**4. Discussion**

**Treatment**

**Group IV**

**Group V**

**Group VI**

**Group VII**

**Group VIII**

**Treatment**

**Group IV**

**Group V**

**Group VI**

**Group VII**

**Group VIII**

**ANOVA F**

**SOD Catalase**

44±2.432 33.3± 1.840\* 42.02± 2.322 59.67± 3.298 (19.52%) (36.53%) (31.11%) (45.95%)

45.6±2.520 35.9± 1.984\* 42.5± 2.349 59.97± 3.315 (24.35%) (47.19%) (34.92%) (47.51%)

55±3.040\* 41.15± 2.274\* 44.05± 2.435 61.96± 3.425\* (52.68%) (68.71%) (47.22%) (57.82%)

60.9±3.366\* 43.13± 2.384\* 45.2± 2.498 62.1± 3.432\* (70.46%) (76.83%) (56.34%) (58.54%)

68±3.759\* 41.83± 2.312\* 50.6± 2.775\* 62.57± 3.458\* (91.86%) (71.50%) (96.03%) (58.54%)

> **ATPase Liver ATPase Kidney (mgPi/100gm/min)**

1721.4± 95.16\* 1814.28±100.29 (60.71%) (4.30%)

1864.28±103.06\* 1907.14±105.43 (83.02%) (18.28%)

1889±104.43\* 2042.85±112.93 (87.09%) (38.71%)

1966.06±108.69\* 2149.99±118.85\* (99.22%) (54.83%)

1969±108.85\* 2107.14 ±116.48 (99.68%) (48.38%)

**Value 22.664@ 12.569@ 4.464@ 2.911@**

Data are mean ± S.E., n=6.ANOVA (F Values at 5% level).# P≤0.05 vs. Control,\*P ≤0.05 vs. ATD,@ Significant

**Group I** 1971±108.95 2449.99±135.44 **Group II** 2114±116.86 2257.14±124.77 **Group III** 1335.71±73.84# 1785.7±98.72#

**ANOVA F Value 6.350@ 4.696@** Data are mean ± S.E., n=6.ANOVA (F Values at 5% level). # P≤0.05 vs. Control,\*P ≤0.05 vs. ATD, @ Significant.

**Table 6.** Therapeutic Effect of *P.amarus* on ATPase against ATD

**Table 5.** Effect of *P.amarus* on SOD, Catalase activity in liver and kidney against anti TB drugs.

**Group I** 70.7±3.908 48.78± 2.696 50.7± 2.802 70.1± 3.875 **Group II** 76±4.201 51.2± 2.830 51.8±2.863 62.14± 3.435 **Group III** 37.52±2.074# 24.39±1.348# 38.1± 2.106# 50.8± 2.808#

288 Pharmacology and Nutritional Intervention in the Treatment of Disease

**(U/min./mg protein) (µ mole of H2O2 oxidised/min/mg protein) Liver Kidney Liver Kidney**

The liver diseases remain one of the serious health problems as variedly exposed to xenobiotics. Modern medicines have little to offer for alleviation of hepatic diseases. Although *P. amarus is* reported to possess varied medicinal properties such as antiviral, anticancer, antioxidant, anti-inflammatory, hepatoprotective (Joshi and Parle, 2007), there is no previous report about the hepatoprotective activity of this plant against anti TB drugs. The present investigation reports the hepatoprotective effect of *P.amarus*. In the present study, hepatotoxicity model in albino rats was successfully produced by administering RIF, INH and PZA.

Effects of administration of Anti TB Drugs and *Phyllanthus amarus* orally on selected biochem‐ ical parameters in rat tissue and blood serum is presented in Table 1-6.

Significant rise above the normal upper limits in the measured serum transaminases in toxicant group on day 60 of the experiment was a biochemical indication of liver injury. Elevated levels of serum enzymes, AST and ALT are indicative of cellular leakage, and loss of functional integrity of cell membrane in liver (Ranawat *et al.,* 2010). Oral administration of *P. amarus* extract at doses (100, 200, 300 and 400 mg/kg) showed significant recoupment in a dose dependent manner (P ≤0.05) (Table1).

The increased level of serum alkaline phosphatase is reliable marker of liver damage, which occurs due to the *de novo* synthesis by the liver cells (Muriel and Escobar, 2003). Serum albumin concentration is affected by hepatic protein and its synthesis is a typical function of normal liver cells (Thirunavukkarasu and Skthisenkaran, 2003). Bilirubin is one of the most frequent clinical test to evaluate the extent of chemically induced hepatotoxicity (Zimmerman, 1973). Toxicants administration caused significant increase in the serum alkaline phosphatase activity. Stabilization of SALP and bilirubin levels by the treatment of *P.amarus* is clear indication of improvement in functional status of liver cells. Results suggested that *P.amarus* at a dose of 400mg/kg, b.w., have protective effect on plasma membrane of hepatocytes. (Table2)

As a measure of renal function status, serum urea, uric acid and creatinine are often regarded as reliable markers (Adebisi *et al*., 2000). Serum creatinine has been used to estimate glomerular filtration rate. Thus, elevations in the serum concentrations of these markers are indicative of renal injury (Adebisi *et al*., 2000; Adewole *et al.,* 2007).The same was observed after toxicant administration. It may be due to dysfunctional and dystrophic changes in the liver and kidney. Experiment has shown that *P.amarus* at different doses, showed significant protective but the highest protection was obsereved at 300 and 400 mg/kg with mere difference indicating normal glomerular filtration rate thereby improved functional status of kidney. (Table 3)

Various biochemical parameters were measured in liver and kidney tissues. The levels of TBARS in liver and kidney tissues of ATD intoxicated rats were significantly elevated when compared to the level of TBARS in control animals. The increased lipid peroxidation results in changes in cellular metabolism of the hepatic and extra hepatic tissues, which ultimately leads to the whole cell deformity and cell death (Arun and Balasubramanian, 2011). The administration of herbal drug *P. amarus* at the different therapeutic doses showed reduction in TBARS level. The standard hepatoprotective drug Silymarin maintained the decreased lipid peroxidation level to the normal limits in the liver.

major parameters showed percent protection level above 50% at dose levels 300 and 400mg/kg. All these properties make *P. amarus*, a novel herb for treating oxidative stress and anti Tb drug associated hepatic toxicity. There is, however, the need of further experiments

Hepatoprotective effect of *Phyllanthus amarus*

http://dx.doi.org/10.5772/57373

291

for chronic study to exercise on excessive and prolonged use of this plant.

**Figure 1.** Hepatocytes of the normal control group showed a normal lobular architecture of the liver (400 x).

**Figure 2.** Hepatocytes of the INH + RIF+ PZA treated group showed liver cell necrosis and inflammation also observed

in the centrilobular region with portal triaditis (400 x).

**6. Histopathological studies**

Concomitant cellular oxidative stress was manifested by reduced GSH levels and in‐ creased lipid peroxidation. The inverse linear relationship between the ROS level and the GSH level indicated that free radical species were generated by exposure to anti TB drugs which reduced intracellular antioxidant levels. The results indicate that, the herbal drug *Phyllanthus amarus* has very good hepatoprotective effect in liver damage. The results were presented in the Table 4.

Superoxide dismutase and Catalase mutually function as important enzymes in elimination of Reactive oxygen species (ROS). SOD is the major attractive metalloprotein in the antioxidant family. The defensive antioxidant enzyme next to SOD is CAT. CAT is an enzymatic antioxi‐ dant widely distributed in all animal tissues, and the highest activity is found in the red cells and liver. Both are the key component of the antioxidant defense system. In the present study, the observed decrease in SOD and CAT activities were presumably associated with the increased oxidative stress caused by these toxicants that might be due to low level of zinc (a metal constituent of the enzyme SOD) in liver tissue (Arun and Balasubramanian, 2011) Therapy at 200-400 mg/kg b.w. reversed the SOD and CAT activity in the liver tissues and protected from free radical induced oxidative stress. These observations are substantiated by author (Gnanadesigan *et al.,* 2011).(Table 5)

ATPase is a membrane bound enzyme. Since, phosphatase is a constituent of all the body tissues; it plays an important role in inorganic pyrophosphates activity. ATPase activity may be considered as a marker for assessing hepatocellular damage induced by hepatotoxic agents. (Table 6)

In our experiment, a concurrent fall was found in ATPase in liver after toxicants exposure. It might be due to dysfunctional and dystrophic changes in the mitochondria and cell membrane permeability. This damage was also very clearly visible in histopathological studies after toxicant administration. These observations are substantiated by other authors (Gao and Zhou, 2005; Krithika and Verma, 2009).The effect of the extracts on ATPase was as pronounced with 100mg/kg as with 200-400mg/kg b.w.

Liver damage induced by toxicant was associated with a variety of biochemical abnormalities following loss of integrity of the cell membrane or interference with normal hepatocytes metabolism and function.

The reason for hepatoprotective effect of the extracts may be due to presence of lignans and flavonoids which might have scavenged the free radical offering hepato protection.
