxidant activity of \textit{D.littoralis}. Thus, the study to assess hepatoprotective activities of \textit{D.littoralis }in paracetamol induced rats. 
    
\section{Materials and Methods}
\textbf{Chemicals}

Paracetamol was purchased from Sigma-Aldrich, the USA utilized in the experiment. All other chemicals and reagents were used AR grade

\textbf{ Gathering and Identification of Plant materials }

The aerial parts of \textit{D.littoralis } Nees. (Acanthaceae) Were gathered from Tirunelveli District, Tamil Nadu, India. Plant identification was made from Survey of Medicinal Plant Unit (SMP), Govt. Siddha Medical College, Palayamkottai, Tirunelveli, Tamil Nadu (Voucher No: 25834).The \textit{D.littoralis }were desiccated under shadowy, segregate, crushed through the grinder\unskip~\citep{634166:14854709}.

\textbf{Preparation of Plant Concentrates }

The pulverized materials were progressively concentrated with PE (40-60\ensuremath{^{0}}C) through hot constant percolation  method in Soxhlet equipment \unskip~\citep{634166:14854694} for twenty-four hours. At that moment, the marc was used to EA (76-780C) for twenty-four hours \& then mark was subjected to methanol for twenty-four hours. The concentrates were concentrated through the rotational evaporator and subjected to solidify drying in a lyophilizer till dry powder was  acquired \unskip~\citep{634166:14948963}.

\textbf{Animals}

Male Wister rats of 17-18 weeks age, weighing 160-185g, were collected from the Central Animal House, MNR College of Pharmacy, Sangareddy, Hyderabad, Telangana, India. The animals were set aside in cages, 2 per cage, with twelve: twelve hours light and dark cycle at 25 \ensuremath{\pm} 2 \text{\textordmasculine}C. The rats were maintained on their particular diets and water ad libitum. The Ethical Committee approved the animal Ethical Committee's clearance of MNR College of Pharmacy, Sangareddy, Hyderabad, Telangana (CPCSEA/COP/05/ 21-01-2019).\textbf{\space }

\textbf{Experimental design}

\textbf{Acute toxicity test}

Albino Wistar rats were separated into six groups, and each group contains six animals (n = 6). Rats fasted for four hours with free access to water only. The various concentrates of \textit{D.littoralis } suspended in normal saline: 0.5\% CMC was administered orally at a dose of 5 mg/kg at first, and mortality was noted for three days. The mortality was observed in 5/6 or 6/6 animals, and then the dose administered was measured as a toxic dose. However, mortality was noted in less than 4 rats. Out of 6 rats, then the same treatment was repeated to confirm the toxic effect. If mortality was not observed, the procedure was repeated for higher doses i.e., 2000mg/kg.

\textbf{ Hepatoprotective activity}

Animals were separated into six groups, and each group contains six animals. 

\textbf{Group I}

Animals served as Control group received with vehicle (0.5\% CMC) for 7 days.

\textbf{Group II}

Animals served as a negative control, received 7 days only 1ml vehicle and paracetamol 2g/kg b.wt. given on the 5\ensuremath{^{th}}-day by orally.

\textbf{Group III}

Animals received Pet. Ether concentrates on \textit{D.littoralis } 200mg/kg b.wt, orally for seven days. On 5\ensuremath{^{th}} day onwards paracetamol 2g/kg b.wt. Administered by orally. 

\textbf{Group IV}

Animals received ethyl acetate concentrates of \textit{D.littoralis } 200mg/kg b.wt, orally for seven days. On 5\ensuremath{^{th}} day onwards paracetamol 2g/kg b.wt. administered by orally. 

\textbf{Group V}

Animals received methanolic concentrates of \textit{D.littoralis } 200mg/kg b.wt, orally for seven days.On 5\ensuremath{^{th}} day onwards paracetamol 2g/kg b.wt. administered by orally. 

\textbf{Group VI}

Animals received 25mg/kg b.wt of Silymarin by oral for seven days and on 5\ensuremath{^{th}} day onwards paracetamol 2g/kg b.wt. administered by orally. 

Groups III, IV, and V rats were orally fed with the various concentrates of \textit{D. littoralis}( PE, EA, and methanol), and Group VI rats were fed with silymarin. Both the \textit{D. littoralis } concentrates and silymarin were suspended in 0.5\% CMC individually and fed to the particular rats through oral intubation. Next day of experiments 8\ensuremath{^{th}} day, all the animals were sacrificed through cervical decapitation \unskip~\citep{634166:14949094}. Blood was collected in test tubes in dry condition and allowed to coagulate at ambient temperature for 30 min. The serum was removed through centrifugation at 2000 rpm for 10 min. The removed serum was utilized for the examination of liver enzymes. 

\textbf{ Liver marker enzymes }

\unskip~\citep{634166:14854706} method was used to determine the SGOT and SGPT and \unskip~\citep{634166:14854700} method were to determine ALP.\unskip~\citep{634166:14854702} method was to determine the total protein (TP) levels, and\unskip~\citep{634166:14854704} method were used to determination of Urea. The\unskip~\citep{634166:14854698} method was used to determination of creatinine and \unskip~\citep{634166:14854696} method was utilized for the estimation of TC.\unskip~\citep{634166:14854703} method was used for the determination of total bilirubin (TB), and \unskip~\citep{634166:14854692}method was used for determination of TG. 

\textbf{Statistical Analysis }

ANOVA conducted the statistical investigation, and groups were compared through Duncan's Multiple Range Test (DMRT)using SPSS Software Package, version 10.0. Results were expressed as means \ensuremath{\pm}standard deviation for six rats in each group. A value of P \ensuremath{\leq } 0.05 was considered to be statistically significant.
    
\section{Results and Discussion}
\textbf{Acute Toxicity}

The results of the critical toxicity study revealed that LD50 values of PE, EA, and methanolic concentrates of \textit{D.littoralis were} better and actually showed the protection of concentrates. Administration of PE, EA, and methanolic foci of \textit{D. littoral} did not change any autonomic or behavioral response in rats. The 0\% mortality for a various concentrates of \textit{D.littoralis } was recorded at 2000mg/kg. Overall results revealed that the LD50 value of 2000mg/kg. Hence the therapeutic dose was considered as 1/10\ensuremath{^{th}} (200mg/kgb.wt.) of the lethal dose for hepatoprotective activity.

\textbf{The activity of various concentrates of \textit{D.littoralis }} \textbf{on average liver weight changes in rats}

Table~\ref{tw-a0771a63ee4b} appeared the activity of various foci of aerial parts of \textit{D.littoralis } on average liver weight changes in normal and paracetamol-induced hepatotoxic rats. The hepatotoxic control group showed increased liver weight. The administration of ethyl acetate concentrates of aerial parts of \textit{D.littoralis } attenuated the liver weight reduced, and silymarin treated group restored liver weight.

\textbf{\textbf{\textbf{\textbf{Effect of various concentrates of \textit{D.littoralis } on liver enzymes and other parameters in paracetamol-induced hepatotoxicity in Wistar rats}}}}

Table~\ref{tw-87a207f48d8e} shows the effect of various concentrates of aerial parts of \textit{D.littoralis } on hepatic marker enzymes in the serum from normal and paracetamol-induced hepatotoxic rats. The hepatotoxic control group showed increased activities of SGPT(176.34\ensuremath{\pm}8.43), SGOT(190.45\ensuremath{\pm}7.65), ALP(278.23\ensuremath{\pm}10.22) and GGT(1.96\ensuremath{\pm}0.14). Administration of PE concentrates of \textit{D.littoralis } showed considerably SGPT(172.12\ensuremath{\pm}6.52), SGOT(185.48\ensuremath{\pm}4.34), ALP(273.65\ensuremath{\pm}6.87)and GGT(1.89\ensuremath{\pm}0.12) in group III rats. Administration of EA concentrates of \textit{D.littoralis } showed considerably SGPT(89.26\ensuremath{\pm}4.76), SGOT(115.63\ensuremath{\pm}4.58), ALP(208.34\ensuremath{\pm}6.21) and GGT(1.48\ensuremath{\pm}0.05)in group IV rats. Administration of methanol concentrates of \textit{D.littoralis } showed considerably SGPT(136.12\ensuremath{\pm}9.65), SGOT(169.65\ensuremath{\pm}4.23), ALP(248.24\ensuremath{\pm}8.38) and GGT(1.64\ensuremath{\pm}0.12) in group V rats. The administration of EA concentrates on \textit{D.littoralis } attenuated the hepatic marker enzymes, and silymarin restored enzyme activities to normal values.

\textbf{\space }Table~\ref{tw-ab741fc74e15} summarized the effect of various concentrates of \textit{D.littoralis } on Creatinine, Urea, and Total Bilirubin in the serum from normal and paracetamol-induced hepatotoxic rats. The hepatotoxic control group showed increased the level of Creatinine, Urea, and Total Bilirubin when compared to a control group of rats. Treatment of EA concentrates on \textit{D.littoralis, }and  a significant reduction in the\textit{\space } Creatinine, Urea, and Total Bilirubin and silymarin restored Creatinine, Urea, and Total Bilirubin to normal values.


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\caption{\boldmath {\textbf{Effect of various concentrates of }\textbf{\textit{D.littoralis }} \textbf{ on average liver weight changes in rats}} }
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\centering 
\begin{tabulary}{\linewidth}{p{\dimexpr.4343\linewidth-2\tabcolsep}p{\dimexpr.5657\linewidth-2\tabcolsep}}
\tbltoprule \rowcolor{kwdboxcolor}\cAlignHack Group & \cAlignHack Final Liver Weight(g/100g) \\
\tblmidrule 
\cAlignHack GroupI &
  \cAlignHack 4.49\ensuremath{\pm}0.34\\
\cAlignHack GroupII &
  \cAlignHack 6.21\ensuremath{\pm}0.40a**\\
\cAlignHack GroupIII &
  \cAlignHack 5.87\ensuremath{\pm}0.12b*\\
\cAlignHack GroupIV &
  \cAlignHack 4.76\ensuremath{\pm}0.62b**\\
\cAlignHack GroupV &
  \cAlignHack 5.17\ensuremath{\pm}0.18b*\\
\cAlignHack Group VI &
  \cAlignHack 4.60\ensuremath{\pm}0.15b**\\
\tblbottomrule 
\end{tabulary}\par 
\begin{tablenotes}\footnotesize 
    
\item{\textbf{\# }Data be articulated as mean\ensuremath{\pm}SEM., \textit{n } = six rats each group.  P values, *P{\textless}0.05; **P{\textless}0.01; ns= not significant; compared to Paracetamol group. One way ANOVA followed by Dunnett's test.\textbf{ \ensuremath{\rightarrow}} Group II compared to Group I; b\textbf{\ensuremath{\rightarrow}} Group II compared to Group III, IV, V, and VI. }
\end{tablenotes}
\end{threeparttable}

\end{table*}

\begin{table*}[!htbp]
\caption{\boldmath {\textbf{Effect of various concentrates of } \textit{\textbf{D.littoralis }} \textit{\textbf{\space }} \textbf{on serum enzymes SGPT, SGOT, ALP and GGT in Paracetamol induced hepatotoxicity on Wistar rats}} }
\label{tw-87a207f48d8e}
\centering 
\begin{threeparttable}

\def\arraystretch{1.1}
\ignorespaces 
\centering 
\begin{tabulary}{\linewidth}{p{\dimexpr.1654\linewidth-2\tabcolsep}p{\dimexpr.21240000000000002\linewidth-2\tabcolsep}p{\dimexpr.2139\linewidth-2\tabcolsep}p{\dimexpr.1972\linewidth-2\tabcolsep}p{\dimexpr.2111\linewidth-2\tabcolsep}}
\tbltoprule \rowcolor{kwdboxcolor}\cAlignHack Group & \cAlignHack SGPT  \mbox{}\protect\newline (IU.L\ensuremath{^{-1}}) & \cAlignHack SGOT  \mbox{}\protect\newline (IU.L\ensuremath{^{-1}}) & \cAlignHack ALP \mbox{}\protect\newline  (IU.L\ensuremath{^{-1}}) & \cAlignHack GGT \mbox{}\protect\newline (IU.L\ensuremath{^{-1}})\\
\tblmidrule 
\cAlignHack Group I~&
  \cAlignHack 65.89\ensuremath{\pm}2.76 &
  \cAlignHack 93.56\ensuremath{\pm}2.43 &
  \cAlignHack 197.23\ensuremath{\pm}3.18 &
  \cAlignHack 1.29\ensuremath{\pm}0.22\\
\cAlignHack Group II &
  \cAlignHack 176.34\ensuremath{\pm}8.43a** &
  \cAlignHack 190.45\ensuremath{\pm}7.65a** &
  \cAlignHack 278.23\ensuremath{\pm}10.22a** &
  \cAlignHack 1.96\ensuremath{\pm}0.14a**\\
\cAlignHack Group III &
  \cAlignHack 172.12\ensuremath{\pm}6.52b** &
  \cAlignHack 185.48\ensuremath{\pm}4.34b** &
  \cAlignHack 273.65\ensuremath{\pm}6.87b** &
  \cAlignHack 1.89\ensuremath{\pm}0.12 b**\\
\cAlignHack Group IV &
  \cAlignHack 89.26\ensuremath{\pm}4.76b** &
  \cAlignHack 115.63\ensuremath{\pm}4.58b*** &
  \cAlignHack 208.34\ensuremath{\pm}6.21b** &
  \cAlignHack 1.48\ensuremath{\pm}0.05b*\\
\cAlignHack Group V &
  \cAlignHack 136.12\ensuremath{\pm}9.65b** &
  \cAlignHack 169.65\ensuremath{\pm}4.23b** &
  \cAlignHack 248.24\ensuremath{\pm}8.38b** &
  \cAlignHack 1.64\ensuremath{\pm}0.12 b**\\
\cAlignHack Group VI &
  \cAlignHack 78.45\ensuremath{\pm}3.89b** &
  \cAlignHack 107.89\ensuremath{\pm}3.88b** &
  \cAlignHack 195.67\ensuremath{\pm}4.54b** &
  \cAlignHack 1.39\ensuremath{\pm}0.08b**\\
\tblbottomrule 
\end{tabulary}\par 
\begin{tablenotes}\footnotesize 
    
\item{\textbf{\# }Statistical information and particulars of group I-VI was similar as in Table~\ref{tw-a0771a63ee4b} \textbf{\space }}
\end{tablenotes}
\end{threeparttable}

\end{table*}
\textbf{\space }


\begin{table*}[!htbp]
\caption{\boldmath {\textbf{Effect of various concentrates of \textit{D.littoralis } on Creatinine, Urea, and Total Bilirubin in Paracetamol induced hepatotoxicity on Wistar rats}} }
\label{tw-ab741fc74e15}
\centering 
\begin{threeparttable}

\def\arraystretch{1.1}
\ignorespaces 
\centering 
\begin{tabulary}{\linewidth}{p{\dimexpr.25\linewidth-2\tabcolsep}p{\dimexpr.25\linewidth-2\tabcolsep}p{\dimexpr.2569\linewidth-2\tabcolsep}p{\dimexpr.2431\linewidth-2\tabcolsep}}
\tbltoprule \rowcolor{kwdboxcolor}\cAlignHack Group & \cAlignHack Creatinine \mbox{}\protect\newline  (millimol \ensuremath{^{-1}}) & \cAlignHack Urea \mbox{}\protect\newline  (millimole \ensuremath{^{-1}}) & \cAlignHack Total Bilirubin (mg/dL)\\
\tblmidrule 
\cAlignHack Group I~&
  \cAlignHack 53.25\ensuremath{\pm}1.76 &
  \cAlignHack 7.62\ensuremath{\pm}0.18 &
  \cAlignHack 1.31\ensuremath{\pm}0.16\\
\cAlignHack Group II &
  \cAlignHack 76.54\ensuremath{\pm}3.25a** &
  \cAlignHack 19.12\ensuremath{\pm}2.14a** &
  \cAlignHack 4.36\ensuremath{\pm}1.32a**\\
\cAlignHack Group III &
  \cAlignHack 72.65\ensuremath{\pm}2.14b* &
  \cAlignHack 17.83\ensuremath{\pm}0.54b** &
  \cAlignHack 4.16\ensuremath{\pm}0.34b*\\
\cAlignHack Group IV &
  \cAlignHack 58.87\ensuremath{\pm}1.45b** &
  \cAlignHack 8.95\ensuremath{\pm}0.28b** &
  \cAlignHack 1.45\ensuremath{\pm}0.14b**\\
\cAlignHack Group V &
  \cAlignHack 66.56\ensuremath{\pm}2.23b* &
  \cAlignHack 11.83\ensuremath{\pm}0.76b** &
  \cAlignHack 2.67\ensuremath{\pm}0.22b*\\
\cAlignHack Group VI &
  \cAlignHack 56.28\ensuremath{\pm}1.88b* &
  \cAlignHack .86\ensuremath{\pm}0.42b** &
  \cAlignHack 1.33\ensuremath{\pm}0.20b**\\
\tblbottomrule 
\end{tabulary}\par 
\begin{tablenotes}\footnotesize 
    
\item{\textbf{\# } Statistical information and particulars of group I-VI was similar as in Table~\ref{tw-a0771a63ee4b} \textbf{\space }}
\end{tablenotes}
\end{threeparttable}

\end{table*}

\begin{table*}[!htbp]
\caption{\boldmath {\textbf{ Effect of various concentrates of \textit{D.littoralis } on TG and TC in paracetamol-induced hepatotoxicity on Wistar rats}} }
\label{tw-ca430eef8b65}
\centering 
\begin{threeparttable}

\def\arraystretch{1.1}
\ignorespaces 
\centering 
\begin{tabulary}{\linewidth}{p{\dimexpr.2348\linewidth-2\tabcolsep}p{\dimexpr.3663\linewidth-2\tabcolsep}p{\dimexpr.3989\linewidth-2\tabcolsep}}
\tbltoprule \rowcolor{kwdboxcolor}\cAlignHack Group & \cAlignHack TG(mg/dl) & \cAlignHack TC(mg/dl)\\
\tblmidrule 
\cAlignHack Group I~&
  \cAlignHack 52.65\ensuremath{\pm}2.34 &
  \cAlignHack 97.63\ensuremath{\pm}2.36\\
\cAlignHack Group II &
  \cAlignHack 80.16\ensuremath{\pm}3.48a** &
  \cAlignHack 192.54\ensuremath{\pm}8.65a**\\
\cAlignHack Group III &
  \cAlignHack 75.56\ensuremath{\pm}3.12b* &
  \cAlignHack 181.32\ensuremath{\pm}5.52b**\\
\cAlignHack Group IV &
  \cAlignHack 55.44\ensuremath{\pm}2.45b** &
  \cAlignHack 109.62\ensuremath{\pm}4.23b**\\
\cAlignHack Group V &
  \cAlignHack 67.55\ensuremath{\pm}2.86b* &
  \cAlignHack 165.43\ensuremath{\pm}5.22b**\\
\cAlignHack Group VI &
  \cAlignHack 53.42\ensuremath{\pm}2.34b*** &
  \cAlignHack 99.76\ensuremath{\pm}3.72b***\\
\tblbottomrule 
\end{tabulary}\par 
\begin{tablenotes}\footnotesize 
    
\item{\textbf{\# } Statistical information and particulars of group I-VI was similar as in Table~\ref{tw-a0771a63ee4b}}
\end{tablenotes}
\end{threeparttable}

\end{table*}

\begin{table*}[!htbp]
\caption{\boldmath {\textbf{Effect of various concentrates of \textit{D.littoralis } on serum (TP and ALB) in paracetamol-induced hepatotoxicity on Wistar rats }} }
\label{tw-d6bc18050ec4}
\centering 
\begin{threeparttable}

\def\arraystretch{1.1}
\ignorespaces 
\centering 
\begin{tabulary}{\linewidth}{p{\dimexpr.2469\linewidth-2\tabcolsep}p{\dimexpr.3525000000000001\linewidth-2\tabcolsep}p{\dimexpr.40059999999999995\linewidth-2\tabcolsep}}
\tbltoprule \rowcolor{kwdboxcolor}\cAlignHack Group & \cAlignHack TP(mg/dl) & \cAlignHack ALB(g/dl)\\
\tblmidrule 
\cAlignHack GroupI &
  \cAlignHack 7.76\ensuremath{\pm}0.24 &
  \cAlignHack 6.56\ensuremath{\pm}0.10\\
\cAlignHack GroupII &
  \cAlignHack 5.12\ensuremath{\pm}0.18a** &
  \cAlignHack 5.06\ensuremath{\pm}0.12a**\\
\cAlignHack Group III &
  \cAlignHack 5.24\ensuremath{\pm}0.30 b** &
  \cAlignHack 5.17\ensuremath{\pm}0.12b*\\
\cAlignHack Group IV &
  \cAlignHack 6.95\ensuremath{\pm}0.24b* &
  \cAlignHack 5.94\ensuremath{\pm}0.13b**\\
\cAlignHack Group V &
  \cAlignHack 5.83\ensuremath{\pm}0.11a** &
  \cAlignHack 5.43\ensuremath{\pm}0.10b*\\
\cAlignHack Group VI &
  \cAlignHack 7.22\ensuremath{\pm}0.18b** &
  \cAlignHack 6.12\ensuremath{\pm}0.10b**\\
\tblbottomrule 
\end{tabulary}\par 
\begin{tablenotes}\footnotesize 
    
\item{\textbf{ \# } Statistical information and particulars of group I-VI was similar as in Table~\ref{tw-a0771a63ee4b}}
\end{tablenotes}
\end{threeparttable}

\end{table*}
Table~\ref{tw-ca430eef8b65} shows the effect of various concentrates of \textit{D.littoralis } on TG and TC in the serum from normal and paracetamol-induced hepatotoxic rats. The hepatotoxic control group showed increased the level of TG as 80.16\ensuremath{\pm}3.48 and TC as 192.54\ensuremath{\pm}8.65 that of group I~rats. Oral administration of PE concentrates showed considerable TG and TC levels in group III rats as 75.56\ensuremath{\pm}3.12 and 181.32\ensuremath{\pm}5.52. Oral administration of ethyl acetate concentrates showed a marked reduction in TG and TC levels in group IV rats as 55.44\ensuremath{\pm}2.45 and 109.62\ensuremath{\pm}4.23. Oral administration of methanol concentrates showed reduced the level of TG and TC in group V rats as67.55\ensuremath{\pm}2.86 and 165.43\ensuremath{\pm}5.22. The administration of EA focuses on \textit{D.littoralis, and} silymarin treated a group of significant rat reduction in the level of TG and TC as compared to PE and methanol concentrates. The various concentrates of \textit{D.littoralis } on TP and albumin in the serum from normal and paracetamol-induced hepatotoxic rats were depicted in Table~\ref{tw-d6bc18050ec4}. The hepatotoxic control group showed a reduction of TP as 5.12\ensuremath{\pm}0.18 and albumin as 5.06\ensuremath{\pm}0.12. Oral administration of PE concentrates showed considerable TP and ALB levels in group III rats as 5.24\ensuremath{\pm}0.30\ensuremath{^{\textbf{\space }}} and 5.17\ensuremath{\pm}0.12. Oral administration of ethyl acetate concentrates showed a marked reduction in TP and ALB levels in group IV rats as 6.95\ensuremath{\pm}0.24 and 5.94\ensuremath{\pm}0.13. Oral administration of methanol concentrates showed reduced the level of TP and ALB  in group V rats as 5.83\ensuremath{\pm}0.11\ensuremath{^{}} and 5.43\ensuremath{\pm}0.10. The administration of EA concentrates on \textit{D.littoralis, } and silymarin treated rats have significantly elevated the level of\textit{\space } TP and albumin as compared to PE and methanol concentrates. 

In the present study shown that a lethal dose of various concentrates of \textit{D.littoralis } was showed the safety of concentrates. Administration of multiple foci of \textit{D.littoralis} in rats did not change any autonomic or behavioral reaction. There was no mortality of various concentrates of \textit{D.littoralis} was recorded at 2000mg/kg. Therefore we have calculated and selected for 200 mg/kg b.wt. \textit{D.littoralis} concentrates was taken for further investigation. 

The liver is profoundly affected primarily by toxic agents. Hence, the liver marker enzymes are very sensitive markers of toxicity and are of great importance in the assessment of hepatic damage.\unskip~\citep{634166:14854690}. Activities of SGPT, SGOT, ALP, GGT, and the level of serum bilirubin are mostly utilized as the most common SGPT, SGOT, ALP, GGT to examine the liver damage. Paracetamol-induced liver damage is considered and also used for the toxic agent of liver toxicity. \unskip~\citep{634166:14854707}. This study, the important elevation of SGPT, SGOT, ALP, GGT, and bilirubin level in rats treated with paracetamol, are suggestive of cellular leakage and defeat of functional integrity of the liver cell membrane. \unskip~\citep{634166:14854708}. This is an agreement with previous studies; we found that the higher dose of paracetamol might be toxic to the hepatocytes \unskip~\citep{634166:14854693,634166:14854699}. 

Administration of EA concentrates on \textit{D.littoralis } in this study suppresses the elevated serum SGPT, SGOT, ALP, GGT, and the bilirubin concentration. Recovery toward normalization of the enzymes following \textit{D.littoralis } treatment suggested that excellent hepatoprotective property and has a specific role in conserving the structural integrity of the hepatocellular membrane, thus inhibiting enzyme leakage into the bloodstream, in addition to restoring of liver injury caused by paracetamol. This activity is in agreement with the generally established sight that serum levels of transaminases come again to standard with the healing of hepatic parenchyma and the renewal of hepatocytes\unskip~\citep{634166:14854686,634166:14854705}. The concentration of Triglyceride and Total cholesterol were elevated in treated with Paracetamol-induced rats as compared to normal rats (group I). The liver is central to the ruling of cholesterol concentration in the body, does not synthesize cholesterol for goes to another cell, but it also separates cholesterol from the body by changing to bile salts and eliminate to bile. Administration of EA concentrates of\textit{ D.littoralis (}200mg/kg b.wt) significantly reduced TG and TC levels when compared with other two concentrates treatment groups. This effect could be the presence of bioactive compounds in the EA concentrates of\textit{ D.littoralis.}
    
\section{ Conclusions}
The current study showed that EA concentrates on \textit{D.littoralis } protects hepatocytes against paracetamol-induced hepatic tissue damage by reducing the activities of hepatic enzymes. From our results, ethyl acetate concentrates of \textit{D.littoralis } had a better hepatoprotective effect compared to that of two foci in paracetamol-induced in rats.
    

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